Subclinical in utero Zika virus infection is associated with interferon alpha sequelae and sex-specific molecular brain pathology in asymptomatic porcine offspring.

Zika virus (ZIKV) infection during human pregnancy may lead to severe fetal pathology and debilitating impairments in offspring. However, the majority of infections are subclinical and not associated with evident birth defects. Potentially detrimental life-long health outcomes in asymptomatic offspring evoke high concerns. Thus, animal models addressing sequelae in offspring may provide valuable information. To induce subclinical infection, we inoculated selected porcine fetuses at the mid-stage of development. Inoculation resulted in trans-fetal virus spread and persistent infection in the placenta and fetal membranes for two months. Offspring did not show congenital Zika syndrome (e.g., microcephaly, brain calcifications, congenital clubfoot, arthrogryposis, seizures) or other visible birth defects. However, a month after birth, a portion of offspring exhibited excessive interferon alpha (IFN-α) levels in blood plasma in a regular environment. Most affected offspring also showed dramatic IFN-α shutdown during social stress providing the first evidence for the cumulative impact of prenatal ZIKV exposure and postnatal environmental insult. Other eleven cytokines tested before and after stress were not altered suggesting the specific IFN-α pathology. While brains from offspring did not have histopathology, lesions, and ZIKV, the whole genome expression analysis of the prefrontal cortex revealed profound sex-specific transcriptional changes that most probably was the result of subclinical in utero infection. RNA-seq analysis in the placenta persistently infected with ZIKV provided independent support for the sex-specific pattern of in utero-acquired transcriptional responses. Collectively, our results provide strong evidence that two hallmarks of fetal ZIKV infection, altered type I IFN response and molecular brain pathology can persist after birth in offspring in the absence of congenital Zika syndrome.

Discrimination learning and judgment bias in low birth weight pigs.

Low birth weight (LBW) is a risk factor for cognitive and emotional impairments in humans. In pigs, LBW is a common occurrence, but its effects on cognition and emotion have received only limited scientific attention. To assess whether LBW pigs suffer from impaired cognitive and emotional development, we trained and tested 21 LBW and 21 normal birth weight (NBW) pigs in a judgment bias task. Judgment bias is a measure of emotional state which reflects the influence of emotion on an animal's interpretation of ambiguous stimuli. Pigs were trained to perform a specific behavioral response to two auditory stimuli, predicting either a positive or negative outcome. Once pigs successfully discriminated between these stimuli, they were presented with intermediate, ambiguous stimuli. The pigs' responses to ambiguous stimuli were scored as optimistic (performance of 'positive' response) or pessimistic (performance of 'negative' response). Optimistic or pessimistic interpretation of an ambiguous stimulus is indicative of a positive or negative emotional state, respectively. We found LBW pigs to require more discrimination training sessions than NBW pigs to reach criterion performance, suggesting that LBW causes a mild cognitive impairment in pigs. No effects of LBW on judgment bias were found, suggesting a similar emotional state for LBW and NBW pigs. This was supported by comparable salivary and hair cortisol concentrations for both groups. It is possible the enriched housing conditions and social grouping applied during our study influenced these results.

Judgement bias in pigs is independent of performance in a spatial holeboard task and conditional discrimination learning.

Biases in judgement of ambiguous stimuli, as measured in a judgement bias task, have been proposed as a measure of the valence of affective states in animals. We recently suggested a list of criteria for behavioural tests of emotion, one of them stating that responses on the task used to assess emotionality should not be confounded by, among others, differences in learning capacity, i.e. must not simply reflect the cognitive capacity of an animal. We performed three independent studies in which pigs acquired a spatial holeboard task, a free choice maze which simultaneously assesses working memory and reference memory. Next, pigs learned a conditional discrimination between auditory stimuli predicting a large or small reward, a prerequisite for assessment of judgement bias. Once pigs had acquired the conditional discrimination task, optimistic responses to previously unheard ambiguous stimuli were measured in the judgement bias task as choices indicating expectation of the large reward. We found that optimism in the judgement bias task was independent of all three measures of learning and memory indicating that the performance is not dependent on the pig's cognitive abilities. These results support the use of biases in judgement as proxy indicators of emotional valence in animals.

Effects of environmental enrichment on cognitive performance of pigs in a spatial holeboard discrimination task.

