Research Project: Immunological and Practical Approaches to Manipulate the Ecological Niches and Reduce Foodborne Pathogens in Poultry

Location: Food and Feed Safety Research

2021 Annual Report

Objectives
Objective 1: Characterize the immune-metabolic signatures of the host:pathogen interactome following Salmonella and Campylobacter infection in the cecum that transforms host defenses from 'disease resistance' to 'disease tolerance' and mitigates long-term persistent intestinal infection in broilers chickens. Objective 2: Determine the intestinal kinome profile and antimicrobial resistance patterns associated with Salmonella and Campylobacter colonization in broiler chickens with high or low concentrations of these foodborne pathogens. Objective 3: Develop new strategies (including immunologic training) to reduce foodborne pathogens by targeting neonatal poultry gut health via stimulating the development and maturation of the intestinal immune system. Sub-objective 3.A: Deliver non-nutritional dietary feed additives to breeder chickens (hens and roosters) to induce /enhance trans-generational trained innate immunity as both a prophylactic and as a therapeutic agent in neonatal chickens against foodborne pathogens. Sub-objective 3.B: Evaluate a novel innate immune therapeutic as an alternative to antibiotics that targets innate immunity based on the ability of the immune system to detect bacterial viability through the recognition of viability-associated microbial associated molecular patterns (vita-MAMPs). Sub-objective 3.C: Develop a novel immunogenic probiotic to improve neonatal poultry gut health and stimulate the development and maturation of the intestinal immune system. Objective 4: Develop intervention strategies and alternatives to antibiotics to reduce the bacterial load of foodborne pathogens during pre-harvest poultry production. Objective 5: Investigate the interaction between yeast and fungi and foodborne bacteria to determine their role as commensals, inhibitors, or their use as alternatives to antibiotics as pre-and probiotics.

Approach
Despite control efforts, foodborne illnesses due to Salmonella continues to impact the consumer. Poultry are commonly identified as a major source of Salmonella. We will take a multi-faceted approach to develop urgently needed new control strategies against Salmonella, and our integrated approaches will identify and evaluate applied and basic pre-harvest approaches to reduce Salmonella in poultry. Based on previous research and collaborations with industry, we will identify and/or modify management practices to reduce foodborne pathogen load and will address environmental conditions associated with higher risks that enable pathogen survival and growth. Direct and indirect modulation of gut microbiome-innate immune interactome will be evaluated to determine which natural host mechanisms can be exploited to strengthen therapeutic benefit. Modulating the innate immune response has considerable potential to induce a profound and rapid cross-protection against multiple serovars of foodborne pathogens. Salmonella have evolved a unique survival strategy in poultry that minimizes host defenses during the initial infection and then exploits and/or induces a dramatic immunometabolic reprogramming in the cecum that alters the host defense to induce a disease tolerance. Therefore, a component of the research will concentrate on modulatory mechanisms (vaccination, pro- and postbiotic) to counter these reprogramming mechanisms and lead to the development of novel immunometabolic therapeutic strategies. This research will enhance the microbiological safety of poultry and reduce potential antimicrobial resistance in animal agriculture and public health.

Progress Report
This is a new project that replaced the expired project 3091-32000-035-00D. Work under this new project in Fiscal Year (FY) 2021 provided the first experimental evidence that segmented filamentous bacteria (SFB), may be a useful probiotic to improve chicken health in early life. SFB are a keystone taxon that intimately binds to the intestine of the chicken. Project work evaluated the probiotic potential of SFB and found that it significantly reduced Salmonella load in chickens in an Immunoglobulin A (IgA) independent manner. IgA is an antibody blood protein that is part of the poultry immune system. Additionally, SFB enhanced several immunometabolic processes. Basically, oral inoculation with SFB significantly improved gut health (Objective 3). Work on Objectives 2 and 3 identified crucial elements of the intestinal mucosal immune system that enhance the ability of birds to fight off pathogenic microbes while maintaining a state of tolerance to the diverse and beneficial intestinal microbes. Under Objective 4, routes by which Salmonella infect turkeys were identified; the findings have significant food safety implications in commercial turkey production. This research is of high interest to the poultry industry given its promise in developing pre-harvest intervention strategies that can be incorporated into commercial turkey production to enhance microbial food safety.

