Heritability and genetic correlations of fear-related behaviour in Red Junglefowl--possible implications for early domestication

Domesticated species differ from their wild ancestors in a number of traits, generally referred to as the domesticated phenotype. Reduced fear of humans is assumed to have been an early prerequisite for the successful domestication of virtually all species. We hypothesized that fear of humans is linked to other domestication related traits. For three generations, we selected Red Junglefowl (ancestors of domestic chickens) solely on the reaction in a standardized Fear of Human-test. In this, the birds were exposed for a gradually approaching human, and their behaviour was continuously scored. This generated three groups of animals, high (H), low (L) and intermediate (I) fearful birds. The birds in each generation were additionally tested in a battery of behaviour tests, measuring aspects of fearfulness, exploration, and sociality. The results demonstrate that the variation in fear response of Red Junglefowl towards humans has a significant genetic component and is genetically correlated to behavioural responses in other contexts, of which some are associated with fearfulness and others with exploration. Hence, selection of Red Junglefowl on low fear for humans can be expected to lead to a correlated change of other behavioural traits over generations. It is therefore likely that domestication may have caused an initial suite of behavioural modifications, even without selection on anything besides tameness.     

Effects of Resource Allocation on Behavioural Strategies: A Comparison of Red Junglefowl (Gallus gallus) and Two Domesticated Breeds of Poultry

Three different breeds of poultry, representing different degrees of domestication, were observed in semi-natural conditions in order to study differences in foraging behaviour, activity levels and social behaviours which could be caused by correlated responses to selection for increased production. The breeds used were: (i) red junglefowl ( Gallus gallus ); (ii) Swedish bantam, which is a domestic breed that has not undergone selection for production traits; and (iii) Hy-Line, a White leghorn laying hybrid, selected mainly for food conversion efficiency. The birds were offered ad libitum food simultaneously from sites where the food was freely available and from sites where the birds had to search and scratch for food which was mixed with wood-shavings. The behaviour was observed three times per day (48 min/d), 3 d per week in eight groups (four birds per group) of each breed between 7 and 18 wks of age. Junglefowl and bantam obtained a significantly higher proportion of their food from the site that required effort. The opposite case was true for the Hy-Line. Overall, bantams performed significantly more foraging behaviour than Hy-Lines. The Hy-Line breed was more inactive and less involved in social interactions than the junglefowl and the bantam. The results support the idea that selection for high production results in modified behavioural strategies. Behaviours that are of high energetic cost, such as extensive foraging and social interactions, were of lower in frequency in the laying hens compared to junglefowl and bantam, allowing the laying hens to save energy that can be reallocated to production traits.     

Domestication effects on behavioural synchronization and individual distances in chickens (Gallus gallus)

Behavioural synchrony (allelomimetic behaviour), and inter-individual distances are aspects of social and anti-predator strategies which may have been affected by domestication. Chickens are known to adjust synchronization and inter-individual distances depending on behaviour. We hypothesized that White Leghorn (WL) chickens would show less synchronized behaviour than the ancestor, the red jungle fowl (RJF). Sixty birds, 15 female and 15 male WL and the same number of RJF (28 weeks old) were studied in groups of three in furnished pens (1 m × 2 m) for 24 consecutive hours per group, following 24 h of habituation. Video tapes covering 4 h per group (dawn, 9-10 am, 1-2 pm and dusk) were analysed. Red junglefowl perched significantly more, but there were no breed effects on the frequency or daily rhythm of any other activities, or on average inter-individual distances. Red junglefowl were more synchronized during perching and a tendency for the same was found for social behaviour. After performance of the two most synchronized behaviours, perching and comfort behaviour, individual distance increased more for RJF than WL. According to this study domestication of chickens appears not to have significantly altered the relative frequencies of different activities or average inter-individual distances, but have caused some changes in behavioural synchronization and maintenance of activity-specific inter-individual distances in chickens. The changes may indicate an adaptive response to captivity and domestication.     

Is domestication driven by reduced fear of humans? Boldness,metabolism and serotonin levels in divergently selected red junglefowl (Gallus gallus)

Domesticated animals tend to develop a coherent set of phenotypic traits. Tameness could be a central underlying factor driving this, and we therefore selected red junglefowl, ancestors of all domestic chickens, for high or low fear of humans during six generations. We measured basal metabolic rate (BMR), feed efficiency, boldness in a novel object (NO) test, corticosterone reactivity and basal serotonin levels (related to fearfulness) in birds from the fifth and sixth generation of the high- and low-fear lines, respectively (44–48 individuals). Corticosterone response to physical restraint did not differ between selection lines. However, BMR was higher in low-fear birds, as was feed efficiency. Low-fear males had higher plasma levels of serotonin and both low-fear males and females were bolder in an NO test. The results show that many aspects of the domesticated phenotype may have developed as correlated responses to reduced fear of humans, an essential trait for successful domestication.     