This study investigated the effects of environmental enrichment on the cognitive performance of female conventional farm (growing) pigs in a spatial holeboard task. Ten pairs of littermates matched for weight were used. From each litter, one piglet was randomly assigned to a barren environment; the other was assigned to an enriched environment from 4 weeks of age. The enriched environment was double the size of the barren environment, had a floor covered with straw, a rooting area filled with peat, and one of the four different enrichment toys which were exchanged daily. Starting at 11 weeks of age, all pigs were tested in a spatial holeboard discrimination task in which 4 out of 16 holes were baited. Furthermore, basal salivary cortisol levels of all pigs were determined after the end of all testing. All pigs were able to acquire the pattern of baited holes (acquisition phase, 40 trials) and the diagonally mirrored pattern (reversal phase, 20 trials). During the acquisition phase, the reference memory performance of the enriched-housed pigs was better than that of their barren-housed littermates, i.e. they reduced visits to the unbaited set of holes. During the reversal phase, enriched-housed pigs had a better general working memory performance than the barren-housed pigs as indicated by reduced revisits to holes already visited during a trial, irrespective of whether they were of the baited or the unbaited set. The enriched-housed pigs also searched for the hidden bait faster during both phases. The environments did not affect basal salivary cortisol levels. In conclusion, environmental enrichment slightly improved the cognitive performance of pigs in a spatial learning task. We hypothesise that the long period of habituation to and testing in the holeboard acted as enrichment that partially reduced the effects of barren housing.

Decision-making under risk and ambiguity in low-birth-weight pigs.

Low birth weight (LBW) in humans is a risk factor for later cognitive, behavioural and emotional problems. In pigs, LBW is associated with higher mortality, but little is known about consequences for surviving piglets. Alteration in hypothalamic-pituitary-adrenal axis function in LBW pigs suggests altered emotionality, but no behavioural indicators have been studied. Decision-making under uncertain conditions, e.g., risk or ambiguity, is susceptible to emotional influences and may provide a means of assessing long-term effects of LBW in piglets. We tested LBW (N = 8) and normal-birth-weight (NBW; N = 8) male pigs in two decision-making tasks. For decision-making under risk, we developed a simple two-choice probabilistic task, the Pig Gambling Task (PGT), where an 'advantageous' option offered small but frequent rewards and a 'disadvantageous' option offered large but infrequent rewards. The advantageous option offered greater overall gain. For decision-making under ambiguity, we used a Judgement Bias Task (JBT) where pigs were trained to make an active response to 'positive' and 'negative' tone cues (signalling large and small rewards, respectively). Responses to ambiguous tone cues were rated as more or less optimistic. LBW pigs chose the advantageous option more often in later blocks of the PGT, and were scored as less optimistic in the JBT, than NBW pigs. Our findings demonstrate that LBW pigs have developed different behavioural strategies with respect to decision-making. We propose that this is guided by changes in emotionality in LBW piglets, and we provide behavioural evidence of increased negative affect in LBW piglets.

Hypothalamic vasotocin and tyrosine hydroxylase levels following maternal care and selection for low mortality in laying hens.

BACKGROUND: Feather pecking and cannibalism are major concerns in poultry farming, both in terms of animal welfare and farm economics. Genetic selection and introduction of (aspects of) maternal care have been suggested as potential interventions to reduce feather pecking in laying hens. Altered brain development has been proposed to reflect welfare states in animals, and can provide more insight into the underlying processes involved in feather pecking. Both vasotocin (the avian homologue of vasopressin) and dopaminergic neural circuitry have roles in control of social behaviors as well as in the stress response, and may be linked to feather pecking. Thus, the hypothalamus of adult laying hens selected for low early mortality (LML), which show low feather pecking, was examined and compared with a control line of adult laying hens selected for production characteristics only (CL). The effect of foster hen rearing on the two genetic lines and their hypothalamic morphology was also investigated. RESULTS: We demonstrated an increase in the number of neurons positive for the rate-limiting enzyme in dopamine production, tyrosine hydroxylase, in the periventricular area of the hypothalamus in the LML hens compared to CL hens. Hen-reared chicks showed more vasotocin -positive neurons in the medial pre-optic area compared to the hens raised without a hen. No correlations were found between behavior in an open field at 5-6 weeks of age, and the histology of the same hens at adulthood. CONCLUSION: The hypothalamic dopaminergic and vasotinergic systems are altered in hens following genetic selection or maternal care, indicating a potential role for these systems in feather pecking.

