Dominance behaviour in a non-aggressive flatfish, Senegalese sole (Solea senegalensis) and brain mRNA abundance of selected transcripts

Elvira Fatsini; Sonia Rey; Zohar Ibarra-Zatarain; Simon Mackenzie; Neil J. Duncan

doi:10.1371/journal.pone.0184283

Abstract

Dominance is defined as the preferential access to limited resources. The present study aimed to characterise dominance in a non-aggressive flatfish species, the Senegalese sole (Solea senegalensis) by 1) identifying dominance categories and associated behaviours and 2) linking dominance categories (dominant and subordinate) with the abundance of selected mRNA transcripts in the brain. Early juveniles (n = 74, 37 pairs) were subjected to a dyadic dominance test, related to feeding, and once behavioural phenotypes had been described the abundance of ten selected mRNAs related to dominance and aggressiveness was measured in the brain. Late juveniles were subjected to two dyadic dominance tests (n = 34, 17 pairs), related to feeding and territoriality and one group test (n = 24, 4 groups of 6 fish). Sole feeding first were categorized as dominant and sole feeding second or not feeding as subordinate. Three social behaviours (i. “Resting the head” on another fish, ii. “Approaching” another fish, iii. “Swimming above another” fish) were associated with dominance of feeding. Two other variables (i. Total time occupying the preferred area during the last 2 hours of the 24 h test, ii. Organisms occupying the preferred area when the test ended) were representative of dominance in the place preference test. In all tests, dominant fish compared to subordinate fish displayed a significantly higher number of the behaviours “Rest the head” and “Approaches”. Moreover, dominant sole dominated the sand at the end of the test, and in the group test dominated the area close to the feed delivery point before feed was delivered. The mRNA abundance of the selected mRNAs related to neurogenesis (nrd2) and neuroplasticity (c-fos) in dominant sole compared to subordinate were significantly different. This is the first study to characterise dominance categories with associated behaviours and mRNA abundance in Senegalese sole and provides tools to study dominance related problems in feeding and reproduction in aquaculture.

Citation: Fatsini E, Rey S, Ibarra-Zatarain Z, Mackenzie S, Duncan NJ (2017) Dominance behaviour in a non-aggressive flatfish, Senegalese sole (Solea senegalensis) and brain mRNA abundance of selected transcripts. PLoS ONE 12(9): e0184283. https://doi.org/10.1371/journal.pone.0184283

Editor: José L. Soengas, Universidade de Vigo, SPAIN

Received: May 1, 2017; Accepted: August 21, 2017; Published: September 6, 2017

Copyright: © 2017 Fatsini et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All data were uploaded to Fisgshare (DOI:10.6084/m9.figshare.4964990).

Funding: This work was supported by Instituto Nacional de Investigación y Tecnología Agraria y Alimentación” (INIA) INIA-FEDER Projects RTA2011-00050 and RTA2014-00048 coordinated by Dr. Neil J. Duncan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist.

Introduction

Behavioural studies in fish have been used as a model background in the field of perceptive ethology/evolutionary psychology. Associating several cognitive traits (memory, kin recognition and learning, among others) with morphology, ecology and a variety of behavioural parameters could provide a general vision for cognitive ethology [1–4]. Moreover, the study of flatfish behaviour has attracted the attention of researchers due to their ecology, life history and adaptations. However, when considered in comparison with pelagic fish species, flatfish behaviour has received little attention. Similarly, studies have tended to focus on aggressive species and few social non-aggressive fish species have been studied. General aspects in the behavioural catalogue of flatfish have been described including feeding behaviour, locomotion, mimicry and spawning behaviour reviewed in Gibson [5]. However, dominance behaviour in flatfish has not been described. The understanding of flatfish dominance behavioural patterns would be very useful to comprehend the biology and ecology of flatfish species and for stock management in aquaculture and natural populations.

Senegalese sole (Solea senegalensis) is a typical flatfish species that undergoes a larval metamorphosis to form a flattened shape adapted for a benthic life. The species inhabits the sediments of the Mediterranean Sea and the Southern European Atlantic coast and in these areas forms an important fishery that is in decline. The combination of declining market supply and high market value has resulted in a rapid increase in Senegalese sole culture in Europe by more than 10 times from 2009 to a production of over 1,200 tons in 2015 [6]. However, at present the culture is unsustainable due to two of the main culture problems [7]. One is the variation in growth that results in large size dispersion during weaning and on-growing which complicates farming practices [8, 9]. The second issue is the complete failure in the spawning of cultured breeders (hatched and reared in captivity), which dictates that currently, the farming of Senegalese sole relies on captures of wild breeders that spawn in captivity. More specifically, cultured males exhibit a dysfunctional behaviour and consequently do not complete the reproductive courtship to fertilise eggs [10–12]. In addition to this problem with cultured males, wild Senegalese sole pairs (male and female) that reproduce in captivity show intra- and inter-annual fidelity and a few families from a few breeders (8–40% of the broodstock) dominate the offspring from a captive wild broodstock [13, 14]. In order to understand the reproductive behavioural dysfunction to control reproduction, the reproductive behaviour has been described (ethogram) [15]. This courtship was characterised as a complex set of behaviours that especially males executed to gain acceptance from a female to spawn in a pair [15]. In this context, cultured males did not engage with females and did not participate in reproductive behaviours [11, 12]. Furthermore, stress coping styles were identified for individual fish and compared between groups with different origins (cultured vs. wild) and reproductive success [16, 17]. However, stress coping styles were not related to origin or reproductive success [17, 18]. All these studies on growth, courtship and behavioural aspects found that Senegalese sole could be considered as a social non-aggressive species [8, 9, 12, 17]. Therefore, under aquaculture conditions Senegalese sole appear to exhibit dominance several aspects as in growth, feeding and reproduction. However, further studies are needed to understand inter-individual interactions in captivity of this species and particularly dominance as both of these problems, variation in growth and failed spawning, may be related to social interactions and dominance.

Dominance was defined as success in competitions over limited resources such as food, specific (or preferred) areas, shelter, mates, spawning areas and offspring [19]. Thus, in general, dominant animals could have better access to food and shelter, lower rates of predation and higher mating success [20] than subordinates. Conversely, subordinate animals might suffer chronic stress, immune depression, reduced disease resistance and lower reproductive success [21]. Aggressive behaviour was related to social hierarchies and competition in fish species such as zebrafish (Danio rerio), Nile tilapia (Oreochromis niloticus) or rainbow trout (Oncorhynchus mykiss) among other species [20, 22–24]. In fish, social hierarchies are often categorised by agonistic behaviours that are habitually registered through feeding contests or territoriality. Several variables could be measured to assess these social hierarchies and one of them is feeding behaviour [25] which has been associated with physiological indicators of stress [26, 27], aggression and mating success [28]. Currently, place preference test is also used for fish with several purposes, such as to observe active implication in reproduction and territorial dominance [29]. For example, Nile tilapia is a tropical fish species that build nests and defends a territory to attract females [30]. Thus, individuals choose between different cubicles depending on the stimulus motivating the visit, shelter or reproduction [29]. The knowledge of the natural preference and the competition for different compartments is essential to avoid a misleading interpretation of results [29, 31]. In addition, animals that acquire the dominant position in early life stages (juvenile) in fitness-related traits might expand this social status to reproductive success [32, 33]. Therefore, the use of feeding and place preference tests adapted to Senegalese sole could provide insights into how dominance is established and expressed in a social flatfish species and such information can provide tools to establish how social interactions lead to growth dispersion and / or suppression of reproductive success.

In addition, previous studies found that the differences in behavioural phenotype were associated to different levels of biological regulation measured in the transcriptome, showing the behaviour might be linked with differential gene transcription [34–36]. These studies are working towards an increased understanding of the behaviour—transcriptome linkage and the present study incorporated some of the identified transcripts, which will be briefly described here. Dominance behaviour has been associated with agonistic behaviour as aggression and some transcripts have been related to this conduct in zebrafish, for example: The serotonin receptor 1 A (5-Hydroxytryptamine receptor 1 A; 5-htr1a), which inhibits serotonin (5-HT) firing, synthesis, release and turnover, has been associated with the control of several behavioural aspects including aggression [37, 38]. The tryptophan hydroxylase 1b (tph1b) that is involved in the synthesis of 5-HT [39] and solute carrier family 6 member 13 (slc6a13) that is a neurotransmitter transporter of gamma—Aminobutyric acid (GABA) were both related to aggression and impulsivity [40–42]. Lastly, the arginine vasopressin-induced protein 1 (avplr1), which is involved in the hypothalamo-neurohypophysial system (HNS) pathway and mineralocorticoid receptor (mr) implicated in hypothalamus-pituitary-interrenal (HPI) axis pathway, were also associated with aggressive behaviour in zebrafish [38]. Similarly, the following transcripts in zebrafish were associated with social status [43], bdnf and c-fos, which are markers of neuroplasticity [44] and nrd2 that was associated with neurogenesis [45]. Finally, ETS translocation variant 5 (etv5) and nuclear receptor subfamily 4, group A, member 2 (nr4a2), which are implicated in the preservation and differentiation of dopaminergic neurons in tilapia (Astatotilapia burtoni) [46]. In addition, all of the mentioned transcripts have been related to social status in zebrafish [37, 38].

In this study, we provide insight into dominance and social interactions of captive Senegalese sole aiming to define dominance behaviour by relating behavioural patterns to feeding response and territory as well as defining mRNA abundance in association to the different dominance related behavioural responses.

Material and methods

Ethics statement

All experimental practises on fish that formed part of this study were in agreement with the Spanish and European regulations on animal welfare (Federation of Laboratory Animal Science Associations, FELASA) and approved by the Animal Ethics Committee of IRTA.

Animal rearing conditions

Senegalese sole juveniles (~ 45 g) were provided by Stolt Sea Farm (Santiago de Compostela, Spain), different fish from the same batch were used for two successive years to conduct different behavioural tests. Fish were transported from A Coruña, where the facilities of Stolt Sea Farm are located, with a specialized transport for aquatic live animals. Fish were maintained at the Research Centre facilities of IRTA, in St. Carles de la Ràpita, North East Spain and distributed in two 10 m³ fiberglass tanks with natural photoperiod (using artificial lighting) and seasonally simulated external temperatures (40°62’82.42”, 0°66’09.37”/ 9–32°C) maintained within the range of 9–20°C with a recirculation system (IRTAMar^®) that recirculated + 400% and renewed 10% of the water daily. Sole were fed ad libitum five days per week with balanced feed (LE-3mm ELITE, Skretting, Co.). A total of six identical 400 L fiberglass tank were used, two tanks were used for acclimation (17–25 fish or 2.5–4.8 kg per tank) and four tanks for the experiments. One week before dyadic tests started, animals were moved from a large holding tank and acclimated to the 400 L fiberglass tanks. All fish were tagged with a passive integrated transponder (PIT) tag (ID—100A, Unique Trovan-Zeuss; Madrid, Spain) and photo identified individually. Individual markings in the photographs were matched with the corresponding PIT Tag for later identification in the video images. The acclimation and experimental tanks were in a recirculation system to control the temperature and water quality in order to reduce environmental variation. During the tests the temperature was maintained at 15°C with a recirculation system (IRTAMar^® described above). The animals were fasted for 48 hours prior to the experiment.

Behavioural studies

Video recording analyses.

Digital cameras (Square black and white CCD camera, model F60B/N80-50G, KT&C Co. Ltd., Korea Technology and Communications Korea, supplied in waterproof housing by Praesentis S.L. Barcelona, Spain) were used to film the dyadic and group dominance tests. One camera per experimental tank was placed above the tank to give a view of the entire tank. Four cameras (one per experimental tank) were connected to a digital video recorder (model XMOTION-304H by Praesentis, S.L). The videos were analysed in two ways, a continuous analysis of 2 hours of the behaviours between the two fish in the dyadic tests and a discrete analysis of behaviours on particular frames in the group tests. The different behaviours (defined in the dominance test section) were registered and their frequency was annotated for further statistical analysis.

Preliminary dyadic test.

