Domesticated species differ from their wild ancestors in a mosaic of traits. Classical domestication theories agree that reactivity to fear and stress is one of the main traits affected. Domesticated species are expected to be less fear and stress prone to than their wild counterparts. To test this hypothesis, we compared the behavioural responses of White Leghorn (WL) chicks to their wild counterparts, Red Junglefowl (RJF) chicks in risk-taking situations. In order to obtain food, the chicks faced an unknown and potentially harmful object at the presence or absence of a social partner. We found that according to our predictions, RJF were more stressed and fearful of the object than the WL. Still, RJF were more explorative than WL. Additionally, the presence of a social partner reduced the fear response in both, but had a stronger effect on RJF. Finally, WL were more food orientated than the RJF. Our results confirmed classical domestication hypotheses of downregulation of the stress system and importance of the social partner in domesticated farm chicken.

Har du undrat varför hundar gärna äter märkliga saker, som gräs eller bajs? Eller varför de rullar sig i stinkande högar och lägger så mycket tid på att leka och sova? På ett lättläst och pedagogiskt sätt tar Per Jensen upp dessa och en rad andra hittills obesvarade gåtor och presenterar, med humor och en stor kärlek till hundar, hur vetenskapen försökt lösa dem.[Bokinfo]

Prosocial behaviour is pronounced in humans and prevalent in some non-human animals, however, the occurrence of the trait in chickens has not yet been investigated. Here, we studied the occurrence of prosociality in four different lines of adult female chickens. To explore the effects of domestication, chickens of the domesticated layer White Leghorn (WL) and the ancestral Red Junglefowl (RJF) were compared. Additionally, to explore the role of tameness, Red Junglefowl selected for high (RJF HF), or low (RJF LF) fear of humans were also studied. The hens were all tested in a prosocial choice task adapted from a previous study conducted on rats. Each individual was first trained to differentiate between a compartment where itself and a companion received food treats simultaneously (representing a prosocial choice), and one where only itself received the treat. Following training, each bird was tested in a free-choice set-up. No occurrence of prosociality was found at group level in any of the lines, however, our results suggest that the trait may occur in some individuals, and that domestication and increased tameness may have increased its prevalence, although alternative explanations such as side bias and social competition cannot be ruled out. Since this study is the first of its kind, further research is required to make any definite conclusions.

A central part of the domestication syndrome is a reduction in relative brain size. In chickens, it has previously been shown that domesticated birds have smaller relative brain mass, but larger relative mass of cerebellum, compared to their ancestors, the Red Junglefowl. It has been suggested that tameness may drive the domestication syndrome, so we examined the relationship between brain characteristics and tameness in 31 Red Junglefowl from lines divergently selected during ten generations for tameness. Our focus was on the whole brain, cerebellum, and the remainder of the brain. We used the isotropic fractionator technique to estimate the total number of cells in the cerebellum and differentiate between neurons and non-neuronal cells. We stained the cell nuclei with DAPI and performed cell counting using a fluorescence microscope. NeuN immunostaining was used to identify neurons. The absolute and relative masses of the brains and their regions were determined through weighing. Our analysis revealed that birds selected for low fear of humans (LF) had larger absolute brain mass, but smaller relative brain mass, compared to those selected for high fear of humans (HF). Sex had a significant impact only on the absolute size of the cerebellum, not its relative size. These findings support the notion that selection for increased tameness leads to an enlargement of the relative size of cerebellum in chickens consistent with comparisons of domesticated and ancestral chickens. Surprisingly, the HF birds had a higher density of neurons in the cerebellum compared to the LF line, despite having a smaller cerebellum overall. These findings highlight the intricate relationship between brain structure and behavior in the context of domestication.

The communicating skills of dogs are well documented and especially their contact-seeking behaviours towards humans. The aim of this study was to use the unsolvable problem paradigm to investigate differences between breed groups in their contact-seeking behaviours towards their owner and a stranger. Twenty-four dogs of ancient breeds, 58 herding dogs, and 17 solitary hunting dogs were included in the study, and their behaviour when presented with an unsolvable problem task (UPT) was recorded for 3 min. All breed groups interacted with the test apparatus the same amount of time, but the herding dogs showed a longer gaze duration towards their owner compared to the other groups and they also preferred to interact with their owner instead of a stranger. Interestingly, the solitary hunting dogs were more in stranger proximity than the other groups, and they also showed a preference to make contact with a stranger instead of their owner. Hence, we found differences in contact-seeking behaviours, reflecting the dog-human relationship, between breed groups that might not only be related to their genetic similarity to wolves, but also due to the specific breeding history of the dogs.

Under commercial conditions, laying hen chicks are exposed to a range of stressful events immediately after hatch. Here, we studied whether environmental enrichment was able to reduce the stress sensitivity of these chicks. A total of 50 hatchery chicks (HC) and 50 control chicks (CC) were raised in enriched pens (E), while 53 HC + 53 CC were raised in standard non-enriched floor pens (NE). HC weighed less on day one, but there was no effect of hatchery treatment at later ages. HC were more pessimistic in a cognitive judgement bias test and emitted more distress calls when socially isolated, indicating that hatchery stress affected behaviour and stress sensitivity. However, enrichment did not affect the behaviour in any of these tests. We found no effects of hatchery stress in a novel environment, but indications that enrichment may have increased fearfulness of HC. The sensitivity of the hypothalamic-pituitary-adrenal (HPA) axis was reduced in HC-E compared to HC-NE, indicating that enrichment buffered the physiological stress sensitivity in HC; however, the opposite pattern was found in CC. In conclusion, our results show complex and somewhat contradictory effects on the ability of enrichment to buffer the consequences of stress in commercial hatcheries.

Red junglefowl (RJF), ancestor of all domesticated chickens, is a highly social, omnivorous bird species, presumably with a capacity for social information sharing. During domestication, birds have been selected to live in large, dynamic groups, and this may have affected their social cognition. Here, we studied social information transfer in female RJF and domesticated White Leghorn (WL) chickens. Individuals were trained to open a puzzle-box feeder by pecking a lid and we then recorded the behaviour towards the same puzzle-box feeder for birds that had either observed the trained individual ("guided") or saw the puzzle-box feeder for the first time ("naive"). WL were considerably faster in approaching the feeder regardless of prior demonstration and pecked more at it. Both breeds were significantly faster to approach the puzzle-box feeder and pecked more after prior demonstration, but the effects were significantly stronger in WL. We conclude that both RJF and WL can utilize social information to address a novel problem, but during domestication this ability appears to have increased. The effects can be an effect of either social learning or stimulus enhancement. Some caution in this conclusion is necessary since we tested relatively few WL. Furthermore, possible confounding explanations include higher fearfulness in RJF and different effects of dominance interactions between demonstrators and observers.

