Impact of natural and artificial prenatal stimulation on the behavioural profile of Japanese quail.

As the sensory systems of vertebrates develop prenatally, embryos perceive many environmental stimuli that can influence the ontogeny of their behaviour. Whether the nature and intensity of prenatal stimuli affect this ontogeny differently remains to be investigated. In this context, this study aimed to analyse the effects of prenatal auditory stimulation (natural stimulation, NS; predator vocalisations or artificial stimulation, AS; metallic sounds) on the subsequent behaviour of young Japanese quail (Coturnix coturnix japonica). For this, behavioural variables recorded during ethological tests evaluating emotional and social reactivity were analysed using a principal component analysis. This analysis revealed significant differences between the behavioural profile of stimulated chicks and that of non-exposed chicks. Indeed, chicks exposed to NS expressed more intense emotional responses in fearful situations, but less neophobia when exposed to a novel environment or object, whereas chicks exposed to AS appeared more sensitive to social isolation. Our results show that the acoustic environment of embryos can influence the way young birds subsequently interact with their social and physical environment after hatching, and face challenges in changing living conditions.

Mother-chick interactions are affected by chicks' sex and brood composition in Japanese quail.

Sex is a trait which leads mothers to invest asymmetrically in their young, especially in species with a strong dimorphism. Authors usually consider that members of the largest sex require the most investment by mothers. In other cases, mothers care preferentially for members of the sex that increase their fitness, e.g. males, in numerous species. In quail, a precocial bird, mother seems to take more care of their males which are more sensitive to their separation. This study investigates the influence of chicks' sex and brood's sexual composition on the maternal behavior of quail. We evaluated three batches of mothers, composed of (a) unisex broods of males (MM), (b) unisex broods of females (FF), and (c) mixed sex broods (Mx). We recorded mother-chick interactions during 11 days of mothering. We found that MM mothers spent more time warming their chicks and FF mothers were more active. Moreover, females rejected more aggressively and earlier their female chicks than their male chicks. Finally, Mx chicks, whatever their sex, interacted more with their mothers. Our results highlight a greater quantity and quality of maternal care towards male. Mothers invest more in male chicks. We discussed results in terms of each sex's needs.

Maternal care affects chicks' development differently according to sex in quail.

Maternal behavior is known to influence the behavioral development of young. Recently, it was demonstrated that maternal behavior also differed according to sex of chicks and brood sex composition. So, here, we explored if these factors influenced behavioral development of quail chicks when they were brooded, and what characteristics of chicks and foster females could best explain this development. We studied three sets of chick pairs brooded by foster females: unisex male, unisex female, and mixed broods. We found that both emotivity profile and sociality depended on the sex: females were more reactive and less social than males. Females' emotivity profile was correlated with brood composition and foster female activity during maternal care. In males, only sociality was correlated with foster females' scores of aggressive rejection. Our results evidence that male and female chicks respond differentially to maternal behavior. This is discussed in terms of ecological and physiological constraints on development according to sex.

Lipids in maternal diet influence yolk hormone levels and post-hatch neophobia in the domestic chick.

We assessed whether the ratio of dietary n-6/n-3 polyunsaturated fatty acids (PUFA) during egg formation engenders transgenerational maternal effects in domestic chicks. We analyzed yolk lipid and hormone concentrations, and HPA-axis activity in hens fed a control diet (high n-6/n-3 ratio) or a diet enriched in n-3 PUFAs (low n-6/n-3 ratio) for 6 consecutive weeks. Their chicks were tested for neophobia during the first week of life. We found higher corticosterone metabolites in droppings of hens fed the diet enriched in n-3 and significantly higher concentrations of yolk progesterone, androstenedione, and estradiol in their eggs compared to controls. Chicks of hens fed the n-3 enriched diet showed a lower body mass at hatch than controls and expressed higher neophobia when exposed to a novel object. These results add support to the hypothesis that the nutritional state of female birds produces variation in yolk hormone levels and engender maternal effects.

3D tracking of animals in the field using rotational stereo videography.

We describe a method for tracking the path of animals in the field, based on stereo videography and aiming-angle measurements, combined in a single, rotational device. In open environments, this technique has the potential to extract multiple 3D positions per second, with a spatial uncertainty of <1 m (rms) within 300 m of the observer, and <0.1 m (rms) within 100 m of the observer, in all directions. The tracking device is transportable and operated by a single observer, and does not involve any animal tagging. As a video of the moving animal is recorded, track data can easily be completed with behavioural data. We present a prototype device based on accessible components that achieves about 70% of the theoretical maximal range. We show examples of bird ground and flight tracks, and discuss the strengths and limits of the method, compared with existing fine-scale (e.g. fixed-camera stereo videography) and large-scale tracking methods (e.g. GPS tracking).

