Tracking and Characterizing Spatiotemporal and Three-Dimensional Locomotive Behaviors of Individual Broilers in the Three-Point Gait-Scoring System.

Gait scoring is a useful measure for evaluating broiler production efficiency, welfare status, bone quality, and physiology. The research objective was to track and characterize spatiotemporal and three-dimensional locomotive behaviors of individual broilers with known gait scores by jointly using deep-learning algorithms, depth sensing, and image processing. Ross 708 broilers were placed on a platform specifically designed for gait scoring and manually categorized into one of three numerical scores. Normal and depth cameras were installed on the ceiling to capture top-view videos and images. Four birds from each of the three gait-score categories were randomly selected out of 70 total birds scored for video analysis. Bird moving trajectories and 16 locomotive-behavior metrics were extracted and analyzed via the developed deep-learning models. The trained model gained 100% accuracy and 3.62 ± 2.71 mm root-mean-square error for tracking and estimating a key point on the broiler back, indicating precise recognition performance. Broilers with lower gait scores (less difficulty walking) exhibited more obvious lateral body oscillation patterns, moved significantly or numerically faster, and covered more distance in each movement event than those with higher gait scores. In conclusion, the proposed method had acceptable performance for tracking broilers and was found to be a useful tool for characterizing individual broiler gait scores by differentiating between selected spatiotemporal and three-dimensional locomotive behaviors.

Undergraduate student attitudes to current poultry industry issues over four semesters: surveying an introductory poultry science course.

Individual background and demographics affect student perceptions of animal production. Understanding how science-based education alters these opinions is a critical aspect of improving university instruction as well as increasing consumer engagement in the poultry industry. The study objectives were to quantify the effects of student background, career interests, and science-based instruction on opinions regarding current issues in the poultry industry. Undergraduate students enrolled in a one semester poultry science course at Iowa State University between 2018 and 2021 were anonymously surveyed at the start and end of the semester as part of a 4-yr study. Students who opted to take the survey answered three demographic questions indicating their 1) livestock experience, 2) sex, and 3) career goals. The body of the survey consisted of 16 "poultry issue statements" where students were directed to mark a vertical dash on a 130 mm horizontal line indicating their level of agreement with each statement. Post-survey collection, the line was separated into 5 sections for discussion: responses within 0%-20% indicated strongly disagree, 21%-40% disagree, 41%-60% neutral, 61%-80% agree, and 81%-100% indicated strongly agree. Responses were analyzed using Proc Mixed in SAS Version 9.4 with a Tukey-Kramer adjustment for all pairwise comparisons using main effects including demographic categories, education (pre- or post-instruction), and year the survey was taken. Responses to various issue statements were affected by students' livestock experience (P < 0.05; 6 out of 16 statements affected), sex (P < 0.05; 5 out of 16 statements), and ultimate career goals (P < 0.05; 4 out of 16 statements). Pre- vs. post-education responses differed significantly in 6 out of 16 statements (P < 0.05), and in 2 out of 16 poultry issue statements, the year of instruction affected student response (P < 0.05). These data indicate that individual student background, sex, and differing career interests impact opinions of current topics in the broiler and layer industries. Further, science-based education as well as the year the course was taken over consecutive semesters significantly altered student opinions.

Individual experience and demographics affect perceptions of animal production. Understanding how science-based education alters these opinions is a critical aspect of improving instruction and increasing consumer engagement in the poultry industry. Undergraduate students enrolled in a poultry science course at Iowa State University between 2018 and 2021 were surveyed at the start and end of the semester as part of a 4-yr study. Students answered three demographic questions and indicated their agreeability with 16 "poultry issue statements." Responses to various issue statements were affected by students' livestock experience (6 out of 16 statements), sex (5 out of 16 statements), and ultimate career goals (4 out of 16 statements). Pre- vs. post-education responses changed in 6 out of 16 statements, and in 2 out of 16 poultry issue statements, the year of instruction affected student response. Individual student background, sex, and career interests impacted opinions on current topics in the broiler and layer industries, including laying hen housing systems, selective poultry breeding, environmental enrichment availability, culling practices, commercial stocking density, purchasing decisions, and more. Science-based instruction with hands-on farm experience as well as the year the course was taken over consecutive semesters significantly altered student opinions.

Age and Staphylococcus aureus Inoculation Route Differentially Alter Metabolic Potential and Immune Cell Populations in Laying Hens.

