The Impact of Early-Life Cecal Microbiota Transplantation on Social Stress and Injurious Behaviors in Egg-Laying Chickens.

Injurious behaviors (i.e., aggressive pecking, feather pecking, and cannibalism) in laying hens are a critical issue facing the egg industry due to increased social stress and related health and welfare issues as well as economic losses. In humans, stress-induced dysbiosis increases gut permeability, releasing various neuroactive factors, causing neuroinflammation and related neuropsychiatric disorders via the microbiota-gut-brain axis, and consequently increasing the frequency and intensity of aggression and violent behaviors. Restoration of the imbalanced gut microbial composition has become a novel treatment strategy for mental illnesses, such as depression, anxiety, bipolar disorder, schizophrenia, impulsivity, and compulsivity. A similar function of modulating gut microbial composition following stress challenge may be present in egg-laying chickens. The avian cecum, as a multi-purpose organ, has the greatest bacterial biodiversity (bacterial diversity, richness, and species composition) along the gastrointestinal tract, with vitally important functions in maintaining physiological and behavioral homeostasis, especially during the periods of stress. To identify the effects of the gut microbiome on injurious behaviors in egg-laying chickens, we have designed and tested the effects of transferring cecal contents from two divergently selected inbred chicken lines on social stress and stress-related injurious behaviors in recipient chicks of a commercial layer strain. This article reports the outcomes from a multi-year study on the modification of gut microbiota composition to reduce injurious behaviors in egg-laying chickens. An important discovery of this corpus of experiments is that injurious behaviors in chickens can be reduced or inhibited through modifying the gut microbiota composition and brain serotonergic activities via the gut-brain axis, without donor-recipient genetic effects.

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.

Cage-Free Pullets Minimally Affected by Stocking Density Stressors.

Management choices during the pullet phase can affect behavior, welfare, and health later in life, but few studies have evaluated the pullet phase, particularly in extensive housing systems. This study was a 2 × 2 factorial randomized complete block design (RCBD) with two strains and two stocking densities. The Lohmann LB-Lite and Lohmann LSL-Lite were housed on the floor at high-stocking density (619-670 cm2/bird) and low-stocking density (1249-1352 cm2/bird), which changed with age from 2 to 16 weeks of age (WOA). Bird-based measures of appearance, blood parameters, organ measurements, and production values were evaluated. Stocking density alone affected (p < 0.05) only relative bursal weight (% of body weight)-3.32% in the low-density versus 3.08% in the high-density group. High-stocking density was correlated with decreased uniformity (high-89.33 ± 0.24%; low-90.41 ± 0.24; p < 0.02) and worse feather coverage in the brown strain. High-stocking density was correlated with greater uniformity (High-90.39 ± 0.24%; Low-88.47 ± 0.24%; p < 0.001) and better feather coverage in the white strain. This study's feed conversion ratio (FCR) was improved by 0.07 in the low-stocking density for both strains. The remaining parameters were affected by strain and age only. Thus, while stocking density effects vary slightly depending on the strain used, cage-free pullets had limited negative effects at both the high and low-stocking densities tested in this study; there were few to no changes in the numerous bird-based welfare parameters tested.

Investigating the effects of jute nesting material and enriched piglet mats on sow welfare and piglet survival.

