The objective was to evaluate the impacts of a complex environment and stocking density on Ross 708 broiler chicken behaviors. Eight pens contained either high complexity (HC) or low complexity (LC) environments, and high (HD) or low (LD) density. Through focal-animal sampling, the frequency and duration of behaviors were recorded continuously for 5 min at two timepoints for one day in weeks 2, 4, and 7. Birds were active for 30% of the observed time, with birds showing more activity in HC compared with LC. Birds in HC pens spent more time preening and foraging than birds in LC pens, which was interpreted as a positive outcome. Dustbathing and play were not impacted by complexity, possibly due to the observation method. Birds were more frequently active at HD compared with LD, but did not spend more time being active, suggesting disturbances. Birds foraged, drank, and ate less frequently in HD compared with LD, presumably because birds had more difficulty accessing resources. Activity and active behaviors reduced as birds aged, while preening frequency increased, possibly due to frustration, but this was not confirmed. Perching was unaffected by age, showing a persistent motivation to perform the behavior. Our results indicate that a complex environment provides positive stimulation for foraging, locomotion, preening, and overall activity. Despite reduced activity, many benefits of the tested environmental complexity and low density persisted as birds aged.

A complex system of neural pathways, collectively known as the microbiota–gut–brain (MGB) axis, interconnects the gut microbiota, the gastrointestinal system, and the brain along with its periphery. Previous studies have demonstrated that modulation of the MGB axis can influence stress-related behaviors such as anxiety. This connection becomes apparent in scenarios like agonistic behavior in laying hens, which is characterized by aggressive head and feather pecks, that can ultimately result in cannibalism and death. The objective was to examine the effects of a dietary synbiotic on agonistic behavior, plasma and brain monoamines, stress parameters, and cecal microbiota counts via modulation of the MGB axis. A total of 396 W36 Hy-Line laying hens were provided at random with a control (CON: basal diet) or treatment (SYN: basal diet supplemented with synbiotic) diet from 50 to 60 weeks old (nine pens/treatment, 22 birds/pen). Blood samples and video recordings (three consecutive days/week) were taken at 50 and 60 weeks. At 60 weeks, three hens/pen were euthanized for brain and cecal microbiota collection. Threatening, fighting, head, body, and feather pecking all occurred less frequently at 60 weeks in the SYN group (p < 0.05). Plasma corticosterone, adrenocorticotropic hormone, dopamine, and serotonin were significantly lower while tryptophan and 5-hydroxyindoleacetic acid were significantly higher in birds from the SYN group (p < 0.05). Significant differences in serotonin, 5-hydroxyindoleacetic acid, dopamine, homovanillic acid, and 3,4-dihydroxyphenylacetic acid were observed in the hypothalamus, hippocampus, and amygdala of the brain. Serotonin and dopamine turnover rates were significantly different in all three regions of the brain (p < 0.05). Cecal counts of Lactobacillus and Bifidobacterium were significantly higher in the SYN group (p < 0.05). Synbiotic supplementation resulted in many significant differences, indicating activation of the serotonergic systems and modulation of both the MGB axis and HPA axis with positive effects on welfare and stress.

Boron supplementation may improve the musculoskeletal health of pullets before entering the lay phase. This study aimed to evaluate the effects of different boron amounts on the performance, muscle deposition, tibia cross-sectional area (CSA) and mineral density (BMD), ash percent, breaking strength, and bone mineralization (bone-specific alkaline phosphatase [BALP] and pro-collagen type 1 n-terminal propeptide [P1NP]) of a white-feathered strain of pullets. A total of 528 Hy-Line W-36 pullets were distributed across 24 pens and fed basal diets containing varying amounts of boron (C: 0 mg/kg; L: 50 mg/kg; M: 100 mg/kg; H: 150 mg/kg) for 17 weeks. Performance parameters (body weight, average daily weight gain/bird, and average daily feed intake/bird) were measured at weeks 4, 7, 10, 13, and 16, while all other measures were taken at 11 and 17 weeks of age. Performance was not impacted by boron supplementation. Pectoralis major weights were higher in H pullets at 11 weeks of age, and we also observed higher pectoralis major, minor, and leg muscle weights in H pullets at 17 weeks of age. Pullets fed the H diet had larger cortical CSA than the other treatment groups at 11 weeks of age. At 17 weeks of age, both the H and M groups had larger cortical CSA than the L and C groups, but the M group had slightly smaller cortical CSA. Pullets fed the H diet had higher BMD values than the other treatment groups at 11 weeks of age. At 17 weeks of age, pullets fed the H diet had the highest total BMD values compared to the other treatment groups, and cortical BMD increased with increasing boron inclusion. Pullets fed the H diet had the highest tibia ash percentages and concentrations of BALP and P1NP. Pullets fed the M and H diets had greater failure load and maximum bending moment than pullets fed the L or C diet at 11 weeks of age, with H pullets having greater stiffness values than other groups. At 17 weeks of age, pullets fed the H diet had greater failure load and maximum bending moment compared to all other treatment groups. Our results suggest that providing boron within the diet at 150 mg/kg can improve the musculoskeletal characteristics of Hy-Line W-36 pullets up to 17 weeks of age, without impacting performance parameters.

