Location: Poultry Production and Product Safety Research
2025 Annual Report
Objectives
1.Implement strategies using plant derived, food-grade phytochemical nanoemulsions for reducing Salmonella and Campylobacter in poultry.
1A.Investigate the efficacy of in-water supplementation of phytochemical nanoemulsions in reducing S. Enteritidis and C. jejuni colonization in broiler chickens.
1B.Reduce Salmonella and Campylobacter on chicken carcasses using phytochemical nanoemulsions applied as a post-harvest intervention at critical control points in processing plants.
1C. Determine the quality, shelf-life and consumer acceptability of chicken meat subjected to the aforementioned interventions.
2. Investigate the potential mechanism(s) of action of phytochemical nanoemulsions against pathogen biofilms and determine efficacy for reducing Salmonella and Campylobacter biofilms in poultry processing plants.
2A. Determine the efficacy of phytochemical nanoemulsions as an antimicrobial wash for eradicating mature S. Enteritidis and C. jejuni biofilm formed on common food contact surfaces.
2B. Determine the efficacy of phytochemical nanoemulsions as an antimicrobial wash for inhibiting S. Enteritidis and C. jejuni biofilm formation on common food contact surfaces and their effect on exopolysaccharide (EPS) production, extracellular DNA (eDNA) production, and quorum sensing.
2C. Investigate the potential mechanism(s) of action of phytochemical nanoemulsions against pathogen biofilm by using transcriptomic and proteomic approaches.
3. Develop vaccine strategies that target multiple pathogens (i.e. Salmonella, Campylobacter, Clostridium, E. coli) utilizing novel Electron-beam technology in poultry.
3A. Test to confirm inactivation of foodborne pathogens in cocktail vaccine consisting of multi-serovars of Salmonella or multiple strains of C. jejuni in broiler chickens.
3B. Determine the efficacy of vaccine consisting of multi- serovars of Salmonella or multiple strains-C. jejuni in reducing colonization and shedding of foodborne pathogens in broiler chickens.
3C. Determine the efficacy of a multi-species cocktail vaccine in reducing colonization and shedding of foodborne pathogens Salmonella enterica, and C. jejuni in broiler chickens.
4. Identify key host neurochemical-microbiota-pathogen interactions across the biogeography of the avian gastrointestinal tract to enhance efficacy of phytochemical and vaccine-based strategies in reducing enteric pathogen colonization.
4A. Determine the ability of heat and cold stressors to influence avian susceptibility to enteric colonization of Salmonella and C. jejuni due to neurochemical production in different regions of the intestinal tract.
4B. Determine functional changes in the microbiome of each region of the avian intestinal tract in response to heat or cold stressors in Salmonella and C. jejuni challenged and unchallenged birds.
4C. Determine the ability of heat and cold stressors to influence efficacies of vaccine and phytochemical modalities on avian susceptibility to enteric foodborne pathogen colonization due to neurochemical production in different...
5.Utilize novel electron-beam technology to reduce pathogen prevalence on poultry products.
5A and 5B, see subobj(s) on uploaded document..
Approach
Food safety is a major priority for the poultry industry, among the foodborne pathogens transmitted through poultry products, Salmonella spp. and Campylobacter are epidemiologically linked to the consumption of contaminated poultry and account for the majority of confirmed cases of bacterial gastroenteritis in the US. Despite substantial progress, they remain as the most common foodborne pathogens transmitted to humans. Antibiotic growth promoters (AGPs) have been an integral part of poultry production contributing significantly to controlling pathogens, reducing infections/mortality and improved growth rate. Their use has been restricted in poultry production amid growing concerns of microbial antimicrobial resistance (AMR). The goal of this project is to use a multi-hurdle approach to develop safe and effective alternatives to antibiotics for controlling foodborne pathogens in conventional and organic poultry sectors. First, we will investigate the ability of phytochemical nanoemulsions to reduce Salmonella and Campylobacter colonization in the poultry intestinal tract, on poultry carcasses, and on food contact surfaces. Mechanism of action will be determined as well as the effect of phytochemical intervention on carcass quality and consumer acceptability. Second, electron-beam-technology will be used to develop a safe and effective vaccine targeting both Salmonella and Campylobacter in the chicken intestinal tract. Finally, comprehensive neurochemical and microbial mapping of the poultry gut will determine the effect of stress-related neurochemicals on pathogen colonization and efficacy of phytochemical and vaccine interventions. This research will lead to innovative non-antibiotic intervention strategies using plant-derived antimicrobials and novel vaccine strategies for reducing colonization of foodborne pathogens, decreasing contamination of poultry products and enhancing the health and overall welfare of poultry.
