Utilizing 3-dimensional models to assess keel bone damage in laying hens throughout the lay cycle.

The global egg industry is rapidly transitioning to cage-free egg production from conventional cages. Hens housed in cage-free systems have an increased prevalence of keel damage that could lead to reduced egg production and compromised well-being. The objective of this study was to determine the effects of dietary supplementation of n-3 fatty acids and vitamin D3 on keel damage in hens housed in multi-tier aviary systems (AV). Brown hens were placed in 4 AV system rooms after rearing at 17 wk of age (woa) with each room containing 576 birds. At 12 woa, rooms were randomly assigned to a dietary treatment of flaxseed oil, fish oil, vitamin D3, or control. Focal birds (36 per treatment) were longitudinally examined for keel damage using quantitative computed tomography (QCT) at nine timepoints from 16 to 52 woa. Three-dimensional digital twins of the keels were created from the QCT scans and visually assessed for damage. An overall keel severity score was recorded as well as the location, direction, and severity of each deviation or fracture. Severity was ranked on a 0 to 5 scale with 0 being no damage and 5 being severe. Damage scores were analyzed utilizing odds ratios with main effects of age and treatment. At 16 woa, 80% of hens had overall keel scores of 0 and 20% had scores of 1. At 52 woa, all hens had damage, with 31% having a score of 1, 61% scored 2 to 3, and 8% scored 4 to 5. Most fractures were not observed until peak lay. Dietary treatments did not affect likelihood of fracture incidences, but younger birds had lower odds of incurring keel fractures than older birds (P < 0.0001). The initial incidences of keel deviations occurred earlier than fractures, with most birds obtaining a keel deviation by 28 woa. Keel damage was not able to be prevented, but the age at which keel fractures and deviations initiate appear to be different, with deviations occurring during growth and fractures during lay.

Genetic background of walking ability and its relationship with leg defects, mortality, and performance traits in turkeys (Meleagris gallopavo).

This study aimed to explore the genetic basis of walking ability and potentially related performance traits in turkey purebred populations. Phenotypic, pedigree, and genomic datasets from 2 turkey lines hatched between 2010 and 2023 were included in the study. Walking ability data, defined based on a scoring system ranging from 1 (worst) to 6 (best), were collected on 192,019 animals of a female line and 235,461 animals of a male line. Genomic information was obtained for 46,427 turkeys (22,302 from a female line and 24,125 from a male line) using a 65K single nucleotide polymorphism (SNP) panel. Variance components and heritability for walking ability were estimated. Furthermore, genetic and phenotypic correlations among walking ability, mortality and disorders, and performance traits were calculated. A genome-wide association study (GWAS) was also conducted to identify SNPs associated with walking ability. Walking ability is moderately heritable (0.23 ± 0.01) in both turkey lines. The genetic correlations between walking ability and the other evaluated traits ranged from -0.02 to -0.78, with leg defects exhibiting the strongest negative correlation with walking ability. In the female line, 31 SNPs were associated with walking ability and overlapped with 116 genes. These positional genes are linked to 6 gene ontology (GO) terms. Notably, genes such as CSRP2, DDX1, RHBDL1, SEZ6L, and CTSK are involved in growth, development, locomotion, and bone disorders. GO terms, including fibronectin binding (GO:0001968), peptide cross-linking (GO:0018149), and catabolic process (GO:0009057), are directly linked with mobility. In the male line, 66 markers associated with walking ability were identified and overlapped with 281 genes. These genes are linked to 12 GO terms. Genes such as RB1CC1, TNNI1, MSTN, FN1, SIK3, PADI2, ERBB4, B3GNT2, and BACE1 are associated with cell growth, myostatin development, and disorders. GO terms in the male line are predominantly related to lipid metabolism. In conclusion, walking ability is moderately heritable in both populations. Furthermore, walking ability can be enhanced through targeted genetic selection, emphasizing its relevance to both animal welfare and productivity.

n-3 essential fatty acid and vitamin D supplementation improve skeletal health in laying hens.

