A two-step Bayesian network approach to identify key SNPs associated to multiple phenotypic traits in four purebred laying hen lines.

When purebred laying hen chicks hatch, they remain at a rearing farm until approximately 17 weeks of age, after which they are transferred to a laying farm. Chicks or pullets are removed from the flocks during these 17 weeks if they display any rearing abnormality. The aim of this study was to investigate associations between single nucleotide polymorphisms (SNPs) and rearing success of 4 purebred White Leghorns layer lines by implementing a Bayesian network approach. Phenotypic traits and SNPs of four purebred genetic White Leghorn layer lines were available for 23,000 rearing batches obtained between 2010 and 2020. Associations between incubation traits (clutch size, embryo mortality), rearing traits (genetic line, first week mortality, rearing abnormalities, natural death, rearing success, pullet flock age, and season) and SNPs were analyzed, using a two-step Bayesian Network (BN) approach. Furthermore, the SNPs were connected to their corresponding genes, which were further explored in bioinformatics databases. BN analysis revealed a total of 28 SNPs associated with some of the traits: ten SNPs were associated with clutch size, another 10 with rearing abnormalities, a single SNP with natural death, and seven SNPs with first week mortality. Exploration via bioinformatics databases showed that one of the SNPs (ENAH) had a protein predicted network composed of 11 other proteins. The major hub of this SNP was CDC42 protein, which has a role in egg production and reproduction. The results highlight the power of BNs in knowledge discovery and how their application in complex biological systems can help getting a deeper understanding of functionality underlying genetic variation of rearing success in laying hens. Improved welfare and production might result from the identified SNPs. Selecting for these SNPs through breeding could reduce stress and increase livability during rearing.

Carvacrol in ovo delivery optimization and flow dynamics in broiler chicken eggs.

In ovo delivery of carvacrol, the primary active compound in oregano essential oil (OEO) has the potential to enhance gut development in broilers. This study aimed to optimize in ovo application of OEO by investigating day and site of injection and delivery of carvacrol to different embryonic tissues. In Experiment 1, 2 d of injection (embryonic day (E) 12 or 17.5) and 3 sites of injection for OEO (air cell, amniotic fluid, or yolk) were evaluated based on hatchability and posthatching performance. Experiment 2 aimed to examine the impact of combining OEO with the nonionic surfactant polysorbate 80 (p80) at ratios to carvacrol of 0:0, 0:1, 0.5:1, and 1:1 on carvacrol concentration in amniotic fluid, blood, and yolk. The concentration of carvacrol was measured at 3, 6, and 9 h after OEO injection either without (0:1) or with (1:1) p80. Injection of OEO on E12 led to a significant lower hatchability compared to E17.5 (P ≤ 0.01; Δ = 9.2%). Injecting OEO into the air cell, amniotic fluid, or yolk at E17.5 did not significantly affect hatchability and posthatching performance. The highest concentrations of carvacrol found in egg tissues were observed when injected together with surfactant at the 1:1 ratio (P ≤ 0.001; 14.45 µM, 16.64 µM, and 124.82 µM, for air cell, amniotic fluid, and yolk, respectively) compared to the 0:0, 0:1 or 0.5:1 ratios. Carvacrol was highest in the amniotic fluid and blood at the first time point (3 h postinjection) and decreased afterward (P ≤ 0.001), whereas the concentration in yolk remained elevated up to 9 h postinjection. In conclusion, the optimization of the in ovo delivery of carvacrol resulted in that early injection (E12) had negative effects on hatchability and should be avoided. The findings also suggest that using a nonionic surfactant was crucial for an effective delivery of carvacrol in ovo and the migration from amniotic fluid to yolk within 3 h. In addition, carvacrol's persistence in yolk may serve as a route for delivery into the gastrointestinal tract via the yolk stalk during the peri-hatching phase, potentially influencing gut development.

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.

Group level and individual activity of broiler chickens hatched in 3 different systems.

Information on the behavior of chickens hatched in different systems is limited and inconsistent across different studies. Changes in broiler activity can be measured automatically and continuously. The aim of this study was to assess the effects of 3 hatching systems on flock activity using a commercial tracking system, and to compare these findings to individual activity measured under experimental conditions. As this experiment was part of a larger study, it was possible to investigate the effects of vaccination on individual activity. In study 1, flock activity was measured in chickens that hatched either conventionally in the hatchery (HH), in a system which provided nutrition in the hatcher (HF), or on-farm (OH). Chickens were reared in 2 batches, in 12 pens/batch (1,155 animals/pen). One camera recorded top-view images of each pen. A daily activity index (moved pixels/total pixels × 100) was calculated by automated image analysis. In study 2, individual activity was measured under experimental conditions using an ultra-wideband (UWB) system. Chickens from the 3 hatching systems were reared in 3 pens (1 pen/treatment, 30 animals/pen). At d14, UWB-tags were attached to 5 chickens/pen, which tracked the distances moved (DM). In study 1, group level activity showed a significant age × hatching system interaction (F8,752= 5.83, P < 0.001). HH and HF chickens showed higher activity levels than OH chickens in wk 1, 4, and 5. In wk 3, higher activity levels were measured in HH compared to HF, and in HF compared to OH pens. In contrast, HH chickens in small groups in study 2 showed lower DM than HF and OH chickens in wk 3 (P < 0.001). DM did not differ between treatments before vaccination, however, thereafter, HH chickens showed longer DM, whereas HF and OH chickens moved less. The results indicate that hatching system affected broiler activity at specific ages. Effects found at flock level could not be reproduced by individual measurements in study 2, although stocking density was comparable.