Approach for New Objective 5: We proposed to utilize an Electron beam to destroy foodborne pathogens and spoilage organisms in poultry meat and poultry meat products. We will determine an E-beam dose to inactivate Salmonella serovars and Campylobacter jejuni on artificially inoculated poultry meat and poultry products; confirm the efficacy of E-beam dose in inactivating pathogens on naturally contaminated poultry meat and poultry product; and evaluate the quality, shelf-life, and consumer acceptability of E-Beam irradiated meat.
Approach for New Subobjectives 5A and 5B. develop research strategies to inactivate foodborne pathogens, Salmonella and Campylobacter, and spoilage bacteria on poultry meat and poultry meat products. We will utilize electron beam (E-Beam) technology to control foodborne pathogens and spoilage organisms in poultry meat and poultry meat products. Inactivation studies will be performed to reduce the pathogen load and spoilage bacteria in meat and meat products. The poultry meat products will also be analyzed for quality, shelf-life, and consumer acceptability of E-Beam-treated meat. Since implementing multiple steps from farm to fork maximizes risk reduction for...See attached for the complete approach.
Progress Report
Under Objective 1: In collaboration with the University of Connecticut, we completed a study evaluating the antimicrobial efficacy of ultrafine ozone bubble (UFOB) water as a natural intervention against Salmonella Enteritidis (SE) on chicken carcasses and assessed its impact on chicken meat quality and sensory attributes. Treating with UFOB water significantly reduced cross-contamination by 50%. Additionally, treatment with UFOB water resulted in ~1 log CFU/sample reduction in SE load on chicken skin compared to the control group (p < 0.05). In a parallel experiment, non-inoculated chicken breast fillets (n = 24/group) were dipped in either DI or UFOB water for 30 min at 4°C and analyzed for meat quality and sensory attributes by a trained panel. No significant differences were observed in pH value, surface color parameters (L*, a*, b*), thaw loss (~8 g), or cook loss (~90 g) following UFOB treatment of samples (p > 0.05). Descriptive sensory analysis indicated no significant differences between control and UFOB (p > 0.05) treated samples in appearance, aroma, flavor, texture, basic taste, or feeling factors, suggesting that UFOB treatment did not negatively affect the sensory quality of chicken meat. These findings demonstrate that UFOB water effectively reduces SE without negatively impacting chicken meat quality or sensory attributes, supporting its potential as a sustainable antimicrobial intervention in poultry processing.
Under Objective 2: We conducted a study, evaluating the effects of plant derived polyphenols (thymol) on biofilm formation of bacteria as well as on inhibition of biofilm formation. The highest dose of thymol (0.5%) used in this study inactivated biofilm within 10 min to below detection limits (~7 log reduction) on both polystyrene and stainless steel surfaces. The lowest dose of 0.125% thymol reduced counts significantly (3-4 logs) when treated for 1 min on polystyrene and 5 min on stainless steel. The results obtained for our study indicate that thymol could be used as an effective treatment for inhibition and inactivation of S. Infantis biofilms on polystyrene and stainless steel surfaces. These are studies are completed and the results will be presented at the Annual Poultry Science Association conference 2025.
Under Objective 3: We conducted studies to evaluate the humoral immune response to Campylobacter colonization in broiler chickens and optimizing eBeam (electron beam) strategies against Campylobacter colonization in broiler chickens. As part of these studies we determined the optimal dose of eBeam to develop a vaccine against Campylobacter for chicken To achieve this, we determined the D10 value (the dose required to reduce the bacterial population by 1 log) and the lethal dose, which is the dose required to completely inactivate the bacteria. Also, treated bacterial cultures were evaluated for membrane integrity (Bac Light staining) and cell viability (Bac Titer-Glo assay). eBeam-treated samples had a similar percentage of cells with intact cell membranes compared to the negative control, as opposed to the formalin control that presented a high percentage of damaged cells. Viability assay revealed that eBeam-treated samples exhibited increased metabolic activity compared to negative control and formalin-treated cells. In conclusion, eBeam treated cells are unable to multiply while maintaining membrane integrity and metabolic activity. These studies are still in progress.