Keel bone fractures and osteoporosis are prevalent and damaging skeletal issues in the laying hen industry. There is a large interest in improving bone quality parameters to reduce or eliminate these conditions, thus improving bird welfare. Both essential fatty acids (EFA) and vitamin D can play a role in bone metabolism. The hypothesis of this study was that birds supplemented with lower n-6:n-3 EFA ratio or vitamin D would have improved bone properties compared to a control diet. A total of 3,520 Lohmann Brown-Lite pullets were used in this study. Pullets were housed on the floor from 0 to 17 wk of age and then moved to an aviary (17-52 wk of age). Starting at 12 wk of age, birds were split into diet treatments-control, flax, fish, or vitamin D diets with n-6:n-3 ratios of 6.750, 0.534, 0.534, and 6.750, respectively. Diets were formulated to be isonitrogenous and isocaloric. Basal vitamin D3 levels were formulated to be 2,760 IU/kg across all diets; for the vitamin D diet, the vitamin D3 level was increased to 5,520 IU/kg. Hens on fish and vitamin D diets had greater bone density, keel bone volume, digital bone mineral content, and keel condition compared to flax and control hens. Additionally, birds fed the vitamin D diet had the heaviest body weights compared to birds fed fish or control diets. Birds fed the flax and vitamin D diets had improved feather coverage across multiple body regions. Feeding an n-3 EFA- or vitamin D-enriched diet decreased mortality by 1.6 to 3.3% compared to the control. The fish and vitamin D diets generated mixed production performance. Compared to the other treatments, the vitamin D diet generated higher case weights but lower hen day percentage throughout the study. When compared to the other treatments, the fish diet had the lowest case weights but had a greater hen day percentage after 36 wk of age. Results indicate that a fish-based EFA and vitamin D supplementation show promise in improving skeletal health but require further investigation.

Osteoimmunology: A Link between Gastrointestinal Diseases and Skeletal Health in Chickens.

Bone serves as a multifunctional organ in avian species, giving structural integrity to the body, aiding locomotion and flight, regulating mineral homeostasis, and supplementing calcium for eggshell formation. Furthermore, immune cells originate and reside in the bone marrow, sharing a milieu with bone cells, indicating a potential interaction in functions. In avian species, the prevalence of gastrointestinal diseases can alter the growth and the immune response, which costs a great fortune to the poultry industry. Previous studies have shown that coccidiosis and necrotic enteritis can dramatically reduce bone quality as well. However, possible mechanisms on how bone quality is influenced by these disease conditions have not yet been completely understood, other than the reduced feed intake. On the other hand, several mediators of the immune response, such as chemokines and cytokines, play a vital role in the differentiation and activation of osteoclasts responsible for bone resorption and osteoblasts for bone formation. In the case of Eimeria spp./Clostridium perfringens coinfection, these mediators are upregulated. One possible mechanism for accelerated bone loss after gastrointestinal illnesses might be immune-mediated osteoclastogenesis via cytokines-RANKL-mediated pathways. This review article thus focuses on osteoimmunological pathways and the interaction between host immune responses and bone biology in gastrointestinal diseases like coccidiosis and necrotic enteritis affecting skeletal health.

Interindividual distances and orientations of laying hens under 8 stocking densities measured by integrative deep learning techniques.

