Report On The Research Activities With . - Loro Parque
Report on the researchactivities with Orcinusorca in Loro ParqueDate:Author:May 23rd 2013F. Javier Almunia Portolés, Ph.D.Deputy Director of Loro Parque Fundación
Reporton researchactivities1 Introduction2 Finished Projects and Scientific Activities3 Active Projects and Scientific Activities1 IntroductionThe scientific research activities of Loro Parqueare based on the Research and ConservationProgram 2010-2015, which has been submittedto the Spanish authorities in compliance with theSpanish Zoo Law (31/2003) and the European ZooDirective (1999/22/CE). This Program is the officialdocument in which the fundamental criteria andguidelines for the research and conservation activitiesare established. These criteria are based on therecommendations of different zoological institutionsrelated with Loro Parque (World Association of zoosand aquariums -WAZA-, European Association ofZoos and Aquarium -EAZA-, Iberian Association ofZoos and Aquariums -AIZA-, European Associationof Aquatic Mammals -EAAM-), and severalbiodiversity conservation organizations relatedwith Loro Parque Fundación (International Unionfor Nature Conservation -IUCN-, Survival SpeciesComission -SSC-, Conservation Breeding SpecialistGroup -CSBG-, etc.).The main goal of this Program is to optimize thefinancial, material and human resources that LoroParque dedicates to the research and conservationactivities, either directly or through Loro ParqueFundación. As no zoological institution can runresearch and conservation projects for every singlespecies in its zoological collection, the Research andConservation Program of Loro Parque establishesthe criteria to distribute the resources among themost appropriate projects and species, based ontheir biodiversity conservation potential and thesuitability of the group of animals hosted at LoroParque.collection. To determine this potential value severalcriteria will be used, such as: status in the IUCN RedList of Threatened Species, availability of breedingprograms (ISB, EEP or ESB), level of knowledgeabout species biometrics, biology or physiology,number of individuals available in the collection,level of husbandry, suitability of the holding facilities,etc.Based on the potential conservation andresearch analysis, a set of research proposals aresubmitted every year to the Board of Directors ofthe Loro Parque Fundación. All the proposals areevaluated in the Advisory Board Annual Meeting,using the most up-to-date scientific knowledge ofthe board of independent members. The Board ofDirectors approve the research and conservationproposal (in situ and ex situ), which are publishedin a final document.In 2012 the LPF Advisory Board decidedto fund 36 projects during 2013 with a total of1.136.036 US , of them 30 were focused on parrotsand terrestrial ecosystems, and 6 of them focused oncetaceans and marine ecosystems. Four out of thesix marine projects are mainly focused on Orcinusorca research activities ex situ, with applicationsfor in situ conservation of this and other cetaceanspecies.The selection of the scientific and conservationactivities is based on an analysis of the potentialvalue for research and conservation (either in situor ex situ) of every single species in the zoologicalReport on the Research Activities with Orcinus orca - 2
2 Finished Projectsandscientific activities2.1 Research on bioacoustic toolsWhen Morgan arrived to Orca Ocean she wasintegrated into the two ongoing research projects,funded by Loro Parque Fundación, that implied exsitu research activities with Orcinus orca. One ofthem, aimed at the development of tools to improvebioacoustic research was finished in 2012. WhenMorgan arrived the project team was acquiringindividually identified sounds in order to establishindividual repertoires. As soon as Morgan couldbe held in the experimental pool (by then holdingPool 2), the setting-up of her individual repertoirewas initiated. As a result, over 250 Morganvocalizations were recorded and archived in 2012LP DialectLP01 LP01.1LP11LP01.2LP11.1LPwaveLP02Figure 1: Spectrogram of one of the sounds attributed to Morgan thatwere registered during the introduction phase.for future analysis. These data were made availableto the researchers of the University of St. Andrewsstudying the evolution of Morgan’s dialect.LP03LP12LP13LP04LP15LP51Figure 2: Acoustic dialect of the orcas in Loro Parque before the introduction of MorganReport on the Research Activities with Orcinus orca - 3LP16LP06
