Effects of light intensity and dual light intensity choice on plasma corticosterone, central serotonergic and dopaminergic activities in birds, Gallus gallus.

Light intensity plays an important role in the regulation of growth, behavior, reproduction, and welfare of avian species. Light intensity preference behavior has been suggested to be involved in welfare of birds. This study aims to investigate the effects of different light intensity and dual light intensity choice (DLIC) lighting program on plasma corticosterone (CORT), and tryptophan hydroxylase 2 (TPH2, the rate-limiting enzyme of serotonin biosynthesis) and tyrosine hydroxylase (TH, the rate-limiting enzyme of dopamine biosynthesis) gene expression in the brainstem of male chickens. Day old broilers were housed in two commercial houses, and placed in 24 pens. All the treatment groups were provided with 23 h light (L) /1 h dark (D) and 30 lx (lx) light intensity during the first week and then 18L:6D (10 lx) from day 7 to 14. Blood and brain were sampled at 14 days of age (10 lx) before the onset of light treatments. On day 15, four treatments (2, 10, 20, and 100 lx), and DLIC treatment (2/20 lx) were initiated. Samples were collected on days 15, 16, 17, 30 and 41. TPH2 expression in the dorsal raphe nucleus (DRN) and caudal raphe nucleus (CRN) of brainstem, and TPH2 and TH expression in ventral tegmental areas (VTN) of the midbrain were determined by qPCR. Results showed that bright light and DLIC lighting program temporarily attenuated plasma CORT, suggesting the short-term stress attenuating effect of bright light and DLIC lighting program. Differential TPH2 expression in the DRN and CRN observed in the DLIC birds indicate a significant effect of DLIC lighting program on the serotonergic activity in the avian brainstem. At the 41 days of age, the significant downregulation of TPH2 and TH expression occurred in the VTA of DLIC treated birds compared to the other group of birds. Taken together, temporal and spatial regulation of TPH2 and TH expression by DLIC lighting program indicate that compensatory regulation of serotonergic and dopaminergic activities might be involved in the light intensity preference behavior of birds, suggesting a possible beneficial effect of the DLIC lighting program on broiler welfare.

Effects of Environmental Enrichments on Welfare and Hepatic Metabolic Regulation of Broiler Chickens.

The aims of this study were to find suitable environmental enrichment (EE) and evaluate the combined effect of two EEs, variable light intensity (VL) lighting program and EH, on mental health and hepatic metabolic regulation in commercial broilers. To find the advantageous EEs for broilers, three different EEs (board, hut, and ramp) were tested in trial 1. EEs were placed and the engagement of birds to EEs, dustbathing behavior, and daily physical activity were observed. Birds treated with huts showed higher engagement than the board- or ramp-treated birds (p < 0.05). The results of dustbathing behavior and daily physical activity indicated that the environmental hut (EH) is the most favorable enrichment for broilers. In the second trial, to test the effect of EHs on mental health and hepatic metabolic conditions, the brain and liver were sampled from the four treatment birds (20 lx_Con, 20 lx_Hut, VL_Con and VL_Hut) on day 42. The lower expression of TPH2 (tryptophan hydroxylase 2) of VL_Hut birds than those of VL_Con and 20 lx_Hut treated birds suggests the combining effect of EHs with the VL lighting program on the central serotonergic homeostasis of broilers. Reduced expressions of TH (tyrosine hydroxylase), GR (glucocorticoid receptor), BDNF (brain-derived neurotrophic factor) of VL_Hut treated birds compared to those of VL_Con and 20 lx_Hut birds suggest lower stress, stress susceptibility, and chronic social stress in VL_Hut treated birds. The expression of CPT1A (carnitine palmitoyl transferase 1) increased over three-fold in the liver of VL_Con birds compared to 20 lx_Con birds (p < 0.05). EHs treatment in VL birds (VL_Hut) significantly decreased CPT1A but not in 20 lx birds (20 lx_Hut). The expression of ACCα (acetyl-CoA carboxylase alpha) was significantly decreased in VL_Con birds compared to 20 lx_Con birds. There was no significant difference in the hepatic FBPase (fructose-1,6-bisphosphatase), GR, and 11ß-HSD1 (11 ß-hydroxysteroid dehydrogenease-1) expression between 20 lx_Con and VL_Con birds, but EHs significantly stimulated GR in 20 lx_Hut birds, and stimulated FBPase and 11ß-HSD1 expression in the VL_Hut birds compared to 20 lx_Con birds, suggesting that the VL lighting program reduced fatty acid synthesis and increased fatty acid ß-oxidation in the broilers' liver and VL_Hut improved the hepatic de novo glucose production. Taken together, the results suggest that the stimulated voluntary activity by EHs in the light-enriched broiler house improved mental health and hepatic metabolic function of broilers and may indicate that the improved hepatic metabolic function contributes to efficient nutritional support for broilers.