Effects of lighted incubation and foraging enrichment during rearing on individual fear behavior, corticosterone, and neuroplasticity in laying hen pullets.

Environmental conditions during incubation and rearing can influence stress responsivity of laying hen pullets throughout their lifespan, and therefore have important implications for their welfare. In this study, a 12:12h green LED light-dark cycle during incubation and larvae provisioning as enrichment during rearing were tested as strategies to optimize early-life conditions and thereby decrease stress responsivity in ISA Brown laying hens. A combination of parameters was measured to indicate neuronal, physiological, and behavioral changes that may affect fear and stress. The proteins calbindin D28k (calbindin1), doublecortin (DCX), and neuronal nuclein protein (NeuN) were quantified after hatch as a proxy for brain plasticity. Plasma and feather corticosterone levels were measured after hatch and at the end of the rearing phase, and fearfulness was investigated through a series of behavioral tests (i.e., voluntary approach, open field, tonic immobility, and manual restraint tests). No effects of light during incubation were found on calbindin1, DCX, or NeuN. Neither of the treatments affected corticosterone levels in blood plasma and feathers. Light-incubated pullets showed less fearfulness towards humans in the voluntary approach test, but not in the other behavioral tests reported in this study. Larvae provisioning had no effect on behavior. Our study showed minor effects of light during incubation and no effects of enrichment during rearing on stress responsivity of laying hen pullets. The small effects may be explained by the enriched rearing conditions for all birds in this experiment (low stocking density, natural daylight, and 24/7 classical music). Given the promising results of lighted incubation in other studies, which were mostly performed in broiler chickens, and evidence regarding the positive effects of enrichment during rearing, the potential of these strategies to improve laying hen welfare needs to be explored further.

Performance of conventional pigs and Göttingen miniature pigs in a spatial holeboard task: effects of the putative muscarinic cognition impairer Biperiden.

BACKGROUND: The pig is emerging as a model species that bridges the gap between rodents and humans in research. In particular, the miniature pig (referred to hereafter as the minipig) is increasingly being used as non-rodent species in pharmacological and toxicological studies. However, there is as yet a lack of validated behavioral tests for pigs, although there is evidence that the spatial holeboard task can be used to assess the working and reference memory of pigs. In the present study, we compared the learning performance of commercial pigs and Göttingen minipigs in a holeboard task. METHODS: Biperiden, a muscarinic M1 receptor blocker, is used to induce impairments in cognitive function in animal research. The two groups of pigs were treated orally with increasing doses of biperiden (0.05 - 20 mg.kg-1) after they had reached asymptotic performance in the holeboard task. RESULTS: Both the conventional pigs and the Göttingen minipigs learned the holeboard task, reaching nearly errorless asymptotic working and reference memory performance within approximately 100 acquisition trials. Biperiden treatment affected reference, but not working, memory, increasing trial duration and the latency to first hole visit at doses ≥ 5 mg.kg-1. CONCLUSION: Both pig breeds learned the holeboard task and had a comparable performance. Biperiden had only a minor effect on holeboard performance overall, and mainly on reference memory performance. The effectiveness needs to be evaluated further before definitive conclusions can be drawn about the ability of this potential cognition impairer in pigs.

Successive and conditional discrimination learning in pigs.

We studied the ability of pigs to discriminate tone cues using successive and conditional discrimination tasks. Pigs (n = 8) were trained in a successive discrimination Go/No-Go task (Experiment 1) to associate a Go-cue with a reward at the end of a runway and a No-Go-cue with the absence of reward. Latency to reach the goal-box was recorded for each cue-type. Learning of a conditional discrimination task was compared between low-birthweight (LBW, n = 5) and normal-birthweight (NBW, n = 6) pigs (Experiment 2) and between conventional farm (n = 7) and Göttingen miniature (n = 8) pigs (Experiment 3). In this active-choice task, one cue signalled a response in a right goal-box was correct and a second cue signalled a response in a left goal-box was correct. Cues were differentially rewarded. The number of sessions to learn the discrimination and number of correct choices per cue-type were recorded. In Experiment 1, four out of eight pigs showed learning on the task, that is, a higher latency to respond to the No-Go-cue, within 25 sessions. In Experiment 2, eight out of 11 pigs learned the discrimination within 46 sessions. LBW learners learned faster than NBW learners. In Experiment 3, all 15 pigs learned the task within 16 sessions. Göttingen miniature pigs learned faster than conventional farm pigs. While some methodological issues may improve the Go/No-Go design, it is suggested that an active-choice task yields clearer and more consistent results than one relying on latency alone.