Domesticated sows are motivated to perform nesting behavior prior to farrowing, and nesting material can reduce piglet cold stress. However, nesting material may not be practical in most production systems due to the potential for clogging slurry systems. Therefore, the study objectives were to assess an alternative nesting material provided prior to farrowing on sow welfare and piglet survival, and to investigate the effect of the entire nesting environment on piglet survival and growth performance. We hypothesized that the provision of jute nesting material would decrease sow stress and farrowing duration, and that nesting mat provision would allow piglets to remain euthermic and improve survival and growth. Sows (N = 20) were randomly assigned to one of two treatments: a farrowing crate with three pieces of 40.6 × 21.6 cm jute nesting material (Nest; n = 10) and two enriched piglet mats made from an acrylic board (28.0 × 86.4 cm) covered with a microfiber material, or a farrowing crate without nesting material (Control; n = 10) and one standard plastic piglet mat (28.0 × 86.4 cm). Jute pieces were attached to the front of the crate to prevent substrate from falling through the slatted floors. Saliva samples were collected to measure cortisol and immunoglobulin A (IgA), on days -1, 0, 1, and 2 relative to farrowing, and a final sample was collected at weaning (day 16.9 ±â€…0.18). Blood was collected from four piglets per litter to measure immunoglobulin G (IgG) at 48 h, day 7, and weaning. Piglet skin temperature (TS) was measured on two piglets per litter using an infrared camera for 3 d after birth at 0800, 1200, 1600, and 2000 h. One piglet was randomly chosen from the heat lamp and nonheat lamp side of the crate to measure TS. Video was continuously coded for observations of jute- and crate-directed interactions. Data were analyzed as a mixed model analysis of variance in SAS 9.4. Nest sows performed less crate-directed behavior than Control sows (P = 0.02). Cortisol tended to be reduced in Nest sows (P = 0.08) when compared to Controls, but no differences in IgA concentrations (P > 0.40) were detected. Nest piglets tended to be heavier on day 7 (P < 0.10), had greater IgG concentrations (P = 0.03), and had greater TS (P = 0.02) versus Controls. No farrowing duration or number of stillbirth differences were observed (P > 0.70). The jute material and piglet nests positively impacted sow welfare and piglet measures but did not translate into improved piglet survival.

Cecal microbiota transplantation: unique influence of cecal microbiota from divergently selected inbred donor lines on cecal microbial profile, serotonergic activity, and aggressive behavior of recipient chickens.

BACKGROUND: Accumulating evidence from human trials and rodent studies has indicated that modulation of gut microbiota affects host physiological homeostasis and behavioral characteristics. Similarly, alterations in gut microbiota could be a feasible strategy for reducing aggressive behavior and improving health in chickens. The study was conducted to determine the effects of early-life cecal microbiota transplantation (CMT) on cecal microbial composition, brain serotonergic activity, and aggressive behavior of recipient chickens. METHODS: Chicken lines 63 and 72 with nonaggressive and aggressive behavior, respectively, were used as donors and a commercial strain Dekalb XL was used as recipients for CMT. Eighty-four 1-d-old male chicks were randomly assigned to 1 of 3 treatments with 7 cages per treatment and 4 chickens per cage (n = 7): saline (control, CTRL), cecal solution of line 63 (63-CMT), and cecal solution of line 72 (72-CMT). Transplantation was conducted via oral gavage once daily from d 1 to 10, and then boosted once weekly from week 3 to 5. At weeks 5 and 16, home-cage behavior was recorded, and chickens with similar body weights were assigned to paired aggression tests between the treatments. Samples of blood, brain, and cecal content were collected from the post-tested chickens to detect CMT-induced biological and microbiota changes. RESULTS: 63-CMT chickens displayed less aggressive behavior with a higher hypothalamic serotonergic activity at week 5. Correspondingly, two amplicon sequence variants (ASVs) belonging to Lachnospiraceae and one Ruminococcaceae UCG-005 ASV were positively correlated with the levels of brain tryptophan and serotonin, respectively. 72-CMT chickens had lower levels of brain norepinephrine and dopamine at week 5 with higher levels of plasma serotonin and tryptophan at week 16. ASVs belonging to Mollicutes RF39 and GCA-900066225 in 72-CMT chickens were negatively correlated with the brain 5-hydroxyindoleacetic acid (5-HIAA) at week 5, and one Bacteroides ASV was negatively correlated with plasma serotonin at week 16. CONCLUSION: Results indicate that CMT at an early age could regulate aggressive behavior via modulating the cecal microbial composition, together with central serotonergic and catecholaminergic systems in recipient chickens. The selected CMT could be a novel strategy for reducing aggressive behavior through regulating signaling along the microbiota-gut-brain axis.

Comparing different environmental enrichments for improving the welfare and walking ability of male turkeys.