Under Objective 4C, we have determined that the humoral immune response drives stress-related neurochemicals that directly affect Campylobacter and Salmonella carriage in poultry. The concentrations of antibodies IgA and IgY, which are part of the humoral immune response elicited by vaccines, were found to modulate neurochemical concentrations that control foodborne pathogen colonization patterns in the chicken gut throughout the production lifecycle of the bird. As a robust humoral immune response is a prerequisite of a successful vaccine, these findings provide an actionable strategy for the development of vaccines that protect poultry from foodborne pathogen carriage under real-world production conditions. Studies are ongoing evaluating the development of a chicken-specific probiotic that can be used to simultaneously target the immune-neurochemical intersection to enhance food safety at the pre-harvest stage.
In addition, we have completed studies evaluating the impact of phytochemicals on controlling the neurochemical stress response in the poultry gut under production-relevant stressors. Cinnamaldehyde and carotenoids were found to alter intestinal concentrations of stress-related neurochemicals in poultry under cold and heat stress conditions, respectively. Manuscripts reporting these results are being prepared for journal submission.
Under Sub-objective 5A, Ground poultry meats were spiked with a cocktail of 5 strains of Salmonella and 5 Campylobacter jejuni. The meats were electron beam treated with different doses: 0 kGy, 1 kGy, 2 kGy, 3 kGy, and 4 kGy, and results revealed that 2 kGy treatment reduced Salmonella and Campylobacter counts by up to 6 logs and aerobic bacterial counts (total bacteria) were below detection limits in samples treated with 4 kGy. In a separate study, ground chicken and turkey meat were purchased from a local grocery store, exposed to eBeam, and evaluated for meat quality and organoleptic properties. The studies are completed, and the data from this study is currently being analyzed for manuscript preparation.
Accomplishments
1. eBeam inactivated bacterial vaccine reduced BCO-lameness in broiler chickens. Vaccine Reduces Chicken Lameness Bacterial-induced lameness, also known as BCO, in broiler chickens is a major animal welfare concern caused by several disease-causing microorganisms, with Staphylococcus being a primary culprit. Our research successfully used electron beam (eBeam) technology to inactivate Staphylococcus bacteria, creating a vaccine. When broiler chicken embryos were vaccinated on day 18, we observed a 50% reduction in lameness in these birds compared to those not vaccinated. This represents a significant breakthrough because eBeam vaccines are more effective than traditional killed vaccines. eBeam technology uniquely preserves crucial parts of the bacteria, called epitopes, which are essential for stimulating the chicken's immune system to build protection. ARS researchers in Fayetteville, Arkansas believe this research is important to chicken farmers, animal welfare groups, and consumers. Chicken farmers care because reducing lameness means healthier flocks, which can lead to lower economic losses and more efficient production. Animal welfare organizations are interested because fewer lame chickens directly translate to improved well-being for the animals. Consumers also benefit from this research as it addresses ethical concerns regarding animal health in the food supply chain. Ultimately, the entire poultry industry is invested in this work because it offers a novel and effective solution to a widespread and costly problem, thereby enhancing both animal welfare and agricultural sustainability.
2. Immune response affects stress-related gut neurochemicals that modulate foodborne pathogens. The humoral immune response causes shifts in stress-related neurochemical concentrations in the gut that modulate foodborne pathogen carriage in poultry. Stress is known to impact vaccine efficacy in poultry, yet it is unknown why this occurs and how this can affect pre-harvest food safety. ARS researchers in Fayetteville, Arkansas demonstrated that the humoral immune response can directly effect changes in poultry stress neurophysiology that are known to modulate foodborne pathogen ability to colonize the intestinal tract. These findings provide industry stakeholders and poultry researchers an actionable platform for the development of vaccines that protect poultry from foodborne pathogen carriage under normal and stress conditions encountered during production.