Toward assessing the role of dietary fatty acids in lamb's neurological and cognitive development.

Understanding and measuring sheep cognition and behavior can provide us with measures to safeguard the welfare of these animals in production systems. Optimal neurological and cognitive development of lambs is important to equip individuals with the ability to better cope with environmental stressors. However, this development can be affected by nutrition with a special role from long-chain fatty acid supply from the dam to the fetus or in lamb's early life. Neurological development in lambs takes place primarily during the first two trimesters of gestation. Through late fetal and early postnatal life, the lamb brain has a high level of cholesterol synthesis. This rate declines rapidly at weaning and remains low throughout adulthood. The main polyunsaturated fatty acids (PUFA) in the brain are ω-6 arachidonic acid and ω-3 docosahexaenoic acid (DHA), which are elements of plasma membranes' phospholipids in neuronal cells. DHA is essential for keeping membrane integrity and is vital for normal development of the central nervous system (CNS), and its insufficiency can damage cerebral functions and the development of cognitive capacities. In sheep, there is evidence that supplying PUFA during gestation or after birth may be beneficial to lamb productive performance and expression of species-specific behaviors. The objective of this perspective is to discuss concepts of ruminant behavior and nutrition and reflect on future research directions that could help to improve our knowledge on how dietary fatty acids (FA) relate to optimal neurological and cognitive development in sheep.

Early-life interventions to prevent feather pecking and reduce fearfulness in laying hens.

Severe feather pecking, the pulling out of feathers of conspecifics, is a major welfare issue in laying hens. Possible underlying causes are fearfulness and lack of foraging opportunities. Because early life is a crucial stage in behavioral development, adapting the incubation and rearing environment to the birds' needs may reduce fearfulness and prevent the development of feather pecking. In a 2 × 2 factorial design study, we investigated whether a green light-dark cycle throughout incubation, which resembles natural incubation circumstances more than the standard dark incubation, and foraging enrichment with live larvae during rearing reduce fearfulness and feather pecking and increase foraging behavior of laying hen pullets from an early age onwards. In this 2-batch experiment, 1,100 ISA Brown eggs were incubated under either 0 h of light/24 h of darkness or 12 h of green LED light/12 h of darkness. After hatching, 400 female chicks (200 per batch) were housed in 44 pens (8-10 chicks per pen). During the entire rearing phase (0-17 wk of age), half of the pens received black soldier fly larvae in a food puzzle as foraging enrichment. We assessed fear of novel objects and humans, feather pecking, plumage condition, foraging behavior, and recovery time after a 3-fold vaccination (acute stressor). A slight increase in the number of foraging bouts was only seen with larvae provisioning (rate ratio 1.19, 95% CI 1.02-1.29, P = 0.008). Neither lighted incubation nor larvae provisioning affected fearfulness, feather pecking, plumage condition or recovery time after vaccination. In conclusion, the present study showed no effects of light during incubation and minor effects of foraging enrichment during rearing on the behavior of laying hen pullets. Further research is recommended on other welfare aspects.

Differential effects of diazepam and MPEP on habituation and neuro-behavioural processes in inbred mice.