Interindividual distances and orientations of laying hens provide quantitative measures to calculate and optimize space allocations for bird flocks. However, these metrics were often measured manually and have not been examined for different stocking densities of laying hens. The objectives of this study were to 1) integrate and develop several deep learning techniques to detect interindividual distances and orientations of laying hens; and 2) examine the 2 metrics under 8 stocking densities via the developed techniques. Laying hens (Jingfen breed, a popular hen breed in China) at 35 wk of age were raised in experimental compartments at 8 different stocking densities of 3,840, 2,880, 2,304, 1,920, 1,646, 1,440, 1,280, and 1,152 cm2•bird-1 (3-10 hens per compartment, respectively), and cameras on the top of the compartments recorded videos for further analysis. The designed deep learning image classifier achieved over 99% accuracy to classify bird's perching status and excluded frames with bird perching to ensure that all birds analyzed were on the same horizontal plane, reducing calculation errors. The YOLOv5m oriented object detection model achieved over 90% precision, recall, and F1 score in detecting birds in compartments and can output bird centroid coordinates and angles, from which interindividual distances and orientations were calculated based on pairs of birds. Laying hens maintained smaller minimum interindividual distances in higher stocking densities. They were in an intersecting relationship with conspecifics for over 90% of the time. The developed integrative deep learning techniques and behavior metrics provide animal-based measurement of space requirement for laying hens.

Automatic detection of brown hens in cage-free houses with deep learning methods.

Computer vision technologies have been tested to monitor animals' behaviors and performance. High stocking density and small body size of chickens such as broiler and cage-free layers make effective automated monitoring quite challenging. Therefore, it is critical to improve the accuracy and robustness of laying hens clustering detection. In this study, we established a laying hens detection model YOLOv5-C3CBAM-BiFPN, and tested its performance in detecting birds on open litter. The model consists of 3 parts: 1) the basic YOLOv5 model for feature extraction and target detection of laying hens; 2) the convolution block attention module integrated with C3 module (C3CBAM) to improve the detection effect of targets and occluded targets; and 3) bidirectional feature pyramid network (BiFPN), which is used to enhance the transmission of feature information between different network layers and improve the accuracy of the algorithm. In order to better evaluate the effectiveness of the new model, a total of 720 images containing different numbers of laying hens were selected to construct complex datasets with different occlusion degrees and densities. In addition, this paper also compared the proposed model with a YOLOv5 model that combined other attention mechanisms. The test results show that the improved model YOLOv5-C3CBAM-BiFPN achieved a precision of 98.2%, a recall of 92.9%, a mAP (IoU = 0.5) of 96.7%, a classification rate 156.3 f/s (frames per second), and a F1 (F1 score) of 95.4%. In other words, the laying hen detection method based on deep learning proposed in the present study has excellent performance, can identify the target accurately and quickly, and can be applied to real-time detection of laying hens in real-world production environment.

Minimal effects of ultraviolet light supplementation on egg production, egg and bone quality, and health during early lay of laying hens.

Chicken vision is sensitive to ultraviolet (UV) light containing the UVA spectrum, while UVB plays a key role in the endogenous production of vitamin D3. However, commercially available light sources are typically deficient in the UV spectrum and thus may not adequately fulfill the lighting requirements of indoor-housed laying hens. We hypothesized that supplementary UVB light may improve egg production and egg quality, and bone health during early lay relative to UVA supplementation or standard control lighting. To investigate the effects of UV light supplementation, an experiment was conducted on 252 ISA Brown hens during 16 to 27 weeks of age. Birds were housed in eighteen pens (14 hens/pen) under three different light treatment groups each with six replications: (i) UVO: standard control lighting with LED white light, (ii) UVA: control lighting plus supplemental daylight with an avian bulb, and (iii) UVA/B: control lighting plus a supplemental full spectrum reptile bulb containing both UVA and UVB wavelengths. Hen-day egg production and egg quality, blood parameters including plasma Ca and P, and serum 25(OH)D3, and hen body weight and external health scoring were measured at different age points; while bone quality was assessed at the end of the experiment at 27 weeks. Data were analyzed in JMP® 16.0 using general linear mixed models with α level set at 0.05. Results showed that UVA and UVA/B supplemented birds reached sexual maturity (50% production) 3 and 1 day earlier, respectively, than control birds. There was a trend for UV lights to increase hen-day egg production (P = 0.06). Among egg quality traits, only eggshell reflectivity and yolk index were affected by UV lights (P = 0.02 and 0.01, respectively); however, most of the egg quality traits changed over age (all P < 0.01). Post-hoc tests showed higher serum 25(OH)D3 in the UVA/B group relative to control hens (P < 0.05); but there was no treatment effect on plasma Ca and P or on bone quality parameters (all P > 0.05). A significant interaction was observed between light treatment and age for the number of comb wounds (P = 0.0004), with the UV supplemented hens showing more comb wounds after 24 weeks. These results demonstrated that supplemental UVA/B light had minimal effects on egg production and egg quality, whereas, UVA/B exposure may increase vitamin D3 synthesis during the early laying period. The optimum duration of exposure and level of intensity needs to be determined to ensure these benefits.

