Research Note: Methods in detecting signs of life after gaseous stun in broilers.

To be compliant with religious requirements for poultry, slaughter facilities using controlled atmosphere stunning may require a reliable method for detecting signs of life. However, the production setting challenges methods utilized in veterinary and field use by generating excess sound and vibration, which can impact the effectiveness of typical measurement tools, such as the stethoscope or doppler ultrasound. In addition, it is required that the process be quick and efficient as to not interfere with the rapid pace of production. The current study assessed various methods including reflexes (pupillary light reflex, nictitating membrane reflex, and pedal reflex), doppler ultrasound, stethoscopes, and pulse oximeter for determination of signs of life in broiler chickens postcontrolled atmosphere stunning in production. Data were analyzed using a generalized linear mixed model in SAS. Each bird was considered an experimental unit (n = 18) and differences between dead on arrival (DOA; n = 6), stunned (n = 10) and sensible (n = 2) birds were compared using contrast statements. Reflexes were seen only in sensible birds. Of the evaluated tools, only the pulse oximeter was consistently capable of detecting heart beats per minute (bpm) within the production environment. Although the values of bpm did not differ between sensible and stunned birds (P = 0.724), DOA birds had a lower bpm than all others (P < 0.001). Although the accuracy of results deserves further investigation and the small sample size of this study posed challenges to our statistical analyses, including low statistical power for the comparison between sensible and stunned groups, the achieved results suggest that the pulse oximeter meets the requirements for a reliable and practical method in detecting signs of life in broilers. With the increasing importance of halal products in a market that prioritizes animal welfare, our results present a promising approach to meet halal certification requirements. Further studies on this topic are strongly encouraged.

Maternal age and maternal environment affect stress reactivity and measures of social behaviour in laying hens.

Maternal effects can shape the phenotypes of offspring, but the extent to which a layer breeder's experience can affect commercial laying hens remains unclear. We aimed to investigate the effects of maternal age and maternal environment on laying hens' behaviour and stress response. In our first experiment (E1), commercial hybrid hens were reared either in aviary or barren brooding cages, then housed in aviary, conventional cages or furnished (enriched) cages, thus forming different maternal housing treatments. Hens from each treatment were inseminated at three ages, and measures of response to manual restraint and social stress were assessed in offspring. In experiment 2 (E2), maternal age effects on offsprings' stress response were further investigated using fertile eggs from commercial breeder flocks at three ages. In E1, maternal age affected struggling and corticosterone during manual restraint, feather pecking and pulling and comb wounds. Additionally, maternal rearing and housing in aviary systems showed positive effects on measures of behaviour and stress response in offspring. Effects of maternal age were not replicated in E2, possibly due to methodological differences or higher tolerance to maternal effects in commercial breeders. Overall, we recommend researchers report parent stock age to increase comparison across studies and thus our understanding of maternal age effects.

Effects of acute stressors experienced by five strains of layer breeders on measures of stress and fear in their offspring.

Stressors experienced by layer breeders during egg production can lead to changes in the egg hormone content, potentially impacting their offspring, the commercial layers. Genetic differences might also affect the offspring's susceptibility to maternal experiences. In this study, we tested if maternal stress affects measures of stress and fear in five strains of layer breeders: commercial brown 1 & 2, commercial white 1 & 2 and a pure line White Leghorn. Each strain was equally separated into two groups: "Maternal Stress" (MS), where hens were subjected to a series of 8 consecutive days of acute psychological stressors, and "Control," which received routine husbandry. Additional eggs from Control were injected either with corticosterone diluted in a vehicle solution ("CORT") or just "Vehicle." Stress- and fear-responses of the offspring were measured in a plasma corticosterone test and a combined human approach and novel object test. While the stress treatments did not  affect the measured endpoints in the offspring, significant strain differences were found. The offspring of the white strains showed a higher physiological response compared to brown strains and the White 2 offspring was the least fearful strain in the human approach test. Our study found that neither the acute psychological stressors experienced by layer breeders nor the egg injections of corticosterone affected the parameters tested in their offspring. Post hoc power analyses suggest that the lack of treatment effects might be due to a small sample size (type II error). Although studies on larger flocks of layers are still needed, our results provide an initial understanding of an important subject, as in poultry production, layer breeders are often subjected to short-term stressors. In addition, our results suggest the dissociation between the physiological and behavioural parameters of stress response in laying hens, showing that increased concentrations of plasma corticosterone in response to stress might not be directly associated with high levels of fear.