Beyond physical and zootechnical characteristics, the process of animal domestication has also altered how domesticated individuals, compared to their wild counterparts, perceive, process, and interact with their environment. Little is known, however, on whether and how domestication altered the perception of conspecific calls on both domesticated and wild breeds. In the present work, we compared the vigilance behavior of domestic and captiveborn wild fowl following the playback of chicken alarm calls and contentment calls (control). The playback tests were performed on four different breeds/lines. We first compared the behavioral reaction of domesticated White Leghorn (WL, a breed selected for egg production) and Red Junglefowl (RJF) hens (ancestor of domestic chickens). We also compared the behavior of Red Junglefowl hens selected for high or low fear of humans (RJF HF and RJF LF, respectively), a proxy to investigate early effects of domestication. Contrary to our expectations, no breed/line reacted accordingly to the calls, as the increase in vigilance behavior after the playback calls was similar for both alarm and contentment calls. Although no call discrimination differences were found, breeds did differ on how they reacted/habituated to the calls. Overall, WL were more vigilant than RJF, and birds from the RJF LF line decreased their vigilance over testing days, while this was not the case for the RJF HF line. These results suggest that birds under commercial-like conditions are unable to discriminate between alarm and contentment calls. Interestingly, domestication and selection for low fear of humans may have altered how birds react to vocal stimuli. It is important to consider that farmed animals may interpret and be affected by the vocalizations of their conspecifics in unexpected ways, which warrants further investigation.

Every year, billions of egg layer chicks around the world are hatched under highly stressful, industrial circumstances. Here, it is investigated how the stressful procedure in the commercial hatchery, including incubation, hatching, processing, and transport affects the chicks with regards to traits relevant for the egg production industry. These traits were compared to those of a control group hatched in a small incubator and handled gently och quietly in a quiet room without any processing and transport. The chicks were weighed at hatch and at eight additional time points: 4 days, 1 week (w), 2 w, 3 w, 5 w, 8 w, 20 w and 25 w of age. Feather pecking was studied at 15 w of age and damages to the feathers and injuries on the comb and wattle were assessed at 25 w of age. From 19 w of age, eggs were collected on three days per week, counted and weighed. Chicks from a commercial hatchery had a lower hatch weight than control chicks (p<0.001). At 20 w of age, the weight of the commercial hatched chicks was still numerically lower, although this did not reach statistical significance. Commercially hatched chicks tended to show more feather pecking behaviour at 15 w of age compared to control chicks (p<0.1), although feather condition at 25 w of age showed the opposite pattern. Regarding production, commercially hatched chickens laid fewer (p<0.05) and smaller (p<0.05) eggs than chicks hatched and handled under calm circumstances. From this experiment, it is concluded that the stressful experience in the commercial hatchery has an overall negative effect on traits relevant for the industry.

Summary:The GBS-MeDIP protocol combines two previously described techniques, Genotype-by-Sequencing (GBS) and Methylated-DNA-Immunoprecipitation (MeDIP). Our method allows for parallel and cost-efficient interrogation of genetic and methylomic variants in the DNA of many reduced genomes, taking advantage of the barcoding of DNA samples performed in the GBS and the subsequent creation of DNA pools, then used as an input for the MeDIP. The GBS-MeDIP is particularly suitable to identify genetic and methylomic biomarkers when resources for whole genome interrogation are lacking.

Play is common in young homeotherm animals and has an important role as a tentative indicator of positive states of welfare. Furthermore, during domestication play is believed to have increased in frequency in several species as part of the domestication syndrome. Here, we studied the ontogeny of play in chickens in two experiments. The first compared the behavioural development between domesticated White Leghorn (WL) laying hen chicks and ancestral Red Junglefowl (RJF) and the second compared the same between WL chicks that had experienced the stress of commercial hatchery routines and a control group, hatched under calm conditions. In both experiments, 10 groups of four chicks each from each of the groups were moved twice per week to an enriched and fully enclosed play arena, starting at day 8 and finishing day 39 or 53 after hatch. In the arena, the frequency of play behaviours was recorded during 30 min and divided into object, locomotory and social play. In experiment one, total play as well as object play was significantly more common in WL whereas locomotor and social play was more common in RJF. In experiment two, total play was significantly more frequent in commercially hatched chicks, despite that none of the sub-categories differed significantly between the groups. In conclusion, domestication as well as early stress does affect the occurrence of play in chickens, but the effects are complex and require further research.

Background Relationships among genetic or epigenetic features can be explored by learning probabilistic networks and unravelling the dependencies among a set of given genetic/epigenetic features. Bayesian networks (BNs) consist of nodes that represent the variables and arcs that represent the probabilistic relationships between the variables. However, practical guidance on how to make choices among the wide array of possibilities in Bayesian network analysis is limited. Our study aimed to apply a BN approach, while clearly laying out our analysis choices as an example for future researchers, in order to provide further insights into the relationships among epigenetic features and a stressful condition in chickens (Gallus gallus). Results Chickens raised under control conditions (n = 22) and chickens exposed to a social isolation protocol (n = 24) were used to identify differentially methylated regions (DMRs). A total of 60 DMRs were selected by a threshold, after bioinformatic pre-processing and analysis. The treatment was included as a binary variable (control = 0; stress = 1). Thereafter, a BN approach was applied: initially, a pre-filtering test was used for identifying pairs of features that must not be included in the process of learning the structure of the network; then, the average probability values for each arc of being part of the network were calculated; and finally, the arcs that were part of the consensus network were selected. The structure of the BN consisted of 47 out of 61 features (60 DMRs and the stressful condition), displaying 43 functional relationships. The stress condition was connected to two DMRs, one of them playing a role in tight and adhesive intracellular junctions in organs such as ovary, intestine, and brain. Conclusions We clearly explain our steps in making each analysis choice, from discrete BN models to final generation of a consensus network from multiple model averaging searches. The epigenetic BN unravelled functional relationships among the DMRs, as well as epigenetic features in close association with the stressful condition the chickens were exposed to. The DMRs interacting with the stress condition could be further explored in future studies as possible biomarkers of stress in poultry species.

