Welfare issues and potential solutions for laying hens in free range and organic production systems: A review based on literature and interviews.

In free-range and organic production systems, hens can make choices according to their needs and desires, which is in accordance with welfare definitions. Nonetheless, health and behavioral problems are also encountered in these systems. The aim of this article was to identify welfare challenges observed in these production systems in the EU and the most promising solutions to overcome these challenges. It is based on a review of published literature and research projects complemented by interviews with experts. We selected EU specific information for welfare problems, however, the selected literature regarding solutions is global. Free range use may increase the risk of infection by some bacteria, viruses and parasites. Preventive methods include avoiding contamination thanks to biosecurity measures and strengthening animals' natural defenses against these diseases which can be based on nutritional means with new diet components such as insect-derived products, probiotics and prebiotics. Phytotherapy and aromatherapy can be used as preventive and curative medicine and vaccines as alternatives to antibiotics and pesticides. Bone quality in pullets and hens prevents keel deviations and is favored by exercise in the outdoor range. Free range use also lead to higher exposure to variable weather conditions and predators, therefore shadow, fences and guard animals can be used to prevent heat stress and predation respectively. Granting a free range provides opportunities for the expression of many behaviors and yet many hens usually stay close to the house. Providing the birds with trees, shelters or attractive plants can increase range use. Small flock sizes, early experiences of enrichment and personality traits have also been found to enhance range use. Severe feather pecking can occur in free range production systems, although flocks using the outdoor area have better plumage than indoors. While many prevention strategies are facilitated in free range systems, the influence of genetics, prenatal and nutritional factors in free range hens still need to be investigated. This review provides information about practices that have been tested or still need to be explored and this information can be used by stakeholders and researchers to help them evaluate the applicability of these solutions for welfare improvement.

Foraging Behavior Shows Individual-Consistency Over Time, and Predicts Range Use in Slow-Growing Free-Range Male Broiler Chickens.

Recent research on free-range chickens shows that individual behavioral differences may link to range use. However, most of these studies explored individual behavioral differences only at one time point or during a short time window, assessed differences when animals were out of their social group and home environment (barn and range), and in specific tests or situations. Therefore, it is yet unclear how different behaviors relate to range use and how consistent these behaviors are at the individual level. To fill this gap, we here aimed to describe the behavioral budget of slow-growing male broiler chickens (S757N) when in their social group and home environment during the whole rearing period (from the second week of life to the twelfth week, before slaughter), and to relate observed behavioral differences to range use. For this, we followed a sample of individuals in two flocks (n = 60 focal chickens out of 200 chickens per flock), over two seasons, during three periods: before range access (from 14 to 25 days old), during early range access (first weeks of range access, from 37 to 53 days old), and during late range access (last weeks of range access, from 63 to 87 days old). By the end of each period, individual tests of exploration and social motivation were also performed, measuring exploration/activity and sociability propensities. Our results show that foraging (i.e., pecking and scratching at the ground) was the only behavior that correlated to range use for all three rearing periods, independent of the season. Foraging was also the only behavior that showed within-individual consistency from an early age and across the three rearing periods. Foraging may, therefore, serve as a useful behavioral predictor of range use in free-range broiler chickens. Our study increases the knowledge of how behaviors develop and relate to each other in a domesticated and intensely selected species, and improves our understanding of the biology of free-range broiler chickens. These findings can, ultimately, serve as a foundation to increase range use and improve chicken welfare.

Microbiota and stress: a loop that impacts memory.

Chronic stress and the gut microbiota appear to comprise a feed-forward loop, which contributes to the development of depressive disorders. Evidence suggests that memory can also be impaired by either chronic stress or microbiota imbalance. However, it remains to be established whether these could be a part of an integrated loop model and be responsible for memory impairments. To shed light on this, we used a two-pronged approach in Japanese quail: first stress-induced alterations in gut microbiota were characterized, then we tested whether this altered microbiota could affect brain and memory function when transferred to a germ-free host. The cecal microbiota of chronically stressed quails was found to be significantly different from that of unstressed individuals with lower α and ß diversities and increased Bacteroidetes abundance largely represented by the Alistipes genus, a well-known stress target in rodents and humans. The transfer of this altered microbiota into germ-free quails decreased their spatial and cue-based memory abilities as previously demonstrated in the stressed donors. The recipients also displayed increased anxiety-like behavior, reduced basal plasma corticosterone levels and differential gene expression in the brain. Furthermore, cecal microbiota transfer from a chronically stressed individual was sufficient to mimic the adverse impact of chronic stress on memory in recipient hosts and this action may be related to the Alistipes genus. Our results provide evidence of a feed-forward loop system linking the microbiota-gut-brain axis to stress and memory function and suggest that maintaining a healthy microbiota could help alleviate memory impairments linked to chronic stress.