A preliminary dyadic test was performed with twelve early juvenile sole (when fish weight was approximately 100 g) according to Huntingford et al., [47] to decide which test and setup was the most appropriate to characterise dominance behaviour in Senegalese sole. Animals were provided with different experimental cues that were visual (animals were separated by a transparent screen through which the animals could see each other), chemical (animals were kept apart by an opaque screen with holes through which the sole could exchange chemical cues) and isolated (sole were separated by an opaque screen without the possibility to see and smell each other). The different tests were performed at different daylight hours (morning or night). When preliminary results (S1 File) were analysed Senegalese sole did not show differences in behaviour due to the different experimental setups, and all fish resumed feeding and ate normally. Therefore, visual and chemical cues between separated fish did not appear to affect feeding. Considering this, the isolation approach was selected to provide a basal condition, where animals had equal status before testing for dominance over a limited resource, which is an approach that has been previously used in other fish species [25, 48].

Dominance tests.

Three different behavioural tests (two in pairs (dyadic) and one in group) were performed for Senegalese sole juveniles to test for dominance. Only one of the behavioural tests, feeding response in a dyadic test, was applied in early juveniles (n = 74; 100.4 ± 10.6 g). All three tests were applied to late juveniles (n = 34; 287.0 ± 30.4 g) and the resting time between tests was 7 days to allow for full recovery, return to basal conditions and to avoid learning and conditioning processes plus 7 days of acclimation to the new setting conditions (see Fig 1 for set-up and time line of the experiments). In all dyadic tests the fish were size-matched and paired (< 10% in weight between animals).

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Fig 1. Experimental chronogram of the different test conducted in Senegalese sole juveniles.

Experimental chronogram of the different dominance behavioural tests conducted on late Senegalese sole juveniles.

https://doi.org/10.1371/journal.pone.0184283.g001

Dyadic tests.

1) Feeding response after a dyadic test. Two size-matched sole taken from different acclimation tanks, to avoid previous competition, were kept apart overnight (19:00 to 8:00 = 13 hours) by a dark polyvinyl chloride (PVC) wall. The wall was removed the following morning at 08:00 and their behaviours were continuously recorded for the first two hours. After two hours, fish were hand-fed with approximately 1% of the total biomass of the experimental tank (two sole) and the individual that ate first was classified as dominant. The individual that ate afterwards or did not eat was classified as subordinate. Behaviours were individually registered and their frequency of occurrence was recorded by video monitoring. The behaviours recorded were: “Approaches”, “Swimming above another” (SAA), “Rest the head” (RTH), “Displacement”, “Burying” and finally order of feeding “Feeding” that was used to define dominance (categorical variable) (Table 1: ethogram of behaviours).
2) Competitive behaviour for a preferred area: place preference test. For the place preference test the settings of the tank were modified. First a bottom was created in the tank by placing twenty white tiling pieces (measuring 24 x 11.5 x 1.5 cm each and the same colour as the bottom of the tank) and second, one tile was removed to provide a space that was filled with sand. The area with sand was the standard size of one fish and so just one fish could comfortably occupy this area. Sole individuals used for this test had an extra week of acclimation to those novel objects: the sand and the tiling. The purpose of this test was to create a preferred area with sand (in nature, sole live buried in sand), which just one fish could occupy and, therefore, dominate that area (see S1 Fig. for set-up details). Previous studies have shown that sole prefer sandy areas [49]. The experimental tank was set up with PVC opaque/grey divisions that separated the two fish from each other and from a third area that contained the sand. The same pairs of sole that were previously used in the feeding test were also used in the place preference test and this enabled the place preference behaviour of the sole to be analysed in relation to the dominant and subordinate status observed in the feeding test. Fish were physically isolated (in separate acclimation tanks) and were introduced into the experimental tank at 19:00 and left overnight. The dividers were removed the following morning at 08:00 (after 13 hours of isolation) allowing the fish to see each other and access the restricted preferred area (sand). The behaviour of the fish in relation to the preferred sandy area was recorded continuously for 24 hours for further video analysis of recorded behaviours (Table 1). Red night lighting was used that allowed recording and observation of the sole behaviour. Red light was from fluorescent lights covered with a red filter. The intensity of the red light was adjusted to approximately 5 lux at the water surface. Previous studies demonstrated that this red light did not affect behaviour or physiological parameters during the night recordings [50]. The variables registered in this test were measured in minutes and regarding to the preferred area (sand). The variables were “Total time” in the preferred area (TT), “Initial time” to occupy the sand (Ti), “Final time” to occupy the sand (Tf) and order: “first” or “last” (categorical variable) (Table 1).

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Table 1. Ethogram of different behaviours registered in Senegalese sole.

https://doi.org/10.1371/journal.pone.0184283.t001

Group test.

3) Dominance in groups. Immediately after the place preference test were finished, four groups of six fish (24 fish in total) were randomly selected and placed in four 400 L experimental tanks. As with previous tests the fish were given a one-week recovery period followed by a one-week acclimation period that allowed the group to acclimate and to establish inter-individual hierarchies (Fig 1). After this period of acclimation, each group of fish was recorded to analyse the social interactions in the group. The different behaviours (Table 1) were recorded for 2 hours (every 5 minutes analysing a total of 24 frames) one hour before fish were fed and one hour after. The social behaviours recorded in the group test were “Rest the head” (RTH) and “Swimming above another” (SAA). The behaviours were registered in counts and different indexes were calculated (actions among animals). The test was performed in four consecutive days and the same behaviours for each of the groups of fish were recorded to test for consistency and repeatability among days. Fish was visually individually identified. An automatic feeder was placed in a corner of each tank and feed was delivered directly to the bottom of the tank through an 18 mm PVC tube to provide a single feed delivery point (see S2 Fig for set up details). Fish were fed with approximately 1% of the total biomass of the experimental tank. This single feed delivery point set-up is known to trigger territoriality and feeding competition among the individuals and dominant fish, at higher positions in the social hierarchical rank, tend to monopolise the feeding point [51–54]. The exact physical position of the fish in relation to the feeding point was recorded every 5 minutes before (1 hour = 12 frames) and every 5 minutes after (1 hour = 12 frames) feeding events (24 frames in total). The positions of the fish were ranked in order of the fish from the feeding point, i.e. the fish closest to the feeding point was ranked 1, second closest was ranked 2 etc. The ranked positions from the analysed frames were used to calculate the indices that were termed: “Position before feeding index” (POSITB index) and “Position after feeding index” (POSITA index) per fish and per day (Table 1). Mean of every index was calculated per fish over the 4 days. In addition, the “Feeding order” (‘pecking order’) for each day of the experimental period was registered to check for consistency over time.

RNA isolation, complementary DNA synthesis and quantitative real-time polymerase chain reaction assay

Thirty of the seventy-four early juveniles were randomly chosen, fifteen fish from each category (dominant/subordinate) and were sacrificed with an overdose of MS-222 (tricaine methanesulfonate; Acros-Organic, New Jersey, USA). Whole brains were extracted, frozen in dry ice and stored at -80°C for molecular analysis. The RNA was extracted using TRI Reagent RNA Isolation Reagent following manufacturer’s instructions (SigmaAldrich). The cDNA was synthesised using 1 μg of total RNA and oligo dT (20) in 20 μl reactions and the SuperScript^® III First-Strand Synthesis SuperMix 50 rxn kit following the manufacturer’s protocol (Invitrogen, Life technologies, USA). Primers were designed using Primer 3 [55] in 3UTR region. Before performing the qPCR, primers were validated by conventional PCR using a cDNA pool from several samples randomly chosen to analyse the primers. MyTaq^™ HS Mix (Bioline) was used to run the conventional PCR with the following conditions: initial activation step at 98°C for 1 min, followed by 35 cycles: denaturation at 95°C for 10s, annealing at Tm (58–60°C) gradient conditions for 15 s and extension at 72°C for 15 s. Primer efficiency was evaluated by serial dilutions to ensure that it was close to 100% performing real time PCR. Target transcripts with implication in neuroplasticity, neurogenesis and brain activation (bdnf, c-fos and nrd2) [43, 44, 56], related to differentiation of dopamine neurons (nr4a2 and etv5) [46, 57], behavioural responses and aggression (5-htr1a, tph1b, avplr1 and slc6a13, mr) [37–39, 42] were analysed by quantitative PCR (qPCR) (see primer design in Table 2). The qPCR was run using a Biometra Optical Thermocycler (Analytik Jena, Goettingen, Germany) in 96-well plates in duplicate 20 μl reaction volumes containing 10 μl of Luminaris Color HiGreen qPCR Master Mix (Thermo Scientific), 1 μl of the primer corresponding to the analysed gene (10 pmol), 3 μl of RNA/DNA water free and 5 μl of cDNA in its corresponding dilution. Furthermore, amplifications were carried out with a systematic negative control (NTC; no template control) containing no cDNA. Standard amplification conditions contained an UDG pre-treatment at 50°C for 2 min, an initial activation step at 95°C for 10 min, followed by 35 cycles: 15 s at 95°C, 30 s at the annealing Tm and 30 s at 72°C. Results were normalised using three housekeeping genes ubiquitin (ubi52), glyceraldehyde-3-phosphate dehydrogenase (gapdh2) and elongase factor 1 alpha (eef1a) and applying a geometric average [58]. The mRNA abundance for each gene was determined using the Pfaffl method [59].

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Table 2. Primers used in this study as possible dominance biomarkers for early Senegalese sole juveniles.

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Statistical analysis

Behaviour.

All means are presented as mean ± standard error of the mean (SEM). In the case of paired tests (early and late juveniles), the coefficient of variation (CV % = SD/mean*100) that represented the inter-individual sole variability were calculated for each category (dominant and subordinate) and compared. All data was tested for normality with the Shapiro-Wilks test and data that failed was corrected with a Log₁₀ transformation. To reduce variables and define behaviours that best represented dominance, a Principal Component Analysis (PCA) with adequacy of Kaiser-Meyer-Olkin test and Bartlett’s test of spherity with Varimax rotation was applied. In the case of early juveniles, a Spearman’s correlation analysis was run for the feeding response test between the variable “Feeding” (if the animals ate or not) and those variables, which were representative in the PCA run for this test. Student’s t-test was performed to compare means of counts of different behaviours of dominant and subordinate fish.

For the group test Kendall’s concordance coefficient (0.43 fair concordances) was calculated for each fish/behaviour index (RTH, SAA, POSITA and POSITB) to check the concordance among the 4 days for the same fish in each group. A “k” means cluster was applied to variables chosen by Kendall’s concordance coefficient for all groups. As there was concordance within the individuals in their behaviours over the 4 days for the index RTH, SAA and POSITB the mean of each index was calculated. Thus, Student’s t-test was applied to check the differences between dominants and subordinates. Statistical analyses were performed using SPSS Statistics 19.0 software (IBM Co., Hong Kong) and GraphPad Prism 6 software (GraphPad Software, Inc.) and P < 0.05 was used to establish significant differences.

q- rtPCR.

The outliers of the different categories (dominant/subordinate) of the corrected ratio of every mRNA were extracted using the Tukey’s test formula (k = 1.5). The data was transformed to Log₁₀ (var + 1) and Student’s t-test was applied to compare mRNA abundance between individuals grouped as dominant or subordinate. The threshold was considered at 0.3 related to pooled control animals simulating the population, where values under that threshold indicated down-regulation and over, up-regulation. The pooled control animals were from the same batch of the experimental sole used for this study and were acclimated to the same tanks as the experimental fish. Nevertheless, these fish were fed normally and were not used for any experimental process to obtain objective data imitating the usual conditions of the facilities. Raw data from both dominance behaviour and mRNA abundance are available in figshare (DOI: 10.6084/m9.figshare.4964990).

Results

Senegalese sole behavioural responses in dominance tests

Senegalese sole were classified as dominant or subordinate according to their feeding response (in dyadic and group feeding tests), the first fish to feed were classified as dominant and fish that fed second or did not feed at all were classified as subordinate. In general, subordinate individuals showed more variability in responses than dominant fish, which indicates that subordinate fish exhibited more variation between individuals.

Comparing the behaviours, “Approaches”, “Swimming above another” (SAA) and “Rest the head” (RTH), the dominant early juveniles (n = 36 of 74) displayed less variability within these behaviours (Approaches = 9.6 ± 1.4 counts; CV = 86.5%; SAA = 5.8 ± 1.2 counts; CV = 119.1%; RTH = 4.5 ± 0.6 counts; CV = 76.5%) than subordinates (n = 38 of 74) (Approaches = 5.5 ± 1.0 counts; CV = 109.2%; SAA = 2.5 ± 0.6 counts; CV = 141.1%; RTH = 2.6 ± 0.5 counts; CV = 105.4%). Whilst, the two behaviours, “Burying” and “Displacement”, presented a similar variability for both dominant and subordinate fish (Burying = 4.4 ± 0.5 counts; CV = 65.8%; Displacement = 1.0 ± 0.2 counts; CV = 150.3% for dominants and Burying = 4.6 ± 0.5 counts; CV = 66.0%; Displacement = 0.5 ± 0.2 counts; CV = 187.6% for subordinates respectively).