Asymmetries,heterosis, and phenotypic profiles of red junglefowl,White Plymouth Rocks,and F<Subscript>1</Subscript> and F<Subscript>2</Subscript> reciprocal crosses

During the domestication of farm animals, humans have manipulated genetic variation for growth and reproduction through artificial selection. Here, data are presented for growth, reproductive, and behavior traits for the red junglefowl, a line of White Plymouth Rock chickens, and their F₁ and F₂ reciprocal crosses. Intra- and intergenerational comparisons for growth related traits reflected considerable additive genetic variation. In contrast, those traits associated with reproduction exhibited heterosis. The role of sexual selection was seen in the evolution of prominent secondary sexual ornaments that lend to female choice and male-male competition. The large differences between parental lines in fearfulness to humans were only mitigated slightly in the intercross generations. Whereas, overall F₁ generation heterosis was not transferred to the F₂, there was developmental stability in the F₂, as measured by relative asymmetry of bilateral traits. Through multigenerational analyses between the red junglefowl and the domestic White Plymouth Rocks, we observed plasticity and considerable residual genetic variation. These factors likely facilitated the adaptability of the chicken to a broad range of husbandry practices throughout the world.     

Tameness correlates with domestication related traits in a Red Junglefowl intercross

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.     

Effects of the domestic thyroid stimulating hormone receptor (TSHR) variant on the hypothalamic-pituitary-thyroid axis and behavior in chicken

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 it’s 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.     

Fear and stress reactions in two species of duck and their hybrid

Susceptibility to fear and stress has often been studied in birds using selection experiments. Crosses between different species of the same genus have also been used to study the type of transmission observed for "fixed action patterns." In this experiment we used a cross between two species (male muscovy and female pekin ducks) to study the genetic transmission of various characteristics of fear reactions and adrenal activity. Males of the three genetic types (two parental species and the hybrid) were submitted to tonic immobility and fear of man tests. Blood was collected for corticosterone assay before and after physiological stress or injection of 5 microg/kg of immediate synacthen (ACTH agonist). The muscovy duck showed lower levels of fear reactions than the pekin duck in the majority of behavioral tests (six of eight). Corticosterone levels were also nearly always the lowest in muscovy ducks (five of six). Generally speaking, the muscovy duck appeared to be less fearful and less susceptible to stress than the pekin duck. The mule duck was similar to one parent, midway between the two, or showed heterosis for fear of man at 10 weeks of age (more fearful than the two parents). Corticosterone levels were midway between the two parents in the mule duck with only one exception, i.e., where the pekin duck was dominant. The existence of heterosis for fear is surprising, as this character is classically considered to be submitted to stabilizing selection. The transmission of corticosterone levels and other fear characteristics is, however, compatible with stabilizing selection. The results are discussed in terms of adaptability.     

Cerebellum size is related to fear memory and domestication of chickens

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.     

Red Junglefowl (Gallus gallus) selected for low fear of humans are larger,more dominant and produce larger offspring

Many traits associated with domestication are suggested to have developed as correlated responses to reduced fear of humans. Tameness may have reduced the stress of living in human proximity and improved welfare in captivity. We selected Red Junglefowl (ancestors of all domestic chickens) for four generations on high or low fear towards humans, mimicking an important aspect of the earliest period of domestication, and tested birds from the third and fourth generation in three different social tests. Growth and plumage condition, as well as size of eggs and offspring were also recorded, as indicators of some aspects of welfare. Birds selected for low fear had higher weight, laid larger eggs and generated larger offspring, and had a better plumage condition. In a social dominance test they also performed more aggressive behaviour and received less of the same, regardless of whether the restricted resource was feed or not. Hence, dominance appeared to increase as a consequence of reduced fear of humans. Furthermore, egg size and the weight of the offspring were larger in the less fearful birds, and plumage condition better, which could be interpreted as the less fearful animals being better adapted to the environment in which they were selected.     