Training level reveals a dynamic dialogue between stress and memory systems in birds.

Chronic stress profoundly affects forms of declarative memory, such as spatial memory, while it may spare non-declarative memory, such as cue-based memory. It is known, however, that the effects of chronic stress on memory systems may vary according to the level of training of an individual was submitted. Here, we investigated, in birds, how chronic stress impact spatial and cue-based memories according to training level. For that, control and chronically stressed Japanese quail were trained in a task that could be solved using spatial and cue-based memory and tested for their memory performance after 5 and 15 training days (initial training and overtraining, respectively) and following an emotional challenge (exposure to an open field). Our results showed that, compared to control quail, chronic stress impacted negatively spatial memory performances in stressed birds after initial training, but these differences were lowered after overtraining. Control birds seemed to shift from spatial to cue-based memory to solve the task across overtraining. However, an emotional challenge before testing reinstated the negative impact of chronic stress on spatial memory performances between the groups, revealing that chronic stress/overtraining did not eliminate the spatial memory and differences caused by stressors can reemerge depending on the individual's immediate psychological state. Contrary to spatial memory, cue-based memory was not affected in chronically stressed birds compared to control birds in any test occasion, confirming its resistance against the negative effects of chronic stress. Altogether these findings reveal a dynamic dialogue between stress, training level, and memory systems in birds.

Working for food is related to range use in free-range broiler chickens.

When animals prefer to make efforts to obtain food instead of acquiring it from freely available sources, they exhibit what is called contrafreeloading. Recently, individual differences in behavior, such as exploration, were shown to be linked to how prone an individual may be to contrafreeload. In this work, our main objective was to test whether and how individual differences in range use of free-range broiler chickens (Gallus gallus domesticus) were related to the individual motivation to contrafreeload. We also verified whether other behavioral variations could relate to range use. To that aim, over three different periods (before range access, first weeks of range access, and last weeks of range access), chickens with different ranging levels (low and high rangers) were submitted to a contrafreeloading test and had different behaviors recorded (such as foraging, resting, locomotion) in their home environment. During the contrafreeloading test, chickens were conditioned to one chamber presenting a foraging substrate and mealworms, while in the other chamber, mealworms were freely available on the floor. During testing trials, chickens had access to both empty chambers, and the time spent in each chamber was quantified. On average, low rangers preferred the chamber where mealworms were easily accessible (without the foraging substrate), while high rangers preferred the chamber where mealworms were accessible with difficulty, showing greater contrafreeloading. Out of ten behaviors recorded in chickens' home environment, foraging was the only one that differed significantly between our two ranging groups, with low rangers foraging, on average, significantly less than high rangers. These results corroborate previous experiences suggesting that range use is probably linked to chickens' exploratory trait and suggest that individual differences in free-range broiler chickens are present even before range access. Increasing our knowledge of individual particularities is a necessary step to improve free-range chicken welfare on the farm.

Spontaneous intake of essential oils after a negative postnatal experience has long-term effects on blood transcriptome in chickens.

Chicks subjected to early stressful factors could develop long-lasting effects on their performances, welfare and health. Free access to essential oils (EO) in poultry farming could mitigate these effects and potentially reduce use of antimicrobial drugs. This study on chicken analyzed long-lasting effects of post-hatch adverse conditions (Delayed group), and the impact of EO intake on blood physiological parameters and transcriptome. Half of the Control and Delayed groups had free access to EO, while the other half had only water for the first 13 days post-hatching. Blood analyses of metabolites, inflammation and oxidative stress biomarkers, and mRNA expression showed sex differences. Long-lasting effects of postnatal experience and EO intake persisted in blood transcriptome at D34. The early adverse conditions modified 68 genes in males and 83 genes in females. In Delayed males six transcription factors were over-represented (NFE2L2, MEF2A, FOXI1, Foxd3, Sox2 and TEAD1). In females only one factor was over-represented (PLAG1) and four under-represented (NFIL3, Foxd3, ESR2 and TAL1::TCF3). The genes showing modified expression are involved in oxidative stress, growth, bone metabolism and reproduction. Remarkably, spontaneous EO intake restored the expression levels of some genes affected by the postnatal adverse conditions suggesting a mitigating effect of EO intake.