The domestic fowl has a different social behavior compared to their ancestor, the red junglefowl. To examine whether selection for tameness has affected their intra-specific social behavior, 32 red junglefowl from two selection lines, one selected for increased tameness and one selected for a high fear of humans for ten generations, were kept in a group of two females and two males each and were observed in a semi-natural undisturbed enclosure. Birds selected for a low fear of humans had more social conflict, and the males from this selection crowed more and were more often observed in low social proximity to others. The high-fear birds spent more time close together with the rest of the group and performed more social, non-aggressive pecking. These results are consistent with known differences between ancestral red junglefowl and domesticated laying hens. Our results show that intra-specific social behavior has been affected as a side-effect of selection for increased tameness. This may have interesting implications for the emergence of the domestication syndrome in chickens.

Although there is evidence to suggest that animal domestication acts as a modulator of spatial orientation, little is known on how domesticated animals, compared to their wild counterparts, orientate themselves when confronted to different environmental cues. Here, using domesticated White Leghorn chicks, and their ancestor, the Red Junglefowl (Gallus gallus), our main objective was to investigate how bird domestication influences the use of distal and local cues, during an orientation task. We also investigated the memory retention of these cues over time, and how persistent/flexible individuals from both breeds were at pecking at unreachable mealworms. Our results showed that the breeds did not differ in their use of distal or local cues, with both showing a marked preference for the use of local cues over distal ones. Over time, individual performance declined, but this was not influenced by the type of cue present during the tests, nor by the breed. Domesticated chicks showed greater signs of persistency compared to their wild conspecifics. In conclusion, domestication did not seem to alter how birds orientate spatially, but may have caused more subtle changes, such as an increase in behavioral persistency, a feature that may be adaptative in human-controlled and homogenous environments.

Red Junglefowl (Gallus gallus) were selected for divergent levels of fear of humans during eight generations, causing the selection lines to differ in fear levels as well as in the proportional brain and cerebellum masses. Birds from the two lines were then crossed to obtain an F3 intercross in order to study the correlations between brain mass and fear learning. We exposed 105 F3-animals individually to a fear habituation and memory test at 8 days of age, where the reactions to repeated light flashes were assessed on 2 consecutive days. After culling, the absolute and relative sizes of each of four brain regions were measured. Stepwise regression was used to analyse the effects of the size of each brain region on habituation and memory. There were no effects of any brain region on the habituation on day one. However, birds with a larger absolute size of cerebellum had significantly reduced reactions to the fearful stimuli on day two, indicating a better memory of the stimuli. No other regions had significant effects. We conclude that increased cerebellum size may have been important in facilitating chicken domestication, allowing them to adapt to a life with humans.

Birds use contact incubation to warm their eggs above ambient temperature required for embryonic development. In contrast, birds in the industry as well as many birds in breeding programs and scientific studies are incubated in conventional incubators that warm eggs via circulating warm air. This means that contact incubated eggs have different thermal properties than eggs incubated in a conventional incubator. In light of previous studies showing that small differences in incubation temperature can affect chicks post-hatching phenotype, we investigated the consequences of incubating Red jungle fowl eggs at the same temperature (37 degrees C) either via contact incubation or warm air incubation. We found that contact incubated chicks had a more robust body composition, were more explorative and had a higher temperature preference early in life, as well as a sex dependent difference in plasma Corticosterone levels pre-hatch (measured in down-feathers) and post-hatch (measured in plasma) compared to chicks incubated in a conventional warm air incubator. While previous studies have demonstrated that embryonic development and post-hatch phenotype is sensitive to small variations in temperature, our study demonstrates for the first time that the way heat is distributed to the egg has a similar magnitude of effect on post-hatch phenotype and highlights the sensitivity of the incubation period in shaping birds post-hatch phenotype.

Domestic chickens are less fearful, have a faster sexual development, grow bigger, and lay more eggs than their primary ancestor, the red junglefowl. Several candidate genetic variants selected during domestication have been identified, but only a few studies have directly linked them with distinct phenotypic traits. Notably, a variant of the thyroid stimulating hormone receptor (TSHR) gene has been under strong positive selection over the past millennium, but its function and mechanisms of action are still largely unresolved. We therefore assessed the abundance of the domestic TSHR variant and possible genomic selection signatures in an extensive data set comprising multiple commercial and village chicken populations as well as wild-living extant members of the genus Gallus. Furthermore, by mean of extensive backcrossing we introgressed the wild-type TSHR variant from red junglefowl into domestic White Leghorn chickens and investigated gene expression, hormone levels, cold adaptation, and behavior in chickens possessing either the wild-type or domestic TSHR variant. While the domestic TSHR was the most common variant in all studied domestic populations and in one of two red junglefowl population, it was not detected in the other Gallus species. Functionally, the individuals with the domestic TSHR variant had a lower expression of the TSHR in the hypothalamus and marginally higher in the thyroid gland than wild-type TSHR individuals. Expression of TSHB and DIO2, two regulators of sexual maturity and reproduction in birds, was higher in the pituitary gland of the domestic-variant chickens. Furthermore, the domestic variant was associated with higher activity in the open field test. Our findings confirm that the spread of the domestic TSHR variant is limited to domesticated chickens, and to a lesser extent, their wild counterpart, the red junglefowl. Furthermore, we showed that effects of genetic variability in TSHR mirror key differences in gene expression and behavior previously described between the red junglefowl and domestic chicken.

Loss of pigmentation is a hallmark of domestication, and dogs offer a unique model for understanding the genetics of fur coloration. The aim of this study was to use dense genetic mapping to map loci underlying variations in color and whiteness in a population of laboratory beagles. A total of 190 beagles with well-defined pedigrees were phenotyped for the amount of white color in six different body parts, including the saddle. All individuals were genotyped on 85,172 informative and valid SNP-markers and the genome-wide associations for the amount of white in each body part were determined. There was a large variation in the amount of white on different parts of the body, and the whiteness was highly correlated within individuals, except for saddle color which was only moderately correlated with overall whiteness. The GWAS showed significant associations with two loci, one on chromosome 5, containing the MC1R gene, and one on chromosome 20, containing the MITF gene. Our results suggest that the variation in loss of pigmentation is largely a function of regulatory variation related to these genes.

Det är svårt att tänka sig ett mänskligt liv utan hundar! Sedan vi levde som jägare och samlare har hunden följt oss människor. Per Jensen, professor i etologi, har skrivit en bok om hundens beteende, språk och tankeförmåga och beskriver pedagogiskt hur kunskapsläget är idag. Ny fakta blir här tillgänglig för en intresserad allmänhet.