Novel 3D-printed Electrodes for Implantable Biopotential Monitoring.

A major bottleneck in the manufacturing process of a medical implant capable of biopotential measurements is the design and assembly of a conductive electrode interface. This paper presents the use of a novel 3D-printing process to integrate conductive metal surfaces on a low-temperature co-fired ceramic base to be deployed as electrodes for electrocardiography (ECG) implants for small animals. In order to fit the ECG sensing system within the size of an injectable microchip implant, the electronics along with a pin-type lithium-ion battery are inserted into a cylindrical glass tube with both ends sealed by these 3D printed composite electrode discs using biomedical epoxy. In the scope of this paper, we present a proof-of-concept in vivo experiment for recording ECG from an avian animal model under local anesthesia to verify the electrode performance. Simultaneous recording with a commercial device validated the measurements, demonstrating promising accuracy in heart rate and breathing rate monitoring. This novel technology could open avenues for the mass manufacturing of miniaturized ECG implants.Clinical relevance- A novel manufacturing process and an implantable system are presented for continuous physiological monitoring of animals to be used by veterinarians, animal scientists, and biomedical researchers with potential future applications in human health monitoring.

Pooling of Laying Hen Environmental Swabs and Efficacy of Salmonella Detection.

ABSTRACT: Environmental testing for Salmonella Enteritidis is required for U.S. shell egg producers with ≥3,000 hens on a farm. The egg producer assumes all costs for the mandatory testing. According to the U.S. Food and Drug Administration (FDA) Egg Rule, either manure scraper or drag swabs can be collected according to published guidelines and requirements. The present study was undertaken to determine the efficacy of Salmonella detection with one-, two-, and four-swab pools of either manure scraper or drag swabs. Resistant isolates of Salmonella serovars Enteritidis (1,000 ppm of streptomycin), Heidelberg (200 ppm of nalidixic acid [NA]), Typhimurium (200 ppm of NA), and Kentucky (200 ppm of NA) were utilized. Low (approximately 8.4 CFU) and high (approximately 84 CFU) levels of inocula were introduced onto a single swab within a pool. Single flocks from each conventional cage (manure scraper swabs) and cage-free barn (drag swabs) were monitored throughout the study at the ages required under the FDA Egg Rule. The highest and most consistent recovery of inoculum was found in single swab samples. For low dose inocula, recovery of isolates was low from single manure scraper swabs (57.9 to 29.2%) and decreased as more swabs were added to the pool. Recovery of isolates from manure scraper swabs was higher for high dose inocula, although Salmonella Heidelberg was outcompeted by the naturally occurring flora and had the lowest rate of recovery among the isolates tested. One- and two-swab pools of drag swabs had similar rates of recovery at both low and high doses for Salmonella Enteritidis, Salmonella Heidelberg, and Salmonella Typhimurium. When Salmonella Enteritidis and Salmonella Kentucky were combined in an inoculum, Salmonella Enteritidis was recovered at a much higher rate than was Salmonella Kentucky for all types of swabs and doses of inocula. Pooling of two drag swabs allowed for similar detection of low and high dose Salmonella, but the pooling of manure scraper swabs decreased detection of low dose Salmonella.