This study investigated age-related changes in turkey welfare measures (wounds, feather quality (FQ), feather cleanliness, and footpad condition (FCON)) and walking ability (gait) as influenced by different types of environmental enrichment (EE). Tom turkeys (n = 420) were randomly assigned to: straw bale (S), platform (P), platform + straw bale (PS), pecking block (B), tunnel (T) or control (C; no enrichment) group. Welfare measures and gait were assessed at 8, 12, 16 and 19 wk and analyzed using PROC LOGISTIC with Firth bias-correction. Better wing FQ with age was observed in turkeys in S and T groups. Turkeys in the S group had better wing FQ at 16 (P = 0.028) and 19 wk (P = 0.011) vs. 8 wk. Wing FQ (P = 0.008) was better at 19 vs. 8 wk for T turkeys. FCON worsened over time for turkeys in all treatment groups except for the S group. FCON was worse at 19 vs.8 wk for P (P = 0.024), PS (P = 0.039), B (P = 0.011), T (P = 0.004) and C (P = 0.014) turkeys and was worse at 19 vs. 12 wk for B (P = 0.038), T (P = 0.015) and C (P = 0.045) turkeys. FCON was worse at 19 vs. 16 wk for T (P = 0.007) and C (P = 0.048) turkeys. FCON was also worse at 16 vs. 8 wk for B (P = 0.046) turkeys. Gait worsened with increasing age in all treatment groups. Gait was worse at 19 wk for S (P < 0.001), P (P < 0.001), PS (P < 0.001) and B turkeys (P < 0.001) vs. earlier ages, while gait in T (P < 0.001) and C turkeys (P < 0.001) worsened starting at 16 wk.

Effects of early-life cecal microbiota transplantation from divergently selected inbred chicken lines on growth, gut serotonin, and immune parameters in recipient chickens.

Recent studies have revealed that fecal microbiota transplantation exerts beneficial effects on modulating stress-related inflammation and gastrointestinal health of the host. The aim of this study was to examine if cecal microbiota transplantation (CMT) presents similar efficiency in improving the health status of egg-laying strain chickens. Chicken lines 63 and 72 divergently selected for resistance or susceptibility to Marek's disease were used as CMT donors. Eighty-four d-old male recipient chicks (a commercial DeKalb XL layer strain) were randomly assigned into 3 treatments with 7 replicates per treatment and 4 birds per replicate (n = 7): saline (control, CTRL), cecal solution of line 63 (63-CMT), and cecal solution of line 72 (72-CMT) for a 16-wk trial. Cecal transplant gavage was conducted once daily from d 1 to d 10, then boosted once weekly from wk 3 to wk 5. The results indicated that 72-CMT birds had the highest body weight and ileal villus/crypt ratio among the treatments at wk 5 (P ≤ 0.05); and higher heterophil/lymphocyte ratios than that of 63-CMT birds at wk 16 (P < 0.05). 72-CMT birds also had higher levels of plasma natural IgG and Interleukin (IL)-6 at wk 16, while 63-CMT birds had higher concentrations of ileal mucosal secretory IgA at wk 5 and plasma IL-10 at wk 16 (P < 0.05), with a tendency for lower mRNA abundance of splenic IL-6 and tumor necrosis factor (TNF)-α at wk 16 (P = 0.08 and 0.07, respectively). In addition, 72-CMT birds tended to have the lowest serotonin concentrations (P = 0.07) with the highest serotonin turnover in the ileum at wk 5 (P < 0.05). There were no treatment effects on the levels of plasma corticosterone and testosterone at wk 16 (P > 0.05). In conclusion, early postnatal CMT from different donors led to different patterns of growth and health status through the regulation of ileal morphological structures, gut-derived serotonergic activities, peripheral cytokines, and antibody production in recipient chickens.

Behavior and welfare impacts of water provision via misting in commercial Pekin ducks.