In commercial egg production, laying hen chicks are exposed to several stressful events during incubation, hatching, and their first hours in life. We have previously shown that hatching and processing are associated with increased corticosterone concentration and further affect behavior and stress sensitivity in a short- as well as long-term perspective. However, it is not known whether these long-term stress effects are caused by the hatchery processing (sex sorting, vaccination, conveying, and loading for transport) or if they are mainly caused by potentially stressful events before processing, during incubation and hatching. In the present study, the aim was to assess the effects of incubation and hatching only, compared to stress effects from the entire hatchery processing. We compared Lohmann LSL chicks incubated, hatched, and processed in a commercial hatchery with chicks incubated and hatched at the same time but not further processed. We studied behavior in a novel arena and during tonic immobility, as well as weight development and corticosterone reaction during a stress challenge. Processed chicks had poorer weight development and were more active in the novel arena test. However, there were no significant differences between the groups in corticosterone reactivity or tonic immobility. When comparing with previous data, both groups had elevated corticosterone concentrations compared to what we had previously reported from chicks hatched under calm and nonstressful conditions. In conclusion, incubation and hatching alone caused long-term stress effects in chickens, but further processing exacerbated these effects to some extent.

Fractures and deviations to the keel bone are common in commercial laying hens, with reported variations in occurrence across strains and breeds. The aetiology is not fully understood, however, modern genetics and selection for efficient egg production has been claimed to be important factors for the keel bone fractures. To explore this further, we investigated keel bones from two different breeds, representing different degrees of selection for egg production: Red jungle fowl (n = 82), and White Leghorn (n = 32), where the latter is a selected laying breed which is the origin for many modern laying hen hybrids. Keel bones from a total of 116 birds, 53 hens and 63 roosters, were examined by necropsy at 80 weeks of age. All birds were raised in modified aviaries in the same holding facility. Overall, 24.5% of the hens had one or more fractures to the keel, with a difference in the prevalence between hens from the two breeds (p&lt;0.01): 10% (95% CI: 3.7-24%) in the Red Jungle fowl hens and 69% (95% CI: 37-90%) in the White Leghorn hens. No roosters, regardless of breed, had keel bone fractures. Mild to moderate keel bone deviations were present in 54% (95% CI: 25-80%) of the hens and 4.7% (95% CI: 0.5-30%) of the roosters, all White Leghorns.

Previously, we found that dogs belonging to the herding breed group, selected for human cooperation, synchronise their long-term stress levels with their owners. The aim of the current study was to investigate features that could influence long-term stress levels in ancient dog breeds, genetically closer to wolves, and dogs specifically selected to work independently of their owner. Twenty-four ancient breed dogs and 18 solitary hunting dogs were recruited and hair samples were obtained from both dogs and owners from which hair cortisol concentration (HCC) was analysed. Additionally, the owners completed lifestyle surveys, the Monash Dog Owner Relationship Scale (MDORS) on human-dog relationship, and both dog and owner personality questionnaires (Dog Personality questionnaire and Big Five Inventory survey). The results from the MDORS indicate that the subscale Perceived cost correlated to the dog HCC of tested breed groups: solitary hunting breeds (chi (2)=4.95, P=0.026, beta =0.055), ancient breeds (chi (2)=2.74, P=0.098, beta =0.027), and herding dogs included from a previous study (chi (2)=6.82, P=0.009, beta=- 0.061). The HCC of the solitary hunting dogs was also related to the owner personality traits Agreeableness (chi (2)=12.30, P&lt;0.001, &lt;beta&gt;=- 0.060) and Openness (chi (2)=9.56, P=0.002, beta =0.048) suggesting a more substantial influence of the owner on the solitary hunting dogs HCC compared to the ancient breeds. No effect of owner HCC on dog HCC was found in either ancient or in solitary hunting breeds. Hence, the long-term stress synchronisation is likely to be a trait in breeds selected for human cooperation. In conclusion, dog HCC is often related to the owners personality, but is primarily influenced by the owner-dog relationship.

Domestication provides an outstanding opportunity for biologists to explore the underpinnings of organismal diversification. In domesticated animals, selective breeding for exaggerated traits is expected to override genetic correlations that normally modulate phenotypic variation in nature. Whether this strong directional selection affects the sequence of tightly synchronized events by which organisms arise (ontogeny) is often overlooked. To address this concern, we compared the ontogeny of the red junglefowl (RJF) (Gallus gallus) to four conspecific lineages that underwent selection for traits of economic or ornamental value to humans. Trait differentiation sequences in embryos of these chicken breeds generally resembled the representative ancestral condition in the RJF, thus revealing that early ontogeny remains highly canalized even during evolution under domestication. This key finding substantiates that the genetic cost of domestication does not necessarily compromise early ontogenetic steps that ensure the production of viable offspring. Instead, disproportionate beak and limb growth (allometry) towards the end of ontogeny better explained phenotypes linked to intense selection for industrial-scale production over the last 100 years. Illuminating the spatial and temporal specificity of development is foundational to the enhancement of chicken breeds, as well as to ongoing research on the origins of phenotypic variation in wild avian species.

Simple Summary Worldwide, billions of laying hen chicks are incubated, hatched and processed in industrial hatcheries every year. When exposed to stress, hormones are incorporated in the feathers of the birds. Here, we measured levels of the stress hormone corticosterone to investigate possible stress during the incubation. Further, animals can perceive their environment either in a positive (optimistic) or a negative (pessimistic) way. We investigated how the early hatchery experiences affects "optimism" and "pessimism". Commercially hatched chicks were exposed to a positive cue, an aversive cue, and ambiguous cues, in order to evaluate the cognitive welfare state of the animals. These chicks were compared to a group of non-stressed animals. Commercially incubated chicks did not have elevated levels of feather corticosterone, which implies that the main part of the stress effects from the hatchery originates from the period around hatch. Latencies to approach ambiguous cues were longer for the stressed chicks, i.e., these showed a more pessimistic-like behaviour. We conclude that the main part of the stress effects in commercially hatched chicks originates in the period around hatch, and further, that these birds show lasting levels of increased pessimism. This strongly indicates a long-time poorer welfare state for the animals. Worldwide, billions of laying hen chicks are incubated and processed under highly industrialised circumstances every year, which, as we have previously shown, has long-lasting effects. Here, we measured corticosterone incorporated in down feathers to investigate possible stress during the incubation and showed that commercially incubated chicks did not have elevated levels of feather corticosterone, which implies that the main part of the stress effects from hatchery originates from the perinatal period and the handling immediately post-hatch. Further, we investigated how the early hatchery stress affects the chicks cognitive welfare state, i.e., "optimism" and "pessimism". We exposed commercially hatched chickens to a positive cue, an aversive cue and ambiguous cues. The birds were tested at 1 and 10 w of age and the behaviour was compared with that of non-stressed chicks. Latencies to approach ambiguous cues were longer for the stressed chicks, both at 1 (p = 0.008) and at 10 (p = 0.020) weeks of age, i.e., these showed a more pessimistic-like behaviour. We conclude that the main part of the stress effects in commercially hatched chicks originates in the perinatal period, and further, that these birds show lasting levels of increased pessimism. This strongly indicates a long-time poorer welfare state for the animals.

