Swimming features in captive odontocetes: Indicative of animals’ emotional state?

Journal Pre-proof Swimming features in captive odontocetes: indicative of animals’ emotional state? Agathe Serres, Hao Yujiang, Wang Ding

PII:

S0376-6357(19)30259-1

DOI:

https://doi.org/10.1016/j.beproc.2019.103998

Reference:

BEPROC 103998

To appear in:

Behavioural Processes

Received Date:

19 June 2019

Revised Date:

30 October 2019

Accepted Date:

4 November 2019

Please cite this article as: Serres A, Yujiang H, Ding W, Swimming features in captive odontocetes: indicative of animals’ emotional state?, Behavioural Processes (2019), doi: https://doi.org/10.1016/j.beproc.2019.103998

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Swimming features in captive odontocetes: indicative of animals’ emotional state?

Agathe Serresa,b,c, Hao Yujianga, Wang Dinga a

Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430050, China

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University of Chinese Academy of Sciences, Beijing 100101, China

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Corresponding author: e-mail: [email protected]

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Different captive odontocetes’ groups exhibited different circular swimming direction biases Finless porpoises and bottlenose dolphins were more active in the morning than at other times of the day Circular swimming was lower when enrichment was provided and higher when potentially stressful events occur or when animals were separated in sub-groups Fast swimming and social swimming were less frequent when enrichment was provided and more frequent when potentially stressful events occurred

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Highlights

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Abstract

Captive welfare studies in odontocete species have been mostly conducted on bottlenose dolphins (Tursiops

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truncatus) while the welfare of many other species’ -including endangered species- remains poorly studied. More research is needed to find and validate potential indicators of welfare for each species and even for each group. Since captive odontocetes spend most of their time swimming, their swimming features are interesting to study in relation to their welfare state. We first analysed the circular swimming direction bias

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in three groups of captive odontocetes (Yangtze finless porpoises: Neophocaena asiaeorientalis asiaeorientalis; East-Asian finless porpoises: N. a. sunameri; and bottlenose dolphins, Tursiops truncatus). Second, we studied the effect of environmental and social factors (i.e., time of the day, delay to training, enrichment, potential perturbation, social grouping, public presence and housing pool) on circular swimming, fast swimming, group swimming, synchronous swimming and contact swimming in the three groups. Yangtze finless porpoises exhibited a clockwise swimming bias while East-Asian finless porpoises

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and bottlenose dolphins swam significantly more in the counter-clockwise direction. Each studied factor significantly impacted the animals’ swimming behaviour slightly differently depending on the group. However, some patterns were common for the three groups: animals seemed to be more active in the morning than at noon and in the afternoon, and enrichment seemed to decrease circular swimming, fast swimming and social swimming (i.e., synchronous, contact and group swimming), while potential perturbations (e.g., pool cleaning, noise) seemed to increase it. In addition, behaviour differed for Yangtze

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finless porpoises and bottlenose dolphins right before the training or when other animals were being trained, suggesting an anticipation of this event or an excited/frustrated state in this context. Social separation also

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impacted these animals’ swimming behaviour with less group swimming but more circular swimming, synchronous swimming and fast swimming when separated. The housing pool had an impact on bottlenose

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dolphins’ behaviour with more circular swimming, more fast swimming and less group swimming when

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having access to a larger space. The effect of the presence of public was unclear and requires further investigation. From our results, we propose that circular swimming, synchronous swimming and contact

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swimming could be useful to monitor animals’ emotional state, but that additional parameters should be added (e.g., swimming speed) since these behaviours can be expressed both in quiet and relaxed contexts

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and in stressful ones. In addition, fast swimming can be a useful indicator of stress for porpoises but might be more ambiguous for bottlenose dolphins that engage in intense social play bouts for instance. Finally, group swimming might be a good behaviour to monitor when wanting to investigate reactions to various conditions or events that can potentially be stressful. We suggest that further research should be conducted

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on other groups of odontocetes to validate our findings. Keywords: bottlenose dolphin; circular swimming; finless porpoise; stress; synchronous swimming; welfare

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Introduction

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Studies on animal welfare have increased in recent years, including studies on odontocete species (Clark, 2013). However, research on positive and negative welfare indicators in odontocetes is still lacking (Clark, 2013; Ugaz et al., 2013; Clegg et al., 2015, 2017a; Brando et al., 2017). In this paper, following recent studies about welfare (Yeates and Main, 2008; Mason and Veasey, 2010; Watters, 2014; Dawkins, 2015; Clegg et al., 2017a), we will use the term “welfare” as a “…balance between positive (reward, satisfaction) and negative (stress) experiences or affective states. The balance may range from positive

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(good welfare) to negative (poor welfare)” (Sprujit et al., 2001). From Webster’s (2005) triangulation principle, it was suggested that welfare should be measured using behaviour, health and cognitive

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experiments. Among these three categories, assessments of behaviour, specifically social interactions, are thought to deliver at least as much information about welfare as physiological or health measures (Waples

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and Gales, 2002; Hill and Broom, 2009; Joseph and Antrim, 2010). Social play for instance was already

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suggested to indicate positive emotional states in a variety of mammal species (Held and Spinka, 2011), and it was shown to vary depending on environmental factors and thus to be a potential welfare indicator

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in bottlenose dolphins (Serres and Delfour, 2017). Conversely, high rates of agonistic interactions have been suggested to be indicative of poor welfare states in mammals (Cunningham, 1988; Ambrose and

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Morton, 2000; Honess and Marin, 2006; Beisner and Isbell, 2011; Usama, 2011; Khan, 2013; Hosey et al., 2016; Salas et al., 2016), including bottlenose dolphins (Clegg et al., 2015). More behavioural indicators such as synchronous swimming or anticipatory behaviours have been suggested to be linked with odontocetes’ emotional state (Clegg et al., 2017a, 2017b, 2018) but still have to be validated in order to

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better assess welfare in captive odontocetes (Brando et al., 2017). Both in the wild and in captivity, odontocetes spend much time swimming. They swim alone or

with partners, with different speeds and in different directions. The large amount of time they spend swimming, and the various ways they swim makes this category of behaviour an interesting one to study in relation to welfare. Studies have reported captive odontocetes as frequently swimming in a repetitive way and following a fixed pattern around the pool (Gygax, 1993; Sobel et al., 1994; Sekiguchi and Koshima,

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2003; Singh, 2005), with no clear link with good or poor welfare established yet (Mason and Latham 2004). Directional biases were found in this circular swimming (bottlenose dolphins: Tursiops truncatus, Sobel et al., 1994; belugas: Delphinapterus leucas, Marino and Stowe, 2007) and it was recommended to investigate the impact of social and environmental changes on this bias (Platto et al., 2017). Odontocetes are frequently observed swimming with partners, and synchronous swimming is often described as an affiliative behaviour in bottlenose dolphins (Connor et al., 2006; Fellner et al., 2006, 2013; Clegg et al., 2017b). However, no

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study has investigated the impact of environmental factors on synchronous swimming in captive groups of odontocetes. Finally, swimming speed has been shown to increase in wild odontocetes exposed to

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disturbances (killer whales: Orcinus orca, Kruse, 1991; bottlenose dolphins: Nowacek et al., 2001), which makes it another worthy parameter to study in relation to welfare in these animals.

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In recent years, because it was noticed that some species react better than others to captivity and

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fare better, comparative studies might be useful to understand species differences (Mason, 2010). Several marine mammal species such as Dall’s porpoises (Phocoenoides dalli) or Fraser’s dolphin (Lagenodelphis

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hosei) have even been declared unsuitable for captivity (for a review, see Mason, 2010). For other species such as killer whales (Orcinus orca), a shorter lifespan in captivity than in the wild and a high frequency

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of abnormal behaviours was reported (Jeff and Ventre, 2015), but the data and analysis have been showed to be flawed (Robeck et al., 2016). Conversely, bottlenose dolphins are successfully kept in captivity, with survival rates and life expectancies being at least as high as those of wild animals (Jaakkola and Willis, 2019). However, to our knowledge, no welfare-oriented study included several odontocete species to

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compare their reactions to captive environments. Bottlenose dolphins (BDs) are the most represented odontocetes in captive facilities worldwide (Pryor and Norris, 1998; Wells and Scott, 1999), and East Asian finless porpoises (Neophocaena asiaeorientalis sunameri) are commonly kept in captivity in Asia (Zhang et al., 2012). Unlike the Yangtze finless porpoise (YFP, N. a. asiaeorientalis), a freshwater species, the East Asian finless porpoise (EAFP), lives in marine environments (Jefferson and Wang, 2011). Both species are classified as endangered with the YFP being critically endangered under the criteria of the IUCN red list of

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threatened species (IUCN, 2013). Despite a captive breeding program started in 1996 for the YFPs in the only facility keeping this species under human care, few calves have been born and have survived in both species (Yang et al 1998, Wang et al 2005, Deng et al in press). Recently, several Chinese aquaria were allowed to keep YFPs for breeding purposes. Unlike BDs, FPs’ welfare indicators were never investigated and the fact these animals have a different morphology and habitat than BDs might result in different responses to similar stimuli. Clegg et al (2017a) suggested that any potential welfare measure should be

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validated by testing it in several situations that could elicit changes in welfare. The purpose of the current study was to analyse the effect of environmental and social factors that might impact individuals’ welfare

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on the swimming features of three groups of odontocetes under human care and to discuss their potential

Material and Methods

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use as welfare indicators.