The trend in late juveniles was similar to that observed in early juveniles, with similar levels of variation in the behaviours and dominant late juveniles (n = 17 of 34) also displayed less variability (Approaches = 18.9 ± 2.6 counts; CV = 56.4%; SAA = 27.3 ± 9.5 counts; CV = 144.3%; RTH = 9.1 ± 1.4 counts; CV = 65.9%) than subordinates (n = 17 of 34) (Approaches = 12.1 ± 2.5 counts; CV = 83.8%; SAA = 12.3 ± 3.6 counts; CV = 120.3%; RTH = 4.1 ± 0.9 counts; CV = 91.5%).

The same pairs from the dyadic feeding tests were used in the place preference test and feeding response defined dominance was used in the place preference test to also define which fish were dominant and subordinate. In the place preference, dominant late juveniles spent more time in the preferred sand area at the end of the test and had less variability (Initial time (Ti) = 25.5 ± 9.2 min; CV = 149.6%; Total time (TT) = 377.1 ± 71.1 min; CV = 77.7%; Final time (Tf) = 60.7 ± 11.4 min; CV = 77.8%) than subordinates (Ti = 33.6 ± 11.7 min; CV = 144.2%; TT = 302.9 ± 93.9 min; CV = 127.8%; Tf = 38.4 ± 11.1 min; CV = 119.6%).

Observed feeding behaviour (MAPs in Senegalese sole).

The feeding behaviour of Senegalese sole was similar to that observed for other flatfish species with a defined “predation cycle or modal action patterns (MAP)”: searching, encountering, capturing and ingesting the food [5, 60]. In the present study, Senegalese sole both early and late juveniles, showed MAPs associated with feeding behaviour at the moment food was introduced into the experimental tank (see S2 File). Therefore, sole classified as dominant interacted first with the food and blocked other interactions. Furthermore, individuals that ate and were considered as dominant sole, interacted with the food in the first 10 minutes of food delivery starting with a “searching” MAP that consisted in slow creeping movements over the bottom, moving the head from side to side in short actions while slowly approaching the food. At the moment an individual detected the food the fish orientated and swam rapidly straight to it and started to protect the food and the area in which the food was delivered (direct observation).

Dominance parameters selection

Individual tests.

In early juveniles, three behaviours, “Approaches”, “Swimming above another” (SAA) and “Rest the head” (RTH) (Table 1) were grouped together (PCA, KMO (0.667), Bartlett’s test (P < 0.001) and X² (133.523); S3 Fig). These three behaviours expressed more than 51% of total variance after the principal component analysis. Spearman’s correlations demonstrated that those selected behaviours were weakly correlated to feeding response (Approaches: r_s = 0.372, P < 0.001; RTH: r_s = 0.358, P = 0.005; SAA: r_s = 0.4, P < 0.001; S4 Fig) In relation to feeding order, animals were classified as dominant or subordinate as described above. There was a pair that did not feed, and in this case both fish were considered subordinates. Consequently, the counts of the three behaviours were significantly different between dominants and subordinates (Approaches: t = 2.675, df = 72, P = 0.01; RTH: t = 2.814, df = 72, P = 0.01; SAA: t = 2.877, df = 72, P = 0.01; Fig 2A). Dominant sole displayed significantly more approaches, resting the head and swimming above another than subordinate sole.

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Fig 2. Frequency of the behaviours analysed in early and late juveniles of Senegalese sole.

Number of movements were counted during the 2-hour test that dominant and subordinate (A) early juveniles and (B) late juveniles Senegalese sole exhibited the three behaviours “Approaches”, “Swimming above another” (SAA) and “Rest the head” (RTH). Data was shown in Mean ± SEM (n = 74 early juveniles; n = 34 late juveniles). An * indicates a significant difference (P < 0.05).

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The association between behaviours and dominance was similar for the late juveniles. The PCA of all the variables from the feeding response and place preference test that were applied to late juveniles extracted three components that explained 72% of the variance of the data (KMO (0.6), Bartlett’s test (P < 0.001) and X² (116.806); S5 Fig). The three behaviours, Approaches, RTH and SAA were grouped together as the first principal component (PC1). These three behaviours were also grouped together for the early juvenile sole establishing that the feeding response test applied to two different groups of sole of different ages and size gave similar consistent results. The second (PC2) and third components (PC3) for the late juveniles were both related to behaviours from the place preference test. The PC2 was formed by the variables; total time each fish occupied the preferred sand area during the last two hours of the test (Final time—Tf) and which fish was in the sand when the test finished at 24 hours (last position—last). PC3 consisted of; the time each fish first occupied the sand during the first two hours (Initial time—Ti), total time each fish was in the sand during the entire 24-hour test (Total time—TT) and which fish was first to enter the sand area (first position—first) (Table 1).

The counts of behaviours “Approaches” (t = 2.036, df = 31.69, P = 0.05) and RTH (t = 2.894, df = 26.30, P = 0.008) were significantly higher in dominants than subordinates in late juveniles. However, SAA (t = 1.083, df = 30.94) was not different between dominants and subordinates (Fig 2B).

The variables that formed the second component were significantly different between dominant and subordinate fish classified by feeding response test, thus sole classified as dominant (in the feeding test) spent more time in the preferred sandy area during the last two hours of the test (Tf: t = 2.186, df = 16, P = 0.044; Fig 3A) and dominated the area when the test finished after 24 hours (X² = 5.674, P = 0.017; Fig 3B). In summary, the dominant fish that ate first spent more time in the preferred area over the last two hours of the test and monopolised the sand, showing that the final position was indicative of a dominant status. All results were analysed from size-matched pairs and, therefore, variability due to unequal size distribution was avoided. Thus dominance parameters were observed in each pair representing pairs of similar weight and length.

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Fig 3. Place preference test in late juvenile sole (n = 74).

(A) Time in minutes during the last two hours of the test that dominant and subordinate Senegalese sole were in the preferred sand area (Tf). (B) The proportion of dominant and subordinate fish that were in the preferred sand area when the test finished (last). An * indicates a significant difference (P < 0.05).

https://doi.org/10.1371/journal.pone.0184283.g003

Group test.

In the group analysis, “Rest the head index” (RTH Index), “Swimming above another index” (SAA Index), “Position of the fish in relation to the feed delivery tube before feed” was delivered (POSITB) and “Feeding Order” (from 1^st to 6^th position) (Table 1) showed agreement (0.43) according to Kendall’s concordance coefficient (W) among the 4 days for each group (S1 Table). The animals in the different groups were analysed as a single population and the Kendall’s concordance coefficient demonstrated that the inter-groups index were consistent. In addition, “k” means cluster classified animals in two clusters that represented dominant and subordinate animals. The two clusters grouped the same animals according to feeding RTH, SAA and POSITB Index indicating concordance of these behaviours in the different groups. Therefore, 12 fish were classified as dominant and 12 as subordinates. Student’s t-test applied in those index to compare the significant differences between dominants and subordinates for different behaviours showed that RTH Index (t = 2.659, df = 10.46, P = 0.015; Fig 4A) and POSITB Index (t = 3.779, df = 21.57, P = 0.001; Fig 4C) were significantly different between dominant and subordinate groups. However, the SAA Index (t = 1.231, df = 19.35; Fig 4B) was not different between dominant and subordinate fish. To summarize, the fish considered dominant more often occupied positions closer to the feed delivery tube even before the food was provided (place preference test was covered with those positions) and rested the head (RTH) in any part of the body more often than the subordinate fish. The group analysis was performed with the four groups as a population consisting of individuals of different sizes (simulating the cohabitation in nature). However, no relationship was found between fish size and the repetition of the different dominance parameters observed. For example, in one of the groups, the weight range was from 195.7 to 310.6 g and the two smallest animals presented more actions in RTH and POSITB and presented more consistency in their data than the largest individual throughout the experimental period.

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Fig 4. Mean behavioural indices for dominant (n = 12) and subordinate (n = 12) late juveniles sole in the group-test according to Kendall’s concordance coefficient.

(A) “Rest the head Index” (RTH Index); (B) “Swimming above another Index” (SAA Index) and (C) “Position before feeding” (POSITB Index). An * indicates a significant difference (P < 0.05).

https://doi.org/10.1371/journal.pone.0184283.g004

Brain mRNA abundance in relation to dominance

Only two of the ten (Table 2) mRNAs tested exhibited significant differences in mRNA abundance between dominant and subordinate sole (Table 3). Both transcripts, c-fos (t = 2.014, df = 20.13, P = 0.041; Fig 5A) and nrd2 (t = 1.861, df = 27.15, P = 0.047; Fig 5B), were down regulated in both dominant and subordinate fish in relation to pooled controls that represent the entire range for the population. This down-regulation was more pronounced in dominant sole than in subordinate individuals. The nr4a2 mRNA presented marginally different abundance (t = 1.987, df = 22.56).

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Table 3. mRNA abundance in Log (var + 1) fold difference for the several mRNAs analysed in dominant (n = 15) and subordinate (n = 15) early juvenile Senegalese sole.

https://doi.org/10.1371/journal.pone.0184283.t003

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Fig 5. mRNA abundance of dominant (n = 15) and subordinate (n = 15) early juvenile sole.

Expression for two genes (A) c-fos and (B) nrd2 shown as mean ± SEM in Log (var + 1) transformation. Dashed line at 0.3 indicates the value of a pooled control simulating the population; values under 0.3 indicate down-regulation. An * denotes a significant difference (P < 0.05).

https://doi.org/10.1371/journal.pone.0184283.g005

Discussion

The present study has shown that Senegalese sole can be categorized for different social status (dominant and subordinate): (a) when competition was observed between pairs and in groups, (b) in relation to both feeding response and place preference tests, and (c) at different juvenile stages, from early juvenile, when fish weight was approximately 100 g, to late juvenile that weighted approximately 300 g (considered as pre-adult). Across the three tests (dyadic feeding, dyadic place preference and group) the same behaviours were associated with social status. Dominant fish compared to subordinate fish displayed a significantly higher number of the behaviours “Rest the head” (RTH) on another fish, “Approach” another fish and “Swim above another” (SAA) fish. This is the first description of behaviours associated to social status in a non-aggressive fish and/or a flatfish species. Other studies on social status in fish have been conducted on round fish that display aggressive behaviours [24–26, 29, 31]. For example, previous studies in rainbow trout demonstrated that after a period of isolation the fish that ate first won the subsequent contest, showed more aggressive behaviour and were subsequently categorized as dominant individuals [25]. This agrees with the present study where both animals were isolated before the dyadic behavioural tests and the sole that ate first exhibited a higher frequency of the behaviours RTH, Approaches and SAA and also dominated preferred spatial zones (sand or feed delivery point).

As mentioned in the introduction two of the main problems for sole culture are a large variation in growth [8, 9] and a complete failure in the spawning of cultured breeders [11, 12]. Therefore, the identification of different social status and associated behaviours in sole have considerable implications in studies directed to tackling effects of dominance in culture and ecological studies for sole and flatfish.

Feeding and growth

A major problem during grow out reported in Senegalese sole is a large variation in growth resulting in a wide dispersion of sizes and the requirement of frequent grading [8, 9]. In the present study, dominant sole clearly had advantages in access to feed and both ate first and dominated the feed source (i.e. space where feed was delivered). Therefore, one theory would be that dominance could explain the size variation observed in cultured stocks and that this study might be fundamental for future research into the dominance of feeding and effects on growth in captive conditions. In agreement with this, aggression in feeding behaviour is the most common method used to establish hierarchies and in different species the dominant fish have often been the larger individuals [61]. However, juvenile Senegalese sole did not show any aggressive behaviour at different stocking densities when fed according to the biomass [8] or in the dominance tests (present study). In the present work, larger body size was not identified as a factor associated to dominant fish in the present study and consequently dominance behavioural parameters specifically RTH, SAA and Approaches were not associated to animal size. These observations would suggest that dominance is probably a contributing factors, but that dominance alone may not be responsible for the growth variation and other factors, such as life strategy, appetite and feeding behaviour, should be considered.