Chicken domestication: an updated perspective based on mitochondrial genomes

Domestic chickens (Gallus gallus domesticus) fulfill various roles ranging from food and entertainment to religion and ornamentation. To survey its genetic diversity and trace the history of domestication, we investigated a total of 4938 mitochondrial DNA (mtDNA) fragments including 2843 previously published and 2095 de novo units from 2044 domestic chickens and 51 red junglefowl (Gallus gallus). To obtain the highest possible level of molecular resolution, 50 representative samples were further selected for total mtDNA genome sequencing. A fine-gained mtDNA phylogeny was investigated by defining haplogroups A–I and W–Z. Common haplogroups A–G were shared by domestic chickens and red junglefowl. Rare haplogroups H–I and W–Z were specific to domestic chickens and red junglefowl, respectively. We re-evaluated the global mtDNA profiles of chickens. The geographic distribution for each of major haplogroups was examined. Our results revealed new complexities of history in chicken domestication because in the phylogeny lineages from the red junglefowl were mingled with those of the domestic chickens. Several local domestication events in South Asia, Southwest China and Southeast Asia were identified. The assessment of chicken mtDNA data also facilitated our understanding about the Austronesian settlement in the Pacific.     

Evolutionary pets: offspring numbers reveal speciation process in domesticated chickens

Since Darwin, the nature of the relationship between evolution and domestication has been debated. Evolution offers different mechanisms of selection that lead to adaptation and may end in the origin of new species as defined by the biological species concept. Domestication has given rise to numerous breeds in almost every domesticated species, including chickens. At the same time, so-called artificial selection seems to exclude mechanisms of sexual selection by the animals themselves. We want to forward the question to the animal itself: With whom do you reproduce successfully? This study focused on the sexual behavior of the domestic chicken Gallus gallus f.dom., particularly the White Crested Polish breed. Experiments on mate choice and the observation of fertilization and hatching rates of mixed-breeding groups revealed breed-specific preferences. In breeding groups containing White Crested Polish and a comparative breed, more purebred chicks hatched than hybrids (number of eggs collected: 1059). Mating was possible in equal shares, but in relation to the number of eggs collected, purebred offspring (62.75%±7.10%, M±SE) hatched to a greater extend compared to hybrid offspring (28.75%±15.32%, M±SE). These data demonstrate that the mechanism of sexual selection is still present in domestic chicken breeds, which includes the alteration of gene frequencies typical for domestication and evolutionary speciation. Due to selection and mate choice we state that breeding in principle can generate new species. Therefore, we see domestication as an evolutionary process that integrates human interests of animal breeding with innate mate choice by the animal.     

Estimation of fear in two lines of the domestic chick: Correlations between various methods

The assessment of fear is a controversial issue and low levels of correlation between different measures have been used to criticise the fear concept. The present study assessed fear levels in individual domestic chicks of each of two lines, flighty and docile, using four commonly employed methods of estimating fear. They were the hole-in-the-wall box, the open field, response to a bell and tonic immobility. On the basis of a wide variety of behavioural responses each chick was ranked for fearfulness in each of the four tests. The degrees of association or correlation between these ranking within lines were then calculated. The significant intra-individual correlations found in both lines provides some support for the use of these tests as methods of estimating fear, at least within the lines used. Differences in the degree or form of fear-responding between the lines are also discussed in terms of reactions to handling and to sudden auditory stimuli.     

Behavioural evaluation of methods for assessing fear responses in weaned pigs

Models of anxiety and fear of novelty were evaluated using correlations and principal component analysis. A total of 84 pigs (LandracexYorkshire) from nine different litters were subjected to a tonic immobility (TI) test at the age of 2.5 weeks, an elevated plus-maze (EPM) at the age of 6 weeks, a light/dark (L/D) exploration test at the age of 7 weeks and an open-field (OF) test at the age of 8 weeks.The first component from the principal component analysis had the highest correlation with number of entries into open arms in the EPM but was also highly correlated to variables from the other three tests confirming a common aversion-related element in the four experimental tests. The second component was negatively correlated with percent entries into and time spent on open arms in the EPM, but positively correlated with the number of entries into closed arms in the same test, number of lines crossed in the OF and time spent in the lit compartment of the L/D test. The last point illustrates a negative relationship between "anxiety" and "activity" in the EPM and OF. To achieve purer measures of fear of novelty and activity in the tests, the components were rotated using the Varimax criterion. The rotated factor pattern demonstrated a simple structure where variables related to "anxiety" or "fear of novelty" (i.e., percent entries into open arms and time spent on open arms of the EPM) had the highest loading on factor 1, whereas variables related to activity (i.e., number of entries into the closed arms in the EPM, number of lines crossed in the OF and time spent in the lit compartment of the L/D test) had the highest loading on factor 2. TI duration loaded more strongly on factor 1 ("fear of novelty") than on factor 2 ("activity"), but did not represent any pure measure of either fear of novelty or activity.In conclusion, all of the test variables were related to one another. Open-arm avoidance represented the purest measure of fear of novelty, whereas entries into closed arms and number of lines crossed in the OF were the purest measures of activity. The EPM appeared to provide the best way to separate the fear of novelty and activity-related elements, indicating that the EPM may be a useful behavioural model of fear of novelty or avoidance in pigs.     