Ducks will access water to maintain feather condition and exhibit natural water-related behaviors such as wet preening. Providing water to ducks commercially is challenging as it may reduce litter and air quality leading to higher duck mortality or illness. This research aimed to measure the behavioral and welfare impacts of water provision via a misting system for commercial Pekin grower ducks in Victoria, Australia. Seven grower flocks were observed (four misted and three nonmisted in open-sided sheds) during May and November 2021. From 26 until 33 d of age, treatment ducks were provided 1 h of misting with shed curtains closed in both treatment and control sheds. At the start and end of the misting application period, external health and welfare measures were taken directly on the ducks via transect walks throughout each shed and catch-and-inspect observations on a sample of 150 ducks from each shed. Video recordings were also made of the misted and nonmisted ducks for 3 h representing time periods prior to, during, and after the 1-h misting across all sheds for all 8 d of the treatment period. Observations were made of all behavior that ducks exhibited at 10-min scan sample intervals across four cameras per shed, totaling 4,198 scans across the seven sheds. General linear mixed models showed the misting application predominantly had impacts on the patterns of behavioral change across the treatment time periods between the misted and nonmisted ducks rather than increasing or decreasing the overall expression of specific behaviors (interaction terms all P ≤ 0.003). The misted ducks increased drinking, tail wagging, and walking, and reduced preening, rooting litter, sitting, and stretching during misting relative to what they showed prior. The nonmisted ducks showed less sitting and more panting during misting relative to prior. Pearson's Chi-square tests showed some differences between the treatment groups in feather cleanliness on the back and wings (both P < 0.0001), likely resulting from pre-existing differences between sheds in blood from pin feathers. Most welfare indicators showed no positive or negative effect of the misting treatment. These results indicate overhead misting does affect duck behavior to some degree without compromising their welfare, but further research with larger water droplet sizes resulting in greater accumulation of surface water or extended durations of misting may lead to greater effects.

Ducks use bathing water for wet preening and feather maintenance. Commercially, it is challenging to provide clean bathing water without compromising litter quality and duck health. Overhead misting may be a mode of water delivery that will wet the ducks' bodies with continuously clean water. This study compared seven grower flocks of Pekin ducks (four misted and three nonmisted treatment flocks) in open-sided sheds during May and November 2021 in Australia. From 26 until 33 d of age, treatment ducks were provided 1 h of misting with shed curtains closed in both treatment and control sheds. External welfare measures were taken directly on the ducks at 26 and 33 d of age. Daily video recordings were made to observe if behaviors differed before, during, or after the 1 h of misting in both treatment and control sheds. Results showed the misting application predominantly affected the way behaviors changed across time between the misted and nonmisted ducks rather than increasing or decreasing the overall expression of specific behaviors. The differences may have in part been related to the curtain closure. Most welfare indicators showed no positive or negative effect of the misting treatment. Larger water droplet sizes may have greater effects on duck behavior.

Assessing the effect of compounds from plantar foot sweat, nesting material, and urine on social behavior in male mice, Mus musculus.

Home cage aggression causes poor welfare in male laboratory mice and reduces data quality. One of the few proven strategies to reduce aggression involves preserving used nesting material at cage change. Volatile organic compounds from the nesting material and several body fluids not only correlate with less home cage aggression, but with more affiliative allo-grooming behavior. To date, these compounds have not been tested for a direct influence on male mouse social behavior. This study aimed to determine if 4 previously identified volatile compounds impact home cage interactions. A factorial design was used with cages equally split between C57BL/6N and SJL male mice (N = 40). Treatments were randomly assigned across cages and administered by spraying one compound solution on each cage's nesting material. Treatments were refreshed after day 3 and during cage change on day 7. Home cage social behavior was observed throughout the study week and immediately after cage change. Several hours after cage change, feces were collected from individual mice to measure corticosterone metabolites as an index of social stress. Wound severity was also assessed after euthanasia. Measures were analyzed with mixed models. Compound treatments did not impact most study measures. For behavior, SJL mice performed more aggression and submission, and C57BL/6N mice performed more allo-grooming. Wound severity was highest in the posterior region of both strains, and the middle back region of C57BL/6N mice. Posterior wounding also increased with more observed aggression. Corticosterone metabolites were higher in C57BL/6N mice and in mice treated with 3,4-dimethyl-1,2-cyclopentanedione with more wounding. These data confirm previous strain patterns in social behavior and further validates wound assessment as a measure of escalated aggression. The lack of observed treatment effects could be due to limitations in the compound administration procedure and/or the previous correlation study, which is further discussed.