Emotionality modulates the impact of chronic stress on memory and neurogenesis in birds.

Chronic stress is a strong modulator of cognitive processes, such as learning and memory. There is, however, great within-individual variation in how an animal perceives and reacts to stressors. These differences in coping with stress modulate the development of stress-induced memory alterations. The present study investigated whether and how chronic stress and individual emotionality interrelate and influence memory performances and brain neurogenesis in birds. For that, we used two lines of Japanese quail (Coturnix japonica) with divergent emotionality levels. Highly (E+) and less (E-) emotional quail were submitted to chronic unpredictable stress (CUS) for 3 weeks and trained in a spatial task and a discrimination task, a form of cue-based memory. E + and E- birds were also used to assess the impact of CUS and emotionality on neurogenesis within the hippocampus and the striatum. CUS negatively impacted spatial memory, and cell proliferation, and survival in the hippocampus. High emotionality was associated with a decreased hippocampal neurogenesis. CUS improved discrimination performances and favored the differentiation of newborn cells into mature neurons in the striatum, specifically in E+ birds. Our results provide evidence that CUS consequences on memory and neural plasticity depends both on the memory system and individual differences in behavior.

Research Note: Role of the hippocampus in spatial memory in Japanese quail.

The Japanese quail is a powerful model to characterize behavioral, physiological, and neurobiological processes in Galliformes. Behavioral tests have already been adapted for quail to assess memory systems, but despite the pivotal role of the hippocampus in this cognitive process, its involvement in spatial memory has not been demonstrated in this species. In this study, lesions were created in the hippocampus of Japanese quail, and both lesioned and control quail were tested for spatial and cue-based learning performances. These hippocampal lesions specifically impacted spatial learning performance, but spared learning performance when birds could solve the task using their cue-based memory. These findings, thus, highlight that the hippocampus plays a crucial role and is essential for spatial declarative memory. Future studies could aim to elucidate the cellular or molecular mechanisms involved in this form of memory.

The quail genome: insights into social behaviour, seasonal biology and infectious disease response.

BACKGROUND: The Japanese quail (Coturnix japonica) is a popular domestic poultry species and an increasingly significant model species in avian developmental, behavioural and disease research. RESULTS: We have produced a high-quality quail genome sequence, spanning 0.93 Gb assigned to 33 chromosomes. In terms of contiguity, assembly statistics, gene content and chromosomal organisation, the quail genome shows high similarity to the chicken genome. We demonstrate the utility of this genome through three diverse applications. First, we identify selection signatures and candidate genes associated with social behaviour in the quail genome, an important agricultural and domestication trait. Second, we investigate the effects and interaction of photoperiod and temperature on the transcriptome of the quail medial basal hypothalamus, revealing key mechanisms of photoperiodism. Finally, we investigate the response of quail to H5N1 influenza infection. In quail lung, many critical immune genes and pathways were downregulated after H5N1 infection, and this may be key to the susceptibility of quail to H5N1. CONCLUSIONS: We have produced a high-quality genome of the quail which will facilitate further studies into diverse research questions using the quail as a model avian species.

Influence of the microbiota-gut-brain axis on behavior and welfare in farm animals: A review.

There is increasing evidence of a pivotal role of the gut microbiota (GUT-M) in key physiological functions in vertebrates. Many studies discuss functional implications of the GUT-M not only on immunity, growth, metabolism, but also on brain development and behavior. However, while the influence of the microbiota-gut-brain axis (MGBA) on behavior is documented in rodents and humans, data on farm animals are scarce. This review will first report the well-known influence of the MGBA on behavior in rodent and human and then describe its influence on emotion, memory, social and feeding behaviors in farm animals. This corpus of experiments suggests that a better understanding of the effects of the MGBA on behavior could have large implications in various fields of animal production. Specifically, animal welfare and health could be improved by selection, nutrition and management processes that take into account the role of the GUT-M in behavior.

Relationship between ranging behavior and spatial memory of free-range chickens.