BACKGROUND: Previous studies have demonstrated a profound lack of habituation in 129P3 mice compared to the habituating, but initially more anxious, BALB/c mice. The present study investigated whether this non-adaptive phenotype of 129P3 mice is primarily based on anxiety-related characteristics. METHODS: To test this hypothesis and extend our knowledge on the behavioural profile of 129P3 mice, the effects of the anxiolyticdiazepam (1, 3 and 5 mg/kg) and the putative anxiolytic metabotropic glutamate receptor 5 (mGlu5R) antagonist 2-methyl-6-(phenylethynyl)pyridine (MPEP, 3, 10 and 30 mg/kg) treatment on within-trial (intrasession) habituation, object recognition (diazepam: 1 mg/kg; MPEP 10 mg/kg) and on the central-nervous expression of the immediate early gene c-Fos (diazepam: 1 mg/kg; MPEP 10 mg/kg) were investigated. RESULTS: Behavioural findings validated the initially high, but habituating phenotype of BALB/c mice, while 129P3 mice were characterized by impaired intrasession habituation. Diazepam had an anxiolytic effect in BALB/c mice, while in higher doses caused behavioural inactivity in 129P3 mice. MPEP revealed almost no anxiolytic effects on behaviour in both strains, but reduced stress-induced corticosterone responses only in 129P3 mice. These results were complemented by reduced expression of c-Fos after MPEP treatment in brain areas related to emotional processes, and increased c-Fos expression in higher integrating brain areas such as the prelimbic cortex compared to vehicle-treated 129P3 mice. CONCLUSIONS: These results suggest that the strain differences observed in (non)adaptive anxiety behaviour are at least in part mediated by differences in gamma-aminobutyric acid- A and mGluR5 mediated transmission.

Cognitive performance of low- and normal-birth-weight piglets in a spatial hole-board discrimination task.

INTRODUCTION: Learning impairments are often seen in children born with low birth weight (LBW). A model with translational value for long-term effects of LBW in humans is needed to further our understanding of how LBW and cognition are related. The similarities between development stages in human infants and piglets, and the high prevalence of LBW piglets make them a naturally occurring potential model in which to study cognitive impairment associated with LBW in humans. RESULTS: Although both groups learned the configurations and rapidly reduced the number of incorrect visits, the LBW piglets showed a transiently retarded acquisition of the first reversal. DISCUSSION: The results of the experiment support the hypothesis that LBW is related to (mild) subsequent cognitive impairments. In the future, piglets may be suitable models for testing the effects of putative therapeutics. METHODS: To examine this potential model, we tested pairs of LBW and NBW (normal-birth-weight) piglets in a spatial hole-board (a matrix with 4 × 4 holes in the floor) task during one acquisition and two reversal phases in their own individual configurations of rewarded holes.

The Relation between Hair-Cortisol Concentration and Various Welfare Assessments of Dutch Dairy Farms.

Many protocols have been developed to assess farm animal welfare. However, the validity of these protocols is still subject to debate. The present study aimed to compare nine welfare assessment protocols, namely: (1) Welfare Quality© (WQ), (2) a modified version of Welfare Quality (WQ Mod), which has a better discriminative power, (3) WelzijnsWijzer (Welfare Indicator; WW), (4) a new Welfare Monitor (WM), (5) Continue Welzijns Monitor (Continuous Welfare Monitor; CWM), (6) KoeKompas (Cow Compass; KK), (7) Cow Comfort Scoring System (CCSS), (8) Stall Standing Index (SSI) and (9) a Welfare Index (WI Tuyttens). In addition, a simple welfare estimation by veterinarians (Estimate vets, EV) was added. Rank correlation coefficients were calculated between each of the welfare assessment protocol scores and mean hair cortisol concentrations from 10 cows at 58 dairy farms spread over the Netherlands. Because it has been suggested that the hair cortisol level is related to stress, experienced over a long period of time, we expected a negative correlation between cortisol and the result of the welfare protocol scores. Only the simple welfare estimation by veterinarians (EV) (ρ = -0.28) had a poor, but significant, negative correlation with hair cortisol. This correlations, however, failed to reach significance after correction of p-values for multiple correlations. Most of the results of the different welfare assessment protocols had a poor, fair or strong positive correlation with each other, supporting the notion that they measure something similar. Additional analyses revealed that the modified Welfare Quality protocol parameters housing (ρ = -0.30), the new Welfare Monitor (WM) parameter health (ρ = -0.33), and milk yield (ρ = -0.33) showed negative correlations with cortisol. We conclude that because only five out of all the parameter scores from the welfare assessment protocols showed a negative, albeit weak, correlation with cortisol, hair cortisol levels may not provide a long term indicator for stress in dairy cattle, or alternatively, that the protocols might not yield valid indices for cow welfare.

Effects of Dark Brooder Rearing and Age on Hypothalamic Vasotocin and Feather Corticosterone Levels in Laying Hens.