In 2018 and 2019, Staphylococcus aureus was isolated from multiple post-molt commercial laying hens with unusually high mortality. A challenge study was conducted to elucidate the role of S. aureus in this disease outbreak and the work herein represents the assessment of immunological responses in laying hens experimentally infected with S. aureus isolates from these cases. A total of 200 laying hens at 22 or 96 weeks of age (100/ age group) were assigned to 1 of 4 experimental inoculation groups (negative control, oral gavage, subcutaneous injection, or intravenous injection) after a 72 h acclimation period. Blood samples were taken prior to inoculation (baseline), 6 h post-inoculation (pi), 24 hpi, 3 dpi, and 7 dpi. Additional spleen samples to further assess systemic immunity were taken at baseline, 3 and 8 dpi. Metabolic phenotypes of peripheral blood mononuclear cells (PBMC) were isolated and assessed by Seahorse metabolic assay. Immune cell profiles in the spleen and PBMC were assessed by multicolor flow cytometry. At baseline, 96-week-old laying hens had 26.7% fewer PBMC-derived T cells compared to 22-week-old birds. Older hens had 28.9% increased helper T cell (TH) populations and 60.5% reduced γδ T cells (P = 0.03 and < 0.0001) which may contribute to variable clinical responses between age groups; however, no age-related differences in metabolic potential were observed. Metabolic outcomes showed that birds remained stressed from transport and re-housing past a 72 h acclimation period and through 24 h- 3 days post-inoculation. Inoculation with S. aureus generally reduced oxidative and glycolytic potentials compared to the control, with the greatest reductions observed in birds inoculated by intravenous injection (P < 0.05). Overall CD3+ T cell populations showed significant reductions in the intravenous group compared to other inoculation routes from 24 hpi to 7 dpi (23.6-39.0%; P ≤ 0.0001). These results suggest that age-related baseline differences in T cell populations and changes to T cell subpopulations and other immune cells due to inoculation route may have an additive effect on S. aureus- induced reductions in metabolic potential; however, further research linking metabolic potential and immune cell profiles is needed.

Evaluation of the metformin initiation rate in veterans with newly identified type 2 diabetes.

BACKGROUND: An estimated 35 million individuals in the United States have diabetes. The American Diabetes Association recommends metformin as first-line pharmacologic treatment. The primary objective of this study was to evaluate the metformin initiation rate in veterans with recently identified type 2 diabetes. METHODS: Veterans with new onset type 2 diabetes were identified using National Veterans Health Administration Data. Retrospective information was obtained from those with a first A1C ≥ 6.5% (48 mmol/mol) between 2013 and 2018. Veterans with at least one additional A1C < 6.5% (48 mmol/mol) documented in the three years prior to the A1C diagnostic for diabetes were included in the analysis. RESULTS: A total of 144,180 veterans were included. Of those, 45,776 (31.7%) were started on metformin within one year of diabetes diagnosis. The median time to metformin initiation was 12 days and median time to initiation of any anti-hyperglycemic was 11 days. Approximately 16,000 veterans were referred for lifestyle interventions within 90 days. CONCLUSION: Metformin initiation occurred in fewer patients than expected given metformin is a generic, well-tolerated medication recommended as first-line pharmacologic treatment option regardless of A1C. Further studies are needed to assess the barriers of initiating metformin at time of diabetes diagnosis.

Mitochondrial and Glycolytic Capacity of Peripheral Blood Mononuclear Cells Isolated From Diverse Poultry Genetic Lines: Optimization and Assessment.