Effects of Maternal Stress on Measures of Anxiety and Fearfulness in Different Strains of Laying Hens.

Maternal stress can affect the offspring of birds, possibly due to hormone deposition in the egg. Additionally, phenotypic diversity resulting from domestication and selection for productivity has created a variety of poultry lines that may cope with stress differently. In this study, we investigated the effects of maternal stress on the behavior of different strains of laying hens and the role of corticosterone as its mediator. For this, fertilized eggs of five genetic lines-two brown (Brown 1 and 2), two white (White 1 and 2), and one pure line White Leghorn-were reared identically as four flocks of 27 birds (24F: 3M) per strain. Each strain was equally separated into two groups: Maternal Stress ("MS"), where hens were subjected to a series of daily acute psychological stressors for 8 days before egg collection, and "Control," which received routine husbandry. Fertile eggs from both treatments were collected at three different ages forming different offspring groups that were treated as replicates; additional eggs from Control were injected either with corticosterone diluted in a vehicle solution ("CORT") or just "Vehicle." Eggs from each replicate were incubated and hatched, and offspring (N = 1,919) were brooded under identical conditions. To measure the effects of maternal stress on anxiety and fear-like behavior, offspring were subjected to a social isolation test (SI) between 5 and 10 days of age and a tonic immobility test (TI) at 9 weeks of age. Compared to Control, MS decreased the number of distress vocalizations emitted by White 2 in SI. No effects of MS were observed in TI, and no effects of CORT were observed in any tests. Overall, brown lines vocalized more in SI and remained in TI for a longer duration than white strains, suggesting genetic differences in fear behavior. Females vocalized more than males in TI and showed a trend toward significance for the same trait in SI. Overall, results suggest that the effects of maternal stress on fearfulness are not directly mediated by corticosterone. Moreover, it highlights behavioral differences across various strains of laying hens, suggesting that fear responses are highly dependent on genotype.

Effects of prenatal stress and genetics on embryonic survival and offspring growth of laying hens.

Early-life exposure to stressors can shape the phenotype of the offspring resulting in changes that may affect their prehatch and posthatch development. This can be modeled indirectly through maternal exposure to stressors (natural model) or by offspring exposure to stress hormones (pharmacological model). In this study, both models were used to investigate the effects of genetic line on hatchability, late embryonic mortality, sex ratio, and body weight until 17 wk of age. To form the parent stock, fertilized eggs of 4 commercial genetic lines - two brown (brown 1 and 2), two white (white 1 and 2), and a pure line White Leghorn - were incubated, hatched, and housed identically in 4 flocks of 27 birds (24 females and 3 males) per strain. Each strain was equally separated into 2 groups: "maternal stress," where hens were subjected to a series of acute psychological stressors (e.g., physical restraint, transportation) for 8 D before egg collection, and "control," where hens received routine husbandry. At 3 maternal ages, fertile eggs from both treatments were collected, and additional eggs from the control group were injected with corticosterone (10 ng/mL egg content) ("CORT"). A "vehicle" treatment was included to account for effects of egg manipulation. Each maternal age comprised a replicate over time. Eggs were incubated and hatched, and the offspring (N = 1,919) were brooded until 17 wk under identical conditions. The results show that prenatal stress interacted with strain to decrease embryonic survival and growth. Among all strains, brown 2 was consistently the most affected line in both prehatch and posthatch development. Our study shows that embryonic survival and offspring growth are mostly affected by the pharmacological model and that strain differences may increase susceptibility to prenatal stress. Moreover, it suggests that the natural stressor model may be useful for quantifying the response of the mother to stressors, whereas the pharmacological model may be useful for quantifying the response of the embryo to increased levels of corticosterone.