Chickens cannot independently thermoregulate at hatch and lack opportunity to behaviorally thermoregulate with a hen in the egg layer industry, thus barns are heated to thermoneutral temperatures. Dark brooders are low-energy-consuming hot plates, which may be environmentally advantageous while providing welfare-enhancing aspects of maternal care (i.e., shelter and separation of active and inactive individuals). Dark brooder use has been demonstrated to decrease injurious pecking and mortality well into the production period of layers. To further understand hen development around lay onset and effects of dark brooders on the brain and HPA-axis, we examined effects of rearing with dark brooders on expression of vasotocin (AVT) in the hypothalamus and corticosterone (CORT) in the feathers of in total 48 layer Isa Warren hens at 16 w and 28 w of age (n = 12 per age and treatment). An age-dependent decreased number of AVT-positive neurons was seen in the medial preoptic area, medial preoptic nucleus, paraventricular nucleus, rostral part (prepeduncular hypothalamus), and lateral preoptic area. Trends to effects of brooder rearing were found in both anteromedial preoptic nucleus and supraoptic nucleus, with dark brooder reared animals showing higher mean counts of AVT-positive neurons in both areas. No interactions between brooder raising and age were observed in AVT-positive neuron count. CORT levels were higher in primary wing feathers from 28 week old hens than in those from 16 week hens. No main effects of rearing with dark brooders or interactions between age and treatment were found on CORT levels. The age-dependent effects seen in the hypothalamus and CORT aids in further understanding of the development of chickens around puberty. The use of brooders tended to increase AVT expression in the anteromedial preoptic nucleus and supraoptic nucleus, an indication that dark brooder rearing may affect physiological responses regulated by these areas. The lack of effect of dark brooders on CORT in feathers is at the least an indication that the use of dark brooders is not stressful; in combination with the benefits of dark brooders on injurious pecking, fearfulness and early mortality, this pleads for the use of dark brooders in on-farm situations.

Spatial memory deficits after vincristine-induced lesions to the dorsal hippocampus.

Vincristine is a commonly used cytostatic drug for the treatment of leukemia, neuroblastoma and lung cancer, which is known to have neurotoxic properties. The aim of this study was to assess the effects of vincristine, injected directly into the dorsal hippocampus, in spatial memory using the spatial cone field discrimination task. Long Evans rats were trained in the cone field, and after reaching training criterion received bilateral vincristine infusions into the dorsal hippocampus. Vincristine-treated animals presented unilateral or bilateral hippocampal lesions. Animals with bilateral lesions showed lower spatial working and reference memory performance than control animals, but task motivation was unaffected by the lesions. Working and reference memory of animals with unilateral lesions did not differ from animals with bilateral lesions and control animals. In sum, intrahippocampal injection of vincristine caused profound tissue damage in the dorsal hippocampus, associated with substantial cognitive deficits.

Evaluation of animal models of neurobehavioral disorders.

Animal models play a central role in all areas of biomedical research. The process of animal model building, development and evaluation has rarely been addressed systematically, despite the long history of using animal models in the investigation of neuropsychiatric disorders and behavioral dysfunctions. An iterative, multi-stage trajectory for developing animal models and assessing their quality is proposed. The process starts with defining the purpose(s) of the model, preferentially based on hypotheses about brain-behavior relationships. Then, the model is developed and tested. The evaluation of the model takes scientific and ethical criteria into consideration.Model development requires a multidisciplinary approach. Preclinical and clinical experts should establish a set of scientific criteria, which a model must meet. The scientific evaluation consists of assessing the replicability/reliability, predictive, construct and external validity/generalizability, and relevance of the model. We emphasize the role of (systematic and extended) replications in the course of the validation process. One may apply a multiple-tiered 'replication battery' to estimate the reliability/replicability, validity, and generalizability of result.Compromised welfare is inherent in many deficiency models in animals. Unfortunately, 'animal welfare' is a vaguely defined concept, making it difficult to establish exact evaluation criteria. Weighing the animal's welfare and considerations as to whether action is indicated to reduce the discomfort must accompany the scientific evaluation at any stage of the model building and evaluation process. Animal model building should be discontinued if the model does not meet the preset scientific criteria, or when animal welfare is severely compromised. The application of the evaluation procedure is exemplified using the rat with neonatal hippocampal lesion as a proposed model of schizophrenia.In a manner congruent to that for improving animal models, guided by the procedure expounded upon in this paper, the developmental and evaluation procedure itself may be improved by careful definition of the purpose(s) of a model and by defining better evaluation criteria, based on the proposed use of the model.