Life strategies with differences in appetite can be compared to stress coping styles that range from a proactive life style where animals take risks to forage for food, suffer higher predation, grow faster and reach puberty at a younger age [17, 62]. In comparison the opposite extreme a reactive life style where animals take less risks, exhibit less foraging for food, higher survival, slower growth and reach puberty at an older age [17, 63]. Proactive and reactive coping styles have been described in all stages of Senegalese sole, larvae [64], juveniles and adults [17] and proactive sole were shown to grow faster and reach puberty earlier than reactive sole [18]. Furthermore, previous studies have shown that benthic larvae of Senegalese sole have more activity than juveniles, and that juveniles present more activity than adult sole under certain conditions [17, 18]. Therefore, different life strategies among individuals may modify changes in activity and feeding to contribute to the size variation observed in cultured stocks.

The feeding behaviour observed during the present study was similar to that described in the common sole (Solea solea) and other species such as turbot where visual and olfactory cues are important in searching for food [65–67]. Performance of the different parts of the predation cycle (search, encounter, capture and ingestion) can vary between the species. In common sole feeding behaviour and boldness (associated in some fish species with aggression and social status such as in cichlid fish [68]) were related to feed intake using isolation and group tests [69] explaining variability in growth where proactive fish appear to have better feeding tactics in captivity. Feeding order in fish could be similar to the pecking order found in other farmed species (poultry, pigs, cows, etc.), pecking order is established at a younger age, and remains stable along their life determining the social status and hierarchies. In addition, pecking order has been associated with different activities such as feeding, drinking, and mating [70, 71]. This could be the explanation for the consistent feeding behaviour in groups among the four days, considering that the individual behaviour can affect the variability of the group in captivity.

Reproduction

As explain in the introduction cultured males (born and reared in captivity) presented a dysfunctional behaviour and consequently did not complete the reproductive courtship to fertilise eggs [10–12]. Wild breeders held in captivity reproduce normally in captive condition, however, in a broodstock a few reproductive pairs show intra- and inter-annual fidelity and dominate the reproductive output [13, 14]. A description of the reproductive behaviour in captive wild breeders has shown that the behaviours Rest the head (RTH) and “Approaches” were essential courtship behaviours that males had to execute towards females to achieve reproductive success [15]. Immediately before spawning, a male was observed to approach the female and rest the head on the female. Once the male was accepted, the female swam from the bottom to initiate the coupled swim to the surface to spawn. Particularly, RTH appeared to be a behaviour that indicated a pair, the female accepting the male and the male protecting or dominating the female from other males. Therefore, the behaviours RTH and “Approaches” indicate dominance in feeding and place preference (present study) and acceptance / dominance of a partner in mate selection and reproduction [15]. Dominance across different resources including mate selection has been described in aggressive round species such as Nile tilapia [30], Ambon damselfish (Pomacentrus amboinensis) [72, 73], Mozambique tilapia (Oreochromis mossambicus) [74] and social mammals such as meerkats [32] or European rabbits [33], where dominance status started in early life stages and were persistent until adult phase. The present study provides the bases for further research to determine if the behaviours RTH and “Approaches” indicate dominance across feed, territory and mate selection.

Place preference test

Place preference test consisted of two areas as choice possibilities, where the animal associates one of those areas as preferred for a particular purpose (hide, shelter, mating, among others) [75, 76]. In our study, there was a clear preference for the area with substrate by all experimental sole. However, the objective of this test was to observe the behaviour of paired Senegalese sole in response to a preferred area (sand) simulating a limited resource. Preference by dominant sole regarding territory was identified taking the different times of every individual dominating the substrate. Therefore, dominant sole spent more time at the end of the test (last two hours of test) and were more often present in the sand at the end of the 24-hour test (last position). The final two hours of the place preference test coincided with the initiation of a daytime resting period, when the dominate sole would occupy the preferred space by burying into the sand to rest. This kind of diurnal activity with daytime burying into sand and resting was first described in common sole [77]. Therefore, subordinate fish spent less time at the end of the test and entered the sand earlier in the test or did not enter at all. This was in agreement with other fish species with different biology and ecology such as Mozambique tilapia where substrate is important in different contexts [78]. Another example of the use of substrate is the Nile tilapia, where animals were isolated and individuals could choose different compartments where the gravel-enriched compartment was the most visited [29].

The movement performed by sole when in contact with the sand was burying. The burying behaviour has three objectives, burying in to rest, hiding and to displace another sole (when the burying is displayed under or on top of another sole) [15]. Kruuk (77) described the same behaviour for common sole having several functions, such as burying to help mimic the sediment (camouflage enables flatfish to avoid predators and remain hidden to prey), to initiate a resting period and to avoid currents. However, this behaviour was not extracted as a representative parameter in feeding response test in early juveniles, it was a key behaviour to determine the time parameters of place preference test in late juveniles. Previous studies demonstrated that habitat preference increased the level of territorial protection or dominance displayed by brown trout (Salmo trutta) [79]. In contests dominant animals defended and displayed more aggression in order to dominate their preferred territory. In our study, when both sole coincided in the sand the sole in the upper position normally the dominant individual rested the head (whatever part of the body) on the sole in the bottom position. This behaviour would not be considered as an aggression as the other animal was not injured, but it could be a harassment tactic. So, the RTH parameter extracted in the feeding response test was also associated with RTH behaviour in the place preference test. However, the situation of both animals coinciding in the sandy area was not usually observed in fact only in two of the seventeen pairs studied. In the case of the group test the place preference was analysed according to the position regarding the feeder supplier area. Intriguingly, the POSITB Index corroborated that animals that dominated the area to which feed was delivered were the animals that ate in the first positions, being stable during the experimental period. This situation is commonly observed in cultivated fish due to domestication, which depending on the fish species is performed in a different manner achieving different tactics according to the food delivery [80].

Brain mRNA abundance in association to dominance behaviour

Several sets of genes related to different behavioural processes were analysed in the present study. However, just two mRNA transcripts related to neurogenesis (nrd2) and neuroplasticity (c-fos) were differently expressed between dominant and subordinate sole. Besides, nr4a2 one gene which encode NR4A2 a receptor associated with differentiation of dopamine neurons [46, 57] was marginally different. These three transcripts presented down-regulation in both categories (dominant and subordinate) compared to control group (see Fig 5 for nrd2 and c-fos) and dominant fish exhibited greater down-regulation than subordinate fish. In some cases, the presence of the same low profile in both dominance categories, dominant and subordinate, results in the adaptive response to slight social stress in comparison to the undisturbed animals [81]. This could be the explanation of why in our study animals of both dominance categories showed down-regulation, therefore, dominant and subordinate sole would present the same pattern of social stress being more pronounced in dominant sole.

In several animal models for social studies such as rodents, social subordination has been associated to reduced neurogenesis [82]. The mRNA transcript related to neurogenesis was nrd2 that is essential for the survival of specific populations of neurons and neuronal differentiation in mice [83, 84]. This transcript has also been related to neural plasticity in zebrafish where the expression was higher in winners than losers in dorsal telencephalic area [45]. Neuroplasticity has also been associated with social interactions in fish where the profile of immediate early genes (IEG), including c-fos, in zebrafish exhibited acute changes in the pattern of expression due to different social status [45, 85]. Another example was found in tilapia (Astatotilapia burtoni); males of this species can change social status between subordinate and dominant in relation to the presence or absence of a larger dominant male in the same territory. This behavioural change between subordinate and dominant was shown to be accompanied at the molecular level with changes in the transcript IEG c-fos [86]. In our study, the pattern of transcript abundance was distinct where dominant and subordinate sole presented lower c-fos abundance than a representative population pool corroborating that neuroplasticity was possibly associated with social behaviour in this species.

In the present study, 5-htr1a jointly with other aggression-related genes such as tph1b, avplr1 and slc6a13, mr presented similar mRNA abundance between dominant and subordinate sole contrarily to other fish species where dominance was determined by aggressive behaviour [38, 87]. However, as previously mentioned Senegalese sole is considered a non-aggressive species and these transcripts, related to social status and aggression in other species, may not have a role in social status in the non-aggressive Senegalese sole.

Conclusions

In conclusion, this is the first study using dyadic contests in a flatfish species to describe and determine social status (dominant/subordinate) when resources were limited. This study reports upon individual differences in dominance behaviour in Senegalese sole for the first time. This fish is considered as a non-aggressive species and accordingly different non-aggressive dominance behavioural parameters (Rest the head, Approaches, and Swim above another) were described in relation to dominance of feeding and territory. Those dominance behaviours were consistent in the different juvenile stages (early and late juveniles) and additionally, those parameters were reliable when applied to sole in groups. Both dominant and subordinate juvenile Senegalese sole exhibited specific and significantly different levels of abundance of two brain mRNAs that used to be associated to neurogenesis and neuroplasticity. Nevertheless, future work would be necessary to completely assess the relationship between brain gene transcription and differences in dominance profiles in this species. These results are highly relevant for the fish farming industry of Senegalese sole in order to tackle problems that appear to be linked to dominance in the culture system and form the bases for work to improve culture conditions by understanding the behavioural profile of these animals. This essential understanding of hierarchical distribution in the population will be linked to methods to ensure future reproductive success.

Supporting information

S1 Fig. Experimental tank set up used for the place preference test (with sand) in pairs.

A Preferred area (sand) and B white tiles forming a false bottom characterized the novel conditions. C Water inlet. D Water outlet.

https://doi.org/10.1371/journal.pone.0184283.s001

(TIF)

S2 Fig. Group experimental tank set up used for feeding response and point feed delivery.

A PVC tube to deliver the food. B Water Inlet. C Water Outlet. Different position areas (1–6) were shown by point lines.

https://doi.org/10.1371/journal.pone.0184283.s002

(TIF)

S3 Fig. Principal component analysis of the different behaviours registered during the “feeding response test” in pairs.

The three variables “Approaches, SAA and RTH” were grouped together and explained the 53% of the variance of the data. KMO (0.667), Bartlett’s test (P < 0.05) and X² (133.523).

https://doi.org/10.1371/journal.pone.0184283.s003

(TIF)

S4 Fig. Spearman’s correlations from the three variables “Approaches, SAA and RTH” with feeding.

(**) correlation was significant P < 0.05.

https://doi.org/10.1371/journal.pone.0184283.s004

(TIF)

S5 Fig. Principal component analysis of the different behaviours registered during the feeding dominance test and place preference test (sand) in pairs.

The three variables “Approaches, SAA and RTH” and the “TF and last” explained the 56% of the variance of the data in two different components. KMO (0.6), Bartlett’s test (P < 0.05) and X² (116.806) (SPSS 19.0 IBM Statistics).

https://doi.org/10.1371/journal.pone.0184283.s005

(TIF)

S1 Table. Classification of the different variables in groups according to Kendall’s concordance coefficient (W) for every group.

(*P < 0.05) level of significance.

https://doi.org/10.1371/journal.pone.0184283.s006

(DOCX)

S1 File. Raw data of preliminary results in dyadic pairs of early Senegalese sole juveniles.

https://doi.org/10.1371/journal.pone.0184283.s007

(XLSX)

S2 File. Modal action patterns related to feeding in late Senegalese sole juveniles.

https://doi.org/10.1371/journal.pone.0184283.s008

(WMV)

Acknowledgments

Thanks are given to J. Celades, P. Cascón, M. Monllaó and X. Ingla for technical support.