Effects of a variable light intensity lighting program on the welfare and performance of commercial broiler chickens.

Our previous variable-light intensity lighting program studies indicate the light intensity preference behavior of broilers for their daily activity including eating and resting. To evaluate the effects of variable-light intensity lighting program on performance and welfare of broilers, four commercial trials were conducted for looking at behaviors, mortality, leg-health, performance, and brain welfare indicator genes including tryptophan hydroxylase 2 and tyrosine hydroxylase (TH), glucocorticoid receptor (GR), brain-derived neurotropic factor (BDNF), and melanopsin (Opn4) gene expression. One-day-old broilers were housed in four commercial broiler houses. Each quadrant (section) of the house was placed with 4,800 chicks. A total of four lighting programs began on day 7 with 5 lux (lx), 20 lx, natural light (NL, 480 lx), and variable light (2-5/40 lx) using LED lights on a 16L:8D photoperiod. In the variable-light house, the number of dustbathing holes was significantly higher than that in natural-light houses and 5-lx and 20-lx houses. Daily physical activities, footpad condition, fear response to novel objects, body weight, feed conversion ratio, and the number of leg-problem induced culled birds were affected by the variable-light intensity lighting program. Expression of tryptophan hydroxylase 2 in the DRN and VTA of variable-light treated birds was lower than that of 5-lx- and 20-lx-treated birds on day 42 (p < 0.05). Higher expression of VTA-TH in 5-lx-treated birds than that in 20-lx-, NL-, and variable-light-treated birds suggests the high stress-susceptibility of 5-lx treated birds. Lower VTA-GR expression in 20-lx- and variable-light-treated birds indicates lower stress than that in NL- and 5-lx-treated birds (p < 0.05). The VTA-BDNF expression of NL-treated birds was 2.5 fold higher than that of 5-lx-, 20-lx-, and variable-light-treated birds (p < 0.05), and variable-light-treated birds showed the lowest level of BDNF expression (p < 0.05), suggesting the chronic social defeat stress in NL-treated birds. The result of VTA-Opn4 expression on day 42 suggests the possible role of VTA-Opn4 in broiler welfare through central light perception. Taken together, the variable-light intensity lighting program increased volunteer natural behaviors and physical activity, which may improve footpad condition and leg health of birds, consequently. Performance data including the increased daily weight gain and the lowered feed conversion ratio and results of brain welfare indicator gene expression showed the beneficial effect of the variable-light intensity lighting program on the performance and welfare of commercial broilers.

Anaerobutyricum and Subdoligranulum Are Differentially Enriched in Broilers with Disparate Weight Gains.

The intestinal microbiota is critically important for animal health and productivity. However, the influence of the intestinal microbiota on animal growth efficiency remains elusive. This current study was aimed at identifying the intestinal bacteria that are associated with the growth rate of broilers in a commercial production setting. Ross 708 broilers with extremely high, medium, and extremely low body weight (BW) were separately selected for each sex from a house of approximately 18,000 chickens on day 42. The cecal content of each animal was subjected to 16S rRNA gene sequencing for microbiota profiling. Our results indicate that a number of bacteria were differentially enriched among different groups of broilers, with several showing a significant correlation (p < 0.05) with BW in both sexes or in a sex-specific manner. Subdoligranulum was drastically diminished in high-BW birds with a strong negative correlation with BW in both males and females. While one Anaerobutyricum strain showed a positive correlation with BW in both sexes, another strain of Anaerobutyricum was positively correlated with BW only in females. These sex-dependent and -independent bacteria could be targeted for improving the growth efficiency and may also be explored as potential biomarkers for the growth rate of broiler chickens.

Impact of experimentally induced bacterial chondronecrosis with osteomyelitis (BCO) lameness on health, stress, and leg health parameters in broilers.