Effects of Maternal Care During Rearing in White Leghorn and Brown Nick Layer Hens on Cognition, Sociality and Fear.

Both genetic background and maternal care can have a strong influence on cognitive and emotional development. To investigate these effects and their possible interaction, White Leghorn (LH) and Brown Nick (BN) chicks, two hybrid lines of layer hen commonly used commercially, were housed either with or without a mother hen in their first five weeks of life. From three weeks of age, the chicks were tested in a series of experiments to deduce the effects of breed and maternal care on their fear response, foraging and social motivation, and cognitive abilities. The LH were found to explore more and showed more attempts to reinstate social contact than BN. The BN were less active in all tests and less motivated than LH by social contact or by foraging opportunity. No hybrid differences were found in cognitive performance in the holeboard task. In general, the presence of a mother hen had unexpectedly little effect on behavior in both LH and BN chicks. It is hypothesized that hens from commercially used genetic backgrounds may have been inadvertently selected to be less responsive to maternal care than ancestral or non-commercial breeds. The consistent and strong behavioral differences between genetic strains highlights the importance of breed-specific welfare management processes.

Stocking Density Affects Stress and Anxious Behavior in the Laying Hen Chick During Rearing.

The recent increases in stocking density, in extreme cases resulting in 'crowding', have a major impact on poultry welfare. In contrast to available research on adult laying hens, there is a gap in the literature studying the rearing phase. The present study investigated the effects of stocking density during the rearing period on the welfare of the laying hen chick. The chicks were housed under one of three crowding conditions, increasing with age: undercrowding (500-1000-1429 cm² per chick), conventional crowding (167-333-500 cm² per chick), or overcrowding (56-111-167 cm² per chick). The parameters evaluated encompassed behavioral and physiological factors related to anxiety and stress. We found that during the first 6 weeks, overcrowded chicks displayed more anxious behavior than undercrowded chicks, and both extreme densities induced higher corticosterone levels compared to chicks housed under conventional crowding. At 10 weeks of age, plasma corticosterone had dropped to the level of conventional crowding group in both groups, whereas feather corticosterone remained high only in the overcrowded group. We conclude that current conventional stocking densities do not seem to impair the welfare state of the laying hen chick, and that a three-fold increase or decrease of density influences corticosterone levels and anxious behavior, but within the adaptive capacity of the chick. Important side notes to this conclusion are that an increase of stocking density did result in a slower rate of adaptation, and that there could be long-term consequences of both the different stocking densities and/or increased costs of adaptation.

Effects of parity and litter size on cortisol measures in commercially housed sows and their offspring.

Breeding sows are regularly exposed to on-farm stressors throughout the duration of their production period. The impact of such stressors may differ for primi- and multiparous sows, as sows could learn to cope with stressors as they gain experience with them. If parity affects stress in sows, it may also impact their prenatal offspring through differential maternal stress. In addition to parity, litter size is another potential factor involved in stress of sows and piglets. Larger litters may be a source of discomfort for gestating sows, while it can result in intra-uterine growth restriction of piglets. In the current study, we aimed to assess whether parity and litter size affect cortisol measures in breeding sows and their offspring. To do this, we measured salivary cortisol concentrations of 16 primiparous and 16 multiparous sows at three time points: 1) while sows were group housed, 2) after sows were separated from the group prior to moving to the farrowing unit and 3) after handling procedures. In addition, hair cortisol concentration was determined for the sows during late gestation and for their low birth weight (n = 63) and normal birth weight (n = 43) offspring on day 3 after birth, to reflect in-utero cortisol exposure. It was expected that if sows adapt to on-farm stressors, the more experienced, multiparous sows would show decreased stress responses in comparison to primiparous sows. However, we found a comparable acute stress response of primi- and multiparous sows to separation from the group. Handling procedures did not influence sows' salivary cortisol concentrations. Sows' hair cortisol concentration was positively correlated with litter size. Future research is needed to assess whether this finding reflects increased stress in sows carrying larger litters. Parity or litter size did not have a direct effect on their offspring's hair cortisol concentration. Larger litters did have a higher occurrence of low birth weight piglets. For these piglets, females had higher neonatal hair cortisol concentrations than males. Overall, our results indicate that breeding sows do not adapt to all on-farm stressors. In addition, litter size may influence HPA axis activity in both sows and piglets.