Long-life partners or sex friends? Impact of parental pair bond on offspring personality.

Previous investigations reported that some traits of parental relationships, including pair-bond duration or mate behavioural compatibility, influence subsequent offspring fitness by acting on their behaviour and growth and thus their early survival. We hypothesized that the development of a pair bond between sexual partners would have a prenatal influence. This study investigated the impact of two pairing managements on the egg characteristics and development of offspring of Japanese quail (Coturnix c. japonica). Thirty males and 30 females were paired either continuously (C; mates together all the time) or non-continuously (NC; pairs met only three times a week for 5 min). Separation-reunion tests evaluated parental pair bond. Egg yolk testosterone and androstenedione levels were evaluated, and the somatic and behavioural development of C and NC chicks was assessed. Our results revealed that members of C pairs were attached to their mates and, although no significant differences in androgen levels could be evidenced between egg sets, a higher proportion of C pairs' eggs were fertilized and their chicks appeared less emotive and more social. Our results revealed that the parental relationship can modulate the behavioural development of their offspring, probably via non-genetic effects, and this could play a major role in the emergence of inter-individual variability.

Motherless quail mothers display impaired maternal behavior and produce more fearful and less socially motivated offspring.

Early maternal deprivation impairs the behavioral development of young individuals. Recently, strong differences between mothered and maternally deprived chicks have been reported concerning their emotionality, sociality, and spatial skills. Here we investigated long-term and cross-generational impacts of maternal deprivation by comparing the characteristics of the non-reproductive and the maternal behavior of 22 mothered and 22 non-mothered adult female Japanese quail (Coturnix c. japonica) and by comparing the behavior of their respective fostered chicks. We reveal that non-brooded mothers were more fearful and less competent in spatial tasks and expressed impaired maternal care, characterized by more aggression towards chicks, higher activity rates, and more abnormal pacing during the first days of the care period. Chicks' behavior was clearly affected by maternal care inducing strong differences in their fearfulness and social motivation. Our results show both long-term and cross-generational impacts of early maternal deprivation in precocial birds.

Trans-generational effects of prenatal stress in quail.

The prenatal environment is a source of phenotypic variability influencing the animal's characteristics. Prenatal stress affects not only the development of offspring, but also that of the following generation. Such effects have been best documented in mammals but can also be observed in birds, suggesting common processes across phylogenetic orders. We found previously that Japanese quail females stressed during laying produced offspring with higher fearfulness, probably related to modulation of testosterone levels in their eggs. Here, we evaluated long-term effects of prenatal stress by analysing reproductive traits of these F(1) offspring and, then, the development of their subsequent (F(2)) offspring. The sexual behaviour of F(1) prenatally stressed (F1PS) males was impaired. F1PS females' eggs contained less yolk and more albumen, and higher yolk testosterone and progesterone levels than did F(1) prenatal control females. The fearfulness of F(2) prenatally stressed quail was greater than that of F(2) prenatal control quail. These F(2) behavioural differences paralleled those evidenced by their parents, suggesting trans-generational transmission of prenatal stress effects, probably mediated by egg compositions of F1PS females.

Prenatal maternal stress is associated with behavioural and epigenetic changes in Japanese quail.

Prenatal maternal stress (PMS) influences many facets of offspring's phenotype including morphology, behaviour and cognitive abilities. Recent research suggested that PMS also induced epigenetic modifications. In the present study, we analysed, in the Japanese quail, the effects of PMS on the emotional reactivity and cognitive abilities of the F1 offspring. We also investigated in the hippocampus, the paraventricular hypothalamic nucleus and subnuclei of the arcopallium/amygdala the level of two histone post-translational modifications, H3K4me2 and H3K27me3, known to be impacted by stress. We found that PMS does not affect F1 quail's learning abilities but increases their emotional reactivity. Moreover, we demonstrated that PMS induced an increased density of H3K27me3 positive cells, in the hippocampus, paraventricular hypothalamic nucleus and dorsal nucleus of the amygdala, but not variations of H3K4me2. As these brain regions are involved in the control of vertebrates' emotional responses, the effect of PMS on the epigenetic mark H3K27me3 could possibly be a mechanism involved in the behavioural effects we observed in F1 quail.

Chicks from stressed females elicit overprotective behaviour in adoptive mother quail.