References

1. Griffin DR. Animal Thinking. Cambridge, MA: Harvard University Press.; 1984.
2. Cheney DL, Seyfarth RM. How monkeys See the World. Chicago: University of Chicago Press; 1991. 377 p.
3. Barkow G. The puzzling paucity of feeding territories among freshwater fish. Mar Behav Physiol. 1992; 22: 157–76.
- View Article
- Google Scholar
4. Dugatkin LA, Wilson DS. Fish behaviour, partner choice experiments and cognitive ethology. Rev Fish Biol Fish 1993; 3: 368–72.
- View Article
- Google Scholar
5. Gibson R, editor. Flatfishes: Biology and Exploitation: Blackwell Science Ltd, a Blackwell Publishing company; 2005.
6. APROMAR. La acuicultura marina de peces en España. Asociación Empresarial de Productores de Cultivos Marino. 2016 (Spain).
7. Morais S, Aragao C, Cabrita E, Conceiçao L, Constenla M, Costas B, et al. New developments and biological insights into the farming of Solea senegalensis reinforcing its aquaculture potential. Rev aquacult. 2014; 6: 1–37.
- View Article
- Google Scholar
8. Salas-Leiton E, Anguis V, Manchado M, Cañavate JP. Growth, feeding and oxygen consumption of Senegalese sole (Solea senegalensis) juveniles stocked at different densities. Aquaculture. 2008; 285: 84–9.
- View Article
- Google Scholar
9. Salas-Leiton E, Anguis V, Martin-Antonio B, Crespo D, Planas JV, Infante C, et al. Effects of stocking density and feed ration on growth and gene expression in the Senegalese sole (Solea senegalensis): potential effects on the immune response. Fish Shellfish Immunol. 2010; 28(2): 296–302. pmid:19909816
- View Article
- PubMed/NCBI
- Google Scholar
10. Martín IE. Advances in the reproductive biology and zootechnics of the Senegalese sole (Solea senegalensis Kaup, 1858). Ph.D. Thesis, Universidad de Cantabria (Spain). 2016.
11. Carazo I, Martin I, Hubbard PC, Chereguini O, Mañanós EL, Canario AVM, et al. Reproductive behaviour, the absence of reproductive behaviour in cultured (G1 generation) and chemical communication in the Senegalese sole (Solea senegalensis). Indian Journal of Science and Technology (9th ISRPF). 2011; 4: 96–7.
- View Article
- Google Scholar
12. Carazo I. Comportamiento reproductivo y fisiología del lenguado senegalés (Solea senegalensis) en cautividad. Ph. D. Thesis, Universidad de Barcelona (Spain). 2013.
13. Martín I, Rasines I, Gómez M, Rodríguez C, Martínez P, Chereguini O. Evolution of egg production and parental contribution in Senegalese sole, Solea senegalensis, during four consecutive spawning seasons. Aquaculture. 2014; 424–425: 45–52.
- View Article
- Google Scholar
14. Porta J, Porta JM, Martínez-Rodríguez G, Álvarez MC. Development of a microsatellite multiplex PCR for Senegalese sole (Solea senegalensis) and its application to broodstock management. Aquaculture. 2006; 265: 159–66.
- View Article
- Google Scholar
15. Carazo I, Chereguini O, Martin I, Huntingford F, Duncan NJ. Reproductive ethogram and mate selection in captive wild Senegalese sole (Solea senegalensis). Span J Agric Res. 2016; 14(4): e0401.
- View Article
- Google Scholar
16. Silva PIM, Martins CIM, Engrola S, Marino G, Øverli Ø, Conceição LEC. Individual differences in cortisol levels and behaviour of Senegalese sole (Solea senegalensis) juveniles: Evidence for coping styles. Appl Anim Behav Sci. 2010; 124: 75–81.
- View Article
- Google Scholar
17. Ibarra-Zatarain Z, Fatsini E, Rey S, Chereguini O, Martin I, Rasines I, et al. Characterization of stress coping style in Senegalese sole (Solea senegalensis) juveniles and breeders for aquaculture. R Soc Open Sci. 2016; 3:160495. pmid:28018634
- View Article
- PubMed/NCBI
- Google Scholar
18. Ibarra-Zatarain Z. The role of stress coping styles in reproduction and other biological aspects in the aquaculture species, Senegalese sole (Solea senegalensis) and gilthead seabream (Sparus aurata). Ph. D. Thesis, Universidad Autónoma de Barcelona (Spain). 2015.
19. Noakes DLG. Social behaviour as it influences fish production In: Gerking SD, editor. Ecology of freshwater fish production. Oxford: Blackwell Scientific Publ., Oxford; 1978. p. 360–82.
20. Grant JWA. Territoriality. In: Godin J-GJ, editor. Behavioural Ecology of Teleost Fishes. United States: Oxford University press Inc., New York; 1997. p. 360.
21. Øverli Ø, Harris CA, Winberg S. Short-term effects of fights for social dominance and the establishment of dominant-subordinate relationships on brain monoamines and cortisol in rainbow trout. Brain Behav Evol. 1999; 54(5): 263–75. pmid:10640786
- View Article
- PubMed/NCBI
- Google Scholar
22. Cardoso P, Volpato GL. Chemical communication, aggression, and conspecific recognition in the fish Nile tilapia. Physiol Behav. 1997; 62(6): 1333–8. pmid:9383122
- View Article
- PubMed/NCBI
- Google Scholar
23. Andersson MA, Höglund E. Linking personality to larval energy reserves in rainbow trout (Oncorhynchus mykiss). PLoS One. 2012; 7(11): e49247. pmid:23155473
- View Article
- PubMed/NCBI
- Google Scholar
24. Ruiz-Gomez ML, Huntingford FA. Boldness and aggressiveness in early and late hatched three-spined sticklebacks Gasterosteus aculeatus. J Fish Biol. 2012; 81(3): 966–76. pmid:22880730
- View Article
- PubMed/NCBI
- Google Scholar
25. Øverli Ø, Korzan WJ, Höglund E, Winberg S, Bollig H, Watt M, et al. Stress coping style predicts aggression and social dominance in rainbow trout. Horm Behav. 2004; 45: 235–41. pmid:15053939
- View Article
- PubMed/NCBI
- Google Scholar
26. Øverli Ø, Kotzian S, Winberg S. Effects of cortisol on aggression and locomotor activity in rainbow trout. Horm Behav. 2002; 42(1): 53–61. pmid:12191647
- View Article
- PubMed/NCBI
- Google Scholar
27. Øverli Ø. Preface: plasticity and diversity in behavior and brain function—important raw material for natural selection? Brain Behav Evol. 2007; 70(4): 215–7. pmid:17914252
- View Article
- PubMed/NCBI
- Google Scholar
28. Korzan WJ, Øverli Ø, Summers CH. Future social rank: forecasting status in the green anole (Anolis carolinensis). Acta Ethol. 2006; 9: 48–57.
- View Article
- Google Scholar
29. Delicio HC, Barreto RE, Normandes EB, Luchiari AC, Marcondes AL. A place preference test in the fish Nile tilapia. J Exp Anim Sci. 2006; 43: 141–8.
- View Article
- Google Scholar
30. Popma T, Masser M. Tilapia life history and biology. SRAC. 1999; 283: 1–4.
- View Article
- Google Scholar
31. Serra EL, Medalha CC, Mattioli R. Natural preference of zebrafish (Danio rerio) for a dark environment. Braz J Med Biol Res. 1999; 32: 1551–3. pmid:10585639
- View Article
- PubMed/NCBI
- Google Scholar
32. English S, Huchard E, Nielsen JF, Clutton-Brock TH. Early growth, dominance acquisition and lifetime reproductive success in male and female cooperative meerkats. Ecol Evol. 2013; 3(13): 4401–7. pmid:24340181
- View Article
- PubMed/NCBI
- Google Scholar
33. Rodel HG, von Holst D. Features of the early juvenile development predict competitive performance in male European rabbits. Physiol Behav. 2009; 97(3–4): 495–502. pmid:19375438
- View Article
- PubMed/NCBI
- Google Scholar
34. Rey S, Boltana S, Vargas R, Roher N, Mackenzie S. Combining animal personalities with transcriptomics resolves individual variation within a wild-type zebrafish population and identifies underpinning molecular differences in brain function. Mol Ecol. 2013; 22: 6100–15. pmid:24118534
- View Article
- PubMed/NCBI
- Google Scholar
35. MacKenzie S, Ribas L, Pilarczyk M, Capdevila DM, Kadri S, Huntingford FA. Screening for coping style increases the power of gene expression studies. PLoS One. 2009; 4(4): e5314. pmid:19390591
- View Article
- PubMed/NCBI
- Google Scholar
36. Driscoll P, Escorihuela RM, Fernandez-Teruel A, Giorgi O, Schwegler H, Steimer T, et al. Genetic selection and differential stress responses. The Roman lines/strains of rats. Ann N Y Acad Sci. 1998; 851: 501–10. pmid:9668644
- View Article
- PubMed/NCBI
- Google Scholar
37. Filby AL, Paull GC, Hickmore TF, Tyler CR. Unravelling the neurophysiological basis of aggression in a fish model. BMC Genomics. 2010; 11: 498. pmid:20846403
- View Article
- PubMed/NCBI
- Google Scholar
38. Filby AL, Paull GC, Searle F, Ortiz-Zarragoitia M, Tyler CR. Environmental estrogen-induced alterations of male aggression and dominance hierarchies in fish: a mechanistic analysis. Environ Sci Technol. 2012; 46(6): 3472–9. pmid:22360147
- View Article
- PubMed/NCBI
- Google Scholar
39. Zhang X, Beaulieu JM, Sotnikova TD, Gainetdinov RR, Caron MG. Tryptophan hydroxylase-2 controls brain serotonin synthesis. Science. 2004; 305(5681): 217. pmid:15247473
- View Article
- PubMed/NCBI
- Google Scholar
40. Miczek KA, Fish EW, De Bold JF, De Almeida RM. Social and neural determinants of aggressive behavior: pharmacotherapeutic targets at serotonin, dopamine and gamma-aminobutyric acid systems. Psychopharmacology. 2002; 163(3–4): 434–58. pmid:12373445
- View Article
- PubMed/NCBI
- Google Scholar
41. Miczek KA, Fish EW, De Bold JF. Neurosteroids, GABAA receptors, and escalated aggressive behavior. Horm Behav. 2003; 44(3): 242–57. pmid:14609546
- View Article
- PubMed/NCBI
- Google Scholar
42. Verri T, Terova G, Romano A, Barca A, Pisani P, Storelli C, et al. The solute carrier (SLC) Family Series in Teleost Fish. In: Saroglia M, Liu ZJ, editors. Functional Genomics in Aquaculture: John Wiley & Sons, Inc.; 2012. p. 219–320.
43. Teles MC, Cardoso SD, Oliveira RF. Social Plasticity Relies on Different Neuroplasticity Mechanisms across the Brain Social Decision-Making Network in Zebrafish. Front Behav Neurosci. 2016; 10: 16. pmid:26909029
- View Article
- PubMed/NCBI
- Google Scholar
44. McClung CA, Nestler EJ. Neuroplasticity mediated by altered gene expression. Neuropsychopharmacology. 2008; 33(1): 3–17. pmid:17728700
- View Article
- PubMed/NCBI
- Google Scholar
45. Mueller T, Wullimann MF. BrdU-, neuroD (nrd)- and Hu-studies reveal unusual non-ventricular neurogenesis in the postembryonic zebrafish forebrain. Mech Dev. 2002; 117(1–2): 123–35. pmid:12204253
- View Article
- PubMed/NCBI
- Google Scholar
46. O'Connell LA, Fontenot MR, Hofmann HA. Neurochemical profiling of dopaminergic neurons in the forebrain of a cichlid fish, Astatotilapia burtoni. J Chem Neuroanat. 2013; 47: 106–15. pmid:23295359
- View Article
- PubMed/NCBI
- Google Scholar
47. Huntingford FA, Metcalfe NB, Thorpe J. Social status ans feeding in Atlantic salmon Salmo salar Parr: the effect of visual exposure to a dominant. Ethology. 1993; 94(3): 201–6.
- View Article
- Google Scholar
48. Øverli Ø, Pottinger TG, Carrick TR, Øverli E, Winberg S. Differences in behaviour between rainbow trout selected for high- and low-stress responsiveness. J Exp Biol. 2002; 205(Pt 3): 391–5. pmid:11854375
- View Article
- PubMed/NCBI
- Google Scholar
49. Hampel M, Ortiz-Delgado JB, Sarasquete C, Blasco J. Effects of sediment sorbed linear alkylbenzene sulphonate on juveniles of the Senegal sole, Solea senegalensis: toxicity and histological indicators. Histol Histopathol. 2008; 23(1): 87–100. pmid:17952861
- View Article
- PubMed/NCBI
- Google Scholar
50. Carazo I, Norambuena F, Oliveira C, Sánchez-Vázquez FJ, Duncan N. The effect of night illumination, red and infrared light, on locomotor activity behaviour and melatonin of Senegalese sole (Solea senegalensis) broodstock. Physiol Behav. 2013; 118: 201–7. pmid:23711567
- View Article
- PubMed/NCBI
- Google Scholar
51. Reebs SG. Aggression in fishes. Université de Moncton, Canada: howfishbehave.ca; 2008 http://www.howfishbehave.ca
52. Rubenstein D. Population density, resource patterning, and territoriality in the Everglades pygmy sunfish. Anim Behav. 1981; 29: 155–72.
- View Article
- Google Scholar
53. Grand TC, Grant JWA. Spatial predictability of food influences its monopolization and defence by juvenile convict cichlids. Anim Behav. 1994; 47: 91–100.
- View Article
- Google Scholar
54. Grant JWA, Girard IL, Breau C, Weir LK. Influence of food abundance on competitive aggression in juvenile convict cichlids. Anim Behav. 2002; 63: 323–30.
- View Article
- Google Scholar
55. Rozen S, Skaletsky H. Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol. 2000; 132: 365–86. pmid:10547847
- View Article
- PubMed/NCBI
- Google Scholar
56. Nyman C, Fischer S, Aubin-Horth N, Taborsky B. Effect of the early social environment on behavioural and genomic responses to a social challenge in a cooperatively breeding vertebrate. Mol Ecol. 2017; 26(12): 3186–203. pmid:28321979
- View Article
- PubMed/NCBI
- Google Scholar
57. Blin M, Norton W, Bally-Cuif L, Vernier P. NR4A2 controls the differentiation of selective dopaminergic nuclei in the zebrafish brain. Mol Cell Neurosci. 2008; 39(4): 592–604. pmid:18822380
- View Article
- PubMed/NCBI
- Google Scholar
58. Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002; 3(7): RESEARCH0034. pmid:12184808
- View Article
- PubMed/NCBI
- Google Scholar
59. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001; 29(9): e45. pmid:11328886
- View Article
- PubMed/NCBI
- Google Scholar
60. Campbell NA. Chapter 50. In: Cummings B, editor. Biology (4 ed). New York 1996.
61. MacLean A, Metcalfe NB, Mitchell DL. Alternative competitive strategies in juvenile Atlantic salmon (Salmo salar): evidence from fin damage. Aquaculture. 2000; 184: 291–302.
- View Article
- Google Scholar
62. Reale D, Garant D, Humphries MM, Bergeron P, Careau V, Montiglio PO. Personality and the emergence of the pace-of-life syndrome concept at the population level. Philos Trans R Soc Lond B Biol Sci. 2010; 365(1560): 4051–63. pmid:21078657
- View Article
- PubMed/NCBI
- Google Scholar
63. Koolhaas JM, Korte SM, De Boer SF, Van Der Vegt BJ, Van Reenen CG, Hopster H, et al. Coping styles in animals: current status in behavior and stress-physiology. Neurosci Biobehav Rev. 1999; 23: 925–35. pmid:10580307
- View Article
- PubMed/NCBI
- Google Scholar
64. Ibarra-Zatarain Z, Morais S, Bonacic K, Campoverde C, Duncan N. Dietary fatty acid composition significantly influenced the proactive-reactive behaviour of Senegalese sole (Solea senegalensis) post-larvae. Appl Anim Behav Sci. 2015; 171: 233–40.
- View Article
- Google Scholar
65. Holmes RA, Gibson RN. Visual cues determining prey selection by the turbot, Scophthalmus maximus L. J Fish Biol. 1986; 29(Suppl. A): 49–58.
- View Article
- Google Scholar
66. Holmes RA, Gibson RN. A comparison of predatory behaviour in flatfish. Anim Behav. 1983; 31: 1244–55.
- View Article
- Google Scholar
67. Livingston ME. Food resource use among five flatfish species (Pleuronectiformes) in Wellington Harbour, New Zealand. New Zeal J Mar Fresh Res. 1987; 21: 281–93.
- View Article
- Google Scholar
68. Clement TS, Parikh V, Schrumpf M, Fernald RD. Behavioral coping strategies in a cichlid fish: the role of social status and acute stress response in direct and displaced aggression. Horm Behav. 2005; 47(3): 336–42. pmid:15708763
- View Article
- PubMed/NCBI
- Google Scholar
69. Mas-Munoz J, Komen H, Schneider O, Visch SW, Schrama JW. Feeding behaviour, swimming activity and boldness explain variation in feed intake and growth of sole (Solea solea) reared in captivity. PLoS One. 2011; 6(6): e21393. pmid:21738651
- View Article
- PubMed/NCBI
- Google Scholar
70. Guhl AM. The social order of chickens. Sci Am. 1956; 194(2): 42–6.
- View Article
- Google Scholar
71. Guhl AM. Social inertia and social stability in chickens. Anim Behav. 1968; 16(2): 219–32. pmid:5691847
- View Article
- PubMed/NCBI
- Google Scholar
72. killen SS, Mitchell MD, Rummer JL, Chivers DP, Ferrari MCO, Meekan MG, et al. Aerobic scope predicts dominance during early life in a tropical damselfish. Funct Ecol. 2014; 28: 1367–76.
- View Article
- Google Scholar
73. McCormick MI. Protogyny in a tropical damselfish: females queue for future benefit. PeerJ. 2016; 4: e2198. pmid:27413641
- View Article
- PubMed/NCBI
- Google Scholar
74. Amorim MCP, Fonseca PJ, Almada VC. Sound production during courtship and spawning of Oreochromis mossambicus: male-female and male-male interactions. J Fish Biol. 2003; 62: 658–72.
- View Article
- Google Scholar
75. Coelho JL, Medalha CC, Mattioli R. Analysis of the effects of CPA and L-histidine on goldfish tested on a conditioned place preference model. Behav Brain Res. 2001; 124(2): 161–5. pmid:11640969
- View Article
- PubMed/NCBI
- Google Scholar
76. Mattioli R, Nelson CA, Huston JP, Spieler RE. Conditioned place-preference analysis in the goldfish with the H1 histamine antagonist chlorpheniramine. Brain Res Bull. 1998; 45(1): 41–4. pmid:9434200
- View Article
- PubMed/NCBI
- Google Scholar
77. Kruuk H. Diurnal periodicity in the activity of the common sole, Solea vulgaris Quensel. Neth J Sea Res. 1963; 2(1): 1–28.
- View Article
- Google Scholar
78. Galhardo L, Almeida O, Oliveira RF. Preference for the presence of substrate in male cichlid fish: effects of social dominance and context. Appl Anim Behav Sci. 2009; 120: 224–30.
- View Article
- Google Scholar
79. Johnsson JI, Carlsson M, Sundström LF. Habitat preference increases territorial defence in brown trout (Salmo trutta). Behav Ecol Sociobiol. 2000; 48: 373–7.
- View Article
- Google Scholar
80. Huntingford FA. Implications of domestication and rearing conditions for the behaviour of cultivated fishes. J Fish Biol. 2004; 65(Supplement A): 122–42.
- View Article
- Google Scholar
81. Berton O, Durand M, Aguerre S, Mormede P, Chaouloff F. Behavioral, neuroendocrine and serotonergic consequences of single social defeat and repeated fluoxetine pretreatment in the Lewis rat strain. Neuroscience. 1999; 92(1): 327–41. pmid:10392854
- View Article
- PubMed/NCBI
- Google Scholar
82. Blanchard RJ, McKittrick CR, Blanchard DC. Animal models of social stress: effects on behavior and brain neurochemical systems. Physiol Behav. 2001; 73(3): 261–71. pmid:11438351
- View Article
- PubMed/NCBI
- Google Scholar
83. Olson JM, Asakura A, Snider L, Hawkes R, Strand A, Stoeck J, et al. NeuroD2 is necessary for development and survival of central nervous system neurons. Dev Biol. 2001; 234(1): 174–87. pmid:11356028
- View Article
- PubMed/NCBI
- Google Scholar
84. Aimone JB, Li Y, Lee SW, Clemenson GD, Deng W, Gage FH. Regulation and function of adult neurogenesis: from genes to cognition. Physiol Rev. 2014; 94(4): 991–1026. pmid:25287858
- View Article
- PubMed/NCBI
- Google Scholar
85. Teles MC, Almeida O, Lopes JS, Oliveira RF. Social interactions elicit rapid shifts in functional connectivity in the social decision-making network of zebrafish. Proc Biol Sci. 2015; 282(1816): 20151099. pmid:26423839
- View Article
- PubMed/NCBI
- Google Scholar
86. Cardoso SD, Teles MC, Oliveira RF. Neurogenomic mechanisms of social plasticity. J Exp Biol. 2015; 218(Pt 1): 140–9. pmid:25568461
- View Article
- PubMed/NCBI
- Google Scholar
87. Pavlidis M, Sundvik M, Chen YC, Panula P. Adaptive changes in zebrafish brain in dominant-subordinate behavioral context. Behav Brain Res. 2011; 225(2): 529–37. pmid:21875623
- View Article
- PubMed/NCBI
- Google Scholar