Early animal domestication may have been driven by selection on tameness. Selection on only tameness can bring about correlated selection responses in other traits, not intentionally selected upon, which may be one cause of the domesticated phenotype. We predicted that genetically reduced fear towards humans in Red Junglefowl, ancestors of domesticated chickens, would be correlated to other traits included in the domesticated phenotype. Fear level was determined by a standardised behaviour test, where the reaction towards an approaching human was recorded. We first selected birds for eight generations for either high or low fear levels in this test, to create two divergent selection lines. An F3 intercross, with birds from the eighth generation as parentals, was generated to study correlations between fear-of-human scores and other unselected phenotypes, possibly caused by pleiotropy or linkage. Low fear-of-human scores were associated with higher body weight and growth rates, and with increased activity in an open field test, indicating less general fearfulness. In females, low fear-of-human scores were also associated with more efficient fear habituation and in males with an increased tendency to emit food calls in a mirror test, indicating increased social dominance. Low fear-of-human scores were also associated with smaller brain relative to body weight, and with larger cerebrum relative to total brain weight in females. All these effects are in line with the changes observed in domesticated chickens compared to their ancestors, and we conclude that tameness may have been a driving factor underlying some aspects of the domesticated phenotype.

Med humor, värme och en stor kärlek till hundar förmedlar Per Jensen det senaste om hur våra fyrbenta vänner fungerar i olika faser av livet och hur vi kan använda de nya rönen för att få en lycklig samvaro.

Plumage colouration in birds is important for a plethora of reasons, ranging from camouflage, sexual signalling, and species recognition. The genes underlying colour variation have been vital in understanding how genes can affect a phenotype. Multiple genes have been identified that affect plumage variation, but research has principally focused on major-effect genes (such as those causing albinism, barring, and the like), rather than the smaller effect modifier loci that more subtly influence colour. By utilising a domestic × wild advanced intercross with a combination of classical QTL mapping of red colouration as a quantitative trait and a targeted genetical genomics approach, we have identified five separate candidate genes (CREBBP, WDR24, ARL8A, PHLDA3, LAD1) that putatively influence quantitative variation in red-brown colouration in chickens. By treating colour as a quantitative rather than qualitative trait, we have identified both QTL and genes of small effect. Such small effect loci are potentially far more prevalent in wild populations, and can therefore potentially be highly relevant to colour evolution.

Epigenetic factors such as DNA methylation act as mediators in the interaction between genome and environment. Variation in the epigenome can both affect phenotype and be inherited, and epigenetics has been suggested to be an important factor in the evolutionary process. During domestication, dogs have evolved an unprecedented between-breed variation in morphology and behavior in an evolutionary short period. In the present study, we explore DNA methylation differences in brain, the most relevant tissue with respect to behavior, between wolf and dog breeds. We optimized a combined method of genotype-by-sequencing (GBS) and methylated DNA immunoprecipitation (MeDIP) for its application in canines. Genomic DNA from the frontal cortex of 38 dogs of 8 breeds and three wolves was used. GBS and GBS-MeDIP libraries were prepared and sequenced on Illuma HiSeq2500 platform. The reduced sample represented 1.18 +/- 0.4% of the total dog genome (2,4 billion BP), while the GBS-MeDIP covered 11,250,788 +/- 4,042,106 unique base pairs. We find substantial DNA methylation differences between wolf and dog and between the dog breeds. The methylation profiles of the different groups imply that epigenetic factors may have been important in the speciation from dog to wolf, but also in the divergence of different dog breeds. Specifically, we highlight methylation differences in genes related to behavior and morphology. We hypothesize that these differences are involved in the phenotypic variation found among dogs, whereas future studies will have to find the specific mechanisms. Our results not only add an intriguing new dimension to dog breeding but are also useful to further understanding of epigenetic involvement.

Simple Summary The keel bone of laying hens is prone to deviations and fractures. Recent studies in the last decade report high prevalence of this welfare concern. The causative factors are not clear; however, selection for efficient egg production has been suggested as a major contributing factor. An important step to shed light on the role of selective breeding as an underlying cause of keel bone damage (KBD) in modern laying hens is to evaluate the keel bones of the ancestor, the red jungle fowl. The aim of this study was therefore to describe the prevalence of KBD in a study group of 29 red jungle hens and roosters by autopsy. No fractures were detected in the roosters, one had a very slight deviation. One of the hens had a fracture to the keel and 10 hens had a very slight deviation. Large scale studies are needed to disentangle the effect of different factors on keel bone damage. Keel bone damage (KBD) is a highly prevalent problem in commercial egg production. KBD consists of two different conditions affecting the keel: Keel bone deviation and keel bone fractures (KBF). Deviations are linked to pressure on the keel, e.g., from perching. The causative factors for KBF are not clear; however, selection for efficient egg production has been suggested as a major contributing factor. An important step to shed light on the role of selective breeding as an underlying cause of KBF in modern laying hens is to evaluate the keel bones of the ancestor, the red jungle fowl. To the authors knowledge, this has never previously been published. The aim of this study was therefore to describe the prevalence of KBD in a study group of red jungle hens and roosters housed in an aviary system. The present study examined 29 red jungle fowls 112 weeks of age post-mortem; 12 hens and 17 roosters. Keel bones were evaluated by external palpation for deviations and fractures. Palpation was followed by autopsy. No fractures were detected in the 17 roosters; one had a very slight deviation. Of the 12 red jungle hens in this pilot study, one had a single fracture and 10 hens had a very slight deviation.