Cellular metabolic preference is a culmination of environment, nutrition, genetics, and individual variation in poultry. The Seahorse XFe24 analyzer was used to generate foundational immune cellular metabolic data in layer, broiler, and legacy genetic strains using fresh chicken peripheral blood mononuclear cells (PBMCs). Baseline mitochondrial respiration [oxygen consumption rate (OCR)] and glycolytic activity [extracellular acidification rate (ECAR)] were determined in modern commercial laying hen (Bovans White) and broiler (Ross 308) lines, as well as the highly inbred lines of Iowa State University (L8, Fayoumi M-15.2, Spanish, Ghs-6), partially inbred broiler line, and advanced intercrosses of broiler by Fayoumi M-15.2 and broiler by Leghorn lines. Commercial broiler vs. Bovans layer and unvaccinated vs. vaccinated Bovans layer immune cell metabolic potential were compared following an in-assay pathway inhibitor challenge. Titrations consistently showed that optimal PBMC density in laying hens and broilers was 3 million cells per well monolayer. Assay media substrate titrations identified 25 mM glucose, 1 mM glutamine, and 1 mM sodium pyruvate as the optimal concentration for layer PBMCs. Pathway inhibitor injection titrations in Bovans layers and broilers showed that 0.5 µM carbonyl cyanide-4 phenylhydrazone (FCCP) and 1 µM oligomycin were optimal. Baseline OCR and ECAR were significantly affected by genetic line of bird (p < 0.05), with the dual-purpose, L8 inbred line showing the highest OCR (mean 680 pmol/min) and the partially inbred broiler line showing the greatest ECAR (mean 74 mpH/min). ECAR metabolic potential tended to be greater in modern layers than broilers (p < 0.10), indicating increased ability to utilize the glycolytic pathway to produce energy. OCR was significantly higher in vaccinated than unvaccinated hens (p < 0.05), while baseline ECAR values were significantly lower in vaccinated Bovans laying hens, showing increased oxidative capacity in activated immune cells. These baseline data indicate that different genetic strains of birds utilized the mitochondrial respiration pathway differently and that modern commercial lines may have reduced immune cell metabolic capacity compared with legacy lines due to intense selection for production traits. Furthermore, the Seahorse assay demonstrated the ability to detect differences in cellular metabolism between genetic lines and immune status of chickens.

Lipid Source and Peroxidation Status Alter Immune Cell Recruitment in Broiler Chicken Ileum.

BACKGROUND: Restaurant oil in poultry diets increases energy content, reduces production costs, and promotes sustainability within the food supply chain. However, variable oil composition and heating temperatures among restaurant oil sources can impact broiler chicken health due to heat-induced lipid modifications. OBJECTIVES: A 21-d experiment was conducted to evaluate ileal morphology, liver cytokine gene expression, and ileal immune cell populations in broilers fed control or peroxidized lipids with varying chain and saturation characteristics. METHODS: Day-old broilers were housed in battery cages (5 birds per cage) and fed diets containing 5% control or peroxidized oils. Eight diets were randomly assigned in a 4 × 2 factorial arrangement consisting of oil source (palm, soybean, flaxseed, or fish) and peroxidation status (control or peroxidized). At day 21, samples were collected for ileal histomorphology [villus height (VH), crypt depth (CrD), and the VH:CrD ratio], and liver cytokine expression (qPCR). Ileum cytokine expression and T-cell markers were analyzed by RNAscope in situ hybridization (ISH). Data were analyzed as a mixed model (SAS 9.4) with fixed effects of lipid source, peroxidation, and lipid × peroxidation interaction. RESULTS: CD3+ T-cells in the ileum decreased 16.2% due to peroxidation (P = 0.001) with 30.3% reductions observed in birds fed peroxidized flaxseed oil (P = 0.01). Peroxidation increased IL6+ and IL1B+ cells by 62.0% and 40.3%, respectively (P = 0.01). Soybean oil increased IFNG+ cells by 55.1% compared with palm oil, regardless of peroxidation status (P = 0.007). Lipid source and peroxidation did not alter ileal histomorphology or liver cytokine expression. CONCLUSIONS: Lipid peroxidation increased ileal IL1B and IL6 in broiler chickens, whereas soybean oil diets increased IFNG. Generally, peroxidation decreased overall CD3+ T-cell populations, suggesting impaired T-cell presence or recruitment. These results identify potential immunomodulatory lipid profiles in restaurant oil while supporting RNAscope-ISH as a method to describe avian tissue-level immune responses.

A novel environmental enrichment device increased physical activity and walking distance in broilers.