Stress and lameness negatively affect the health, production, and welfare of broilers. Bacterial chondronecrosis with osteomyelitis (BCO) is a leading cause of stress and lameness in commercial broilers. External changes in skin temperature related to changes in blood flow can be detected with infrared thermography (IRT), offering a noninvasive tool to assess the health of animals. This study compared physiological and noninvasive measures of stress and lameness in clinically healthy and lame male broiler chickens between 25 and 56 d. Birds were raised in pens within separate environmental chambers containing either litter flooring (sound) or wire flooring, with the latter established to induce BCO lameness (lame). Physiological and noninvasive measures of stress and lameness were collected: body weight, (BW), relative bursa weight, core body temperature, corticosterone (CORT) concentrations in serum and feathers, surface temperatures of the head (eye and beak) and leg (hock, shank, and foot) regions by infrared thermography (IRT), leg blood oxygen saturation (leg O2), and BCO lesion severity scores of tibial head necrosis (THN) and femoral head necrosis (FHN). Lame birds exhibited greater FHN and THN lesion severities, core body temperatures, and serum CORT (P < 0.05), but had lower BW, relative bursa weight, leg O2, and IRT surface temperatures of the beak, hock, shank, and foot compared with sound birds (P < 0.05). The difference in THN lesion severity between sound and lame birds decreased with age. Linear relationships between leg O2 with IRT leg surface temperatures were positive and negative between leg O2 with BCO lesion severity (P < 0.05). There were negative correlations between serum CORT with hock, shank and foot temperatures (P < 0.001), indicating that BCO is stressful. These results indicate that birds lame from BCO are stressed, have reduced oxygen saturation of blood in their legs, and that IRT surface temperatures can be used as noninvasive indicators of stress and lameness in broilers.

Broiler stress responses to light intensity, flooring type, and leg weakness as assessed by heterophil-to-lymphocyte ratios, serum corticosterone, infrared thermography, and latency to lie.

Stress and leg weakness are detrimental to broiler production, health, and welfare. Traditional methods to evaluate stress may be stressful to the bird because they are invasive and require handling and restraint. Two studies examined the effects of light intensity and flooring on the following in broilers: 1) traditional methods for assessing stress using heterophil-to-lymphocyte ratios and serum corticosterone (CORT) concentrations, 2) noninvasive measures of stress from infrared thermography (IRT) eye and beak surface temperatures, and 3) latency-to-lie (LTL) test times of birds tested individually and in groups of 5. Day-of-hatch male broiler chicks were placed into 6 pens (N = 120 chicks/pen). At 1 wk, pens were allocated to 3 light intensity treatments (2, 5, or 10 lux). At 4 wk, half of the birds from each pen were moved to a pen with wire flooring and the same light intensity. At 1, 4, 5, and 8 wk, blood samples were collected and IRT images of the heads of 5 clinically healthy broilers from each pen were captured. In study 2, IRT images of the heads of birds that became lame in the wire flooring pens were taken. There were no treatment effects on the LTL times of birds tested in groups or individually (P > 0.05). On day 56 in study 1, birds on wire flooring had elevated heterophil-to-lymphocyte ratios and CORT concentrations (P ≤ 0.002) and depressed IRT eye and beak temperatures (P < 0.0001). In both studies, there were negative correlations between CORT concentrations and IRT beak surface temperatures (P < 0.05). Lame birds had lower IRT eye and beak surface temperatures than sound birds (P ≤ 0.004), and the IRT beak surface temperatures of lame birds were lower than their eye surface temperatures (P = 0.004) in study 2. These studies indicate that the IRT surface temperatures of the eye, and more distinctly of the beak, can be used as sensitive noninvasive indicators of stress.

The utility of infrared thermography for evaluating lameness attributable to bacterial chondronecrosis with osteomyelitis.

Bacterial chondronecrosis with osteomyelitis (BCO) is a leading cause of lameness in broilers. Infrared thermography (IRT) is a noninvasive technique for measuring infrared radiation from an object and can be used to evaluate clinical health. Two replicated studies compared the effect of light intensity on broilers grown on a wire flooring model that experimentally increased their susceptibility to and incidence of BCO lameness. Day-of-hatch male broiler chickens were placed into 6 pens on wood shavings litter, and at 1 wk one of 3 light intensity treatments (2, 5, or 10 lux) was allotted. At 4 wk half of the population from each pen was moved to a pen with wire flooring and the same light intensity. At 1, 4, 5, and 8 wk, an IRT image of the legs of 5 clinically healthy broilers from each pen was taken. The right and left proximal femora and tibiae of sound and lame broilers were scored for femoral head necrosis (FHN) and tibial head necrosis (THN) lesion severity. There were minimal effects of light intensity and flooring. In Study 1, but not Study 2, broilers on wire flooring weighed less on day 38 (P = 0.007) and days 57 to 58 (P = 0.003) compared to those on litter. The proportion of broilers that became lame on wire flooring was 52% in Study 1 and 14% in Study 2. The proportion of sound broilers from litter and wire flooring pens with subclinical signs of BCO in their right or left proximal growth plates was over 45% for FHN and 92% for THN, and lame broilers had more severe (P < 0.0001) FHN and THN compared to sound broilers. IRT surface temperatures of the hock joint, shank, and foot were consistently lower (P < 0.0001) in broilers that became lame when compared to sound. Therefore, IRT surface temperatures of broiler leg regions may be useful for detecting lesions attributed to BCO.

