Reproductive success of a marine teleost was correlated with proactive and reactive stress‐coping styles

The present study investigated the relationship between reproductive success and stress‐coping styles in gilthead seabream Sparus aurata in captivity. To characterise stress‐coping styles, a total of 22 breeders were submitted to three different individual‐based tests, one group‐based test and post‐handling glucocorticoid quantification. To assess spawning participation, a microsatellite analysis was performed on a total of 2698 larvae, which allowed each offspring to be assigned unambiguously to a single parental couple. Overall, S. aurata showed defined proactive and reactive behavioural traits. Proactive breeders exhibited higher levels of activity and risk taking and lower glucocorticoid blood levels than reactive breeders. The stress‐coping style traits were consistent over time and context (different tests). Breeders that contributed to a higher number of progeny exhibited proactive behaviours, while those showing low progeny contribution exhibited reactive behaviour. Therefore, breeders with a high proportion of progeny (> 20%) had significantly higher activity and risk taking and lower cortisol than breeders with low progeny contribution (< 20%). In addition, males were more proactive than females and males exhibited significantly higher activity, risk taking and lower cortisol than females. This study is the first to establish in S. aurata breeders: (a) a relationship between stress‐coping styles and spawning success; (b) a relationship between stress‐coping styles and gender; and (c) the existence of proactive and reactive traits at the adult stage.info:eu-repo/semantics/acceptedVersio

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

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. &ldquo;Resting the head&rdquo; on another fish, ii. &ldquo;Approaching&rdquo; another fish, iii. &ldquo;Swimming above another&rdquo; 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 &ldquo;Rest the head&rdquo; and &ldquo;Approaches&rdquo;. 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

The presence of wild Senegalese sole breeders improves courtship and reproductive success in cultured conspecifics

The life cycle of Senegalese sole (Solea senegalensis) is not closed in captivity due to a reproductive dysfunction related to the lack of participation of cultured male breeders in the courtship. To discern a possible solution to this social reproductive dysfunction, the main objective of this study was to determine the effect of the presence of spawning wild Senegalese sole breeders on the courtship and reproduction of cultured breeders. Three experimental groups were formed: Control group (n = 10) formed with only cultured sole; groups M1 and M2 constituted of mixed-origin sole (10 cultured and 8 or 9 wild breeders). All cultured breeders were from the same stock, which had never successfully reproduced, whilst the wild broodstock had spawned viable eggs in captivity. All groups were held in the same captive conditions and spawning, and behaviour were recorded for each group over four spawning seasons. All spawns were collected, and the parameters registered were floating and non-floating eggs, fertilization and hatching rates. In addition, parental analysis was made of larvae from viable spawns. Behaviour was studied with video recordings to compare locomotor activity and courtship behaviours including the "Follow" behaviours where sole swim after each other in a procession. Fertilized spawns were obtained from the mixed-origin groups (M1 and M2) including spawns involving a cultured male. The cultured males also participated actively in the "Follow" behaviours with the courting wild sole and this participation of cultured males increased significantly over the four years of the experiment. Male percentage participation in the "Follow" behaviours was positively correlated (R = 0.81) to participation in spawning to indicate the importance of increasing cultured male participation in the "Follow" behaviours. A total of seven spawns were obtained from a cultured male that fertilized eggs from one cultured female and two wild females. The same cultured female also spawned with a wild male. No viable spawns were obtained from the Control group and locomotor activity and courtship behaviour counts were significantly lower than in the experimental mixedorigin groups. This is the first report of cultured male breeders participating in reproductive behaviour and spawning, which could be associated with social learning processes, mate selection and dominance where cultured males copied the behaviour and spawning of wild Senegalese sole breeders.Spanish National Institute for Agronomic Research (INIA)-FEDER [RTA201100050, RTA2014-00048]FCT - Foundation for Science and TechnologyPortuguese Foundation for Science and Technology [UID/Multi/04326/2019]info:eu-repo/semantics/submittedVersio

Linking stress coping styles with brain mRNA abundance of selected transcripts for Senegalese sole (Solea senegalensis) juveniles

