Maternal stress effects across generations in a precocial bird.

Prenatal maternal stress (PMS) is known to shape the phenotype of the first generation offspring (F1) but according to some studies, it could also shape the phenotype of the offspring of the following generations. We previously showed in the Japanese quail that PMS increased the emotional reactivity of F1 offspring in relation to (i) a variation in the levels of some histone post-translational modification (H3K27me3) in their brains and (ii) a modulation of the hormonal composition of the eggs from which they hatched. Here, we wondered whether PMS could also influence the behaviour of the second (F2) and third (F3) generation offspring due to the persistence of the specific marks we identified. Using a principal component analysis, we found that PMS influenced F2 and F3 quail profiles with subtle differences between generations. It increased F2 neophobia, F3 fearfulness and F3 neophobia but only in females. Interestingly, we did not find any variations in the level of histone post-translational modification in F3 brains and we observed inconsistent modulations of androstenedione levels in F1 and F2 eggs. Although they may vary over generations, our results demonstrate that PMS can have phenotypical effects into the third generation.

Facial blushing and feather fluffing are indicators of emotions in domestic fowl (Gallus gallus domesticus).

The study of facial expressions in mammals provided great advances in the identification of their emotions and then in the comprehension of their sentience. So far, this area of research has excluded birds. With a naturalist approach, we analysed facial blushing and feather displays in domestic fowl. Hens were filmed in situations contrasting in emotional valence and arousal level: situations known to indicate calm states (positive valence / low arousal), have rewarding effects (positive valence / high arousal) or induce fear-related behaviour (negative valence / high arousal). Head feather position as well as skin redness of comb, wattles, ear lobes and cheeks varied across these situations. Skin of all four areas was less red in situations with low arousal compared to situations with higher arousal. Furthermore, skin redness of the cheeks and ear lobes also varied depending on the valence of the situation: redness was higher in situations with negative valence compared to situations with positive valence. Feather position also varied with the situations. Feather fluffing was mostly observed in positively valenced situations, except when hens were eating. We conclude that hens have facial displays that reveal their emotions and that blushing is not exclusive to humans. This opens a promising way to explore the emotional lives of birds, which is a critical step when trying to improve poultry welfare.

Captive Blue-and-yellow macaws (Ara ararauna) show facial indicators of positive affect when reunited with their caregiver.

In mammals, human-animal bonding is recognized as a source of positive affect for companion or farm animals. Because this remains unexplored in birds, we investigated captive parrots' perspective of the human-animal relationship. We used a classical separation-reunion paradigm and predicted that variations in parrots' facial displays and behaviours would indicate their appraisal of the relationship. The test was divided into three phases of two minutes each: the bird was placed in an unfamiliar environment with a familiar caregiver (union), then the bird was left alone (separation) and finally, the caregiver returned (reunion). The test was repeated 10 times for each bird and video recorded in order to analyze their behaviour. The data show significantly higher crown and nape feather heights, higher redness of the skin and higher frequency of contact-seeking behaviours during the union and reunion phases than during the separation phase during which they expressed long distance contact calls. We observed the expression of eye pinning during the union and reunion phases in one out of five macaws. We argue that variation in facial displays provides indicators of parrot's positive appraisal of the caretaker presence. Our results broaden the scope for further studies on parrots' expression of their subjective feelings.

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.

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.

Treatment of gaseous emissions from tire manufacturing industry using lab-scale biofiltration pilot units.

Continuously seeking the improvement of environmental protection, the limitation of exhaust emissions is of significance for the tire manufacturing industry. The aim of this study is to assess the potential of biofiltration for the treatment of such gaseous emissions. This work highlights that biofiltration is able to remove both hydrophilic and hydrophobic compounds within a single pilot unit of biofiltration. Due to Ethanol/Alkanes ratios (95/5 and 80/20), high performance levels were observed for low EBRT (16 and 12 s). After twenty days of stable running, the dynamic of stratification patterns could be explained as a result of species coexistence mechanisms. While its impact on performance has not been observed under stable operating conditions, the use of an adsorbent support such as granular activated carbon (GAC) could be relevant to promote system stability in the face of further perturbations, such as transient regimes, that are problematic in full-scale industrial applications.