[ref1] 1. Griffin DR. Animal Thinking. Cambridge, MA: Harvard University Press.; 1984.

[ref2] 2. Cheney DL, Seyfarth RM. How monkeys See the World. Chicago: University of Chicago Press; 1991. 377 p.

[ref3] 3. Barkow G. The puzzling paucity of feeding territories among freshwater fish. Mar Behav Physiol. 1992; 22: 157–76.
View Article
Google Scholar

[4] View Article

[5] Google Scholar

[ref4] 4. Dugatkin LA, Wilson DS. Fish behaviour, partner choice experiments and cognitive ethology. Rev Fish Biol Fish 1993; 3: 368–72.
View Article
Google Scholar

[7] View Article

[8] Google Scholar

[ref5] 5. Gibson R, editor. Flatfishes: Biology and Exploitation: Blackwell Science Ltd, a Blackwell Publishing company; 2005.

[ref6] 6. APROMAR. La acuicultura marina de peces en España. Asociación Empresarial de Productores de Cultivos Marino. 2016 (Spain).

[ref7] 7. Morais S, Aragao C, Cabrita E, Conceiçao L, Constenla M, Costas B, et al. New developments and biological insights into the farming of Solea senegalensis reinforcing its aquaculture potential. Rev aquacult. 2014; 6: 1–37.
View Article
Google Scholar

[12] View Article

[13] Google Scholar

[ref8] 8. Salas-Leiton E, Anguis V, Manchado M, Cañavate JP. Growth, feeding and oxygen consumption of Senegalese sole (Solea senegalensis) juveniles stocked at different densities. Aquaculture. 2008; 285: 84–9.
View Article
Google Scholar

[15] View Article

[16] Google Scholar

[ref9] 9. Salas-Leiton E, Anguis V, Martin-Antonio B, Crespo D, Planas JV, Infante C, et al. Effects of stocking density and feed ration on growth and gene expression in the Senegalese sole (Solea senegalensis): potential effects on the immune response. Fish Shellfish Immunol. 2010; 28(2): 296–302. pmid:19909816
View Article
PubMed/NCBI
Google Scholar

[18] View Article

[19] PubMed/NCBI

[20] Google Scholar

[ref10] 10. Martín IE. Advances in the reproductive biology and zootechnics of the Senegalese sole (Solea senegalensis Kaup, 1858). Ph.D. Thesis, Universidad de Cantabria (Spain). 2016.

[ref11] 11. Carazo I, Martin I, Hubbard PC, Chereguini O, Mañanós EL, Canario AVM, et al. Reproductive behaviour, the absence of reproductive behaviour in cultured (G1 generation) and chemical communication in the Senegalese sole (Solea senegalensis). Indian Journal of Science and Technology (9th ISRPF). 2011; 4: 96–7.
View Article
Google Scholar

[23] View Article

[24] Google Scholar

[ref12] 12. Carazo I. Comportamiento reproductivo y fisiología del lenguado senegalés (Solea senegalensis) en cautividad. Ph. D. Thesis, Universidad de Barcelona (Spain). 2013.

[ref13] 13. Martín I, Rasines I, Gómez M, Rodríguez C, Martínez P, Chereguini O. Evolution of egg production and parental contribution in Senegalese sole, Solea senegalensis, during four consecutive spawning seasons. Aquaculture. 2014; 424–425: 45–52.
View Article
Google Scholar

[27] View Article

[28] Google Scholar

[ref14] 14. Porta J, Porta JM, Martínez-Rodríguez G, Álvarez MC. Development of a microsatellite multiplex PCR for Senegalese sole (Solea senegalensis) and its application to broodstock management. Aquaculture. 2006; 265: 159–66.
View Article
Google Scholar

[30] View Article

[31] Google Scholar

[ref15] 15. Carazo I, Chereguini O, Martin I, Huntingford F, Duncan NJ. Reproductive ethogram and mate selection in captive wild Senegalese sole (Solea senegalensis). Span J Agric Res. 2016; 14(4): e0401.
View Article
Google Scholar

[33] View Article

[34] Google Scholar

[ref16] 16. Silva PIM, Martins CIM, Engrola S, Marino G, Øverli Ø, Conceição LEC. Individual differences in cortisol levels and behaviour of Senegalese sole (Solea senegalensis) juveniles: Evidence for coping styles. Appl Anim Behav Sci. 2010; 124: 75–81.
View Article
Google Scholar

[36] View Article

[37] Google Scholar

[ref17] 17. Ibarra-Zatarain Z, Fatsini E, Rey S, Chereguini O, Martin I, Rasines I, et al. Characterization of stress coping style in Senegalese sole (Solea senegalensis) juveniles and breeders for aquaculture. R Soc Open Sci. 2016; 3:160495. pmid:28018634
View Article
PubMed/NCBI
Google Scholar

[39] View Article

[40] PubMed/NCBI

[41] Google Scholar

[ref18] 18. Ibarra-Zatarain Z. The role of stress coping styles in reproduction and other biological aspects in the aquaculture species, Senegalese sole (Solea senegalensis) and gilthead seabream (Sparus aurata). Ph. D. Thesis, Universidad Autónoma de Barcelona (Spain). 2015.