Beyond the genetic links between mother and young, prenatal effects experienced by offspring can modulate their mother's behaviour and stress. Here we investigated the consequences of stress-induced prenatal maternal effects on quail chicks' behaviour and maternal behaviour. We evaluated how adoptive quail mothers (unstressed) raised chicks of females that had either been stressed (PS) or were non-stressed (NPS) during laying. We demonstrated previously that our social stress procedure during laying increases levels of egg yolk steroid and chicks' emotional reactivity during the postnatal period. We found that, compared to NPS mothers, PS mothers remained closer to their chicks, were aggressed and neglected them less, emitted more maternal vocalisations and called their chicks more when they were separated. Moreover, PS chicks made more requests on their mother. Our results show that mothers responded to chicks' behavioural changes caused by prenatal stress by adapting their maternal behaviour. As exemplified here, for the first time in birds, prenatal environment can affect postnatal maternal behaviour by changing their offspring's behaviour.

Human behaviour at the origin of maternal effects on offspring behaviour in laying hens (Gallus gallus domesticus).

Regular visual presence of humans is known to reduce chickens' human-generated stress responses. Here we questioned whether, more than mere visual presence, human behaviour affects laying hen behaviour and subsequently their offspring's behaviour. We hypothesized that human behaviour triggers maternal effects via variations in yolk hormone levels. For five consecutive weeks, two groups of hens were exposed to the same durations of human presence (30 min twice a day, five days a week) but the behaviour of the human differed between groups. The first group (H+) was exposed to predictable arrival of the experimenter, slow movements combined with static presence, stroking during handling and human voice. Whereas the second group of hens (H-) was exposed to unpredictable arrival of the experimenter which remained silent, in motion, and did not provide stroking during handling. At the end of the treatment, we evaluated egg quality and offspring behaviour. We found that avoidance of the experimenter by H+ hens but not by H- hens decreased significantly. Fertility rates and concentrations of yolk progesterone and estradiol in H+ hens' eggs were higher than in H- hens' eggs. Fear of humans, neophobia or the capacity to solve a detour task did not differ significantly between H+ and H- chicks. Social discrimination tests showed that H+ chicks but not H- chicks typically preferred a familiar conspecific to a stranger. These results show that, with the same duration in the presence of the birds, humans through their behaviour engender variations in fertility rates, yolk hormone levels and transgenerational effects on social skills. Rarely explored, our data suggest that maternal effects influence filial imprinting. These data have broad implications for laboratory, commercial systems and conservatory programs where the inevitable presence of humans could trigger maternal effects on offspring phenotype.

Habituation to humans affects yolk steroid levels and offspring phenotype in quail.

In the field as well as in the laboratory, human-generated stress responses are reduced in adult animals previously habituated to humans in comparison to non-habituated individuals. In birds, yolk steroid levels vary with maternal environment and condition. We tested the hypothesis that the experience of female birds with humans could affect yolk steroids levels and offspring phenotype. Two groups of Japanese quail, one habituated to humans (H) and a second non-habituated (NH), were exposed daily to brief human disturbances. We analysed egg quality, offspring growth, and offspring emotional reactivity. NH females produced eggs with less androgens (testosterone and androstenedione) and more immunoreactive progesterone compared to birds habituated to humans. NH females produced eggs with less yolk, heavier shell and chicks hatching later and being smaller as compared to habituated individuals. A lower emotional reactivity was found in young of NH females compared to young of H females. Thus, human disturbance of the mother triggered different effects on chick phenotype depending on previous experience of mother birds with humans. In addition, we describe for the first time the influence of environmental stimuli on yolk immunoreactive progesterone levels. Our results show that a relatively minor difference in behavioral habituation may have substantial effects on eggs and offspring. This has obvious implications for keeping and handling laboratory animals, for conservation biology and for animal welfare.

Incubation temperature affects the expression of young precocial birds' fear-related behaviours and neuroendocrine correlates.

The influence of embryonic microclimate on the behavioural development of birds remains unexplored. In this study, we experimentally tested whether chronic exposure to suboptimal temperatures engendered plasticity in the expression of fear-related behaviours and in the expression of the corticotropin-releasing factor in the brains of domestic chicks (Gallus g. domesticus). We compared the neurobehavioural phenotypes of a control group of chicks incubated in an optimal thermal environment (37.8 °C) with those of a group of experimental chicks exposed chronically in ovo to suboptimal temperatures (27.2 °C for 1 hour twice a day). Chronic exposure to a suboptimal temperature delayed hatching and decreased growth rate and experimental chicks had higher neophobic responses than controls in novel food and novel environment tests. In addition, experimental chicks showed higher expression of corticotropin-releasing factor than did controls in nuclei of the amygdala, a structure involved in the regulation of fear-related behaviours. In this study, we report the first evidence of the strong but underappreciated role of incubation microclimate on the development of birds' behaviour and its neurobiological correlates.