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

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

Human Face Recognition in Horses: Data in Favor of a Holistic Process.

Recent studies have demonstrated that horses can recognize humans based simply on visual information. However, none of these studies have investigated whether this involves the recognition of the face itself, or simply identifying people from non-complex external clues, such as hair color. To go beyond this we wanted to know whether certain features of the face were indispensable for this recognition (e.g., colors, hair or eyes). The 11 horses in this study had previously learned to identify four unfamiliar faces (portrait view and in color) presented repeatedly on a screen. We thus assessed whether they were able to identify these same faces spontaneously when they were presented in four other conditions: profile view, black and white, eyes hidden, changed hairstyle. The horses' performances remained higher than chance level for all the conditions. In a choice test under real conditions, they then approached the people whose face they had learned more often than unknown people. In conclusion, when considering all the individuals studied, no single facial element that we tested appears to be essential for recognition, suggesting holistic processing in face recognition. That means horses do not base their recognition solely on an easy clue such as hair color. They can also link faces from photographs with people in real life, indicating that horses do not process images of faces as simple abstract shapes.

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

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

Bill covering and nape feather ruffling as indicators of calm states in the Sulphur-crested cockatoo (Cacatua galerita).

Parrots are highly social birds that are recognized for their primate-like cognitive abilities but their way to express emotions remain overlooked. Herein we explored potential facial indicators of emotions in cockatoos. We predicted that facial feather ruffling is an indicator of a cockatoo's emotional state and hypothesized that specific facial feather positions would be present more during positive valence and low arousal situations. We observed feather position on the crest, cheek and nape during the daily routine of a group of five captive, non-breeding, Sulphur-crested cockatoos. The data show that cheek and nape feather ruffling occurred significantly more during activities associated with low arousal levels and positive valence such as maintenance behaviours, positive and quiet social contact and resting. Our data suggest that ruffling feathers over the bill (i.e. cheek feather ruffling) and nape ruffling may provide visual indicators of calm/relaxed states in cockatoos. Subtle movement of facial feathers may be an effective close-ranged visual signal to communicate birds' affective states or their intention to engage in specific activities. This work provides a novel approach to assessing the positive welfare of captive cockatoos and to understanding emotional communication in non-mammalian species.

Object and food novelty induce distinct patterns of c-fos immunoreactivity in amygdala and striatum in domestic male chicks (Gallus gallus domesticus).

Avoidance of novelty, termed neophobia, protects animals from potential dangers but can also impair their adaptation to novel environments or food resources. This behaviour is particularly well described in birds but the neurobiological correlates remain unexplored. Here, we measured neuronal activity in the amygdala and the striatum, two brain regions believed to be involved in novelty detection, by labelling the early gene c-fos following chicks exposure to a novel food (NF), a novel object (NO) or a familiar food (FF). NF and NO chicks showed significantly longer latencies to touch the food, less time eating and emitted more fear-vocalizations than control chicks. Latency to touch the food was also longer for NO than for NF chicks. Significantly higher densities of c-fos positive cells were present in all the nuclei of the arcopallium/amygdala of NF and NO chicks compared to FF chicks. Also, NO chicks showed higher positive cell densities than NF chicks in the posterior amygdaloid, the intermediate and the medial arcopallium. Exposure to novel food or object induced a similar increase in c-fos expression in the nucleus accumbens and the medial striatum. Our data provide evidence activation of the arcopallium/amygdala is specific of the type of novelty. The activation of striatum may be more related to novelty seeking.

High social motivation induces deficits in maternal behaviour but not plasticity of the subventricular zone in Japanese quail (Coturnix japonica).