Modern broilers are selected for fast growth and a large proportion of breast tissue, contributing to a top-heavy phenotype, leg disorders, and inactivity as birds reach market weight. Therefore, the objective was to motivate broilers to move through environmental enrichment. A total of 1,200 Ross 308 broilers were housed in pens of 30 for 6 wk: 600 birds were exposed to a novel laser enrichment device (LASER) and 600 were control. Each device projected 2 randomly moving red laser dots onto the floor 4 times/day for 4-min "laser periods." Seven LASER and 7 control pens, with 5 focal birds/pen (n = 70), were randomly selected to be video-recorded day 0 to 8 and once weekly for the remainder of the trial. Videos were analyzed to measure broiler time-budget and behaviors such as latency to feed and distance walked during laser periods. Focal birds were gait scored weekly on-farm. A test of the human-approach paradigm was carried out on weeks 1 and 6 on all pens. LASER birds were more active on days 0, 1, 3, 4, 5, 7, and 8, moving 254% more on day 7 (P ≤ 0.05). Time spent active was increased in LASER treatment by 114% on week 2; 157% on week 3; 90% on week 4; and 82% on week 5. LASER birds spent more time at the feeder on days 0, 1, 2, 5, 8, and on weeks 1 and 5, with 84% more time at feeder than control on day 5 (P ≤ 0.05). LASER birds walked further during laser periods on day 0 to 8, reaching 646.5 cm greater (day 1), and on weeks 2, 3, 4, and 5, with an increase of 367.5 cm on week 2 (P ≤ 0.05). Over week 1 to 6, 60.54 ± 7.4% of focal birds in the laser treatment were at the feeder during or within 5 min following laser periods. The laser enrichment device was successful in stimulating broiler physical activity and feeding, and did not negatively impact walking ability.

Development and Validation of Broiler Welfare Assessment Methods for Research and On-farm Audits.

Required auditing of on-farm broiler welfare in the United States has increased; however, a lack of validated tools exists for assessment of enrichment. National Chicken Council (NCC) guidelines were used on a subset of 300 Ross 308 broilers out of 1200 to validate and adapt welfare measures. Half of the broilers were exposed to environmental enrichment, hence these measures were used to evaluate the enrichment within the context of behavior and welfare, although the nature of the enrichment is not described in detail here as the aim is to serve solely as a description and validation of methods using a subset of example data. Birds were recorded in repeated 4-min periods to quantify behavior and walking distance. Outcomes were categorized to improve auditing and make recommendations to producers and researchers. Bone mineral density, content, and breaking strength were successful in determining numerical differences. Quantifying lameness using an enclosed walkway and measuring footpad dermatitis weekly are recommended on-farm. We recommend including additional measures not required by the NCC: monitoring breast condition in the flock and including a behavior component with a scoring ethogram.

Composition and inclusion of probiotics in broiler diets alter intestinal permeability and spleen immune cell profiles without negatively affecting performance1.

Probiotic feed additives with potential to enhance performance, health, and immunity have gained considerable popularity in commercial broiler production. The study objectives were to measure broiler performance, gut integrity, and splenic immune cell profiles in birds fed one of two probiotics at two inclusion levels. Nine hundred sixty Ross 708 broilers (12 per pen) were randomly assigned to no additive control, 0.05% or 0.10% LactoCare (Lactobacillus reuteri), or 0.05% or 0.10% LactoPlan (Lactobacillus plantarum) dietary treatments for 6 wk. On day 27, a 20-pen subset was utilized for a fluorescein isothiocyanate dextran (FITC-d) assay, where half of the pens were subject to a 12-h feed restriction (FR) pregavage. Serum collected from blood drawn 1-h postgavage was analyzed for relative fluorescence of FITC-d absorbed across the intestinal barrier as a gut leakiness indicator. On day 42, spleens from eight birds per treatment were collected for immune cell profile analysis by multicolor flow cytometry. Although performance outcomes were not affected by dietary treatment, FITC-d absorption post-FR was increased 57% in the 0.05% LactoPlan treatment, and was decreased by 12.6% in the 0.05% LactoCare diet, 12% in the 0.10% LactoCare diet, and 22% in the 0.10% LactoPlan diet compared with the control. This indicates a positive impact in barrier integrity maintenance due to 0.05% and 0.10% LactoCare and 0.10% LactoPlan diet following a challenge. Immune cell profiles varied between the two probiotic compositions, with an approximately 50% reduction in splenic innate immune cells (monocyte/macrophage+) in birds fed LactoPlan (P < 0.0001) and greater overall percentages of CD45+ leukocytes and CD3+ T cells in birds fed 0.10% LactoCare (P < 0.0001). LactoPlan diets shifted splenic T-cell populations in favor of CD8α + cytotoxic T cells (TC; P = 0.007), while higher inclusions (0.10%) of either probiotic increased the percentage of activated CD4+ helper T cells (TH; P < 0.0001). These results indicate that compositionally different probiotics had varying effects on the gut permeability and splenic immune cell profiles in broiler chickens, particularly at higher inclusion rates, but observed changes to underlying physiology did not negatively impact performance outcomes. The ability of a probiotic to alter gut permeability and immune cell profile, therefore, may depend on the compositional complexity of the product as well as inclusion rate.