A critical review of advancement in scientific research on food animal welfare-related air pollution.

Air pollution generates hazardous pollutants that have resulted in safety, health, and other welfare issues of food animals. This paper reviewed scientific research advancement in food animal welfare-related air pollution based on 219 first-hand research publications in refereed journals (referred to as "RPs") over the past nine decades. Scientific studies in this area began in the 1930s. The number of RPs has increased significantly with each decade from the 1960s to the 1980s, then decreased until the 2010s. Twenty-six countries have contributed to this multidisciplinary research. About 52% of the studies were conducted in the U.S. and U.K. Research activities have surged in China since the 2010s. On-farm discoveries in air toxicity that resulted in animal death or injury were all from observational studies. About 75% of the studies were experimental and conducted primarily under laboratory conditions. Ammonia (NH3) was the main pollutant in 59% of the RPs, followed by dust, hydrogen sulfide (H2S), bacteria and endotoxins, carbon dioxide (CO2), carbon monoxide (CO), silo gas, sulfur dioxide (SO2), and odor. Approximately 23% of RPs reported multiple pollutants in the same study. The most intensively studied animal species were poultry (broilers, hens, turkeys, ducks, and eggs and embryos in 44% of the RPs) and pigs (also 44%), followed by cattle, and sheep and goats. Scientific investigations in this area were driven by the research focuses in the areas of animal agriculture and industrial air pollution. Some major research teams played important roles in advancing scientific research. However, research in this area is still relatively limited. There is a great need to overcome some technical challenges and reverse the trend of decreasing research activities in North America and Europe.

Who's the Boss? Assessing Convergent Validity of Aggression Based Dominance Measures in Male Laboratory Mice, Mus Musculus.

Aggression among group housed male mice continues to challenge laboratory animal researchers because mitigation strategies are generally applied at the cage level without a good understanding of how it affects the dominance hierarchy. Aggression within a group is typically displayed by the dominant mouse targeting lower ranking subordinates; thus, the strategies for preventing aggression may be more successful if applied specifically to the dominant mouse. Unfortunately, dominance rank is often not assessed because of time intensive observations or tests. Several correlates of dominance status have been identified, but none have been directly compared to home cage behavior in standard housing. This study assessed the convergent validity of three dominance correlates (urinary darcin, tube test score, preputial gland to body length ratio) with wound severity and rankings based on home cage behavior, using factor analysis. Discriminant validity with open field measures was assessed to determine if tube test scores are independent of anxiety. Cages were equally split between SJL and albino C57BL/6 strains and group sizes of 3 or 5 (N = 24). Home cage behavior was observed during the first week, and dominance measures were recorded over the second. After controlling for strain and group size, darcin and preputial ratio had strong loadings on the same factor, which was a significant predictor of home cage ranking showing strong convergent validity. Tube test scores were not significantly impacted by open field data, showing discriminant validity. Social network analysis revealed that despotic power structures were prevalent, aggressors were typically more active and rested away from cage mates, and the amount of social investigation and aggression performed by an individual were highly correlated. Data from this study show that darcin and preputial ratio are representative of home cage aggression and provide further insight into individual behavior patterns in group housed male mice.

Compounds from plantar foot sweat, nesting material, and urine show strain patterns associated with agonistic and affiliative behaviors in group housed male mice, Mus musculus.