An evaluation of methods for measuring stress in broiler chickens.

There are potential advantages for using noninvasive methods instead of conventional approaches for measuring corticosterone (CORT) as a metric of stress. Two studies compared blood heterophil: lymphocyte (H:L) ratios, serum CORT, cecal content CORT (Study 1), colon content CORT (Study 2), and feather CORT concentrations for broiler chicks receiving drinking water with or without added CORT. On day 28, male broilers (N = 140) were provided tap water (Control group) or water containing 20 mg/L of CORT (CORT group) for up to 72 h. Body weight (BW), blood, feather, cecal, and colon content samples were collected from 10 birds from each treatment group at 0, 6, 12, 24, 30, 48, and 72 h. Data were analyzed with a JMP Pro ANOVA. Mean comparisons were analyzed on significant treatment effects using post hoc t-tests to compare control and CORT measures within each sampling time point. Pearson's pairwise correlations for all data were performed. A treatment by time interaction affected all measures in both studies. In both studies, CORT treatment reduced body weight and increased H:L ratios at later time points. In study 1 cecal content CORT concentrations were elevated with CORT treatment after 24 h, whereas in study 2 colon content CORT concentrations were elevated after 6 h. CORT treatment increased body and primary feather CORT and primary feather CORT increased with time in control groups in both studies. Serum CORT exhibited the lowest variability compared with all other sample sources used for determining CORT. Estimates of CORT by different methods were positively correlated. These results indicate that serum CORT concentrations most reliably reflect the CORT status of broilers, and that feather CORT concentrations demonstrated potential for estimating stress in broilers.

Effects of bacitracin methylene disalicylate and diet change on gastrointestinal integrity and endotoxin permeability in the duodenum of broiler chicken.

OBJECTIVE: To determine the effect of bacitracin methylene disalicylate (BMD) and feed changes on gastrointestinal integrity, endotoxin permeability, and morphometric parameters in the duodenum of broilers. RESULTS: Birds were raised on a starter diet without growth promoting antibiotics for 31 days then switched to a grower diet. Four of the pens including 50 g/ton of BMD while 4 pens remained antibiotic free. Eight birds per treatment were sampled prior to the feed change and at 3 and 7 days following the feed change. Gastrointestinal integrity and endotoxin permeability in the duodenum were determined using a modified Ussing Chamber and an adjacent section fixed in 10% formalin for morphometric analysis. Data were analyzed using Proc Glimmix of SAS with the model fitting BMD treatment, time, and the interaction of BMD treatment and time as fixed effects. Intestinal integrity increased at d 3 and 7 compared to prior to the feed change and addition of BMD (P > 0.001) and villus height was decreased with BMD supplementation (P = 0.049). All other tested effects similar (P > 0.1). In conclusion, the practice of changing feed had a greater effect on intestinal health than addition of BMD. However, the factors driving these differences 42 are unclear.

Symposium: Animal welfare challenges for today and tomorrow.

The increasing separation of the public from production agriculture means there is often a lack of knowledge among consumers about current production practices and a perception that increased productivity and economic efficiency are necessarily associated with a decline in animal welfare. A symposium was organized to present information about animal welfare issues and the challenges they pose for both scientists and the poultry and allied industries. Companion papers provide information about understanding public attitudes and physiological/immunological approaches to welfare assessment, while this paper outlines current and future challenges to egg and meat production and industry responses to those challenges. For broiler chickens, increases in growth rate result in corollary increases in metabolic heat generation and water consumption, leading to the need for continuing improvements in housing, ventilation, and litter management. Stocking densities, lighting programs, muscle myopathies, and use of antibiotics are also areas that require research attention. In the layer industry, the key challenge is housing, with the industry undergoing a shift from conventional cage housing to alternatives like enriched colonies or cage-free. While these alternative systems have hen welfare advantages, there are also welfare disadvantages that require the development of mitigation strategies, and it is also essential to address associated issues including economic, environmental, egg safety, and worker health impacts. Concerns on the horizon include euthanasia of surplus male chicks and spent hens as well as beak-trimming. The humaneness of slaughter methods is an important welfare and consumer confidence issue, and the current regulations for poultry slaughter in the USA are discussed and compared to those for livestock. The poultry and allied industries, including retailers, are responding to these concerns by consulting with experts, developing science-based animal care standards and auditing programs, strengthening training and oversight programs, promoting research, and improving communication channels. In future, intensifying multi-disciplinary research efforts and developing mechanisms to improve communication between scientists and stakeholders, including the public, will be critical to addressing these issues.