The present study conforms to the 'Guidelines for the use of animals in research' as published in Animal

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Behaviour (1991, 41, 183–186).

2.1 Subjects, Housing and Group Composition

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The observations were conducted from early September 2017 to late October 2018. Five YFPs were observed in Baiji Dolphinarium, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan (Table 1). When all individuals were housed together, YFPs were kept in a kidney- shaped pool with a length of 20m, width of 7m and depth of 3.5m, linked by a corridor to a round pool with a 10m diameter and 3.5m

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depth. These two pools were separated by a gate allowing animals to see each other when separated. A third pool (13m diameter and 3.2m depth), not connected to the two others, was used from February 2017 to house the female, F7, and the male, Taotao, until F7 gave birth (after birth, she was alone in this pool with her calf and Taotao was moved back in the two-pool complex). For group management reasons (i.e. management of pregnant females), the social grouping changed several times during the data collection period (Figure 1). Since the three females gave birth during summer 2018, three calves were also present

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during certain periods of the data collection (two of them were only present for less than two weeks after their birth and the third one was present from its birth until the end of the data collection, Figure 1). Four EAFPs and five BDs were observed in Haichang Polar Ocean World, Wuhan (Table 1). EAFPs were always kept together in a 13.75m length, 8m width, 5.8m depth rectangular pool. BDs were kept in a three-pool complex, with two 8.86m diameter, 5m deep wide round pools (“small pools”) connected to the main pool of 27.44m long, 12m wide, and 6m deep (“big pool”). Depending on the observation sessions, animals had

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access to one, two or all pools. On January 16th, a new female arrived in the facility and the other female was placed with her starting January 23rd. When males and females were separated, females were kept in

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one of the round pools and males in the other round pool and/or in the main pool. On two occasions, the social grouping changed for a few days (Figure 1). The female, Beila, was absent from several morning

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observations because of a medical treatment administered in the medical pool during one month.

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YFPs were subject to four to six training sessions a day with no public presentation, but occasional visitors were allowed to watch animals both from the surface and from underwater windows. YFPs were

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fed between 3 and 3.5kg of thawed (Basilewsky) and/or live fish per day during training sessions. EAFPs were not trained but had three feeding sessions a day with a total feed of between 2.5 and 3kg of thawed

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fish (capelin, herring, squid, mackerel, greasy back shrimp, loach) per day, sometimes including live fish. BDs participated in three training sessions and two public presentations a day (up to five on particular days), within which they were fed between 10 and 13 kg of thawed fish (capelin, herring, squid, mackerel). Animals were provided human-made objects (i.e., toys) or live fish (for YFPs and EAFPs) at times decided by caretakers, and caretakers frequently interacted with BDs and YFPs outside of training sessions.

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All pools were frequently cleaned by divers and/or caretakers scrubbing the upper part of the pools’ walls.

2.2 Data Collection

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The data collection was conducted over 14 months for YFPs, and 12 months for EAFPs and BDs. Data were collected two days a week for each group, with no breaks during the data collection period (no week without data collection during the data collection period). A one-month preliminary ad libitum pilot study was conducted to identify and become familiar with each individual and, based on the literature, to build a common ethogram for the three species (Table 2). For the formal research protocol, each group was monitored minimum three times a day (in early

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morning, at noon and in the early afternoon), between training sessions/public presentations/feedings. Observation sessions consisted of 15 min video and voice recordings, using two to six cameras to monitor

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each group depending on the pool configuration. For YFPs, two underwater and two overhead monitoring cameras were used for the kidney shaped pool, one underwater camera for the connected round pool and

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two underwater and one overhead cameras for the disconnected round pool. For the EAFPs, two Xiaoyi 4K

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cameras were placed in front of two underwater windows. For BDs, two Xiaoyi 4K cameras were placed in front of a bubble shaped window situated five meters deep in the main pool and three other Xiaoyi 4K

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cameras were used to monitor this pool and the other pools from a bridge above. The position of the observation bridge and the small size and depth of round pools enabled the recording of behaviour from the surface only. Approximately 90% of every pool was covered by cameras with a good enough quality to be

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analysed. A complementary direct observation with a voice recorder or with the cameras’ audio recording was always conducted synchronously with the video recording to ensure identification of each individual and to narrate events for easier analysis.

During every data collection day, environmental data were noted. This data consisted of time of the

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day, delay to training (only for YFPs and BDs), social housing (all animals together: “altogether”, group divided in subgroups: “separated”, individual kept alone: “alone”), pools in which animals were observed (only for BDs, since their access to pools changed within and between days), presence of visitors, presence of enrichment, and any unusual event that occurred (“perturbation”, Table 3).

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Data were collected during 142 days for YFPs, 100 days for EAFPs and 100 days for BDs. In total, every YFP individual was monitored 135 hours (540 recording sessions), 76 hours for EAFPs (304 recording sessions), and 80 hours for BDs (320 recording sessions).

2.3 Analysis Videos were visually analysed to record all occurrences of previously defined swimming categories

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for each individual using incident sampling (Altmann, 1974, Table 2). Individuals could be engaged in several categories at the same time (e.g., group swimming and synchronous swimming, synchronous

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swimming and fast swimming, etc).

Statistical analysis was performed using R 3.5.2. The swimming direction (frequency and number of

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circles in each direction) was analysed for each species by fitting negative binomial generalized linear

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mixed-effects models (GLMMs) using the glmmADMB package (“glmmadmb()” function from the glmmADMB package, Skaug et al., 2016) to determine the prevailing direction for each species. Because

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we observed an obvious difference in swimming direction between him and others, the male EAFP, Xiaozhuang, was analysed separately from other EAFPs. In each model, the frequency or the number of

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circles were used as response variables and the direction was used as the predictor. The individual ID and the date were included in models as random factors for the three groups except for the analysis of Xiaozhuang (only the date).

For further analysis (i.e., analysis of the effect of environmental and social factors), to avoid a pool

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size bias, an estimation of the distance swam in each direction was calculated using the number of circles and the perimeter of each pool (YFPs: kidney shaped pool: 54m, connected round pool: 31m, non-connected round pool: 41m; EAFPs: 36m; BDs: big pool: 65m, small pools: 27m). The effect of environmental parameters (time of the day, delay to training, enrichment, potential perturbation, separation, presence of visitors and housing pool, Table 3) on the distance swam (clockwise distance and counter-clockwise distance) was analysed using negative binomial GLMMs fitted with the glmmADMB package. For each

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model, the distance swam was included as the response variable and the environmental parameters as predictors. The date and individual ID were included as random factors. Collinearity was tested for each model by checking predictors’ variance inflation factor (VIF) and collinear variables were removed or modified: for BDs, the enrichment variable that originally included five levels (types of enrichment) was transformed in a two-level variable (presence/absence of enrichment). For the analysis of the effect of environmental on the frequency of each swimming category (fast

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swimming, group swimming, synchronous swimming and contact swimming), negative binomial GLMMs were fitted with the lme4 package (“glmer.nb()” function, Bates et al., 2015). For each model, the frequency

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of each behaviour was included as the response variable and the environmental parameters as predictors. The date and individual ID were included as random factors. For each model, collinearity was tested by

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checking predictors’ VIF, and if existing, collinear variables (VIF>10) were removed from models: for

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YFPs, the public variable was removed from models including fast swimming and contact swimming as response variable and the social grouping variable was transformed into a two-level variables

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(altogether/separated) from models including synchronous swimming, contact swimming and group swimming as response variables, and for EAFPs, the enrichment and the perturbation variables were

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transformed into two-level variables. Wald chi-square tests were used to extract p values from all models. Pairwise tests were also conducted using Wald chi-square tests ran on models with data subsetting. Fitted means presented in tables and figures were extracted from models and back-transformed if needed (when

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using the glmmADMB package).