In fish, proactive and reactive individual stress copying styles (SCS) have been used to resolve variation in molecular expression data. Stress coping styles have been previously described in several stages of Solea senegalensis by validating for the species the use of standard behavioural screening tests. The present study aimed to link behavioural SCS tests with brain transcript abundance in early Senegalese sole juveniles in order to observe the natural variation in a molecular pathway in this species. A total of 50 juveniles were subjected to three individual behavioural (Restraining, New environment and Confinement) and one group (Risk-taking) screening tests. The fish were classified in SCS categories by applying a hierarchical cluster to the variable “Total activity” (the total activity time that the fish was moving in each individual test). Three categories were defined, proactive, intermediate and reactive sole. Six transcripts were chosen and tested, one related to basic metabolism (gapdh-2), three to feeding behaviour (per1, igf-Ia, pparß) and two to the stress response (crh-BP and hsp90aa) in 30 juveniles (10 individuals per SCS category) using rt-qPCR to observe differences in the abundance of those transcripts among SCS. Four transcripts were differentially expressed (DETs) among them. The transcript gapdh-2 showed up-regulation for proactive and intermediate SCS sole while reactive individuals showed down-regulation. Target mRNAs per1, igf-Ia and pparß, showed different levels of up-regulation for proactive and reactive fish while intermediates were highly down-regulated. Surprisingly no differences in stress related transcripts were observed. Correlations were found between variation in coping styles and variation in the abundance of mRNAs involved in important biological functions in Senegalese sole. These results are the first evidence of the relationship between the behavioural individual variation and the fluctuation in brain transcripts abundance in Senegalese sole.info:eu-repo/semantics/acceptedVersio

Characterization of stress coping style in Senegalese sole (Soleasenegalensis) juveniles and breeders for aquaculture

The aim of this work was to characterize stress coping styles of Senegalese sole (Solea senegalensis) juveniles and breeders and to select an operational behavioural screening test (OBST) that can be used by the aquaculture industry to classify and select between behavioural phenotypes in order to improve production indicators. A total of 61 juveniles and 59 breeders were subjected to five individual behavioural tests and two grouping tests. At the end of the individual tests, all animals were blood sampled in order to measure cortisol, glucose and lactate. Three tests (restraining, new environment and confinement) characterized the stress coping style behaviour of Senegalese sole juveniles and breeders and demonstrated interindividual consistency. Further, the tests when incorporated into a principal components analysis (PCA) (i) identified two principal axes of personality traits: ‘fearfulness-reactivity’ and ‘activity-exploration’, (ii) were representative of the physiological axis of stress coping style, and (iii) were validated by established group tests. This study proposed for the first time three individual coping style tests that reliably represented proactive and reactive personalities of Senegalese sole juveniles and breeders. In addition, the three proposed tests met some basic operational criteria (rapid testing, no special equipment and easy to apply and interpret) that could prove attractive for fish farmers to identify fish with a specific behaviour that gives advantages in the culture system and that could be used to establish selection-based breeding programmes to improve domestication and production.info:eu-repo/semantics/publishedVersio

Senegalese sole (Solea senegalensis) coping styles are consistent over time: behavioural and physiological responses during ontogenesis

Individuals differ in how they cope with stressful situations along a behavioural continuum, being proactive and reactive at the extremes of this continuum. Proactive individuals are usually bold, highly active and take risks, while reactive organisms are generally shy, exhibit low activity and avoid risky situations. Definitions of stress coping styles state that proactive and reactive traits are consistent over time and across contexts. The present study evaluated the individual differences in stress coping style, physiological changes and reproductive status in Senegalese sole juveniles and breeders over three and two-years, respectively. To determine stress coping style, the fish were subjected to three individual (restraining, new environment, confinement) and one group screening test (risk taking). Both groups were tested on three occasions, juveniles were tested each year and adults were tested in the first year and twice (spring and autumn) in the second year. On the third year, a proportion of the juveniles initiated puberty and the reproductive status of all individuals was assessed and compared with their behavioural responses. Results demonstrated individual differences that were consistent with proactive and reactive traits in juveniles and breeders. Significant intra-individual repeatability and consistency of juveniles and breeder's behavioural responses were observed over time and across situations. In addition, glucocorticoid levels (cortisol) were consistent for individuals. Another result to highlight was that juveniles that past puberty and initiated gametogenesis had significant higher activity, risk predisposition and lower plasma cortisol levels compared to fish that remained immature (did not initiate puberty)

Characterization of the different behaviours exhibited by juvenile flathead grey mullet (Mugil cephalus Linnaeus, 1758) under rearing conditions