Production animals are constantly subjected to early adverse environmental conditions that influence the adult phenotype and produce epigenetic effects. CpG dinucleotide methylation in red blood cells (RBC) could be a useful epigenetic biomarker to identify animals subjected to chronic stress in the production environment. Here we compared a reduced fraction of the RBC methylome of chickens exposed to social isolation to non-exposed. These experiments were performed in two different locations: Brazil and Sweden. The aim was to identify stress-associated DNA methylation profiles in RBC across these populations, in spite of the variable conditions to which birds are exposed in each facility and their different lineages. Birds were increasingly exposed to a social isolation treatment, combined with food and water deprivation, at random periods of the day from weeks 1-4 after hatching. We then collected the RBC DNA from individuals and compared a reduced fraction of their methylome between the experimental groups using two bioinformatic approaches to identify differentially methylated regions (DMRs): one using fixed-size windows and another that preselected differential peaks with MACS2. Three levels of significance were used (P &lt;= 0.05, P &lt;= 0.005, and P &lt;= 0.0005) to identify DMRs between experimental groups, which were then used for different analyses. With both of the approaches more DMRs reached the defined significance thresholds in BR individuals compared to SW. However, more DMRs had higher fold change values in SW compared to BR individuals. Interestingly, ChrZ was enriched above expectancy for the presence of DMRs. Additionally, when analyzing the locations of these DMRs in relation to the transcription starting site (TSS), we found three peaks with high DMR presence: 10 kb upstream, the TSS itself, and 20-40 kb downstream. Interestingly, these peaks had DMRs with a high presence (&gt;50%) of specific transcription factor binding sites. Three overlapping DMRs were found between the BR and SW population using the most relaxed p-value (P &lt;= 0.05). With the most stringent p-value (P &lt;= 0.0005), we found 7 and 4 DMRs between treatments in the BR and SW populations, respectively. This study is the first approximation to identify epigenetic biomarkers of long-term exposure to stress in different lineages of production animals.

Brain size reduction is a common trait in domesticated species when compared to wild conspecifics. This reduction can happen through changes in individual brain regions as a response to selection on specific behaviours. We selected red junglefowl for 10 generations for diverging levels of fear towards humans and measured brain size and composition as well as habituation learning and conditioned place preference learning in young chicks. Brain size relative to body size as well as brainstem region size relative to whole brain size were significantly smaller in chicks selected for low fear of humans compared to chicks selected for high fear of humans. However, when including allometric effects in the model, these differences disappear but a tendency towards larger cerebra in low-fear chickens remains. Low-fear line chicks habituated more effectively to a fearful stimulus with prior experience of that same stimulus, whereas high-fear line chicks with previous experience of the stimulus had a response similar to naive chicks. The phenotypical changes are in line with previously described effects of domestication.

Domestication is one of the strongest examples of artificial selection and has produced some of the most extreme within-species phenotypic variation known. In the case of the chicken, it has been hypothesized that DNA methylation may play a mechanistic role in the domestication response. By inter-crossing wild-derived red junglefowl with domestic chickens, we mapped quantitative trait loci for hypothalamic methylation (methQTL), gene expression (eQTL) and behaviour. We find large, stable methylation differences, with 6,179 cis and 2,973 trans methQTL identified. Over 46% of the trans effects were genotypically controlled by five loci, mainly associated with increased methylation in the junglefowl genotype. In a third of eQTL, we find that there is a correlation between gene expression and methylation, while statistical causality analysis reveals multiple instances where methylation is driving gene expression, as well as the reverse. We also show that methylation is correlated with some aspects of behavioural variation in the inter-cross. In conclusion, our data suggest a role for methylation in the regulation of gene expression underlying the domesticated phenotype of the chicken.

Domesticated animals share a unique set of morphological and behavioral traits, jointly referred to as the domesticated phenotype. Striking similarities amongst a range of unrelated domesticated species suggest that similar regulatory mechanisms may underlie the domesticated phenotype. These include color pattern, growth, reproduction, development and stress response. Although previous studies have focused on the brain to find mechanisms underlying domestication, the potential role of the pituitary gland as a target of domestication is highly overlooked. Here, we study gene expression in the pituitary gland of the domesticated White Leghorn chicken and its wild ancestor, the Red Junglefowl. By overlapping differentially expressed genes with a previously published list of functionally important genes in the pituitary gland, we narrowed down to 34 genes. Amongst them, expression levels of genes with inhibitory function on pigmentation (ASIP), main stimulators of metabolism and sexual maturity (TSHB and DIO2), and a potential inhibitor of broodiness (PRLR), were higher in the domesticated breed. Additionally, expression of 2 key inhibitors of the stress response (NR3C1, CRHR2) was higher in the domesticated breed. We suggest that changes in the transcription of important modulatory genes in the pituitary gland can account not only for domestication of the stress response in domestic chickens, but also for changes in pigmentation, development, and reproduction. Given the pivotal role of the pituitary gland in the regulation of multiple shared domesticated traits, we suggest that similar changes in pituitary transcriptome may contribute to the domesticated phenotype in other species as well.

In commercial egg production, chicks are exposed to a potentially stressful procedure during their first day of life. Here, we investigated how this procedure affects the chickens in a short-as well as long-term perspective by conducting two behaviour tests and measuring corticosterone (CORT) and sex hormone levels at different time points. These results were compared with a group of control chickens from the same hatchery and incubator that did not go through the commercial hatchery routine. Chickens were continuously weighed, egg production data was collected and feather scoring was performed. We found that chicks have a significant increase in CORT during the hatchery process, which implies they are exposed to stress. During first weeks of life, these chicks were more fearful, had a higher CORT reactivity during restraint and weighed more than control chicks. Later in life, hatchery treated chickens had more feather damages and injuries on combs and wattles, a faster onset of egg laying and higher levels of estradiol. We conclude that processing at the commercial hatchery was a stressful event with short-and long-term effects on behaviour and stress reactivity, and potentially also positive effects on production. The results are relevant for a large number of individuals, since the chicken is by far the globally most common farm animal.

The evolution of gene expression in mammalian organ development remains largely uncharacterized. Here we report the transcriptomes of seven organs (cerebrum, cerebellum, heart, kidney, liver, ovary and testis) across developmental time points from early organogenesis to adulthood for human, rhesus macaque, mouse, rat, rabbit, opossum and chicken. Comparisons of gene expression patterns identified correspondences of developmental stages across species, and differences in the timing of key events during the development of the gonads. We found that the breadth of gene expression and the extent of purifying selection gradually decrease during development, whereas the amount of positive selection and expression of new genes increase. We identified differences in the temporal trajectories of expression of individual genes across species, with brain tissues showing the smallest percentage of trajectory changes, and the liver and testis showing the largest. Our work provides a resource of developmental transcriptomes of seven organs across seven species, and comparative analyses that characterize the development and evolution of mammalian organs.