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Results

3.1 Swimming Laterality YFPs swam significantly more frequently and swam significantly more circles in the clockwise

direction than in the counter-clockwise direction (Table 4). EAFPs swam significantly more frequently and swam significantly more circles in the counter-clockwise direction than in the clockwise direction. The

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male Xiaozhuang swam significantly more frequently and swam significantly more circles in the clockwise direction than in the counter-clockwise direction. Finally, BDs swam significantly more frequently and swam significantly more circles in the counter-clockwise direction than in the clockwise direction.

3.2 Effect of environmental and social parameters 3.2.1 Circular swimming

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The three groups exhibited significant circular swimming distance differences depending on environmental and social parameters (Table 5). The time of the day significantly impacted YFPs’ counter-

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clockwise circular swimming with a higher distance swam in the morning than at noon and in the afternoon (Table 5a). The distance YFPs swam in clockwise direction was significantly lower before training than

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away from it, it was also lower when toy(s) were provided or when humans were interacting with animals

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than when no enrichment was provided. The distance YFPs swam in clockwise direction was significantly higher when noisy events occurred than when no perturbation occurred, and the distance YFPs swam in

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counter-clockwise direction was significantly higher when pool cleaning occurred than when no perturbation occurred. The distance YFPs swam in clockwise direction was significantly higher when

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animals were separated or alone than when altogether, and higher when alone than when separated, and the distance YFPs swam in counter-clockwise direction was significantly higher when housed alone than when altogether or separated. The distance YFPs swam in clockwise direction was significantly lower when many visitors were present than when a few or no visitors were, and the distance YFPs swam in counter-clockwise direction was significantly higher when few visitors were present than when many or none were.

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The time of the day significantly impacted EAFPs’ counter-clockwise circular swimming with a

higher distance swam in the morning than at noon and in the afternoon, and in the afternoon than at noon (Table 5b). The distance EAFPs swam in clockwise direction was significantly lower when new object(s) or live fish were provided than when no enrichment was, and the distance swam in counter-clockwise direction was significantly lower when toy(s) or live fish were provided than when no enrichment was

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provided. The distance EAFPs swam in clockwise direction was significantly higher when noisy events, pool cleaning or other perturbations occurred than when no perturbation occurred. The time of the day significantly impacted BDs’ clockwise circular swimming with a lower distance swam in the afternoon than in the morning and at noon (Table 5c). The distance BDs swam in counter- clockwise direction was significantly lower when toy(s) were provided or when humans were interacting with animals with toy(s) than when no enrichment was provided. The distance BDs swam in

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counter-clockwise direction was significantly higher when noisy events occurred than when no perturbation occurred. The distance BDs swam in counter-clockwise direction was significantly higher when animals

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were altogether than when separated. The distance BDs swam in clockwise direction was significantly lower when housed in the small pool than when housed in the big pool or when having access to both pools,

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and the distance BDs swam in counter-clockwise direction was significantly higher when having access to

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both pools than when housed in the small pool or in the big pool and higher when housed in the small pool

3.2.1 Other swimming behaviours

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than in the big pool.

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The three groups exhibited significant differences in their swimming behaviour depending on environmental and social parameters (Table 6). YFPs engaged in fast swimming significantly less in the morning than at noon and in the afternoon, they engaged in synchronous swimming the most in the morning, followed respectively by at noon and in the afternoon (Table 6a). Contact swimming was significantly less

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frequent at noon than in the morning and in the afternoon, and group swimming was significantly more frequent in the morning than at noon. Synchronous swimming and group swimming were significantly less frequent for YFPs before training than away from it. The frequency of fast swimming and contact swimming was significantly lower when live fish was provided than when no enrichment was provided, and the frequency of synchronous swimming was significantly lower when toy(s) or live fish were provided or when humans were interacting with animals (with or without toy(s)) than when no enrichment was

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provided. The frequency of group swimming was significantly lower when new object(s) were provided or when humans were interacting with animals (with or without toy(s)) than when no enrichment was provided. The frequency of contact swimming was significantly higher during pool cleaning than when no perturbation occurred. The frequency of fast swimming, synchronous swimming and group swimming was significantly higher during pool cleaning or other perturbations than when no perturbation occurred. The frequency of synchronous swimming and group swimming was also significantly higher when noisy events

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occurred (and when social events occurred for group swimming) than when no perturbation did. The frequency of fast swimming and synchronous swimming was significantly higher, and the frequency of

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group swimming significantly lower when YFPs were separated than when not. The frequency of synchronous swimming was the highest when few visitors were present, followed respectively by when no

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visitors were present and when many were, and the frequency of group swimming was lower when no

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visitors were present than when few or many visitors were present.

EAFPs engaged in synchronous swimming and group swimming significantly less at noon than in

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the morning and in the afternoon, and they engaged in synchronous swimming and group swimming significantly less when enrichment was provided than when not (Table 6b). They engaged in fast swimming,

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synchronous swimming, contact swimming and group swimming significantly more often when a perturbation occurred than when not. EAFPs engaged in fast swimming significantly more often when no visitors were present than when few were, and in contact swimming significantly the most when many visitors were present, followed respectively by when few visitors were present and by when no visitors

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were. They engaged in group swimming significantly less when many visitors were present than when few or none were.

BDs engaged in fast swimming significantly less at noon than in the morning and in the afternoon, and

in synchronous swimming less in the afternoon than in the morning and at noon (Table 6c). They engaged in group swimming the most in the morning, followed respectively by at noon and in the afternoon. Fast swimming was significantly more frequent and synchronous swimming was significantly less frequent for

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BDs before training than away from it or when training other animals. The frequency of group swimming was significantly lower when toy(s) were provided or when humans were interacting with animals with toy(s) than when no enrichment was provided. The frequency of fast swimming was significantly higher during pool cleaning, social events or other perturbations than when no perturbation occurred. Synchronous swimming, contact swimming and group swimming were more frequent during pool cleaning than when no perturbation occurred. The frequency of group swimming was significantly lower when BDs were

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separated than when not. The frequency of fast swimming was the highest when having access to both pools, followed respectively by when housed in the large or the small pool. Synchronous swimming and contact

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swimming were significantly more frequent when housed in the large pool than when having access to both pools. Group swimming was significantly less frequent when having access to both pools than when housed

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in the large or the small pool. The frequency of contact swimming was significantly higher when few

Discussion

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visitors were present than when not.

First, this study highlighted differences in circular swimming direction bias between groups of

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odontocetes under human care. Second, the circular distance swam, and the frequency of fast swimming, synchronous swimming, contact swimming and group swimming were influenced by environmental and social factors in each group.

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4.1 Swimming laterality

Several studies on captive dolphins have reported a bias in swimming direction, which was

described to be mostly counter-clockwise (Caldwell et al., 1965; Ridgway, 1990; Sobel, 1994). However, other BDs and belugas were observed with the opposite swimming lateralization (Marino and Stowe, 1997a, b). Here, different groups exhibited different biases: BDs and EAFPs (except Xiaozhuang) were mostly swimming counter-clockwise while YFPs were mostly swimming clockwise. This group of YFPs has

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already been reported swimming in clockwise more than in counter-clockwise direction (Platto et al., 2017). The direction of swimming places one of the eyes towards the walls and windows and thus towards any events outside the pool which could be of importance for the animals. This bias could result from hemispheric dominance playing a role in the treatment of visuospatial information (Kilian et al., 2000). Animals may prefer to watch things with one eye more than with the other, which would then explain the swimming direction bias. However, the fact that all individuals in one group exhibit the same bias and that

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different direction biases are found in different groups might suggest a social factor. Animals born later or arriving later could simply follow former ones, resulting in a same bias for all individuals. In this study, the

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swimming laterality of one EAFP male was analysed separately from others because it exhibited a clockwise direction bias while other EAFPs were mostly swimming counter-clockwise. This individual was

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the oldest one, and was present in the facility before the three younger EAFPs. The newest members of the

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social group might already have had a counter-clockwise bias when they arrived and did not change to follow the already present male. The exception was the EAFP male, considered the lowest ranked (Serres

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et al., in press), and that, unlike other individuals, was displaying abnormal/stereotypical behaviours (e.g., rubbing its belly for long periods of time on the floor at the exact same place, rubbing against a wall at the

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exact same place after each circle it swam, floating at the surface at the exact same place for long periods of time). This male might have been experiencing a different psychological state than others, which could be another explanation for his difference.