Selection for tonic immobility duration does not affect the response to novelty in quail

Genetic selection on a single fear test, the tonic immobility test, seems to result in selection on fearfulness, i.e. the propensity to exhibit fear responses, whatever the fear tests used. However, the conception of fear as a single variable has been challenged by the recognition that fear is multidimensional. This study was designed to test whether genetic selection on a classic index of fear in birds, tonic immobility duration, is accompanied by changes in the response to a single dimension of fear – novelty.Two lines of quail divergently selected for long (LTI) or short (STI) duration of tonic immobility were exposed to a novel object in their home cage. Quail of both lines showed typical fear reactions in response to novelty but there was no difference between lines. We conclude that genetic selection for tonic immobility duration does not affect all dimensions of fear, notably not novelty. Further studies are needed to investigate the dimensions of fear on which the two lines of quail could have been selected.     

Postnatal development of conditioned reflex behavior: comparison of the periods related to the onset of the different forms of hippocampal plasticity in rats

Fear conditioning, escape and active avoidance reactions in two-way avoidance paradigm were compared in rats of different ages. Fear conditioning, but not escape and active avoidance reactions could be acquired on the 16-17th postnatal days, and the acquisition was more effective than in adults. Escape behavior matured beginning from the 18th postnatal day reaching the adult level within the 3d-4th postnatal weeks. Maturation of the mechanisms of Pavlovian (fear reaction) and instrumental (escape reaction) conditioning did not facilitate the acquisition of two-way avoidance until the 4th postnatal week, young animals displayed low acquisition in this period. The maturation of these memory processes is proposed to be related to developmental stages of different mechanisms of hippocampal plasticity.     

The origin and genetic diversity of Chinese native chicken breeds

The first 539 bases of mitochondrial DNA D-loop region of six Chinese native chicken breeds (Gallus gallus domesticus) were sequenced and compared to those of the red junglefowl (Gallus gallus), the gray junglefowl (Gallus sonneratii), the green junglefowl (Gallus varius) and Lafayette's junglefowl (Gallus lafayettei) reported in GenBank, and the phylogenetic trees for the chickens were constructed based on the D-loop sequences. The results showed that the four species of the genus Gallus had great differences among each other, the G. g. domesticus was closest to the red junglefowl in Thailand and its adjacent regions, suggesting the Chinese domestic fowl probably originated from the red junglefowl in these regions. The two subs pecies of Thailand, G. g. gallus and G. g. spadiceus, should belong to one subspecies because of their resemblance. In the case of native breeds, there existed a great difference between the egg breeds and general purpose breeds, which suggested different maternal origins of the two types.     

Genome‐wide scans to detect positive selection in Large White and Tongcheng pigs

Due to the direction, intensity, duration and consistency of genetic selection, especially recent artificial selection, the production performance of domestic pigs has been greatly changed. Therefore, we reasoned that there must be footprints or selection signatures that had been left during domestication. In this study, with porcine 60K BeadChip genotyping data from both commercial Large White and local Chinese Tongcheng pigs, we calculated the extended haplotype homozygosity values of the two breeds using the long‐range haplotype method to detect selection signatures. We found 34 candidate regions, including 61 known genes, from Large White pigs and 25 regions comprising 57 known genes from Tongcheng pigs. Many selection signatures were found on SSC1, SSC4, SSC7 and SSC14 regions in both populations. According to quantitative trait loci and network pathway analyses, most of the regions and genes were linked to growth, reproduction and immune responses. In addition, the average genetic differentiation coefficient F_ST was 0.254, which means that there had already been a significant differentiation between the breeds. The findings from this study can contribute to further research on molecular mechanisms of pig evolution and domestication and also provide valuable references for improvement of their breeding and cultivation.