2.2 Genetic comparison between speciesof Marine MammalsShort after the arrival of Morgan to LoroParque, the Museum of Natural History ofCopenhagen requested good quality geneticmaterial to perform a comparison between differentspecies of marine mammals. As they already usedMorgan’s genetic samples in a previous stage ofthe project (when they obtained the samples todetermine Morgan’s original pod), they wantedto use genetic material from the same individualto continue the project. Loro Parque requested thenecessary CITES permits to export the blood samples,and sent them to the research team, which receivedand processed them accordingly. Morgan’s geneticmaterial was sequenced and provided a very highquality genome. Despite that the researchers arestill working on the publication of the comparativegenetic adaptations in different marine mammalspecies, the genome sequence obtained fromMorgan has been published as reference forOrcinus orca as public domain: 2.3 Audiogram of Orcinus orcaDuring 2012 Loro Parque requested a groupof international experts to evaluate the hearingcapacity of Morgan, as the trainers inferred thatshe was inconsistent in responding to the soundbridge (whistle). The research team suggested theuse of Auditory Evoked Potential (AEP) as it allowsa quick assessment of hearing capabilities, andhas been extensively used with dolphins and othersmall cetaceans. In order to get the most scientificinformation possible from this hearing evaluationnot only Morgan, but also the other four orcas, weretrained to perform the auditory evoked potentialmeasurements underwater, and the other two weremade available to be measured out of the water.As a result, the basic audiometric functionsof six killer whales were assessed, becoming thelargest test using Auditory Evoked Potential in thisspecies ever. This methodology was used only oncebefore in 1999 with two killer whales to describethe audiogram of the species. Before this researchwas performed, all the scientific information on thehearing capacity of the species was based just onthree individuals.The results of the test revealed a lack of clickevoked response in Morgan, which confirmedthat this animal suffers from a hearing deficit. Themagnitude and frequency range over which thehearing deficit occurs couldn’t be specified withFigure 3: Morgan desensitization training session. From left to right and top to down: 1 attaching dummy suction cups, 2 giving signal to putnose on target, 3 correct position and 4 incorrect position.Report on the Research Activities with Orcinus orca - 4
Figure 4: Left: The research team measuring the AEP in water, this experimental arrangement is more complex and difficult to train, but givesbetter results as the sound reaches the animal in a more natural way. Right: Measuring AEP in a beached individual using a jawphone.the methodology used in the study. Unfortunately,although AEP measurements allow a quickassessment of hearing capabilities, due to the sizedependent loss of the electrophysiological signal, thismethodology was revealed to be inappropriate todetect the lower hearing threshold of Orcinus orca.As a consequence, the research team submitted anew proposal to perform a behavioural audiogramof the same animals during 2013, the proposalwas approved by the Advisory Board Meeting andpartial funding granted by Loro Parque Fundación.Figure 5: Results of AEP measurements in water (left) and beached (right), show a total absence of brain response to the soundstimulus in Morgan.Report on the Research Activities with Orcinus orca - 5
3 Activeprojects andscientific activitiesCurrently, there are five active projects andone proposal, all of them focused on ex situ researchactivities with the group of Orcinus orca hosted atLoro Parque, thus all of them include Morgan.3.1 Pollution markers in Europeanbottlenose dolphinsThis project is mainly focused on the effectsof pollution in the pathology of the wild dolphinpopulations in Europe. The research team collectedbiopsies of wild and stranded dolphins around theCanary Islands simultaneously with other researchersin the Mediterranean and North Atlantic. The groupof dolphins hosted at Loro Parque were used as areference group, to measure the biodynamics andbio-accumulation of pollutants in a group of dolphinswith a controlled diet.When the research team started the samplingat Loro Parque, the opportunity to obtain bloodsamples from other marine mammals with the samediet came up. As a consequence, the researchersinitiated a parallel data collection to study apoorly understood issue: the relationship betweenconcentrations of Persistent Organic Pollutants(POPs) in blubber and those in blood of marinemammals, as a better indicator of organ exposureto pollutants.Morgan’s blood samples were obtainedregularly (as for the rest the group of orcas,dolphins and sea lions) and shipped to the researchteam in order to be analysed and