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4.2 Effect of environmental and social parameters 4.2.1 Circular swimming In all groups, the circular swimming distance was higher in the morning and/or at noon than in the

afternoon. It has already been shown that captive odontocetes were less active in the afternoon (Serres et al., 2017; Delfour and Aulagnier, 1997). This diurnal pattern should be taken into account when wanting to monitor behaviour, especially when attempting to observe behavioural changes: observations have to be

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conducted at the same time every day to avoid an activity level bias. The choice of the best time to conduct observations might be discussed in each facility depending on the specific goal of the behavioural monitoring. For YFPs, the distance swam in their preferred direction was also lower right before training than at other times, which might be explained by their display of anticipatory behaviours (e.g., spy-hops) and by their tendency to stay close to the trainers’ office before training. Enrichment is often used for captive odontocetes (Kuczaj et al., 2002; Delfour and Beyer, 2012;

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Clark, 2013a; Eskelinen et al., 2015; Perez et al., 2017; Serres and Delfour, 2017). As enrichment efficiency and consequences on animals’ behaviour might depend on species and even groups (Mellen and Sevenich,

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MacPhee 2001; Kuczaj et al., 2002; Eskelinen et al., 2015), studying each captive group in order to optimize the use of enrichment is needed. It was suggested that behavioural monitoring should be conducted as an

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essential part of captive programs to provide information on animals’ welfare state and responses to

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enrichment (Watters et al., 2009; Brando et al., 2017). Here, the distance animals swam in their preferred direction was lower for the three groups with the presence of enrichment: toy(s) and human(s) for YFPs

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and BDs, and toy(s) and live fish for EAFPs. Studies reported captive odontocetes as more or less frequently swimming in a repetitive way and following a fixed pattern around the pool (Gygax, 1993; Sobel et al.,

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1994; Sekiguchi and Koshima, 2003; Singh, 2005; Miller et al., 2011). This kind of repetitive circular swimming could be categorized as stereotypical because it is fulfilling the definition of such behaviours (Mason, 1991), however most researchers argue that studies are missing to categorize it as such (Brando et al., 2017; Clegg et al., 2017a). Additionally, even if novel enrichment was already shown to reduce the

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frequency of circular swimming in a young bottlenose dolphin male (Bahe, 2014), and if belugas showed less solitary and circular swimming when enrichment was present (Hill and Ramirez, 2014). no clear link was found between circular swimming and good or poor welfare state (Mason and Latham ,2004). The fact the presence of enrichment decreased circular swimming at least highlights the fact that enrichment has the benefit of diversifying the animals’ activities (i.e., if not engaged in circular swimming, they are doing something else).

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Oppositely, the distance swam in the animals’ preferred direction was higher when potential perturbations –noise for YFPs and BDs, and pool cleaning, noise and other events for EAFPs- occurred than when no perturbation occurred. In addition, for YFPs, the distance swam in their not preferred direction was higher during pool cleaning than when no perturbation occurred. Platto et al. (2017) and Ugaz et al. (2013) suggested that more tests including different stimuli should be conducted to determine if group dynamics or environmental factors were influencing animals’ circular swimming frequency and swimming

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direction bias. Oppositely to the presence of enrichment, the distance animals swam in their preferred direction was higher when potential perturbations –noise for YFPs and BDs, and pool cleaning, noise and

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other events for EAFPs- occurred than when no perturbation occurred. Animals might have reacted to these events by swimming more and/or faster. In addition, for YFPs, the distance swam in their not preferred

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direction was higher during pool cleaning than when no perturbation occurred. This increase in circular

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swimming in the direction animals usually use less could be a sign of a change in their emotional state, and indicate stress (Barnard et al., 2015). A change in swimming direction has already been observed in a

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captive beluga following a novel stimulus (Marino and Stowe, 1997b). Perturbations faced by YFPs were not always new: divers or noise were occurring frequently, but animals did not seem to get used to it. A recent review pointed out the need to investigate how husbandry and/or management decisions

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affect captive odontocetes’ social life, including separation of individuals (Brando et al., 2017). For YFPs and BDs, the distance swam in their preferred direction was higher when separated in subgroups, and for YFPs, the distance swam in their not-preferred direction was the highest when housed alone. When

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separated or housed alone, animals have fewer partners to interact with and, therefore, a lower diversity of behaviours to express (i.e., less social behaviours), which can be an explanation for this increased circular swimming distance. A higher level of circular swimming could thus be a sign of a lack of stimuli, but for these social animals, it could also reflect a change in their emotional state when separated from others. The presence of visitors has been shown to impact several species’ behaviour in captivity (Mallapur et al., 2005; Wells, 2005; Davey, 2006, 2007). Negative behavioural indicators in primates have been shown

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to increase with noise and density of visitors (Wells, 2005; Cooke and Schillaci, 2007), but these kind of studies have never been conducted on odontocete species. Since it is thought that the acoustic perception threshold and sensitivity to noise of a species influences its responses to the presence of visitors (Heffner and Heffner 2007) and since odontocetes are well known to be sensitive to noise (Buckstaff, 2004; David, 2006), they are good candidates to study the impact of visitors who are often a source of noise. However, captive odontocetes are sometimes observed interacting with visitors through underwater windows (Trone

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et al., 2005; Brando et al., 2017), revealing a potential enriching property of the presence of public. Nonetheless, the density of visitors’ effect on their behaviour was rarely studied. The only significant

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visitors’ effect found in this study was for YFPs: the distance they swam in their preferred direction was the lowest when many visitors were present. The presence of public might have increased vigilance or

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anticipatory behaviours, which could explain this decrease in circular swimming in the prevalent direction

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when many visitors were present. Animals were also observed interacting with people through underwater windows when visitors were present. In addition, when few people were present, it was often people from

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the research base whereas when many people were present, they were usually unfamiliar. YFPs might discriminate between familiar and unfamiliar people and may find unfamiliar people more interesting,

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which would result in less swimming behaviours. For the distance swam in the not-preferred direction, the effect of public was bell shaped, with animals being engaged in counter-clockwise swimming when few visitors were present. The presence of few people often implied noise and proximity with the pool, whereas thanks to the instruction given to large groups of visitors, their presence usually did not last long nor implied

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high levels of noise. These differences could be a potential factor that played a role in this bell-shape pattern. Finally, the distance BDs swam in their preferred direction was the highest when having access to

both pools. This pattern might be explained by the fact that fast swimming was significantly more frequent when having access to both pools or when housed in the large pool than when housed in the small pool and more frequent when having access to both pools than when housed in the big pool - their higher speed allowing them to swim a longer distance. In addition, the distance they swam in their not-preferred direction

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was the lowest when housed in the small pool. We often observed BDs swimming in this direction during intense social interactions (e.g., social play, agonistic interactions), which happened more often when having access to the large pool.

4.2.2 Other swimming behaviours Like what was observed for circular swimming, the frequency of most swimming behaviours was

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higher in the morning than at other times of the day for the three groups. This pattern might again be due to the higher activity levels in the morning in captive odontocetes.

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Social swimming (synchronous swimming and group swimming) was less frequent for YFPs and BDs, and fast swimming was more frequent for BDs before training than away from it (or during the training

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of other animals for BDs). This pattern can be explained by the fact animals were often displaying

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anticipatory behaviours (e.g., spy-hops) and were staying close to the trainers’ office (YFPs) or to the beach (BDs) before the training. In addition, for BDs, fast swimming was often accompanied by jumping or other

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aerial or noisy behaviours (e.g., noisy acoustic emissions, tail slaps on the water surface) before training or when other animals were being trained. These behaviours, including fast swimming could be indicative of

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an excited or frustrated emotional state in reaction to this context. The presence of enrichment seemed to decrease social swimming in the three groups (contact swimming and synchronous swimming for YFPs, synchronous swimming and group swimming for EAFPs, and group swimming for BDs). Among types of enrichment, live fish, toy(s) and humans for YFPs, and

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toy(s) and humans for BDs affected these behaviours. The impact of the presence of humans interacting with animals on their behaviours is congruent with the fact BDs are attracted by the interaction with humans outside of training sessions and are more likely to play with toys when humans are involved (Delfour and Beyer, 2012; Eskelinen et al., 2015). Toys and live fish also seemed to be effective in affecting the animals’ behaviour. Individual preferences for particular items have been shown in BDs (Delfour and Beyer, 2012; Eskelinen et al., 2015), a deeper analysis on the effect of each type of enrichment on each individual’s

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behaviour might be useful. In most facilities, live fish is rarely provided to captive odontocetes as part of a routine management, or as enrichment (Brando et al., 2017), a deeper investigation on the effect of this kind of enrichment would also be useful to determine the best way to use it. Oppositely, fast swimming and social swimming were more frequent when perturbations occurred for all three groups. This pattern is congruent with the fact that wild odontocetes have been reported to react to disturbances (boats) by getting closer to each other (killer whales: DeNardo, 1998, bottlenose dolphin:

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Nowacek et al., 2001) and swimming quicker (killer whales: Kruse, 1991, bottlenose dolphins: Nowacek et al., 2001). Synchronous swimming has been often studied in dolphins (Connor et al 2006; Sakai et al

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2010) and is thought to indicate social affiliation (Connor et al., 2006; Clegg et al., 2017a) and to be linked with positive emotions (Clegg et al., 2017b). Synchronous swimming was observed in captive BDs when

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new items were introduced in the pool (McBride and Hebb, 1948). Social support, expressed here by social

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swimming, might be an important factor helping odontocetes to cope with stress (Waples and Gales, 2012; Fellner et al., 2013), with strong inter-individual bonds increasing animals’ survivorship in dolphins (Frere

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et al., 2010; Stanton and Mann, 2012). It was suggested that synchronous swimming could serve both as an affiliative behaviour and as a reaction to disturbances in dolphins (Hastie et al., 2003; Senigaglia et al.,

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2012). Thus, it could be an ambiguous cue to indicate good or poor welfare in this species. Until now, no study confirmed if this behaviour could be used as an indicator of positive emotions in odontocete species (Connor et al., 2006; Holobinko and Waring, 2010; Kuczaj et al., 2013). Synchronous swimming should thus be studied further to be validated or not as a welfare indicator (Clegg et al, 2017b). Regarding group

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swimming, it might be a strategy to increase vigilance and to decrease the risk of predation (Norris and Schilt, 1987; Fellner et al., 2006; Senigaglia and Whitehead, 2011). This behaviour could be an answer to potentially acute stressful environmental changes, suggesting its usefulness to measure the effect of management decisions or unusual events that occur in captive facilities on the emotional state of these animals. In all groups, almost all kinds of social swimming recorded here were higher when a potential perturbation occurred and lower with the presence of enrichment, suggesting their link with animals’

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emotional state. Baseline behavioural pattern should always be assessed prior to attempting to use such behaviours to monitor stress or welfare. Among potential perturbation types, noise and pool cleaning were the events that provoked the highest behavioural change in YFPs and BDs. Odontocetes are sensitive to noise which can affect their behaviour (Buckstaff, 2004; David, 2006). They have been shown to change their swimming direction (Miller et al., 2015), increase their swimming speed and avoid the noise source (Perry, 1998), which is congruent with our results. The reaction to divers or caretakers cleaning the pool

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might be due to the presence of something (the diver or the brush) in the water. Even though these cleaning events occurred frequently and for years in each facility, animals did not seem to adapt and still reacted

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strongly. These kinds of reaction should be studied further to find ways to reduce the acute response of the animals, that could be a sign of negative stress.

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The frequency of fast swimming and synchronous swimming was higher for YFPs when separated,

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and the frequency of group swimming was lower for YFPs and BDs when separated than when not. We observed that the reaction to a potential perturbation changed with the social grouping: when not separated,

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YFPs mostly swam in group (not particularly fast) when a perturbation occurred, while, when separated, they usually swam synchronously and/or fast when the same kind of perturbation occurred. We hypothesize

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that the separation increased the reaction to perturbation in these animals. Being separated lowers the group effect that helps individuals to cope with stress (Waples and Gales, 2012; Fellner et al., 2013), therefore animals could react in a stronger way, but more data is needed to validate this hypothesis. When many visitors were present, YFPs engaged more often in group swimming, EAFPs and BDs in

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contact swimming, which suggests that this presence could be a kind of stressful event resulting in animals getting closer to each other. However, the frequency of synchronous swimming was the highest for YFPs when few visitors were present and fast swimming and group swimming were the highest when no public was present for EAFPs. These unclear patterns might once again be explained by the features of the visitors, and we suggest that in further studies, more parameters such as visitors’ noise or activity should be recorded to obtain results that can be interpreted better.

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Finally, for BDs, the frequency of fast swimming was the highest, and group swimming the lowest when having access to both pools or when housed in the large than when housed the small pool. The access to a larger space probably allowed them to swim faster, especially when chasing each other during social play or agonistic interactions (Serres et al., 2019). Regarding group swimming, it is possible that being housed in a very small pool that increases proximity between animals also increased the frequency of such

5

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behaviour.

Conclusion

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All swimming features investigated in this study were modulated by environmental and/or social factors. We first suggest that circular swimming could be used as a tool to measure the efficiency of enrichment,

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enrichment being designed to increase the animals’ behavioural diversity and to prevent boredom. Secondly,

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the circular swimming direction should be studied further to investigate if swimming in the not-preferred direction could provide information about in captive odontocetes’ emotional state. Social swimming as well

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as fast swimming seem to decrease with enrichment and to increase during potential perturbations, suggesting their use to monitor the impact of routine or unusual events on the animals. Among social swimming behaviours, we suggest that synchronous swimming and contact swimming should be used very

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carefully when wanting to monitor animals’ welfare state because they can both be displayed in a relaxed bonding context and in a stressful context. The swimming speed might be a useful parameter to differentiate between these contexts: animals swim faster when facing stressful events. Group swimming and fast

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swimming might less ambiguous tools to use to monitor animals’ emotional state, especially for FPs that rarely engage in intense social interactions involving fast swimming and that can reflect a positive state (e.g. social play). We suggest that monitoring swimming behaviours as we did in this study can further be standardized to be utilized as a tool. Facilities could establish baseline levels for each species and for each animal to monitor their responses to environmental and social changes as a group and as individuals.

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The differences observed depending on the time of the day should be taken in account when using behaviour to evaluate welfare. A low frequency of a certain behaviour at a certain time of the day is potentially “normal”, while the same frequency at another time of the day potentially indicates a change of the animal’s state. The fact that each group reacted slightly differently to the environmental and social factors we analysed confirmed that even closely related species can differ in terms of response to stress and welfare indicators (Mason, 2010). Moreover, as we mentioned here with case of the EAFP male,

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Xiaozhuang, all individuals do not behave the same and may likewise not react similarly to environmental or social events. Each animal’s personality might play a role in the behaviours it displays and in its response

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to environmental changes (Kuczej et al., 2012; Birgersson et al., 2014). Welfare assessments using behavioural tools should follow a study of each individual’s behaviour to be able to detect behavioural

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changes for each animal. We finally suggest that more studies should be conducted to validate the potential

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indicators we analysed here in other groups and that more environmental factors such as noise level should be tested. The behaviour and welfare of FPs should particularly be focused since more individuals have

Acknowledgements

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been recently allowed to be kept in Chinese aquaria for breeding purposes.

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We are grateful to the caretakers’ teams of Baiji dolphinarium and Wuhan Haichang Polar Ocean World who allowed us to conduct our observations. Hunter Doerksen revised the English language. A financial contribution for Ph.D. students was supplied by the CAS-TWAS President’s fellowship. This work was

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supported by the Ocean Park Conservation Foundation Honk Kong (AW01-1819).

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Serres, A., Delfour, F., 2017. Environmental changes and anthropogenic factors modulate social play in captive bottlenose dolphins (Tursiops truncatus). Zoo Biol. 36(2), 99-111.

-p

Singh, S., 2005. Welfare assessment of captive bottlenose dolphins (Tursiops truncatus) in captivity.

re

Master’s thesis, Universidad Nacional Autónoma de México: México.

Skaug, H., Fournier, D., Bolker, B., Magnusson, A., & Nielsen, A. (2016). Generalized Linear Mixed

lP

Models using 'AD Model Builder'. R package version 0.8.3.3.

Sobel, N., Supin, A.Y., Myslobodsky, M.S., 1994. Rotational swimming tendencies in the dolphin

ur na

(Tursiops truncatus). Behav. Brain Res. 65, 41-45.

Spruijt, B.M., van den Bos, R., Pijlman, F.T., 2001. A concept of welfare based on reward evaluating mechanisms in the brain: anticipatory behaviour as an indicator for the state of reward systems. Appl. Anim. Behav. Sci. 72, 145-171.

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Stanton, M.A., Mann, J., 2012. Early social networks predict survival in wild bottlenose dolphins, PLoS One. 7, 1–6.

The paper, 2018. https://www.thepaper.cn/newsDetail_forward_2324329 (Accessed 18 June 2019). Trone, M., Kuczaj, S., Solangi, M., 2005. Does participation in Dolphin–Human Interaction Programs affect bottlenose dolphin behaviour? Appl. Anim. Behav. Sci. 93(3), 363-374.