Excessive home cage aggression often results in severe injury and subsequent premature euthanasia of male laboratory mice. Aggression can be reduced by transferring used nesting material during cage cleaning, which is thought to contain aggression appeasing odors from the plantar sweat glands. However, neither the composition of plantar sweat nor the deposits on used nesting material have been evaluated. The aims of this study were to (1) identify and quantify volatile compounds deposited in the nest site and (2) determine if nest and sweat compounds correlate with social behavior. Home cage aggression and affiliative behavior were evaluated in 3 strains: SJL, C57BL/6N, and A/J. Individual social rank was assessed via the tube test, because ranking may influence compound levels. Sweat and urine from the dominant and subordinate mouse in each cage, plus cage level nest samples were analyzed for volatile compound content using gas chromatography-mass spectrometry. Behavior data and odors from the nest, sweat, and urine were statistically analyzed with separate principal component analyses (PCA). Significant components, from each sample analysis, and strain were run in mixed models to test if odors were associated with behavior. Aggressive and affiliative behaviors were primarily impacted by strain. However, compound PCs were also impacted by strain, showing that strain accounts for any relationship between odors and behavior. C57BL/6N cages displayed the most allo-grooming behavior and had high scores on sweat PC1. SJL cages displayed the most aggression, with high scores on urine PC2 and low scores on nest PC1. These data show that certain compounds in nesting material, urine, and sweat display strain specific patterns which match strain specific behavior patterns. These results provide preliminary information about the connection between home cage compounds and behavior. Salient compounds will be candidates for future controlled studies to determine their direct effect on mouse social behavior.

Willingness to pay for whole turkey attributes during Thanksgiving holiday shopping in the United States.

Although whole turkeys served at Thanksgiving are the ubiquitous kickoffs to the US winter holiday season, much remains unknown about shopping behaviors for holiday food items. Given the once-a-year purchase of the whole turkey for most households, collecting data about demand and preferences necessitated the collection of data during the week before Thanksgiving, while turkey shopping was at the forefront of consumers' minds. Despite a self-reported confidence in cooking turkeys, many respondents indicated they thawed frozen meat using improper methods. Ninety-five percent of respondents indicated that they consumed meat; 89% of respondents who consumed meat or had someone in the household who did, indicated they had purchased turkey products. Positive willingness to pay (WTP) was found for all attributes of whole turkeys studied: free range, fed a vegetarian diet, hormone use not permitted, and antibiotic use not permitted. Mean estimated WTP for free range ranged from $0.37/lb for industry verified free range to $0.74/lb for USDA verified free range; although those 2 estimates were not statistically different from each other, they were both statistically different from zero. The statistically significant estimated mean WTP for hormone use not permitted ranged from $0.85/lb for industry verification to $1.35 for USDA verification but were again not statistically different from each other. Mean WTP estimates, which were statistically significant but not different from one another for antibiotic use not permitted, ranged from $0.62/lb for industry certification to $0.72 for retailer certified. Turkeys certified to be fed a vegetarian diet had a mean WTP estimate of $0.39/lb for retailer verification to $0.60/lb for USDA verification; those mean WTP estimates were not statistically different from each other but were each statistically different from zero. Social desirability bias, which can be defined as the relative over-reporting of one's own goodness, was detected with respect to self-reported holiday eating and healthfulness statements. Relationships were found between social desirability bias, gender, and age for holiday eating statements using a seemingly unrelated regression.

Poultry in the classroom: effectiveness of an online poultry-science-based education program for high school STEM instruction.

Public knowledge of the poultry industry is limited, yet attitudes towards the industry shape consumer behavior and influence the attractiveness of poultry careers and future study. This study assessed an online learning program contextualizing STEM learning within poultry science which was designed to increase poultry knowledge and interest. High school student participants (n = 169) across 16 classes (n = 12 teachers) in Indiana completed seven 30-min online modules during the fall 2018 semester. This case study used a mixed-methods, sequential explanatory design. Student knowledge and interest in poultry and teacher perceptions of the program were examined using quantitative measures. The study's qualitative portion assessed perceptions of the learning experience and comprised open-ended student and teacher survey questions and a teacher focus group. Qualitative data suggested that students' poultry knowledge, including awareness of the industry and related careers, was improved following the modules. Paired t-tests showed moderate to large effect sizes regarding increases in content quiz scores following each module (P < 0.001, Cohen's d = 0.45 to 0.80). Students' mean interest in poultry was initially low and remained statistically similar upon completion of the program (M = 1.84, 1.87; p = 0.67). Student and teacher statements indicated that design features of the program enhanced interest for some students. Other students did not perceive the program and its poultry science topics to be interesting or relevant to their lives. Results from this study can be used to inform the creation of effective poultry learning resources to contribute to workforce development and enhance the industry's public image.