[ref19] 19. Noakes DLG. Social behaviour as it influences fish production In: Gerking SD, editor. Ecology of freshwater fish production. Oxford: Blackwell Scientific Publ., Oxford; 1978. p. 360–82.

[ref20] 20. Grant JWA. Territoriality. In: Godin J-GJ, editor. Behavioural Ecology of Teleost Fishes. United States: Oxford University press Inc., New York; 1997. p. 360.

[ref21] 21. Øverli Ø, Harris CA, Winberg S. Short-term effects of fights for social dominance and the establishment of dominant-subordinate relationships on brain monoamines and cortisol in rainbow trout. Brain Behav Evol. 1999; 54(5): 263–75. pmid:10640786
View Article
PubMed/NCBI
Google Scholar

[46] View Article

[47] PubMed/NCBI

[48] Google Scholar

[ref22] 22. Cardoso P, Volpato GL. Chemical communication, aggression, and conspecific recognition in the fish Nile tilapia. Physiol Behav. 1997; 62(6): 1333–8. pmid:9383122
View Article
PubMed/NCBI
Google Scholar

[50] View Article

[51] PubMed/NCBI

[52] Google Scholar

[ref23] 23. Andersson MA, Höglund E. Linking personality to larval energy reserves in rainbow trout (Oncorhynchus mykiss). PLoS One. 2012; 7(11): e49247. pmid:23155473
View Article
PubMed/NCBI
Google Scholar

[54] View Article

[55] PubMed/NCBI

[56] Google Scholar

[ref24] 24. Ruiz-Gomez ML, Huntingford FA. Boldness and aggressiveness in early and late hatched three-spined sticklebacks Gasterosteus aculeatus. J Fish Biol. 2012; 81(3): 966–76. pmid:22880730
View Article
PubMed/NCBI
Google Scholar

[58] View Article

[59] PubMed/NCBI

[60] Google Scholar

[ref25] 25. Øverli Ø, Korzan WJ, Höglund E, Winberg S, Bollig H, Watt M, et al. Stress coping style predicts aggression and social dominance in rainbow trout. Horm Behav. 2004; 45: 235–41. pmid:15053939
View Article
PubMed/NCBI
Google Scholar

[62] View Article

[63] PubMed/NCBI

[64] Google Scholar

[ref26] 26. Øverli Ø, Kotzian S, Winberg S. Effects of cortisol on aggression and locomotor activity in rainbow trout. Horm Behav. 2002; 42(1): 53–61. pmid:12191647
View Article
PubMed/NCBI
Google Scholar

[66] View Article

[67] PubMed/NCBI

[68] Google Scholar

[ref27] 27. Øverli Ø. Preface: plasticity and diversity in behavior and brain function—important raw material for natural selection? Brain Behav Evol. 2007; 70(4): 215–7. pmid:17914252
View Article
PubMed/NCBI
Google Scholar

[70] View Article

[71] PubMed/NCBI

[72] Google Scholar

[ref28] 28. Korzan WJ, Øverli Ø, Summers CH. Future social rank: forecasting status in the green anole (Anolis carolinensis). Acta Ethol. 2006; 9: 48–57.
View Article
Google Scholar

[74] View Article

[75] Google Scholar

[ref29] 29. Delicio HC, Barreto RE, Normandes EB, Luchiari AC, Marcondes AL. A place preference test in the fish Nile tilapia. J Exp Anim Sci. 2006; 43: 141–8.
View Article
Google Scholar

[77] View Article

[78] Google Scholar

[ref30] 30. Popma T, Masser M. Tilapia life history and biology. SRAC. 1999; 283: 1–4.
View Article
Google Scholar

[80] View Article

[81] Google Scholar

[ref31] 31. Serra EL, Medalha CC, Mattioli R. Natural preference of zebrafish (Danio rerio) for a dark environment. Braz J Med Biol Res. 1999; 32: 1551–3. pmid:10585639
View Article
PubMed/NCBI
Google Scholar

[83] View Article

[84] PubMed/NCBI

[85] Google Scholar

[ref32] 32. English S, Huchard E, Nielsen JF, Clutton-Brock TH. Early growth, dominance acquisition and lifetime reproductive success in male and female cooperative meerkats. Ecol Evol. 2013; 3(13): 4401–7. pmid:24340181
View Article
PubMed/NCBI
Google Scholar

[87] View Article

[88] PubMed/NCBI

[89] Google Scholar

[ref33] 33. Rodel HG, von Holst D. Features of the early juvenile development predict competitive performance in male European rabbits. Physiol Behav. 2009; 97(3–4): 495–502. pmid:19375438
View Article
PubMed/NCBI
Google Scholar

[91] View Article

[92] PubMed/NCBI

[93] Google Scholar

[ref34] 34. Rey S, Boltana S, Vargas R, Roher N, Mackenzie S. Combining animal personalities with transcriptomics resolves individual variation within a wild-type zebrafish population and identifies underpinning molecular differences in brain function. Mol Ecol. 2013; 22: 6100–15. pmid:24118534
View Article
PubMed/NCBI
Google Scholar

[95] View Article

[96] PubMed/NCBI

[97] Google Scholar

[ref35] 35. MacKenzie S, Ribas L, Pilarczyk M, Capdevila DM, Kadri S, Huntingford FA. Screening for coping style increases the power of gene expression studies. PLoS One. 2009; 4(4): e5314. pmid:19390591
View Article
PubMed/NCBI
Google Scholar

[99] View Article

[100] PubMed/NCBI

[101] Google Scholar

[ref36] 36. Driscoll P, Escorihuela RM, Fernandez-Teruel A, Giorgi O, Schwegler H, Steimer T, et al. Genetic selection and differential stress responses. The Roman lines/strains of rats. Ann N Y Acad Sci. 1998; 851: 501–10. pmid:9668644
View Article
PubMed/NCBI
Google Scholar

[103] View Article

[104] PubMed/NCBI

[105] Google Scholar

[ref37] 37. Filby AL, Paull GC, Hickmore TF, Tyler CR. Unravelling the neurophysiological basis of aggression in a fish model. BMC Genomics. 2010; 11: 498. pmid:20846403
View Article
PubMed/NCBI
Google Scholar

[107] View Article

[108] PubMed/NCBI

[109] Google Scholar

[ref38] 38. Filby AL, Paull GC, Searle F, Ortiz-Zarragoitia M, Tyler CR. Environmental estrogen-induced alterations of male aggression and dominance hierarchies in fish: a mechanistic analysis. Environ Sci Technol. 2012; 46(6): 3472–9. pmid:22360147
View Article
PubMed/NCBI
Google Scholar

[111] View Article

[112] PubMed/NCBI

[113] Google Scholar

[ref39] 39. Zhang X, Beaulieu JM, Sotnikova TD, Gainetdinov RR, Caron MG. Tryptophan hydroxylase-2 controls brain serotonin synthesis. Science. 2004; 305(5681): 217. pmid:15247473
View Article
PubMed/NCBI
Google Scholar

[115] View Article

[116] PubMed/NCBI

[117] Google Scholar

[ref40] 40. Miczek KA, Fish EW, De Bold JF, De Almeida RM. Social and neural determinants of aggressive behavior: pharmacotherapeutic targets at serotonin, dopamine and gamma-aminobutyric acid systems. Psychopharmacology. 2002; 163(3–4): 434–58. pmid:12373445
View Article
PubMed/NCBI
Google Scholar

[119] View Article

[120] PubMed/NCBI

[121] Google Scholar

[ref41] 41. Miczek KA, Fish EW, De Bold JF. Neurosteroids, GABAA receptors, and escalated aggressive behavior. Horm Behav. 2003; 44(3): 242–57. pmid:14609546
View Article
PubMed/NCBI
Google Scholar

[123] View Article

[124] PubMed/NCBI

[125] Google Scholar

[ref42] 42. Verri T, Terova G, Romano A, Barca A, Pisani P, Storelli C, et al. The solute carrier (SLC) Family Series in Teleost Fish. In: Saroglia M, Liu ZJ, editors. Functional Genomics in Aquaculture: John Wiley & Sons, Inc.; 2012. p. 219–320.

[ref43] 43. Teles MC, Cardoso SD, Oliveira RF. Social Plasticity Relies on Different Neuroplasticity Mechanisms across the Brain Social Decision-Making Network in Zebrafish. Front Behav Neurosci. 2016; 10: 16. pmid:26909029
View Article
PubMed/NCBI
Google Scholar

[128] View Article

[129] PubMed/NCBI

[130] Google Scholar

[ref44] 44. McClung CA, Nestler EJ. Neuroplasticity mediated by altered gene expression. Neuropsychopharmacology. 2008; 33(1): 3–17. pmid:17728700
View Article
PubMed/NCBI
Google Scholar

[132] View Article

[133] PubMed/NCBI

[134] Google Scholar

[ref45] 45. Mueller T, Wullimann MF. BrdU-, neuroD (nrd)- and Hu-studies reveal unusual non-ventricular neurogenesis in the postembryonic zebrafish forebrain. Mech Dev. 2002; 117(1–2): 123–35. pmid:12204253
View Article
PubMed/NCBI
Google Scholar

[136] View Article

[137] PubMed/NCBI

[138] Google Scholar

[ref46] 46. O'Connell LA, Fontenot MR, Hofmann HA. Neurochemical profiling of dopaminergic neurons in the forebrain of a cichlid fish, Astatotilapia burtoni. J Chem Neuroanat. 2013; 47: 106–15. pmid:23295359
View Article
PubMed/NCBI
Google Scholar

[140] View Article

[141] PubMed/NCBI

[142] Google Scholar

[ref47] 47. Huntingford FA, Metcalfe NB, Thorpe J. Social status ans feeding in Atlantic salmon Salmo salar Parr: the effect of visual exposure to a dominant. Ethology. 1993; 94(3): 201–6.
View Article
Google Scholar

[144] View Article

[145] Google Scholar

[ref48] 48. Øverli Ø, Pottinger TG, Carrick TR, Øverli E, Winberg S. Differences in behaviour between rainbow trout selected for high- and low-stress responsiveness. J Exp Biol. 2002; 205(Pt 3): 391–5. pmid:11854375
View Article
PubMed/NCBI
Google Scholar

[147] View Article

[148] PubMed/NCBI

[149] Google Scholar

[ref49] 49. Hampel M, Ortiz-Delgado JB, Sarasquete C, Blasco J. Effects of sediment sorbed linear alkylbenzene sulphonate on juveniles of the Senegal sole, Solea senegalensis: toxicity and histological indicators. Histol Histopathol. 2008; 23(1): 87–100. pmid:17952861
View Article
PubMed/NCBI
Google Scholar

[151] View Article

[152] PubMed/NCBI

[153] Google Scholar

[ref50] 50. Carazo I, Norambuena F, Oliveira C, Sánchez-Vázquez FJ, Duncan N. The effect of night illumination, red and infrared light, on locomotor activity behaviour and melatonin of Senegalese sole (Solea senegalensis) broodstock. Physiol Behav. 2013; 118: 201–7. pmid:23711567
View Article
PubMed/NCBI
Google Scholar

[155] View Article

[156] PubMed/NCBI

[157] Google Scholar

[ref51] 51. Reebs SG. Aggression in fishes. Université de Moncton, Canada: howfishbehave.ca; 2008 http://www.howfishbehave.ca