Different personalities may lead to different ways of processing environmental information; however, the relationship between personality and cognition is not fully understood as studies on diverse species present contrasting results. As there is great within-flock variability of outdoor ranging behavior in free-range broiler chickens, we tested whether and how ranging behavior impacts on individual spatial memory abilities. The experiment was conducted on one flock (n = 200) reared in the same conditions throughout the study, to simulate on-farm situations. As the ranging behavior was stable over time, we compared two distinct groups of male chickens: one that visited the range more (High rangers) and one that was more prone to staying in the poultry house (Low rangers). To test the spatial memory, individuals (n = 30) went through two main phases in an arena with 8 cups. For the familiarization phase, individuals were submitted to one trial per day, for seven days, to a situation where all eight cups were baited with mealworms. Animals had to reach a criterion of 5 cups visited out of 8 to advance to the next phase. For the spatial test, only four cups were baited and systematically placed at the same location. This last phase comprised two trials per individual per day, for nine days. During these two phases, latency to visit cups and the number of visits and revisits of all cups were recorded. Low ranger chickens took less time to attain the pre-established threshold of visiting 5 cups out of 8, over the familiarization phase. During the spatial test, the latency to visit four cups decreased between the within-day trials for low ranger chickens and increased for high ranger chickens. Moreover, in the within-day trial analysis, low ranger chickens exhibited an improvement on spatial memory and better spatial memory compared to high ranger chickens. Different speed-accuracy trade-offs may explain these differences between low and high ranger chickens and the way individuals interact and solve the task. Our study strengthens the scientific evidence relating consistent individual differences in behavior, with the ranging behavior of free-range chickens, and cognitive performance during a spatial memory task.

Effects of gut microbiota transfer on emotional reactivity in Japanese quails (Coturnix japonica).

The interaction between the gut microbiota (GM) and the brain has led to the concept of the microbiota-gut-brain axis but data for birds remain scarce. We tested the hypothesis that colonization of germ-free chicks from a quail line selected for a high emotional reactivity (E+) with GM from a line with low emotional reactivity (E-) would reduce their emotional behaviour in comparison with germ-free chicks from an E+ line colonized with GM from the same E+ line. The GM composition analysis of both groups revealed a shift in terms of microbial diversity and richness between day 21 and day 35 and the GM of the two groups of quails were closer to each other at day 35 than at day 21 at a phylum level. Quails that received GM from the E- line expressed a lower emotional reactivity than quails colonized by GM from the E+ line in tonic immobility and novel environment tests carried out during the second week of age. This result was reversed in a second tonic immobility test and an open-field run 2 weeks later. These behavioural and GM modifications over time could be the consequence of the resilience of the GM to recover the equilibrium present in the E+ host, which is in part driven by the host genotype. This study shows for the first time that a GM transfer can influence emotional reactivity in Japanese quails, supporting the existence of a microbiota-gut-brain axis in this species of bird.

A multiplatform metabolomic approach to characterize fecal signatures of negative postnatal events in chicks: a pilot study.

BACKGROUND: Negative experiences in early life can induce long-lasting effects on the welfare, health, and performance of farm animals. A delayed placement of chicks in rearing houses has negative effects on their performance, and results in fecal-specific odors detectable by rats. Based on this observation, the volatile organic compounds (VOCs) and metabolites from the feces of 12-day-old chickens were screened for early markers of response to negative events using gas-chromatography and liquid-chromatography coupled with mass spectrometry (GC-MS, LC-HRMS). RESULTS: The low reproducibility of solid-phase micro-extraction of the VOCs followed by GC-MS was not suitable for marker discovery, in contrast to liquid extraction of metabolites from freeze-dried feces followed by GC-MS or LC-HRMS analysis. Therefore, the fecal metabolome from 12-day-old chicks having experienced a normal or delayed placement were recorded by GC-MS and LC-HRMS in two genotypes from two experiments. From both experiments, 25 and 35 metabolites, respectively explaining 81% and 45% of the difference between delayed and control chickens, were identified by orthogonal partial least-squares discriminant analysis from LC-HRMS and GC-MS profiling. CONCLUSION: The sets of molecules identified will be useful to better understand the chicks' response to negative events over time and will contribute to define stress or welfare biomarkers. .

Spontaneous Intake and Long-Term Effects of Essential Oils After a Negative Postnatal Experience in Chicks.