Prior research suggests exercise during pullet rearing can mitigate lay-phase bone fractures by strengthening muscles, enhancing balance, and increasing bone mass. This study aimed to confirm that Hy-Line brown pullets with multi-tier perches show increased activity and improved musculoskeletal health. Pullets (n = 810) were randomly allocated to housing systems, either with multi-tier perches (P; n = 15 pens) or without (NP; n = 15 pens), spanning from 0–17 weeks of age. At 5, 11, and 17 weeks, individual birds were meticulously monitored for activity using accelerometers over three consecutive days (n = 90 randomly selected birds/week). At 11 and 17 weeks, 60 birds underwent euthanasia and computed tomography (CT) scans to ascertain tibiotarsal bone mineral density and cross-sectional area measurements. Post-CT scanning, birds were dissected for muscle size, tibiotarsal breaking strength, and tibiotarsal ash percentage measurements. Additionally, serum concentrations of bone-specific alkaline phosphatase and procollagen type 1 N-terminal propeptide were assessed as markers of bone formation (n = 90 birds/week). Pullet group P exhibited heightened vertical activity (P0.05) during weeks 5, 11, and 17 compared to group NP. Tibiotarsal bones of P pullets demonstrated superior total and cortical bone mineral density at week 11, alongside increased cortical bone cross-sectional areas and heightened total and cortical bone mineral densities at week 17 (P<0.05) compared to NP pullets. At week 11, P pullets displayed larger leg muscles, including triceps, pectoralis major and minor, and leg muscles at week 17 (P<0.05) compared to NP pullets. Notably, at both weeks, P pullets’ tibiae exhibited greater breaking strengths, higher ash percentages, and elevated concentrations of bone-specific alkaline phosphatase and procollagen type 1 N-terminal propeptide compared to NP pullets (P<0.05). The study findings underscore the benefits of providing multi-tier perches for pullets, serving as a valuable tool for enhancing bird activity and musculoskeletal health preceding the lay phase.

Perches can enhance laying hen welfare, but their effectiveness might be age-dependent. We investigated early and late perch access effects on anxiety and fear in pullets through attention bias (AB) and tonic immobility (TI) tests. Pullets (n = 728) were raised with or without multi-level perches: CP (continuous perch access: 0–37 weeks), EP (early perch access: 0–17 weeks), LP (late perch access: 17–37 weeks), and NP (no perch access). AB was conducted in weeks 21 and 37 (n = 84/week), and TI was performed in weeks 20, 25, and 37 (n = 112/week). CP hens fed quicker than EP, LP, and NP in AB at weeks 21 and 37 (p ≤ 0.05). CP and NP feeding latencies were stable, while EP and LP fed faster at week 37 (p ≤ 0.05). CP had the shortest TI at week 20 (p < 0.05). CP and LP had the shortest TI in weeks 25 and 37 (all p ≤ 0.05). Unlike NP, CP reduced anxiety and fear. Adding perches during laying (LP) raised anxiety at week 21, adapting by week 37, and removing pre-laying perches (EP) worsened fear at weeks 20 and 25 and anxiety at week 21, recovering by week 37. Adding or removing perches prior to the lay phase increased fear and anxiety, an effect that disappeared by week 37 of age. Our study indicates that continuous perch access benefits animal welfare compared to no perch access at all.

Laying hens can experience a progressive increase in bone fragility due to the ongoing mobilization of calcium from bones for eggshell formation. Over time, this escalates their susceptibility to bone fracture, which can reduce their mobility and cause pain. The provision of perches as an exercise opportunity could potentially enhance bone strength, but the timing of exposure to perches during the birds’ development may modulate its impact. The objective of this study was to investigate the enduring impacts of perch provision timing on the musculoskeletal health of laying hens. A total of 812 pullets were kept in different housing conditions (seven pens/treatment, 29 birds/pen) with either continuous access to multi-tier perches from 0 to 40 weeks of age (CP), no access to perches (NP), early access to perches during the rearing phase from 0 to 17 weeks of age (EP), or solely during the laying phase from 17 to 40 weeks of age (LP). At weeks 24, 36, and 40 of age (n = 84 birds/week), three birds per pen were monitored for individual activity level, and blood samples were collected from a separate set of three birds per pen to analyze serum levels of tartrate-resistant acid phosphatase 5b (TRACP-5b) and C-terminal telopeptide of type I collagen (CTX-I) as markers of bone demineralization. At 40 weeks of age, three birds per pen (n = 84) were euthanized for computed tomography scans to obtain tibial bone mineral density (BMD) and cross-sectional area (CSA) with further analysis including muscle deposition, tibial breaking strength, and tibial ash percent. During week 24, hens from CP, EP, and LP pens had the highest overall activity compared to hens from NP pens (p < 0.05) with no differences between treatments for overall activity level during weeks 36 or 40 (p > 0.05). During weeks 24, 36, and 40, hens from CP and LP pens showed greater vertical and less horizontal activity compared to hens from EP and NP pens (p < 0.05). TRACP-5b and CTX-I concentrations did not differ between treatments at week 24 of age (p > 0.05). Hens from CP pens had the lowest TRACP-5b and CTX-I concentrations at 36 weeks of age with EP and LP hens showing intermediate responses and NP hens having the highest concentration (p < 0.05). At 40 weeks of age, CP hens had the lowest TRACP-5b and CTX-I concentrations compared to NP hens (p < 0.05). Total bone CSA did not differ between treatments (p > 0.05), but CP had greater total BMD than NP (p < 0.05) with no differences between EP and LP treatments. CP and LP hens had larger biceps brachii, pectoralis major, and leg muscle groups as well as greater tibial breaking strengths than EP and NP treatments (p < 0.05). CP hens had higher tibial ash percentages compared to EP, LP, and NP (p < 0.05). Our results indicate that providing continuous perch access improves the musculoskeletal health and activity of laying hens at 40 weeks of age compared to no access and that late access to perches has a beneficial impact on activity, muscle deposition, and bone strength.