Contamination of eggs by Salmonella Enteritidis in experimentally infected laying hens of four commercial genetic lines in conventional cages and enriched colony housing.

Human illness caused by the consumption of eggs contaminated with Salmonella Enteritidis is a continuing international public health concern. This pathogen is deposited inside the edible contents of eggs as a consequence of its ability to colonize reproductive tissues in infected hens. Conditions in the housing environment can influence the persistence and transmission of avian Salmonella infections, but the food safety ramifications of different poultry management systems are not entirely clear. The present study assessed the deposition of S. Enteritidis inside eggs laid by groups of experimentally infected laying hens of 4 commercial genetic lines (designated as white egg lines W1 and W2 and brown egg lines B1 and B2). Groups of hens from each line were housed at 555 cm2 of floor space per bird in both conventional cages and colony units enriched with access to perches and nesting areas. All hens were orally inoculated with 5.75 × 107 cfu of a 2-strain S. Enteritidis mixture, and the internal contents of eggs laid 5 to 24 D post-inoculation were cultured to detect the pathogen. No significant differences in egg contamination frequencies were found between the 2 housing systems for any of the hen lines. Contaminated eggs were laid between 7 and 21 D post-inoculation at an overall frequency of 2.47%, ranging from 0.25 to 4.38% for the 4 hen lines. The frequency of S. Enteritidis recovery from egg samples was significantly (P < 0.05) lower for line B2 than for any of the other lines, and the egg contamination frequency for line W1 was significantly greater than for line W2. The overall incidence of contamination among white eggs (3.38%) was significantly higher than among brown eggs (1.56%). These results demonstrate that S. Enteritidis deposition inside eggs can vary between genetic lines of infected laying hens, but housing these hens in 2 different systems did not affect the production of contaminated eggs.

Colonization of internal organs by Salmonella Enteritidis in experimentally infected laying hens of four commercial genetic lines in conventional cages and enriched colony housing.

The prevalence of Salmonella Enteritidis in commercial egg-laying flocks is a prominent public health concern because contaminated eggs cause human illness. Deposition of this pathogen inside eggs results from bacterial colonization of reproductive tissues in infected hens. Environmental conditions can influence avian Salmonella infections, but the food safety consequences of different poultry housing systems remain uncertain. The present study assessed the invasion of internal organs by Salmonella Enteritidis in groups of experimentally infected laying hens of four commercial genetic lines (designated as white egg lines W1 and W2 and brown egg lines B1 and B2). Groups of hens from each line were housed at 555 cm2 of floor space per bird in both conventional cages and colony units enriched with access to perches and nesting areas. All hens were orally inoculated with 5.75 × 107 colony-forming units of a two-strain Salmonella Enteritidis mixture. At 6 to 7 d post-inoculation, hens were euthanized, and samples of liver, spleen, ovary, oviduct, and intestinal tract were removed for bacteriologic culturing. The frequency of Salmonella Enteritidis recovery from intestinal samples was significantly (P < 0.05) greater for the two white egg lines combined than for the two brown egg lines combined in both conventional cage (72.2% vs. 50.0%) and enriched colony housing systems (66.7% vs. 37.5%). The frequency of intestinal Salmonella Enteritidis isolation from line B1 was significantly higher from hens in conventional cages (47.2%) than in enriched colonies (22.2%), but no differences were observed for other hen lines. Line W1 yielded more positive intestinal samples than either brown egg line in conventional cages, and line B2 had fewer positive intestinal samples than all other lines in enriched colonies. There were no significant differences between hen lines or housing systems in Salmonella Enteritidis isolation from other internal organs. These results demonstrate that Salmonella Enteritidis colonization of the intestinal tract can vary between genetic lines of egg-laying hens and that some lines are subject to housing system influences on Salmonella susceptibility.