Daily laying time in free-living European starlings: solar noon, a potential synchronizer.

Reproduction is generally controlled by important temporal constraints involving complex adaptive mechanisms. Birds, in temperate zones, present marked breeding seasonality as well as marked daily organization of reproductive behavior, especially laying. Intra-specific variability and determinants of this pattern have been investigated mainly in domestic non-passerine birds. The present study analyzed the daily temporal organization of laying in a free-living species, the European starling, Sturnus vulgaris. Breeding in a starling colony was monitored for five consecutive years using a non-invasive method (infrared video camera) to precisely estimate laying times. European starlings present a marked daily laying rhythm, ovipositions occurring only during a morning species-specific temporal window. Inside this laying window, time intervals between successive eggs varied greatly among females. Contrary to many species, the light/dark cycle did not appear to control laying time in European starlings, but daily variations of the ultraviolet composition of the solar spectrum appeared to be a possible Zeitgeber of this behavior.

Stable individual profiles of daily timing of migratory restlessness in European quail.

Temporal characteristics of migratory behavior in birds are usually studied at the species and population levels, and rarely at the individual level. Variations among species and populations of the seasonal onset of migratory behavior have been widely investigated, but very little is known about its daily organization or whether birds are conservative in their behavior. The determination of intra- and inter-individual variability is important for the study of genetic variations and can reveal the existence of different adaptation capacities within populations. This laboratory study analyzed intra- and inter-individual variability of daily initiation and time course of nocturnal restlessness in partial-migrant European quail (Coturnix coturnix coturnix). Thirty-five quail were selected randomly from a captive stock, and their spring activity was recorded under natural daylenghs. Eighteen of the thirty-five quail presented behavioral profiles of migrant birds. Migrant birds initiated their nocturnal activity punctually, and the time courses of the nocturnal activity of 88% of them revealed intra-individual stability over six consecutive nights. All birds initiated their nocturnal activity after sunset and civil twilight, and they were more active at the beginning than the middle or end of the night, suggesting that their drive to migrate could be synchronized with particular skylight conditions. For the first time, stable individual profiles in the daily time course of migratory restlessness are shown. These results support previous findings concerning biological rhythms of quail and raise questions concerning the timing of migratory behavior.

Brood size can influence maternal behaviour and chick's development in precocial birds.

Mothers have a crucial influence on offspring development. Variations of maternal behaviour can be due to numerous parameters, for instance costs are related to the size of a brood/litter, which in turn can influence the level of mothers' investment in each offspring. Here we investigated the influence of brood size on the behaviour of Japanese quail mothers and chicks during the mothering period and on offspring development. We compared two types of broods: small broods of three chicks (N=9) and large broods of six chicks (N=9). Behavioural tests assessed chicks' social and emotional traits. Mothers of large broods emitted more maternal vocalisations at the beginning of the mothering period, but at the end they assumed more non-covering postures and trampled chicks more than mothers of small broods. Chicks in large broods huddled up more whereas chicks in small broods rested alone more frequently. Moreover, the social motivation of chicks in large broods was higher than that of chicks in small broods, although their emotional reactivity levels were similar. Our results evidence the importance of brood size for maintaining family cohesion and the influence of brood size on chicks' interactions with their siblings. We evaluated the influence of mothers and siblings on chicks' behavioural development.

Effect of a cranium-directed daily illumination cycle on the laying rhythm in Japanese quail.

This study tests the effect on the laying rhythm of a light cycle reaching directly the encephala via a diode in Japanese quail maintained in constant darkness. In DD, all the birds expressed their free-running laying rhythm (period close to 25 h). When the diode is switched on 14 h per 24 h cycle, females showed the same organization as in LD with the same laying time. Thus, a photoperiodic cycle, where light was only perceived through the skull of the female quail, could synchronize its laying rhythm. This device with a LED is an interesting alternative solution to eye-patching or blinding of birds.

Cyclic melatonin synchronizes the circadian rhythm of feeding activity in Japanese quail, Coturnix c. japonica.