Maternal behaviour develops differently depending on the characteristics of an individual, such as age or emotional reactivity. Social motivation, defined as the propensity to establish social contact, has received little attention in relation to maternal behaviour in birds. In addition, the transition to motherhood is a time of plasticity in the brain of the new mother in mammals. However, it remains to be determined how maternal brain plasticity is affected in avian species. The present study investigated how a the social motivation of a mother alters maternal behaviour and brain plasticity of the Japanese quail (Coturnix japonica). Adult females from lines selected for high and low social motivation were exposed to chicks for 11 days. After maternal care testing, and at matched time points in controls, the brains of females were perfused for assessment of doublecortin-immunoreactive staining, a marker of neurogenesis, in the subventricular zone (SVZ), a neurogenic niche. The results obtained showed that high socially motivated female quail spent significantly less time performing maternal behaviour when exposed to chicks compared to low socially motivated females. Moreover, the warming of chicks by high socially motivated females involved less covering postures and mothers were more rejecting of chicks. Interestingly, the plasticity indicators in the SVZ did not differ between low and high socially motivated females and were not associated with differences in maternal caregiving when using doublecortin-immunoreactive staining. Thus, high social motivation in this avian species does not favour maternal behaviour and this level of motivation to the mother is not related to changes in neuroplasticity in the SVZ of the female quail.

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.

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

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

Facial display and blushing: Means of visual communication in blue-and-yellow macaws (Ara Ararauna)?

Mainly recognized for their cognitive performance, the visual communication system and, particularly, the potential function of facial displays in parrots remain thus far unexplored. Here, we provide the first descriptive study of facial display use in captive blue-and-yellow macaws. We observed the feather position (sleeked or ruffled) on the crown, nape and cheek at the group level during the macaws' daily routine and individually while interacting with a familiar animal caretaker. In the latter context, blushing was also assessed on the bare skin of the cheek. Group level observations showed that crown, nape and cheek feathers ruffling was more frequent in activities requiring no locomotion than in activities requiring locomotion. With the animal caretaker, crown ruffling was significantly more frequent when the caretaker was actively engaging with the parrot than during a control phase with no mutual interaction. In addition, a significantly higher proportion of naïve observers judged blushing as being present on photographs taken during the mutual interaction phase than during the control phase. We thus showed significant variations in facial displays and bare skin colour based on the birds' social context and activity. Our results broaden the scope for further studies to determine whether parrots' faces provide visual social signals.

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

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

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.

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.

Genetic Determinism of Fearfulness, General Activity and Feeding Behavior in Chickens and Its Relationship with Digestive Efficiency.

The genetic relationships between behavior and digestive efficiency were studied in 860 chickens from a cross between two lines divergently selected on digestive efficiency. At 2 weeks of age each chick was video-recorded in the home pen to characterize general activity and feeding behavior. Tonic immobility and open-field tests were also carried out individually to evaluate emotional reactivity (i.e. the propensity to express fear responses). Digestive efficiency was measured at 3 weeks. Genetic parameters of behavior traits were estimated. Birds were genotyped on 3379 SNP markers to detect QTLs. Heritabilities of behavioral traits were low, apart from tonic immobility (0.17-0.18) and maximum meal length (0.14). The genetic correlations indicated that the most efficient birds fed more frequently and were less fearful. We detected 14 QTL (9 for feeding behavior, 3 for tonic immobility, 2 for frequency of lying). Nine of them co-localized with QTL for efficiency, anatomy of the digestive tract, feed intake or microbiota composition. Four genes involved in fear reactions were identified in the QTL for tonic immobility on GGA1.

Yolk hormones influence in ovo chemosensory learning, growth, and feeding behavior in domestic chicks.

In this study, we assessed whether prenatal exposure to elevated yolk steroid hormones can influence in ovo chemosensory learning and the behavior of domestic chicks. We simulated a maternal environmental challenge by experimentally enhancing yolk progesterone, testosterone, and estradiol concentrations in hen eggs prior to incubation. The embryos from these hormones-treated eggs (HO) as well as sham embryos (O) that had received the vehicle-only were exposed to the odor of fish oil (menhaden) between embryonic Days 11 and 20. An additional group of control embryos (C) was not exposed to the odor. All chicks were tested following hatching for their feeding preferences between foods that were or were not odorized with the menhaden odor. In the 3-min choice tests, the behavior of O chicks differed significantly according to the type of food whereas C and HO chicks showed no preference between odorized and non-odorized food. Our result suggests weaker response in HO chicks. In addition, HO chicks showed impaired growth and reduced intake of an unfamiliar food on the 24-h time scale compared to controls. Our data suggest that embryonic exposure to increased yolk hormone levels can alter growth, chemosensory learning, and the development of feeding behaviors.