Identifying the molecular mechanisms of animal behaviour is an enduring goal for researchers. Gaining insight into these mechanisms enables us to gain a greater understanding of behaviour and their genetic control. In this paper, we perform Quantitative Trait Loci (QTL) mapping of tonic immobility behaviour in an advanced intercross line between wild and domestic chickens. Genes located within the QTL interval were further investigated using global expression QTL (eQTL) mapping from hypothalamus tissue, as well as causality analysis. This identified five candidate genes, with the genes PRDX4 and ACOT9 emerging as the best supported candidates. In addition, we also investigated the connection between tonic immobility, meat pH and struggling behaviour, as the two candidate genes PRDX4 and ACOT9 have previously been implicated in controlling muscle pH at slaughter. We did not find any phenotypic correlations between tonic immobility, struggling behaviour and muscle pH in a smaller additional cohort, despite these behaviours being repeatable within-test.

This study reveals, for the first time, an interspecific synchronization in long-term stress levels. Previously, acute stress, has been shown to be highly contagious both among humans and between individuals of other species. Here, long-term stress synchronization in dogs and their owners was investigated. We studied 58 dog-human dyads and analyzed their hair cortisol concentrations (HCC) at two separate occasions, reflecting levels during previous summer and winter months. The personality traits of both dogs and their owners were determined through owner-completed Dog Personality Questionnaire (DPQ) and human Big Five Inventory (BFI) surveys. In addition, the dogs activity levels were continuously monitored with a remote cloud-based activity collar for one week. Shetland sheepdogs (N = 33) and border collies (N = 25), balanced for sex, participated, and both pet dogs and actively competing dogs (agility and obedience) were included to represent different lifestyles. The results showed significant interspecies correlations in long-term stress where human HCC from both summer and winter samplings correlated strongly with dog HCC (summer: N = 57, chi(2) = 23.697, P amp;lt; 0.001, beta = 0.235; winter: N = 55, chi(2) = 13.796, P amp;lt; 0.001, beta = 0.027). Interestingly, the dogs activity levels did not affect HCC, nor did the amount of training sessions per week, showing that the HCC levels were not related to general physical activity. Additionally, there was a seasonal effect in HCC. However, although dogs personalities had little effects on their HCC, the human personality traits neuroticism, conscientiousness, and openness significantly affected dog HCC. Hence, we suggest that dogs, to a great extent, mirror the stress level of their owners.

The domestication of the fowl resulted in a large diversity of integumental structures in chicken breeds. Several integumental traits have been investigated from a developmental genetics perspective. However, their distribution among breeds and their developmental morphology remain unexplored. We constructed a discrete trait-breed matrix and conducted a disparity analysis to investigate the variation of these structures in chicken breeds; 20 integumental traits of 72 chicken breeds and the red junglefowl were assessed. The analyses resulted in slight groupings of breed types comparable to standard breed classification based on artificial selection and chicken type use. The red junglefowl groups together with bantams and European breeds. We provide new data on the red junglefowl and four chicken breeds, demonstrating where and when variation arises during embryonic development. We document variation in developmental timing of the egg tooth and feather formation, as well as other kinds of developmental patterning as in the anlagen of different type of combs. Changes in epithelial-mesenchymal signaling interactions may drive the highly diverse integument in chickens. Experimental and comparative work has revealed that the cranial neural crest mesenchyme mediates its interactions with the overlying epithelium and is the likely source of patterning that generates diversity in integumental structures.

DNA methylation in CpGs dinucleotides is associated with high mutability and disappearance of CpG sites during evolution. Although the high mutability of CpGs is thought to be relevant for vertebrate evolution, very little is known on the role of CpG-related mutations in the genomic diversification of vertebrates. Our study analysed genetic differences in chickens, between Red Junglefowl (RJF; the living closest relative to the ancestor of domesticated chickens) and domesticated breeds, to identify genomic dynamics that have occurred during the process of their domestication, focusing particularly on CpG-related mutations. Single nucleotide polymorphisms (SNPs) and copy number variations (CNVs) between RJF and these domesticated breeds were assessed in a reduced fraction of their genome. Additionally, DNA methylation in the same fraction of the genome was measured in the sperm of RJF individuals to identify possible correlations with the mutations found between RJF and the domesticated breeds. Our study shows that although the vast majority of CpG-related mutations found relate to CNVs, CpGs disproportionally associate to SNPs in comparison to CNVs, where they are indeed substantially under-represented. Moreover, CpGs seem to be hotspots of mutations related to speciation. We suggest that, on the one hand, CpG-related mutations in CNV regions would promote genomic flexibility in evolution, i.e., the ability of the genome to expand its functional possibilities; on the other hand, CpG-related mutations in SNPs would relate to genomic specificity in evolution, thus, representing mutations that would associate with phenotypic traits relevant for speciation.

The production of food can have large impacts on sustainable development in relation to various socio-ecological dimensions, like climate change, the environment, animal welfare, livestock epidemiology, and the economy. To achieve a sustainable food production system in Sweden, an integrated approach that considers all five of these dimensions, and all parts of the food production chain, is necessary. This paper systematically reviewed the literature related to food production in Sweden, especially in association with resource distribution and recycling logistics, and identified potential sustainability interventions and assessed their effects according to the five dimensions. Participation of stakeholders across the food production chain contributed with the focus of the literature search and subsequent synthesis. In general, there were synergies between the sustainability interventions and their effect on climate change and the environment, while there often were trade-offs between effects on the economy and the other dimensions. Few interventions considered effects on animal welfare or livestock epidemiology and few studies dealt with resource distribution and recycling logistics. This indicates that there is a need for future research that considers this in particular, as well as research that considers the whole food production chain and all dimensions at once, and investigates effects across multiple scales.