3. Developed consensus methodology for microbiota sequencing to help safeguard poultry food safety. ARS researchers in Fayetteville, Arkansas developed a consensus methodology for next-generation sequencing technologies that serve as critical tools in safeguarding poultry food safety, these technologies are routinely used to trace and identify foodborne pathogenic bacteria in poultry production chain. Best practices for next generation sequencing in poultry are needed to ensure reproducibility and accuracy in practice. An interdisciplinary team uniting leading industry, academic, and government poultry researchers and stakeholders have developed an evidence-based set of guidelines that will inform the effective use by stakeholders of next—generation technologies used to identify foodborne pathogens in poultry.
Review Publications
Lyte, J.M., Assumpcao, A., Caputi, V., Ashwell, C.W., Seyoum, M., Honakaer, C.F., Daniels, K., Lyte, M., Siegel, P.B., Taylor, R. 2024. Co-evolution of the humoral immune and serotonergic systems in chickens selected for high or low blood antibody titer response to sheep red blood cells. Poultry Science. 104/104699. https://doi.org/10.1016/j.psj.2024.104699.
Perera, R., Pillai, S.D., Alrubaye, A., Jesudhasan, P. 2025. Leveraging electron beam (eBeam) technology for advancing the development of inactivated vaccines. Vaccines. 13(2). Article 179. https://doi.org/10.3390/vaccines13020179.
Lyte, J.M., Seyoum, M., Assumpcao, A., Caputi, V., Ashwell, C.M., Honaker, C.F., Daniels, K., Lyte, M., Siegel, P.B., Taylor, R.L. 2025. Multigenerational selection for high or low antibody response to sheep red blood cells modulates the chicken cecal microbiome and its relationship to the immune and serotonergic systems ¿. Poultry Science. 104. https://doi.org/10.1016/j.psj.2025.104943.
Rosell-Cardona, C., Leigh, S., Knox, E., Tirelli, E., Lyte, J.M., Martinez-Herrero, S., O'Driscoll, C.M., Goodson, M.S., Kelley-Loughnane, N., Aburto, M.R., Cryan, J.F., Clarke, G. 2025. Acute stress-induced alterations in short-chain fatty acids: Implications for intestinal and blood brain barriers. Brain Behavior and Immunity-Health. https://doi.org/10.1016/j.bbih.2025.100992.
Lyte, J.M., Seyoum, M.M., Ayala, D., Kers, J.G., Caputi, V., Johnson, T., Zhang, L., Rehberger, J., Zhang, G., Dridi, S., Hale, B., De Oliveira, J.E., Grum, D., Smith, A., Kogut, M.H., Ricke, S.C., Ballou, A., Potter, B., Proszkowiec-Wegla, M.K. 2025. Do we need a standardized 16S rRNA gene amplicon sequencing analysis protocol for poultry microbiota research? Poultry Science. 104(7). Article e105242. https://doi.org/10.1016/j.psj.2025.105242.
Anthney, A.P., Alharbi, K.S., Perera, R., Do, A., Asnayanti, A., Onyema, R., Reichelt, S., Meuter, A., Jesudhasan, P., Alrubaye, A. 2025. Evaluating the effectiveness of probiotic and multi-valent vaccination strategies in mitigating BCO lameness using a hy-brid challenge model. Animals. https://doi.org/10.3390/ani15040570.
Assumpcao, A., Jesudhasan, P., Arsi, K., Alharbi, K.S., Asnayanti, A., Trieu, A.D., Read, Q.D., Perera, R., Shwani, A., Hasan, A., Pillai, S.D., Anderson, R.C., Donoghue, A.M., Rhoads, D., Alrubaye, A. 2024. Electron beam-killed Staphylococcus vaccine reduced lameness in broiler chickens. Vaccine. https://doi.org/10.3390/vaccines12111203.
Baskaralingam, V., Jesudhasan, P. 2024. Vaccines in Aquaculture Development, Production, and Applications. Book Chapter. 1st edition. Cambridge, MA: Eelsevier. p. 300.