to track anypossible variation of the pollution markers. During2013 the research team is obtaining biopsy samplesto evaluate the toxicological parameters in theblubber of the three species and compare themwith the plasmatic concentrations. The measurementof the toxic concentrations in plasma, blubber andfood should shed light on the bioaccumulationmechanisms in the three species.3.2 Evolution of Morgan’s DialectWhen Morgan arrived to Loro Parque aproposal to evaluate the changes on her dialectwas received from the University of St. Andrews(Scotland). This was the same research team thatmade the initial comparison of Morgan’s dialectwith the records of pod dialects from the three majororca populations in the North Atlantic.The proposal tested the acoustic evolution ofMorgan’s dialect, as there is a hypothesis about theevolution of the orcas’ dialect during their entirelifetime. If this hypothesis proved to be correct,Morgan’s dialect should be modified with theincorporation of calls specific to the Orca Oceangroup. Similarly, the rest of the animals in the groupwould be integrating sounds from Morgan into theirdialect.A researcher from St. Andrews Universityvisited Loro Parque two weeks during 2012 tocollect sounds and make observations on Morgan’sdialect. Complimentarily, Loro Parque Fundaciónshared all the identified sounds of Morgan that wereregistered from the on-going bioacoustics researchsince she arrived to Loro Parque, in order to providea broader set of data. A complete description of thegroup’s dialect previous to Morgan’s incorporationwas also provided by Loro Parque, so the researchteam could verify if any eventual new call inMorgan’s dialect derived from there.The preliminarily analysis of Morgan’s dialectduring 2012 did not show any incorporation ofsounds from the Orca Ocean group. Given thatby the end of 2012 the audiogram of Morganconfirmed that she had a significative hearing deficit(that could even be total deafness) the experimentcan not be considered valid, and the hypothesisof the lifetime evolution of vocal dialect in Orcinusorca can not be rejected.Alternatively, the research team from St.Andrews University submitted a continuationproposal to test the lifetime evolution of vocaldialects in Orcinus orca during 2013 using a differentapproach. The objective is to analyze the individualdialects of the six orcas at Loro Parque except forMorgan. As the auditory capacity of five animalshas been verified, the analysis of the evolution oftheir dialects after the introduction of Morgan intothe group can be used to test the hypothesis. During2013 researchers from the University of St. Andrewswill be visiting Loro Parque to collect sounds andmake observations of the dialect of every individualexcept Morgan. The recordings will be analyzedto find incorporations of Morgan’s calls into theindividual dialects of the rest of the group. In orderto provide the research team with a broader dataset to analyze, Loro Parque will make available tothe researchers all the identified calls registeredand archived as a result of its in- house bioacousticresearch activities.Report on the Research Activities with Orcinus orca - 6
Figure 6: Voluntary blood extraction of an orca at Loro Parque3.3 Biometric information derived frombioacoustic analysis of Orcinus orcacallsThis project is aimed to find the way to obtainbiometric data from the bioacoustic signals of theorcas. This project is based on the technologydeveloped by the University of La Laguna and LoroParque Fundación in a previous 6 year projectfinished in 2011. Thanks to this technology, anindividual repertoire for each of the seven orcas atLoro Parque is being established.The research team is working in theparametrization of the calls as the stage prior toobtaining information related to the biometry ofthe animal producing the sound. The animals arealso carefully measured and weighed on a regularbasis to collect the accurate biometric information.The bioacoustic records of the group (includingMorgan) have been made available to researcherswhen required, after presentation of an adequatescientific proposal.The hydrophone system at Orca Ocean isbeing improved to digitalize the sound signals beforeany amplification. That will provide the top qualitymeasurements needed to parameterize the signalsand, subsequently find biometric characteristics fromthe bioacoustic signals. If this methodology finallyworks, it would mean a very positive outcome inthe medium range non-invasive assessment of thebody shape of the wild killer whales, which couldbe helpful in the conservation of the species.3.4 Behavioral audiogram ofOrcinus orcaAs the Auditory Evoked Potential (AEP)methodology was revealed to be inappropriateto detect the lower hearing threshold of Orcinusorca, a new proposal to perform a behaviouralaudiogram of the same animals during 2013 wassubmitted to the Advisory Board Meeting of LoroParque Fundación.In this new research the hearing capacity ofMorgan and other four or five individuals will beaccurately established. This methodology impliesan intensive training of the animals to make themrespond consistently to a discriminative stimulus(sd). The stimulus will initially be visual and acousticand gradually shifted to only acoustic. Once theanimals respond consistently to the acoustic sd,Report on the Research Activities with Orcinus orca - 7