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Ugaz, C., Valdez, R.A., Romano, M.C., Galindo, F., 2013. Behavior and salivary cortisol of captive dolphins (Tursiops truncatus) kept in open and closed facilities. J. Vet. Behav.: Clinical Applications and Research 8, 285-290. Usama, A.A., 2011. The effects of cage enrichment on agonistic behaviour and dominance in male laboratory rats (Rattus norvegicus). Res. Vet. Sci. 90(2), 346-351. Wang, D., Hao, Y., Wang, K., Zhao, Q., Chen, D., Wei, Z. Zhang, X., 2005. The first Yangtze finless

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porpoise successfully born in captivity. Env. Sci. Poll. Res. 12, 247-250. Waples, K.A., Gales, N.J., 2002. Evaluating and minimising social stress in the care of captive bottlenose

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dolphins (Tursiops aduncus). Zoo Biol. 21, 5-26.

Watters, J.V., 2014. Searching for behavioral indicators of welfare in zoos: Uncovering anticipatory

-p

behavior. Zoo Biol. 33, 251-256.

re

Webster, J., 2005. Challenge and response. in: Kirkwood, J.K., Hubrecht, R.C., Roberts. E.A. (Eds.), Animal welfare: limping towards Eden. Oxford, RU: Blackwell Publishing Ltd.

lP

Wells, R.S., Scott, M.D., 1999. Bottlenose dolphin: Tursiops truncatus (Montagu, 1821). in: Ridgway, S.H., Harrison, R. (Eds.), Handbook of Marine Mammals: The Second Book of Dolphins and Porpoises. Academic Press: San Diego, CA, pp. 137-182.

ur na

Wells, D.L., 2005. A note on the influence of visitors on the behaviour and welfare of zoo-housed gorillas. Appl. Anim. Behav. Sci. 93, 13–17.

Yang, J., Zhang, X.F., Yukiko, H., Asami, F., 1998. Observation of parturition and related behaviors of

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finless porpoise (Neophocaena phocaenoides) in Enoshima aquarium, Japan. Chinese Journal of Oceanology and Limnology 29, 41-46. Yeates, J.W., Main, D.C.J., 2008. Assessment of positive welfare: a review. The Vet. J., 175, 293-300. Zhang, P., Sun, S., Yao, Z., Zhang, X., 2012. Historical and current records of aquarium cetaceans in China. Zoo Biol. 31(3), 336-349.

30

31

of

ro

-p

re

lP

ur na

Jo

Table 1. Catalogue of studied individuals’ features (YFP: Yangtze finless porpoise, EAFP: East-Asian finless porpoise, BD: bottlenose dolphin)

Name

Species

Age

Weight

Length

(year)

(Kg)

(m/h)

Sex

Facility

YFP

M

8

NA

157

Baiji dolphinarium, IHB

F7*

YFP

F

8

NA

145

Baiji dolphinarium, IHB

F9*

YFP

F

8

NA

145

Baiji dolphinarium, IHB

Taotao

YFP

M

14

NA

156

Baiji dolphinarium, IHB

Yangyang*

YFP

F

11

NA

147

Baiji dolphinarium, IHB

Xiaomeng

EAFP

F

4

33

1.43

Haichang Wuhan Polar Ocean park

Xiaomi

EAFP

M

7

31

1.60

Xiaoxi

EAFP

M

4

41.5

1.49

Xiaozhuang

EAFP

M

7

48

Ailun

BD

M

13

280

Beila

BD

F

11

Jiesen

BD

M

14

R*

ro

-p

Haichang Wuhan Polar Ocean park

re

Haichang Wuhan Polar Ocean park Haichang Wuhan Polar Ocean park

2.74

Haichang Wuhan Polar Ocean park

250

2.52

Haichang Wuhan Polar Ocean park

290

2.69

Haichang Wuhan Polar Ocean park

lP

1.70

ur na

Luoke

of

Duoduo

BD

M

13

260

2.70

Haichang Wuhan Polar Ocean park

BD

F

15

260

2.55

Haichang Wuhan Polar Ocean park

Jo

*pregnant females

32

Table 2. Catalogue of behaviours and interactions used for the video analysis Behavioural

Behaviour and description

Analysed parameter

category Swimming pattern Circular swimming

Individual

is

swimming

in Frequency, number of circles,

direction. One circle is at least

ro

the size of three third of the pool

of

clockwise or counter-clockwise distance swam

-p

and at least three third of it is close to a wall. Circular

swimming

re

Clockwise swimming

individual

swimming

with Frequency, number of circles, in distance swam

Counter-clockwise

Circular

swimming

with Frequency, number of circles,

individual swimming in counter- distance swam

ur na

swimming

lP

clockwise direction.

clockwise direction.

Fast swimming

Individual is swimming fast, Frequency making waves on the surface, or

Jo

making riddles on its skin

Social

Any swimming event involving several individuals

swimming

Synchronous swimming

Two

or

more

individuals Frequency

swimming more or less close to

33

each other (no more than one body length), with synchronous swimming movements Contact swimming

Two

individuals

swimming Frequency

close to each other with a part of their body in contact (pectoral

was made with the genital parts

ro

of one individual, it was not

of

fin, belly-back). If the contact

recorded as contact swimming At

least

three

individuals Frequency

-p

Group swimming

re

swimming in the same direction with a distance of less than one

Jo

ur na

lP

body length between them

34

day

training

Morning

Away

(from 8am to

training

to Social grouping

Perturbation

Altogether

None

from

11:30am)

11:30am

to

2pm)

Before

training Separated

(not

alone,

gate

(recording

Noise (construction work

ending less than five before

YFP

minutes the

between

groups

allowing

visual

noise or loud people

Toy(s) (balls)

underwater windows or next to the pool: employees or visitors)

acoustic

ur na

training)

Low (<5 persons in front of

noise)

and

Visitors

None

re

(from

Housing pool

None

lP

Noon

Enrichment

ro

Delay

Species

-p

Time of the

of

Table 3. Environmental and social factors’ features

NA

contact)

Afternoon

Alone

between

5pm)

individual

Jo

(from 2pm to

(gate single

Pool

cleaning

and/or

(diver caretaker

Human(s)

(caretakers

interacting with animals High (>5 persons in front of

and

scrubbing

from

the

outside

of

training underwater windows or next to

others

allowing

visual

and

surface using long handle

sessions at the surface of

brushes)

from

the pool: visitors)

acoustic contact)

windows)

35

underwater

after

separation

or

Live fish

-p

ro

reunion of groups)

Human(s) + toy(s)

of

Social perturbation (right

New object in the water

re

Other (shoal of small fish

(soundtrap,

stretcher,

in the pool, water level experiments’ material,

unusually high or low)

lP

NA

ur na

Morning

(from 8am to 11:00am) EAFP

new toy)

None

None

NA

Noon

None

NA

(from

Pool cleaning (diver or

to

Jo

11:00am

Low (<15 visitors in front of Toy(s) (ball, Soundtrap)

small boat)

underwater windows)

1pm)

36

of

Afternoon Human(s)

(public

4pm)

interacting

through

High (>15 visitors in front of underwater windows)

ro

(from 1pm to

underwater windows)

work,

(construction

-p

Noise

New object (Soundtrap,

microphone

filtration items)

re

speakers)

lP

Other (water unusually high, unknown person

Morning

ur na

sampling water)

Away

(from 8am to 11:30am) Noon

(from

Jo

BD

training

11:30am 1.45pm)

to

from

Small

Altogether

None

None

frequent

None

housing pool)

Separated

(gate

allowing

visual

Large

(public

Low (1-8 persons next to the

Toy(s) (balls, buoys, Pool cleaning (divers)

and

(most

acoustic

presentation ropes + buoys) pool)

contact)

37

pool, usually employees)

Before

(from 2pm to

(recording

Human(s)

1:45pm)

ending less than

interacting with animals

ro

before

minutes

outside

the

re

training

of other animals

Jo

ur na

when separated)

lP

(belugas or other group of dolphins

of

training

sessions)

training) During

(caretakers

-p

five

training

of

Afternoon

38

Human(s) + toy(s)

Access to both pools

of ro

Noise

(public rehearsal,

-p

presentation

Jo

ur na

lP

re

construction work)

39

of ro -p re lP

ur na

Table 4. Fitted means and standard-errors of the number of circles and frequency of swimming in each direction per observation session (15 min) for each species, and outputs from Wald chi-square tests ran on GLMMs. Species

Parameter

Number of circles

Jo

YFPs

Frequency

Number of circles

EAFPs

Xiaozhuang (EAFP)