Aim of study: To describe the common behaviour of flathead grey mullet (Mugil cephalus) under rearing conditions. Area of study: Tepic, Mexico. Material and methods: Behaviours exhibited by mullets were videorecorded with submersible cameras installed inside of three tanks. A total of 690 min per day (07:30 - 18:30 h) were recorded per tank during a week. Afterwards, the different behaviours exhibited by juvenile M. cephalus were described, identified and characterized in an ethogram and grouped into two categories: a) locomotion, including three different observed behaviours (resting, swimming and fast swimming) and b) feeding, including three behaviours (surface feeding, bottom feeding and rubbing). Each of the behavioural variables were quantified. Main results: M. cephalus is a species with a constant locomotion associated to feeding, since fish showed continuous movement during most of day light period. On the contrary, fish exhibited reduced movement during dark periods. Mullets were observed to be a non-aggressive fish species under conditions of the present study, since the absence of dominance and aggression towards conspecifics was observed, which suggested a high predisposition for adaptation to captivity. Finally, behavioural frequencies of grey mullet juveniles were similar among the three tanks for most of the behavioural variables analysed (p>0.05) except for the variable bottom feeding (p=0.02). Research highlights: Results from this study could be of interest for the aquaculture industry to optimize rearing techniques and welfare for the production of grey mullet.info:eu-repo/semantics/publishedVersio

Paired spawning with male rotation of meagre Argyrosomus regius using GnRHa injections, as a method for producing multiple families for breeding selection programs

Weekly gonadotropin-releasing hormone agonist (GnRHa) injections were used to induce spawning in paired male and female meagre (Argyrosomus regius) with a weekly rotation of the males, in order to produce a large number of families, as a method to facilitate selective breeding programs. Two different broodstocks were used (HCMR and IRTA), with females of mean weights of 11.7 ± 2.6 kg and 20.0 ± 1.8 kg, and males of 10.2 ± 1.2 kg and 15.1 ± 1.0 kg, respectively. A single GnRHa injection of 15 μg kg−1 was administered to each selected female, and 7.5 or 15 μg kg−1 to each male to induce spawning. In the subsequent weeks, maturity was checked and fish were induced as above, but males (n = 18) were rotated to form a different pair with the selected females (n = 21). Experiments finished when all paired combinations had been completed or a fish lost maturity status and could not be induced further. A total of 56 families were produced with a mean number of eggs from each family of 87,666 ± 11,244 eggs kg−1. There was a decline in the fecundity, number of spawns and percentage of pairs that spawned successfully after consecutive weekly GnRHa injections. Relative fecundity declined significantly from 134,495 ± 25,557 eggs kg−1 female body weight after the first injection, to 44,252 ± 17,638 eggs kg−1 after the fourth injection. However, there were no differences amongst weeks in egg fertilization success, hatching success or larval survival to 5 days post hatch. The decrease in fecundity and spawning success was attributed to a loss of maturity observed in the females, which may be related to differences in mate selection strategies between male and female meagre. The study demonstrated that paired spawning with male rotation was a successful method that can be used for breeding programs to produce a limit of three families per female or as a scaling up step to produce large numbers of offspring from a limited number of selected pairs.info:eu-repo/semantics/acceptedVersio

Exploring the relationship between stress coping styles and sex, origin and reproductive success, in Senegalese sole (Solea senegalensis) breeders in captivity

Individual animals commonly adopt different stress coping styles that have been shown to impact reproductive success and differ between sexes (female/male) and origin (wild/hatchery). Hatchery reared Senegalese sole (Solea senegalensis) exhibit a behavioural reproductive dysfunction and a complete failure to spawn viable eggs. Hence, the present study examined whether 1) reproductive success of Senegalese sole was linked to coping styles and 2) behavioural differences exist in relation to sex or origin. A total of 198 breeders held in two research institutions were submitted to three individual tests (restraining, new environment and confinement) and one grouping test (risk taking). In addition, a blood sample to quantify cortisol, glucose and lactate levels was obtained from each individual after completing the individual tests. Senegalese sole breeders showed individual differences in behaviour across the different behavioural tests that were consistent with proactive and reactive coping styles traits. However, the most striking result was that reproductive success, sex and origin of Senegalese sole was not biased to any specific coping style. Indeed, the behavioural responses were similar and consistent between fish grouped by reproductive success, sex and origin. This study presented information that contrasts with different studies on dominant aggressive species and indicated that social non-aggressive species such as Senegalese sole follow a cooperative strategy that favours equal opportunities between stress coping styles and sexes. Therefore, results suggest that maintaining both coping styles strategies are fundamental for a sustainable breeder population approach