long and detailed testing will be initiated by playingdifferent sound intensities at multiple frequencies inorder to determine the hearing threshold of everyindividual. If Morgan fails to shift from the visual/acoustic discriminative stimulus to the acoustic sd,that would be an indication of total deafness.The information resulting from this study willmean a major improvement in the actual knowledgeof the auditory capacity of the species that comesfrom a study on a subadult male 40 years ago, andone more recent with two females over 20 years old.All the animals tested in this experiment are under17 years old, which will give a better approach tothe auditory capacity of young specimens of Orcinusorca. A detailed description of the auditory capacityof the species is a critical knowledge to be achievedin order to better protect the wild populations fromthe noise pollution.3.5 Bioacoustical characterization ofdifferent behavioural situations in OrcinusorcaThe Advisory Board Meeting of Loro ParqueFundación approved a proposal from the Universityof La Laguna and the Free University of Berlin tostudy the prevalence of different types of callsrelated with behaviour in Orcinus orca. It has beensuggested that there is a relation between thedifferent proportional call type usage by the samepods and their behaviour, despite that few studieshave focused on the characterization of call usageand behaviour.The goal of this proposal is to perform anethological observation of the group of orcas atLoro Parque, while simultaneously recording thesounds that they produce underwater. A randomfocal observation schedule has been set up, andfocal individuals are randomly selected and videorecorded for 15 minutes. Complementarily, avideo recording covering the whole main pool(See Anex I) will be also made to provide furtherad libitum or focal observations. The ethologistsfrom both universities will analyse the video andaudio recordings in order to determine if there aresignificant variations in the proportional call typeusage in different behavioural situations.3.6 Biometrics proposalIn the beginning of 2013 Loro Parquereceived a proposal from the University of St.Andrews to perform a research project to measurethe dimensions of the killer whales in Loro Parqueusing a high-frequency scanning sonar. Each whalewill be scanned by the sonar as it swims past,and measurements taken from the sonar imagewill be related to hand-measurements taken fromthat individual. The sonar uses sound frequenciescentered at 375 and 610 kHz. The results shouldlead to a peer-reviewed publication on the feasibilityof this method to remotely measure dimensions ofwild whales. This methodology would mean a verypositive outcome in the non-invasive assessment ofthe body shape of the wild killer whales, whichcould help in the management of its endangeredsubpopulations and, thus in the conservation of thespecies.Loro Parque has expressed its willingness tocooperate in this research as long as it does notcompromise the welfare of the animals. CurrentlyLoro Parque is gathering advice from differentexperts in acoustics and cetacean hearing toensure that the sonar will not affect the animals.If the welfare of the animals can be guaranteed,the experiments will be performed during the lastquarter of 2013.Report on the Research Activities with Orcinus orca - 8
Annex IReport on the Research Activities with Orcinus orca - 9
visited Loro Parque two weeks during 2012 to collect sounds and make observations on Morgan’s dialect. Complimentarily, Loro Parque Fundación shared all the identifiedsounds of Morgan that were registered from the on-going bioacoustics research since she arrived to Loro
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Le genou de Lucy. Odile Jacob. 1999. Coppens Y. Pré-textes. L’homme préhistorique en morceaux. Eds Odile Jacob. 2011. Costentin J., Delaveau P. Café, thé, chocolat, les bons effets sur le cerveau et pour le corps. Editions Odile Jacob. 2010. 3 Crawford M., Marsh D. The driving force : food in human evolution and the future.