[ref52] 52. Rubenstein D. Population density, resource patterning, and territoriality in the Everglades pygmy sunfish. Anim Behav. 1981; 29: 155–72.
View Article
Google Scholar

[160] View Article

[161] Google Scholar

[ref53] 53. Grand TC, Grant JWA. Spatial predictability of food influences its monopolization and defence by juvenile convict cichlids. Anim Behav. 1994; 47: 91–100.
View Article
Google Scholar

[163] View Article

[164] Google Scholar

[ref54] 54. Grant JWA, Girard IL, Breau C, Weir LK. Influence of food abundance on competitive aggression in juvenile convict cichlids. Anim Behav. 2002; 63: 323–30.
View Article
Google Scholar

[166] View Article

[167] Google Scholar

[ref55] 55. Rozen S, Skaletsky H. Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol. 2000; 132: 365–86. pmid:10547847
View Article
PubMed/NCBI
Google Scholar

[169] View Article

[170] PubMed/NCBI

[171] Google Scholar

[ref56] 56. Nyman C, Fischer S, Aubin-Horth N, Taborsky B. Effect of the early social environment on behavioural and genomic responses to a social challenge in a cooperatively breeding vertebrate. Mol Ecol. 2017; 26(12): 3186–203. pmid:28321979
View Article
PubMed/NCBI
Google Scholar

[173] View Article

[174] PubMed/NCBI

[175] Google Scholar

[ref57] 57. Blin M, Norton W, Bally-Cuif L, Vernier P. NR4A2 controls the differentiation of selective dopaminergic nuclei in the zebrafish brain. Mol Cell Neurosci. 2008; 39(4): 592–604. pmid:18822380
View Article
PubMed/NCBI
Google Scholar

[177] View Article

[178] PubMed/NCBI

[179] Google Scholar

[ref58] 58. Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002; 3(7): RESEARCH0034. pmid:12184808
View Article
PubMed/NCBI
Google Scholar

[181] View Article

[182] PubMed/NCBI

[183] Google Scholar

[ref59] 59. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001; 29(9): e45. pmid:11328886
View Article
PubMed/NCBI
Google Scholar

[185] View Article

[186] PubMed/NCBI

[187] Google Scholar

[ref60] 60. Campbell NA. Chapter 50. In: Cummings B, editor. Biology (4 ed). New York 1996.

[ref61] 61. MacLean A, Metcalfe NB, Mitchell DL. Alternative competitive strategies in juvenile Atlantic salmon (Salmo salar): evidence from fin damage. Aquaculture. 2000; 184: 291–302.
View Article
Google Scholar

[190] View Article

[191] Google Scholar

[ref62] 62. Reale D, Garant D, Humphries MM, Bergeron P, Careau V, Montiglio PO. Personality and the emergence of the pace-of-life syndrome concept at the population level. Philos Trans R Soc Lond B Biol Sci. 2010; 365(1560): 4051–63. pmid:21078657
View Article
PubMed/NCBI
Google Scholar

[193] View Article

[194] PubMed/NCBI

[195] Google Scholar

[ref63] 63. Koolhaas JM, Korte SM, De Boer SF, Van Der Vegt BJ, Van Reenen CG, Hopster H, et al. Coping styles in animals: current status in behavior and stress-physiology. Neurosci Biobehav Rev. 1999; 23: 925–35. pmid:10580307
View Article
PubMed/NCBI
Google Scholar

[197] View Article

[198] PubMed/NCBI

[199] Google Scholar

[ref64] 64. Ibarra-Zatarain Z, Morais S, Bonacic K, Campoverde C, Duncan N. Dietary fatty acid composition significantly influenced the proactive-reactive behaviour of Senegalese sole (Solea senegalensis) post-larvae. Appl Anim Behav Sci. 2015; 171: 233–40.
View Article
Google Scholar

[201] View Article

[202] Google Scholar

[ref65] 65. Holmes RA, Gibson RN. Visual cues determining prey selection by the turbot, Scophthalmus maximus L. J Fish Biol. 1986; 29(Suppl. A): 49–58.
View Article
Google Scholar

[204] View Article

[205] Google Scholar

[ref66] 66. Holmes RA, Gibson RN. A comparison of predatory behaviour in flatfish. Anim Behav. 1983; 31: 1244–55.
View Article
Google Scholar

[207] View Article

[208] Google Scholar

[ref67] 67. Livingston ME. Food resource use among five flatfish species (Pleuronectiformes) in Wellington Harbour, New Zealand. New Zeal J Mar Fresh Res. 1987; 21: 281–93.
View Article
Google Scholar

[210] View Article

[211] Google Scholar

[ref68] 68. Clement TS, Parikh V, Schrumpf M, Fernald RD. Behavioral coping strategies in a cichlid fish: the role of social status and acute stress response in direct and displaced aggression. Horm Behav. 2005; 47(3): 336–42. pmid:15708763
View Article
PubMed/NCBI
Google Scholar

[213] View Article

[214] PubMed/NCBI

[215] Google Scholar

[ref69] 69. Mas-Munoz J, Komen H, Schneider O, Visch SW, Schrama JW. Feeding behaviour, swimming activity and boldness explain variation in feed intake and growth of sole (Solea solea) reared in captivity. PLoS One. 2011; 6(6): e21393. pmid:21738651
View Article
PubMed/NCBI
Google Scholar

[217] View Article

[218] PubMed/NCBI

[219] Google Scholar

[ref70] 70. Guhl AM. The social order of chickens. Sci Am. 1956; 194(2): 42–6.
View Article
Google Scholar

[221] View Article

[222] Google Scholar

[ref71] 71. Guhl AM. Social inertia and social stability in chickens. Anim Behav. 1968; 16(2): 219–32. pmid:5691847
View Article
PubMed/NCBI
Google Scholar

[224] View Article

[225] PubMed/NCBI

[226] Google Scholar

[ref72] 72. killen SS, Mitchell MD, Rummer JL, Chivers DP, Ferrari MCO, Meekan MG, et al. Aerobic scope predicts dominance during early life in a tropical damselfish. Funct Ecol. 2014; 28: 1367–76.
View Article
Google Scholar

[228] View Article

[229] Google Scholar

[ref73] 73. McCormick MI. Protogyny in a tropical damselfish: females queue for future benefit. PeerJ. 2016; 4: e2198. pmid:27413641
View Article
PubMed/NCBI
Google Scholar

[231] View Article

[232] PubMed/NCBI

[233] Google Scholar

[ref74] 74. Amorim MCP, Fonseca PJ, Almada VC. Sound production during courtship and spawning of Oreochromis mossambicus: male-female and male-male interactions. J Fish Biol. 2003; 62: 658–72.
View Article
Google Scholar

[235] View Article

[236] Google Scholar

[ref75] 75. Coelho JL, Medalha CC, Mattioli R. Analysis of the effects of CPA and L-histidine on goldfish tested on a conditioned place preference model. Behav Brain Res. 2001; 124(2): 161–5. pmid:11640969
View Article
PubMed/NCBI
Google Scholar

[238] View Article

[239] PubMed/NCBI

[240] Google Scholar

[ref76] 76. Mattioli R, Nelson CA, Huston JP, Spieler RE. Conditioned place-preference analysis in the goldfish with the H1 histamine antagonist chlorpheniramine. Brain Res Bull. 1998; 45(1): 41–4. pmid:9434200
View Article
PubMed/NCBI
Google Scholar

[242] View Article

[243] PubMed/NCBI

[244] Google Scholar

[ref77] 77. Kruuk H. Diurnal periodicity in the activity of the common sole, Solea vulgaris Quensel. Neth J Sea Res. 1963; 2(1): 1–28.
View Article
Google Scholar

[246] View Article

[247] Google Scholar

[ref78] 78. Galhardo L, Almeida O, Oliveira RF. Preference for the presence of substrate in male cichlid fish: effects of social dominance and context. Appl Anim Behav Sci. 2009; 120: 224–30.
View Article
Google Scholar

[249] View Article

[250] Google Scholar

[ref79] 79. Johnsson JI, Carlsson M, Sundström LF. Habitat preference increases territorial defence in brown trout (Salmo trutta). Behav Ecol Sociobiol. 2000; 48: 373–7.
View Article
Google Scholar

[252] View Article

[253] Google Scholar

[ref80] 80. Huntingford FA. Implications of domestication and rearing conditions for the behaviour of cultivated fishes. J Fish Biol. 2004; 65(Supplement A): 122–42.
View Article
Google Scholar

[255] View Article

[256] Google Scholar

[ref81] 81. Berton O, Durand M, Aguerre S, Mormede P, Chaouloff F. Behavioral, neuroendocrine and serotonergic consequences of single social defeat and repeated fluoxetine pretreatment in the Lewis rat strain. Neuroscience. 1999; 92(1): 327–41. pmid:10392854
View Article
PubMed/NCBI
Google Scholar

[258] View Article

[259] PubMed/NCBI

[260] Google Scholar

[ref82] 82. Blanchard RJ, McKittrick CR, Blanchard DC. Animal models of social stress: effects on behavior and brain neurochemical systems. Physiol Behav. 2001; 73(3): 261–71. pmid:11438351
View Article
PubMed/NCBI
Google Scholar

[262] View Article

[263] PubMed/NCBI

[264] Google Scholar

[ref83] 83. Olson JM, Asakura A, Snider L, Hawkes R, Strand A, Stoeck J, et al. NeuroD2 is necessary for development and survival of central nervous system neurons. Dev Biol. 2001; 234(1): 174–87. pmid:11356028
View Article
PubMed/NCBI
Google Scholar

[266] View Article

[267] PubMed/NCBI

[268] Google Scholar

[ref84] 84. Aimone JB, Li Y, Lee SW, Clemenson GD, Deng W, Gage FH. Regulation and function of adult neurogenesis: from genes to cognition. Physiol Rev. 2014; 94(4): 991–1026. pmid:25287858
View Article
PubMed/NCBI
Google Scholar

[270] View Article

[271] PubMed/NCBI

[272] Google Scholar

[ref85] 85. Teles MC, Almeida O, Lopes JS, Oliveira RF. Social interactions elicit rapid shifts in functional connectivity in the social decision-making network of zebrafish. Proc Biol Sci. 2015; 282(1816): 20151099. pmid:26423839
View Article
PubMed/NCBI
Google Scholar

[274] View Article

[275] PubMed/NCBI

[276] Google Scholar

[ref86] 86. Cardoso SD, Teles MC, Oliveira RF. Neurogenomic mechanisms of social plasticity. J Exp Biol. 2015; 218(Pt 1): 140–9. pmid:25568461
View Article
PubMed/NCBI
Google Scholar

[278] View Article

[279] PubMed/NCBI

[280] Google Scholar

[ref87] 87. Pavlidis M, Sundvik M, Chen YC, Panula P. Adaptive changes in zebrafish brain in dominant-subordinate behavioral context. Behav Brain Res. 2011; 225(2): 529–37. pmid:21875623
View Article
PubMed/NCBI
Google Scholar

[282] View Article

[283] PubMed/NCBI

[284] Google Scholar

Figures

Abstract

Introduction

Material and methods

Ethics statement

Animal rearing conditions

Behavioural studies

Video recording analyses.

Preliminary dyadic test.

Dominance tests.

Dyadic tests.

Group test.

RNA isolation, complementary DNA synthesis and quantitative real-time polymerase chain reaction assay

Statistical analysis

Behaviour.

q- rtPCR.

Results

Senegalese sole behavioural responses in dominance tests

Observed feeding behaviour (MAPs in Senegalese sole).

Dominance parameters selection

Individual tests.

Group test.

Brain mRNA abundance in relation to dominance

Discussion

Feeding and growth

Reproduction

Place preference test

Brain mRNA abundance in association to dominance behaviour

Conclusions

Supporting information

S1 Fig. Experimental tank set up used for the place preference test (with sand) in pairs.

S2 Fig. Group experimental tank set up used for feeding response and point feed delivery.

S3 Fig. Principal component analysis of the different behaviours registered during the “feeding response test” in pairs.

S4 Fig. Spearman’s correlations from the three variables “Approaches, SAA and RTH” with feeding.

S5 Fig. Principal component analysis of the different behaviours registered during the feeding dominance test and place preference test (sand) in pairs.

S1 Table. Classification of the different variables in groups according to Kendall’s concordance coefficient (W) for every group.

S1 File. Raw data of preliminary results in dyadic pairs of early Senegalese sole juveniles.

S2 File. Modal action patterns related to feeding in late Senegalese sole juveniles.

Acknowledgments

References