Frequency Number of circles

Direction clockwise counter-clockwise clockwise counter-clockwise clockwise counter-clockwise clockwise counter-clockwise clockwise

Fitted mean±SE 19.199±0.126 0.308±0.146 6.204±0.127 0.173±0.148 0.986±0.081 14.267±0.091 0.838±0.042 7.55±0.044 5.945±0.084

40

Wald chi-square output χ² = 3588.20 p < 0.001*** χ² = 4679.60, p < 0.001*** χ² = 5818.30, p < 0.001*** χ² = 2392.30, p < 0.001*** χ² = 56.70, p < 0.001***

χ² = 46.32, p < 0.001*** χ² = 5482.4, p < 0.001*** χ² = 2414.3, p < 0.001***

-p

Frequency

of

Number of circles BDs

2.801±0.13 4.201±0.068 2.342±0.111 0.337±0.131 26.244±0.136 0.257±0.081 4.235±0.08

ro

Frequency

counter-clockwise clockwise counter-clockwise clockwise counter-clockwise clockwise counter-clockwise

re

Table 5. Fitted means and standard-errors of the distance swam per observation session (15 min) in each direction for YFPs (a), EAFPs (b) and BDs (c), and outputs from Wald chi-square tests ran on GLMMs. Letters and asterisks denote significant pairwise differences (p < 0.05): for parameters

lP

with less than four levels, levels sharing the same letters do not differ significantly, and for parameters with more than four levels, levels owning and asterisk significantly differ from the absence of stimuli level (“none”). Clockwise swimming distance

Parameter Time of the day to

Level

Fitted mean±SE

Wald chi-square output

Morning Noon Afternoon Away from training Before training None Toy(s) Human(s) Human(s) and toy(s) Fish None

729.423±0.55 672.663±0.552 667.691±0.542 719.468±0.547 205.855±0.504 746.005±0.557 499.245±0.586 465.426±0.584 550.388±0.633 387.972±0.514 671.665±0.56

a a a a b

Jo

Delay training

ur na

(a)

Enrichment

Perturbation

* *

χ² = 2 3.779, p = 0.151

χ² = 17.7804, p < 0.001***

χ² = 3.413, p= 0.637

χ² = 10.852, p = 0.028 *

41

Counter-clockwise swimming distance Fitted Wald chi-square output mean±SE 5.364±3.201 a 1.491±3.185 b χ² = 11.816, p = 0.003** 0.857±3.199 b 2.364±3.198 a χ² = 0.222, p = 0.638 0.019±2.876 a 2.624±3.235 1.583±3.395 χ² = 3.650, p= 0.601 0.387±3.405 0.194±3.503 0.156±2.928 1.762±3.241 χ² = 31.446, p < 0.001***

(b)

Time of the day

Enrichment

Visitors

χ² = 33.584, p < 0.001***

Morning Noon Afternoon None Toy(s) Human(s) Fish New object None Pool cleaning Noise Other None Few

28.867±1.953

a

Many

27.122±1.94

a

Level

9.546±3.157 5.032±3.409 0.582±3.254 3.316±3.119 1.162±3.262 2.287±3.216 13.58±3.146 1.365±3.204 8.996±3.305 0.916±3.106

of ro

-p

χ² = 71.281, p < 0.001 ***

Clockwise swimming distance Fitted Wald chi-square output mean±SE 21.954±1.976 a 33.604±1.948 a χ² = 5.996, p = 0.0699 42.214±1.942 a 26.642±2.172 36.808±2.156 39.192±2.204 χ² = 11.390, p= 0.023 * 2.938±1.764 * 1.179±2.051 * 24.008±2.022 50.173±2.076 * χ² = 18.2048, p = 0.001** 65.673±1.954 * 431.728±2.332 * 41.198±1.983 a

Jo

Perturbation

a b c a a b

ur na

Parameter

*

re

Visitors

547.698±0.532 1329.009±0.563 605.531±0.562 705.304±0.552 308.42±0.568 923.4±0.557 1712.526±0.527 710.415±0.558 747.35±0.554 301.455±0.521

lP

Social grouping

Pool cleaning Noise Social event Other Altogether Separated Alone None Few Many

χ² = 1.393, p = 0.498

42

*

a a b a b a

χ² = 19.281, p < 0.001***

χ² = 4.844, p = 0.089

Counter-clockwise swimming distance Fitted Wald chi-square output mean±SE 627.222±0.658 a 401.004±0.656 b χ² = 10.408, p = 0.005 ** 491.056±0.651 c 628.885±0.716 482.798±0.708 * 445.627±0.713 χ² = 3.808, p = 0.433 560.186±0.6 * 259.815±0.737 488.997±0.684 470.077±0.689 χ² = 12.233, p = 0.162 538.2±0.723 745.278±0.713 515.907±0.662 a 494.9±0.655

a

471.376±0.648

a

χ² = 0.686, p = 0.709

to

Enrichment

Perturbation

Social grouping Housing pool

Few

Wald chi-square output

872.092±0.237 824.062±0.238 778.945±0.229 826.155±0.231

a a a a

854.61±0.249

a

of

Fitted mean±SE

8.53±1.61 10.494±1.589 4.642±1.863 4.701±1.414 7.713±1.58 12.076±1.455 6.139±2.019 11.545±1.712 4.891±1.825 7.388±1.435 8.21±1.569 2.355±1.632 57.772±1.47 24.613±1.571 8.053±1.561

a a a b b a

8.167±1.498

a

χ² = 1.535, p = 0.674

χ² = 1.648, p = 0.800

χ² = 0.037, p = 0.848 χ² = 39.176, p < 0.001 ***

χ² = 0.001, p = 0.977

887.775±0.243 684.331±0.252 1033.278±0.255 446.56±0.212 816.498±0.239 793.113±0.216 1222.076±0.291 882.244±0.258 838.994±0.276 1065.351±0.216 790.858±0.236 865.588±0.238 633.272±0.228 1210.814±0.244 825.613±0.235 840.069±0.226

*

χ² = 3.979, p = 0.137

χ² = 0.126, p = 0.722

χ² = 81.295, p < 0.001***

*

*

χ² = 5.728, p = 0.220

a b a b c a

χ² = 11.532, p < 0.001***

a

χ² = 74.670, p < 0.001***

χ² = 0.0481, p = 0.826

Jo

Visitors

Counter-clockwise swimming distance

ro

Delay training

Morning Noon Afternoon Away from training Before training or training others None Toy(s) Human(s) Human(s) and toy(s) None Pool cleaning Noise Social event Other Altogether Separated Small Large Both None

-p

Time of the day

re

Level

ur na

Parameter

Clockwise swimming distance Fitted Wald chi-square output mean±SE 9.651±1.598 a 11.488±1.532 a χ² = 9.736, p = 0.008 ** 3.982±1.562 b 8.831±1.53 a χ² = 3.274, p = 0.070 3.027±1.7 a

lP

(c)

Table 6. Fitted means and standard-errors of the frequency of swimming behaviours per observation session (15 min) for YFPs (a), EAFPs (b) and BDs (c), and outputs from Wald chi-square tests ran on GLMMs. Letters and asterisks denote significant pairwise differences (p < 0.05): for

43

owning and asterisk significantly differ from the absence of stimuli level (“none”).

Enrichment

Perturbation

Social grouping

Visitors

a

1.337±0.784

0.081±0.03

a

0.273±0.203

a

0.064±0.055

a

0.269±0.061 0.387±0.148 0.47±0.174

χ² = 2.812, p = 0.094

χ² = 12.958, p = 0.024

0.109±0.076 0.082±0.041 0.18±0.054

*

4.165±1.584

*

χ² = 153.845, p < 0.001***

0.149±0.062 0.107±0.042 0.635±0.233 0.123±0.042 0.314±0.094 0.261±0.127 NA NA NA

* a b ab NA NA NA

ro

0.249±0.074

-p

to

Group swimming Fitted Wald chi-square mean±SE output 0.392±0.294 a χ² = 20.273, p 0.18±0.136 b < 0.001*** 0.238±0.179 ab

χ² = 20.344, p < 0.001***

NA

a

re

Delay training

Morning Noon Afternoon Away from training Before training None Toy(s) Human(s) Human(s) and toy(s) Fish None Pool cleaning Noise Social event Other Altogether Separated Alone None Few Many

Contact swimming Fitted Wald chi-square mean±SE output 0.092±0.037 a χ² = 11.544, p 0.045±0.019 b = 0.003** 0.121±0.049 a