The postnatal period is critical for broiler chicks as they are exposed to potentially stressful environmental changes in the hatchery and during transportation to the rearing houses. The ability of broiler chicks to spontaneously drink essential oils (EO) to mitigate the effects of a negative postnatal experience was tested. Chicks were placed in the rearing facility either immediately (C group), or after a 24 h-delay period (D group) to mimic a delay in transportation possible under commercial conditions. In experiment 1, each group had access to either water only or to water and one EO (cardamom, marjoram, or verbena) from D1 to D13. Verbena EO intake was higher in the D group than in the C group from D1 to D6 and cardamom EO intake was lower in the D group than in the C group from D6 to D13. In experiment 2, half of the groups had access to water only and the other half had both water and the three EO simultaneously. Chicks from D and C groups chose the EO similarly except for cardamom EO with a lower intake being observed in the D than in the C group from D6 to D12. The delayed placement of the D group reduced chicken growth until 34 days of age and temporarily increased the feed conversion ratio, but did not affect their welfare or the prevalence of health disorders. The EO intake did not mitigate the growth reduction in D group chicks, but did mitigate the reduced Pectoralis major muscle yield. In conclusion, chicks were able to make spontaneous choices regarding EO intake according to their postnatal experience when EO were presented individually, but not when presented simultaneously as in our experimental design. The EO intake only partially mitigated the decrease in chicken performance after the negative postnatal experience.

Facial expression and oxytocin as possible markers of positive emotions in horses.

Behavioural and physiological markers of discrete positive emotions remain little investigated in animals. To characterise new markers in horses, we used tactile stimulations to induce emotional situation of contrasting valence. In the Gentle grooming group (G, N = 13) horses were gently groomed during 11 sessions on the body areas they appreciated the most. Horses in the Standard grooming group (S, N = 14) were groomed using a fixed procedure, reported to induce avoidance reactions in some horses. At session 11, G horses expressed significantly more contact-seeking behaviours than S horses, which expressed significantly more avoidance behaviours. This result suggests positive emotions in G horses and negative emotions in S horses. Blood cortisol, oxytocin, heart rate and heart rate variability never differed between before and after the grooming session. However, after the 11 sessions, basal oxytocin levels were lower in the G than in the S group. This difference was unexpected, but supports studies showing that a low level of basal oxytocin could be a marker of better well-being. Analyses of facial expressions during grooming revealed significant differences between groups. These expressions appear to be more sensitive than behavioural indicators because they alone enabled differentiating emotions according to the group when horses were re-exposed to neutral grooming one year after the treatment.

A trait for a high emotionality favors spatial memory to the detriment of cue-based memory in Japanese quail.

Recent studies provided evidence that a personality trait such as a trait for a high or a low emotionality can either promotes or impairs learning and memory performances. This variability can be partly explained because this trait may have opposite effect on memory performances depending on the memory system involved. The present study investigated in Japanese quail the relationships between emotionality and two forms of memory, spatial- memory and cue-based memory. We showed that birds with a high emotionality trait (E+), compared with birds with a low emotionality trait (E-), reached slowly but more accurately the spatial location of a rewarded cup in an arena that contains 8 identical cups. Then a second cohort of E + and E- quails was trained to solve a dual spatial/cued task in which they could use either their spatial or cue-based memory. Whereas E + birds predominantly solved the task using their spatial memory, E- birds preferentially used their cue-based memory. These findings demonstrate that a trait for a high emotionality, can influence spatial memory performances but also contributes to favor the selection of this form of memory.

Are there facial indicators of positive emotions in birds? A first exploration in Japanese quail.

The positive aspect of emotions, like pleasure, remains overlooked in birds. Our aim was to contribute to the exploration of facial indicators of positive emotions. To observe contrasting emotional expressions, we used two lines of Japanese quail divergently selected on their inherent fearfulness: a fearful line (long tonic immobility duration: LTI) and a less fearful line (short tonic immobility duration: STI). To induce positive emotions, we gave individual quail the opportunity to perform a rewarding behaviour, dustbathing, in an unfamiliar cage. More STI than LTI quail expressed dustbathing and latencies to dustbathe were significantly shorter in STI than LTI quail. This result indicated that the lines of quail differed in their fearfulness of the situation. We observed crown feather height, throat feather angle and pupil surface before (control) and during dustbathing. We found significant increases in crown feather height, pupil area and angle of throat feathers between the control and the dustbathing phases in STI quail, and pupil area correlated positively with crown feather height. In LTI quail, the angle of throat feathers increased during dustbathing, but the other parameters did not differ. We argue that variation in crown feather height and pupil area may provide indications of positive emotions in Japanese quail.