Domestic animals are adapted to conditions vastly different from those of their wild ancestors, and this is particularly true for their diets. The most numerous of all domestic species, the chicken, originated from the Red Junglefowl (RJF), a native of subtropical forests in Southeast Asia. Surprisingly however, in domestic chicken breeds, a common haplotype of the beta-carotene oxygenase 2 (BCO2) gene, which is involved in carotenoid metabolism, is introgressed from a related species, the Gray Junglefowl, and has been under strong selective pressure during domestication. This suggests that a hybridization event may have conferred a fitness advantage on chickens carrying the derived allele. To investigate the possible biological function of the introgressed BCO2 allele in chicken, we introgressed the ancestral BCO2 allele into domestic White Leghorn chickens. We measured gene expression as well as carotenoid accumulation in skin and eggs of chickens carrying either the ancestral or the derived BCO2 allele. The derived haplotype was associated with down-regulation of BCO2 in skin, muscle, and adipose tissue, but not in liver or duodenum, indicating that carotenoid accumulation occurred in the tissues with reduced gene expression. Most importantly, we found that hens with the derived BCO2 genotype were capable of allocating stored carotenoids to their eggs, suggesting a functional benefit through buffering any shortage in the diet during egg production. Nevertheless, it is of interest that loss of function mutations in BCO2 gene are prevalent in other domesticates including cows, rabbits, and sheep, and, given the importance of carotenoids in development, reproduction, and immunity, it is possible that derived BCO2 alleles may provide a general mechanism in multiple domestic species to deal with higher demand for carotenoids in an environment with carotenoid shortage in the diet.

Behavior and welfare are closely dependent on the underlying genetics, yet so far this connection has been poorly investigated. Using domestication as a model, this chapter outlines the fundamentals of genetic inheritance, and explains some central concepts such as linkage, pleiotropy, epistasis, and heritability. It then outlines the essential approaches to finding single genes associated with specific behavioral- and welfare-related traits. These are split into top-down and bottom-up approaches, depending on whether the phenotype or genotype is the starting point for analysis. Finally, the novel field of epigenetics and its importance for welfare science are covered.

The domesticated phenotype is a set of behavioural, morphological and physiological traits that is common for domesticated species. Previous research has found that selection for tameness only can give rise to correlated selection responses that resemble the domesticated phenotype. It has therefore been suggested that tameness may drive the domesticated phenotype as correlated traits. We selected Red Junglefowl for divergent levels of fear of human for eight generations and assessed possible correlated selection responses in other behaviours in semi-natural settings. Behavioural studies were carried out on birds from generations six to eight, and at different ages, in order to study possible effects on general activity, social behaviour and male courtship behaviour. We found that the differences between selection lines changed with age. Adult low fear birds were generally more active and high fear males showed a more intense courtship behaviour. Our study shows that several behaviours can be modified through correlated selection responses by selection on reduced fear of humans only, emphasising the putative role of tameness as a driver of domestication related phenotypes.

Background: Domestication of animals leads to large phenotypic alterations within a short evolutionary time-period. Such alterations are caused by genomic variations, yet the prevalence of modified traits is higher than expected if they were caused only by classical genetics and mutations. Epigenetic mechanisms may also be important in driving domesticated phenotypes such as behavior traits. Gene expression can be modulated epigenetically by mechanisms such as DNA methylation, resulting in modifications that are not only variable and susceptible to environmental stimuli, but also sometimes transgenerationally stable. To study such mechanisms in early domestication, we used as model two selected lines of red junglefowl (ancestors of modern chickens) that were bred for either high or low fear of humans over five generations, and investigated differences in hypothalamic DNA methylation between the two populations. Results: Twenty-two 1-kb windows were differentially methylated between the two selected lines at p amp;lt; 0.05 after false discovery rate correction. The annotated functions of the genes within these windows indicated epigenetic regulation of metabolic and signaling pathways, which agrees with the changes in gene expression that were previously reported for the same tissue and animals. Conclusions: Our results show that selection for an important domestication-related behavioral trait such as tameness can cause divergent epigenetic patterns within only five generations, and that these changes could have an important role in chicken domestication.

Background: The genetics underlying body mass and growth are key to understanding a wide range of topics in biology, both evolutionary and developmental. Body mass and growth traits are affected by many genetic variants of small effect. This complicates genetic mapping of growth and body mass. Experimental intercrosses between individuals from divergent populations allows us to map naturally occurring genetic variants for selected traits, such as body mass by linkage mapping. By simultaneously measuring traits and intermediary molecular phenotypes, such as gene expression, one can use integrative genomics to search for potential causative genes. Results: In this study, we use linkage mapping approach to map growth traits (N = 471) and liver gene expression (N = 130) in an advanced intercross of wild Red Junglefowl and domestic White Leghorn layer chickens. We find 16 loci for growth traits, and 1463 loci for liver gene expression, as measured by microarrays. Of these, the genes TRAK1, OSBPL8, YEATS4, CEP55, and PIP4K2B are identified as strong candidates for growth loci in the chicken. We also show a high degree of sex-specific gene-regulation, with almost every gene expression locus exhibiting sex-interactions. Finally, several trans-regulatory hotspots were found, one of which coincides with a major growth locus. Conclusions: These findings not only serve to identify several strong candidates affecting growth, but also show how sex-specificity and local gene-regulation affect growth regulation in the chicken.

The identification of genes affecting sociality can give insights into the maintenance and development of sociality and personality. In this study, we used the combination of an advanced intercross between wild and domestic chickens with a combined QTL and eQTL genetical genomics approach to identify genes for social reinstatement, a social and anxiety-related behavior. A total of 24 social reinstatement QTL were identified and overlaid with over 600 eQTL obtained from the same birds using hypothalamic tissue. Correlations between overlapping QTL and eQTL indicated five strong candidate genes, with the gene TTRAP being strongly significantly correlated with multiple aspects of social reinstatement behavior, as well as possessing a highly significant eQTL.

Domestication of animals offers unique possibilities to study evolutionary changes caused by similar selection pressures across a range of species. Animals from separate genera tend to develop a suite of phenotypic alterations referred to as "the domesticated phenotype". This involves changes in appearance, including loss of pigmentation, and alterations in body size and proportions. Furthermore, effects on reproduction and behaviour are typical. It is hypothesized that this recurring phenotype may be secondary effects of the increased tameness that is an inevitable first step in the domestication of any species. We first provide a general overview of observations and experiments from different species and then review in more detail a project attempting to recreate the initial domestication of chickens. Starting from an outbred population of Red Junglefowl, ancestors of all modem chickens, divergent lines were selected based on scores in a standardized fear-of-human test applied to all birds at 12 weeks of age. Up to the eighth selected generation, observations have been made on correlated effects of this selection on various phenotypes. The fear score had a significant heritability and was genetically correlated to several other behavioural traits. Furthermore, low-fear birds were larger at hatch, grew faster, laid larger eggs, had a modified metabolism and increased feed efficiency, had modified social behaviour and reduced brain size. Selection affected gene expression and DNA-methylation in the brains, but the genetic and epigenetic effects were not specifically associated with stress pathways. Further research should be focused on unraveling the genetic and epigenetic mechanisms underlying the correlated side-effects of reduced fear of humans.