χ² = 11.146, p < 0.001***

χ² = 1.519, p = 0.218

0.41±0.279

b

0.027±0.026

a

0.061±0.054

b

1.475±0.865 0.756±0.464 0.585±0.359

* *

0.087±0.033 0.108±0.064 0.043±0.027

*

0.295±0.22 0.378±0.303 0.054±0.044

*

0.592±0.397

*

0.064±0.064

*

0.571±0.381 1.014±0.596

*

2.848±1.719

*

lP

Time of the day

Level

Synchronous swimming Fitted Wald chi-square mean±SE output 1.443±0.849 a χ² = 5.724, p 1.136±0.669 b = 0.057 1.29±0.761 c

Jo

Parameter

Fast swimming Fitted Wald chi-square mean±SE output 0.168±0.053 a χ² = 17.025, p 0.375±0.118 b < 0.001*** 0.218±0.07 b

ur na

(a)

of

parameters with less than four levels, levels sharing the same letters do not differ significantly, and for parameters with more than four levels, levels

3.785±2.283

χ² = 47.507, p < 0.001***

0.223±0.202 0.005±0.006 0.056±0.022 0.257±0.133

χ² = 93.704, p < 0.001***

*

1.253±0.749 1.658±0.992 0.348±0.206 2.162±1.268 NA 1.218±0.714 1.795±1.062 0.625±0.388

χ² = 8.596, p = 0.126

* χ² = 21.075, p < 0.001***

0.109±0.057

44

χ² = 311.006, p < 0.001*** χ² = 28.207, p < 0.001***

0.159±0.078 0.066±0.028 0.083±0.032 NA 0.072±0.027 0.095±0.041 0.127±0.082

χ² = 34.116, p < 0.001***

0.271±0.254 0.155±0.116

0.106±0.051 * a b NA a b c

χ² = 9.614, p = 0.002**

a a NA a a a

χ² = 0.780, p = 0.377 χ² = 1.739, p = 0.419

0.988±0.77

*

0.686±0.534

*

0.544±0.418

*

0.64±0.497 0.94±0.288 0.658±0.187 NA 0.203±0.151 0.502±0.38 0.637±0.526

* a b NA a b b

χ² = 99.255, p < 0.001***

χ² = 6.248, p = 0.044* χ² = 30.049, p < 0.001***

Perturbation

Present None Few Many

Visitors

(c) Level

Delay training

to

Enrichment

Perturbation

Morning Noon Afternoon Away from training Before training or training others None Toy(s) Human(s) Human(s) and toy(s) None Pool cleaning

of

0.318±0.248

b

p < 0.001***

2.601±0.401

b

< 0.001***

0.065±0.053 0.19±0.146 0.51±0.41

a b c

χ² = 25.051, p < 0.001***

1.37±0.243 1.266±0.132 0.907±0.184

a a b

χ² = 6.596, p = 0.037*

Synchronous swimming Fitted Wald chi-square mean±SE output 2.543±0.021 a χ² = 11.387, 2.439±0.02 a p = 0.003** 2.016±0.015 b

Contact swimming Fitted Wald chimean±SE square output 0.087±0.06 a χ² = 3.221, 0.075±0.052 a p = 0.199 0.058±0.041 a

Group swimming Fitted Wald chi-square mean±SE output 0.062±0.055 a χ² = 27.684, p 0.05±0.044 b < 0.001*** 0.026±0.023 c

0.336±0.068

2.429±0.018

0.078±0.053

0.045±0.04

ur na

Time of the day

Group swimming Fitted Wald chi-square mean±SE output 1.639±0.231 a χ² = 52.376, p 0.719±0.098 b < 0.001*** 1.736±0.252 a 1.804±0.284 a χ² = 1.082, p = 0.298 1.116±0.106 a 1.015±0.098 a χ² = 21.553, p

Fast swimming Fitted Wald chimean±SE square output 0.392±0.083 a χ² = 10.778, 0.279±0.06 b p = 0.005** 0.429±0.092 a

0.645±0.176

a

b

0.355±0.072 0.383±0.095 0.294±0.087

Jo

Parameter

ro

Enrichment

Morning Noon Afternoon Absent Present Absent

Contact swimming Fitted Wald chi-square mean±SE output 0.156±0.121 a χ² = 4.942, p 0.154±0.12 a = 0.085 0.229±0.178 a 0.281±0.221 a χ² = 8.030, p = 0.005** 0.153±0.118 b 0.148±0.114 a χ² = 14.096,

-p

Time of the day

Level

Synchronous swimming Fitted Wald chimean±SE square output 3.149±1.881 a χ² = 11.272, 2.546±1.517 b p = 0.004** 3.903±2.332 a 4.16±2.495 a χ² = 4.468, p 2.873±1.703 b = 0.035* 2.68±1.589 a χ² = 27.024, p < 5.443±3.264 b 0.001*** 3.225±1.941 a χ² = 0.240, p 3.078±1.827 a = 0.887 2.963±1.79 a

re

Parameter

Fast swimming Fitted Wald chimean±SE square output 0.25±0.082 a χ² = 5.758, 0.391±0.121 a p = 0.056 0.402±0.128 a 0.399±0.135 a χ² = 1.066, p = 0.302 0.324±0.097 a χ² = 92.499, 0.242±0.073 a p < 1.254±0.397 b 0.001*** 0.525±0.18 a χ² = 7.926, 0.285±0.087 b p = 0.019 * 0.363±0.136 ab

lP

(b)

χ² = 9.573, p = 0.002**

χ² = 1.0534, p = 0.788

0.378±0.113 0.309±0.063 0.7±0.19

1.642±0.014

a b

2.312±0.018 2.759±0.022 2.654±0.021

χ² = 8.934, p = 0.003**

χ² = 4.120, p = 0.390

1.903±0.013 *

χ² = 22.0334, p < 0.001***

2.211±0.017 4.195±0.029

0.042±0.032

a a

0.077±0.053 0.054±0.039 0.127±0.098

χ² = 4.856, p = 0.183

0.045±0.036 *

45

χ² = 12.830, p = 0.012*

0.078±0.053 0.169±0.125

χ² = 2.750, p = 0.097

*

a

0.046±0.043

a

0.053±0.047 0.03±0.027 0.076±0.069

*

0.009±0.009

*

0.038±0.034 0.19±0.172

*

χ² = 0.009, p = 0.925

χ² = 56.868, p < 0.001*** χ² = 47.125, p < 0.001***

re

χ² = 1.787, p = 0.181

46

of

0.018±0.021 0.043±0.033 0.036±0.031 0.039±0.03 0.083±0.057 0.073±0.05 0.097±0.068 0.043±0.031 0.076±0.052 0.066±0.048

χ² = 0.444, p = 0.505

ro

χ² = 180.964, p < 0.001***

a a ab a b a a

χ² = 8.141, p = 0.017*

-p

χ² = 0.281, p = 0.596

lP

Visitors

* * a a a b c a a

2.028±0.016 2.485±0.02 2.813±0.022 2.202±0.015 2.38±0.019 2.275±0.018 2.726±0.02 1.982±0.016 2.4±0.019 2.166±0.015

ur na

Housing pool

0.296±0.12 0.595±0.175 0.666±0.223 0.317±0.092 0.363±0.074 0.206±0.044 0.512±0.114 1.691±0.379 0.371±0.075 0.297±0.072

Jo

Social grouping

Noise Social event Other Altogether Separated Small Large Both None Few

χ² = 1.955, p = 0.376

a a ab a b a a

χ² = 14.754, p = 0.005** χ² = 3.408, p = 0.065 χ² = 6.716, p = 0.035* χ² = 0.236, p = 0.627

0.021±0.021 0.052±0.05 0.1±0.093 0.25±0.239 0.033±0.029 0.045±0.04 0.063±0.056 0.021±0.019 0.036±0.032 0.11±0.098

*

χ² = 18.115, p < 0.001*** a a b a b

χ² = 21.396, p < 0.001 *** χ² = 33.063, p < 0.001***

of ro -p

Figure 1. Social grouping and social events during the observation period. All = all animals together; M/F

re

= males and females separated; D/O = Duoduo separated from other YFPs; G1 = Duoduo alone, Yangyang

lP

with F9, F7 with Taotao; G2 = Yangyang, F9 and Duoduo together, F7 with Taotao; G3 = Yangyang alone, F9 alone, Duoduo alone, F7 with Taotao; G4 = Yangyang alone, F9 with Duoduo, F7 with Taotao; G5 = Yangyang alone, F7 alone, F9, Duoduo with Taotao; G6: F7 alone, all others together; G7 = F7 alone,

Jo

ur na

Yangyang with Taotao, F9 with Duoduo, G8 = F7 alone, Yangyang with F9, Duoduo with Taotao.

47