Absence of Gut Microbiota Reduces Emotional Reactivity in Japanese Quails (Coturnix japonica).

Background: Recent studies have demonstrated an effect of the gut microbiota on brain development and behavior leading to the concept of the microbiota-gut-brain axis. However, its effect on behavior in birds is unknown. The aim of the present study was to determine the effect of the absence of gut microbiota on emotional reactivity in birds by comparing germ-free (GF) quails to those colonized (COL) with gut microbiota. Material and Methods: From hatching, the quails of both groups GF (n = 36) and COL (n = 36) were reared in sterile isolators. The COL quails were colonized at day 2 by introducing a pool of droppings from conventional adult females into the drinking water and feed. The quails were weighed individually on day 2, 6, and 14. From day 8, emotional reactivity was assessed in each group in the isolators through several behavioral tests. Results: GF quails showed a considerable decrease in emotional reactivity demonstrated by spending less time in tonic immobility during the tonic immobility test (242 s ± 31 vs. 331 s ± 32, p ≤ 0.05), traveling a shorter distance (3,897 cm ± 242 vs. 4,827 cm ± 278, p ≤ 0.05) at a lower velocity (6.55 cm/s ± 0.4 vs. 8.1 cm/s ± 0.5, p ≤ 0.05) during the social separation test and spending more time near an object at the beginning of the novel object test (33.7 s ± 6.4 vs. 18.5 s ± 4.1, p ≤ 0.05). No difference in growth was found between the 2 groups. Conclusion: For the first time, this study demonstrates that the absence of gut microbiota reduces emotional reactivity in Japanese quails in situations of fear and social perturbation without influence on growth.

The influence of a probiotic supplementation on memory in quail suggests a role of gut microbiota on cognitive abilities in birds.

The gut microbiota is involved in host behaviour and memory in mammals. Consequently, it may also influence emotional behaviour and memory in birds. Quail from two genetic lines with different fearfulness (LTI: long tonic immobility, n=37; STI: short tonic immobility, n=32) were either or not supplemented with a probiotic (Pediococcus acidilactici) from hatching. Emotional reactivity was measured in a tonic immobility test (d6 and 7 of age) and two open-field tests (d13-15; d22-24). Memory was measured in a test rewarded with mealworms, where quail had to remember the cups previously visited (d34-36). Quail endured a 5-days stress period from days 17 to 21 to help revealing the potentially beneficial effect of the probiotic. As expected, STI quail were less fearful compared to the LTI quail (p<0.05). Probiotic supplementation had no effect on most measures of emotional reactivity (p>0.05), except in the tonic immobility test where supplemented STI quail had lower immobility duration (p=0.0001). Regarding the memory test, the two lines had similar performances. Quail fed with probiotics made fewer errors (p=0.040). There was no significant correlation between traits of emotional reactivity and of memory. In conclusion, the supplementation with Pediococcus acidilactici as a probiotic, affected a specific trait of emotional reactivity in STI quail, and improved memory in both lines, whichstrengthens the idea that the influence of gut microbiota on the host behaviour and memory seen in mammals is shared by birds.

Accounting for linkage disequilibrium in genome scans for selection without individual genotypes: The local score approach.

Detecting genomic footprints of selection is an important step in the understanding of evolution. Accounting for linkage disequilibrium in genome scans increases detection power, but haplotype-based methods require individual genotypes and are not applicable on pool-sequenced samples. We propose to take advantage of the local score approach to account for linkage disequilibrium in genome scans for selection, cumulating (possibly small) signals from single markers over a genomic segment, to clearly pinpoint a selection signal. Using computer simulations, we demonstrate that this approach detects selection with higher power than several state-of-the-art single-marker, windowing or haplotype-based approaches. We illustrate this on two benchmark data sets including individual genotypes, for which we obtain similar results with the local score and one haplotype-based approach. Finally, we apply the local score approach to Pool-Seq data obtained from a divergent selection experiment on behaviour in quail and obtain precise and biologically coherent selection signals: while competing methods fail to highlight any clear selection signature, our method detects several regions involving genes known to act on social responsiveness or autistic traits. Although we focus here on the detection of positive selection from multiple population data, the local score approach is general and can be applied to other genome scans for selection or other genomewide analyses such as GWAS.