Skip to main content
ARS Home » Southeast Area » Fayetteville, Arkansas » Poultry Production and Product Safety Research » Research » Research Project #439695

Research Project: Multi-hurdle Approaches for Controlling Foodborne Pathogens in Poultry

Location: Poultry Production and Product Safety Research

2022 Annual Report

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 regions of the intestinal tract. 5. Utilize novel electron-beam technology to reduce pathogen prevalence on poultry products.

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.

Progress Report
Under Sub-objective 1B, we conducted studies to investigate the efficacy of phytochemicals, namely, ginger oil and garlic oil, in reducing Salmonella in postharvest poultry. The antimicrobial efficacy of these phytochemicals against Salmonella (S) Infantis on chicken skin samples was tested at the temperature of the scalding tank (54°C) for 2 minutes. There was a significant reduction (> 2 Log CFU/sample reduction) of S. Infantis surviving on the surface of the groups treated with ginger oil (1%) or garlic oil (1%) or their combinations. Studies evaluating the mechanisms of action(s) using sub-inhibitory concentration (SIC) on adhesion, quorum sensing, and gene expression analyses are underway. Under Sub-objective 2C, we have completed a study to investigate the efficacy of phytochemicals in reducing Campylobacter (C) jejuni and the response of the host (broiler chickens using multi-omics analysis. We profiled biologically relevant molecules (protein, lipids, metabolites, and exosomes) that mediate C. jejuni colonization in the intestinal tract of broilers. Based on Principal Component Analysis (PCA) and differential expression analysis, a total of 1216 analytes (275 compounds, 7 inorganics, 407 lipids, 527 proteins) were identified. The manuscript is currently under preparation. Under Sub-objectives 3A and 3B, we have completed a study investigating the efficacy of an electron beam-killed vaccine consisting of multi-serovars of Salmonella. We delivered the killed vaccine to Day-18 embryos, which induces immunity against the pathogen and eventually prevents colonization and shedding. The manuscript is currently under preparation. Under Sub-objectives 4A and 4B, we have conducted studies utilizing chickens to understand how pre-harvest heat or cold stressors affect the intestinal synthesis of stress-related neurochemicals that have been demonstrated to mediate host susceptibility to C. jejuni, Salmonella spp. and other foodborne pathogen colonization. Temperature-based stressors caused increased production of norepinephrine and serotonin in the chicken gut. In these same studies we have also utilized shallow shotgun whole genome sequencing of the gut microbiome, and identified compositional and metabolic changes in the bacterial microbiome that result from host exposure to cold stress; heat stressed chicken gut microbiome samples are currently undergoing sequencing. The manuscript is currently being prepared for submission to a peer-reviewed journal. These findings provide novel insight into how pre-harvest stressors may increase foodborne pathogen carriage by impacting poultry gut neurochemistry and the microbiome.

1. Quantitative map of neurochemical concentrations along the entire broiler chicken intestinal tract was completed. Little was known regarding the distribution and concentrations of neurochemicals in the broiler chicken intestinal tract represented a critical gap in knowledge as neurochemicals have been shown to mediate host-microbe interaction and bacterial growth. In addition, ARS researchers in Fayetteville, Arkansas, discovered that as gut neurochemistry exhibits plasticity to host stress, investigations into how pre-harvest forms of stress may increase intestinal carriage of foodborne pathogens first require an understanding of which neurochemicals are present at what concentrations in which regions of the avian intestinal tract. Our findings have been published in Poultry Science, thereby providing poultry researchers with the first neurochemical biogeography of the complete broiler chicken intestinal tract.

Review Publications
Lyte, J.M., Martinez, D.A., Robinson, K., Donoghue, A.M., Daniels, K.M., Lyte, M. 2022. A neurochemical biogeography of the broiler chicken intestinal tract. Poultry Science. 101(30). Article 101671.
Diviccaro, S., Caputi, V., Cioffi, L., Giatti, S., Lyte, J.M., Caruso, D., O'Mahony, S.M., Melcangi, R.C. 2021. Exploring the impact of the microbiome on neuroactive steroid levels in germ-free animals. International Journal of Molecular Sciences. 22(22). Article 12551.
Lyte, J.M., Koester, L., Daniels, K., Lyte, M. 2022. Distinct cecal and fecal microbiome responses to stress are accompanied by sex-and diet-dependent changes in behavior and gut serotonin. Frontiers in Neuroscience. 16. Article 827343.
Van De Wouw, M., Vigano, G.M., Lyte, J.M., Boehme, M., Gual, A., Walsh, A.M., Crispie, F., Clarke, G., Dinan, T.G., Cotter, P.D., Cryan, J.F. 2021. Kefir ameliorates specific microbiota-gut-brain axis impairments in a mouse model relevant to autism spectrum disordor. Brain Behavior and Immunity. 97:119-134.
Mancinnelli, A.C., Mattioli, S., Twining, C., Dal Bosco, A., Donoghue, A.M., Arsi, K., Chaittelli, D., Angelucci, E., Castellini, C. 2022. Poultry meat and eggs as an alternative source of n-3 long-chain polyunsaturated fatty acids for human nutrition. Nutrients. 14(9). Article 1969.
Wagle, B., Donoghue, A.M., Jesudhasan, P. 2021. Select phytochemicals reduce Campylobacter jejuni in postharvest poultry and modulate the virulence attributes of C. jejuni. Frontiers in Microbiology. 12. Article 725087.
Gupta, A., Bansal, M., Liyanage, R., Upadhyay, A., Rath, N., Donoghue, A.M., Sun, X. 2021. Sodium butyrate modulates chicken macrophages proteins essential for Salmonella Enteritidis invasion. PLoS ONE. 16(4):e0250296.
Upadhyaya, I., Arsi, K., Fanatico, A., Wagle, B.R., Shrestha, S., Upadhyay, A., Coon, C.N., Owens, C.M., Mallman, B., Donoghue, A.M. 2022. Impact of feeding bigheaded carp fish meal on meat quality and sensory attributes in organic broiler chickens. Journal of Applied Poultry Research. 31(1). Article 100224.