In Japanese quail, we can observe the circadian rhythm of feeding activity in constant conditions, especially in birds from selected lines. In order to try to test the importance of melatonin as hormonal output for the circadian system, we gave a 24-h period cycle of exogenous melatonin to some of these birds when they were free running. We used castrated males firstly in order to cancel the known effect of steroids on circadian organisation. Secondly, as castrated birds generally expressed a very short periodicity, it allowed us to check induced synchronisation more easily. We maintained ten castrated males in constant dim light. We divided the experiment into five successive phases. The birds received a 24-h period cycle of melatonin (M phase) or of control solution with only the alcoholic solvent (C phase) as a drink. Before and after each one of these two phases, we gave water continually to drink (W1, W2 and W3 phases). Thus, the successive phases were W1-M-W2-C-W3. We measured intake of liquids and plasma melatonin concentrations to check melatonin ingestion. We automatically recorded individual feeding activity by infrared detectors, and analysed this by spectral analysis. At the beginning of the experiment, eight birds showed a rhythmic feeding activity, with a mean period of 22.9 +/- 0.2 h, and the two others an arrhythmic circadian activity. During the 24-h period cycle of exogenous melatonin, for the rhythmic birds, the circadian period became approximately 24 h (23.9 +/- 0.2 h), the inactive phase corresponding to the period of melatonin availability. During the W2 and C phases, the circadian period was similar to that expressed during the W1 phase. Moreover, when birds only drink water, we found a significant positive relationship between the clarity of the circadian rhythm and the ratio, between the melatonin level of the inactive phase and that of the active phase. These facts support the hypothesis of the role of this hormone in the regulation of the circadian system, at least for feeding activity, in quail.

Daily temporal organization of laying in Japanese quail: variability and heritability.

In birds, many behavioral and physiological processes that occur during reproduction show daily rhythms in response to environmental temporal constraints. In this study, the individual daily organization of laying and its genetic determinant in Japanese quail (Coturnix coturnix japonica) were analyzed. For this purpose, the oviposition time of 102 randomly chosen females, maintained in long-day photoperiodic conditions (LD 14 h:10 h) for 1 mon was observed and extreme phenotypes selected. Laying is characterized by two parameters: oviposition interval and laying hour. The birds showed a specific time of laying during 24 h. All eggs were laid in the afternoon between 6.5 and 14 h after lights on (HALO). Two laying profiles were determined: 20% of females with an oviposition interval greater than 24 h (24.7 +/- 0.2 h) (the "delayed" profile) laid progressively later each day until a pause day. The remaining 80% of the females laid at the same time each day, with few pause days and an oviposition interval close to 24 h (24.0 +/- 0.2 h) (the "stable" profile). Among the females, showing this last profile, an intra-individual stability and an inter-individual variability of laying hour was established. Two extreme laying phenotypes were then determined: the "early" phenotype (E) for females laying on average between 7.5 and 9.5 HALO and the "late" phenotype (L) for females laying between 12.5 and 14HALO. In order to study the genetic basis of the laying hour, three females of each extreme phenotype were selected and crossed with two different males. The E and L females produced 57 F1E and 42 F1L daughters, respectively. F1 females displayed both laying profiles. However, the proportion of females displaying a "delayed" profile was higher in the L line (50%) than in the E line (29.8%). For the "stable" daughters, artificial selection induced an advance in laying hour of 4.7% for the E line and a delay of 4.7% for the L line. Realized heritability was estimated at 0.5. Moreover, the laying hour of the daughters was correlated positively to that of the mothers (N = 61; r = 0.45). These results support the notion of heritability of oviposition time in Japanese quail.

Circadian rhythms of oviposition and feeding activity in Japanese quail: effects of cyclic administration of melatonin.

The aim of these experiments was to test the effect of a cyclic administration of melatonin, by mimicking the daily rhythm of hormone levels, on the circadian organization of two distinct functions in quail: oviposition and feeding activity. Laying and feeding rhythms under photoperiodic conditions and constant darkness (DD) were investigated. Under DD, where the two rhythms were free running, a daily rhythm of melatonin was administered. In LD 14 h:10 h, two different individual profiles of laying were established, with stable females laying at the same time each day and delayed females laying progressively later each day. For feeding activity, all birds were clearly synchronized to the photoperiodic cycle. In DD, the laying birds showed a free-running rhythm of oviposition with a period longer than 24 h for both profiles but the delayed profile females had a longer period than stable profile females. In comparison, the free-running period of feeding rhythm of the same birds was shorter than 24 h. A cyclic administration of melatonin had no effect on laying rhythm, which continued to free-run in DD, whereas feeding activity was synchronized as soon as the first cycle of melatonin was administered. From these results, it seems that two different circadian systems drive each of the two types of behavior separately. Melatonin could be the main synchronizer for the temporal control of feeding behavior, but it does not play a part in the control of oviposition in Japanese quail.