Accomplishments
1. Infection routes of foodborne pathogens in turkeys. As with other poultry species reared commercially for food production, growing turkeys are susceptible to colonization by a number of bacterial species that can cause food poisoning in humans if the meat products are contaminated during processing. The turkey industry is in critical need of effective strategies to assure that turkey food products reaching the consumer are microbiologically safe. ARS researchers at College Station, Texas, working closely with key turkey industry scientists and managers, established the various routes by which the growing birds can be infected by Salmonella, one of the more important problematic bacteria. This accomplishment is important because understanding how growing turkeys can be infected by dangerous microorganisms is critical to the development of new technology and/or management strategies to assure microbiological safety of turkey food products. The industry and relevant U.S. regulatory agencies are well aware of and highly supportive of this work.

2. Beneficial bacteria protect young chickens. Harmful bacteria such as Salmonella, Campylobacter, and Listeria are well known to infect/colonize the intestinal tract of commercially grown poultry, and ultimately can cause food poisoning in people if the bacteria contaminate meat products during processing. There is ongoing need for new technology that can be economically implemented by the poultry industry to minimize or prevent such infections. ARS researchers at College Station, Texas, discovered that segmented filamentous bacteria (SFB), a class of harmless microorganisms, are effective when administered orally to young chickens in preventing subsequent colonization by Salmonella. The apparent mode of action is that SFB binds intimately with the intestinal wall of the living chickens and thus occupies the space or binding sites that Salmonella or other harmful bacteria might otherwise occupy. This accomplishment is important because it represents discovery of a new approach to bird health and food safety that may be implementable in commercial poultry production in a cost-effective manner.

Review Publications
Infante-Rodriguez, F.M., Domínguez Muñoz, M.A., Montaño-Gómez, M.F., Hume, M.E., Anderson, R.C., Manríquez-Nuñez, O.M., López-Acevedo, E.A., Bautista-Martinez, Y., Salinas-Chavira, J. 2020. Effect of protein concentrations in the diet on productive performance, carcass characteristics, and meat chemical composition of broiler chickens in the dry subtropics. Nova Scientia. 12(25). https://doi.org/10.21640/ns.v12i25.2585.
Redweik, G.A., Kogut, M.H., Arsenault, R.J., Mellata, M. 2020. Oral treatment with ileal spores triggers immunometabolic shifts in chicken gut. Frontiers in Veterinary Science. 7: Article 629. https://doi.org/10.3389/fvets.2020.00629.
Lee, A., Bortoluzzi, C., Pilla, R., Kogut, M.H. 2020. A role for the microbiota in the immune phenotype alteration associated with the induction of disease tolerance and persistent asymptomatic infection of Salmonella in the chicken. Microorganisms. 8(12). Article 1879. https://doi.org/10.3390/microorganisms8121879.
Bortoluzzi, C., Layahe, L., Perry, F., Arsenault, R.J., Santin, E., Korver, D., Kogut, M.H. 2021. A protected complex of biofactors and antioxidants improved growth performance and modulated the immunometabolic phenotype of broiler chickens undergoing early life stress. Poultry Science. 100(7). Article 101176. https://doi.org/10.1016/j.psj.2021.101176.
Jesudhasan, P., Bhatia, S.S., Sivakumar, K.K., Praveen, C., Genovese, K.J., He, L.H., Droleskey, R.E., McReynolds, J.L., Byrd II, J.A., Swaggerty, C.L., Kogut, M.H., Nisbet, D.J., Pillai, S.S. 2021. Controlling the colonization of Clostridium perfringens in broiler chickens by an electron-beam-killed vaccine. Animals. 11(3). Article 671. https://doi.org/10.3390/ani11030671.
Dal Pont, G.C., Belote, B.L., Lee, A., Bortoluzzi, C., Eyng, C., Sevastiyanova, M., Khadem, A., Santin, E., Farnell, Y.Z., Gougoulias, C., Kogut, M.H. 2021. Novel models for chronic intestinal inflammation in chickens: Intestinal inflammation pattern and biomarkers. Frontiers in Immunology. 12. Article 676628. https://doi.org/10.3389/fimmu.2021.676628.
Moreno-Mendoza, Y., López-Villarreal, K.D., Hernández-Martínez, C.A., Rodríguez-Tovar, L.E., Hernández-Coronado, A.C., Soto-Domínguez, A., Hume, M.E., Méndez-Zamora, G. 2021. Effect of moringa leaf powder and agave inulin on performance, intestinal morphology, and meat yield of broiler chickens. Poultry Science. 100(2):738-745. https://doi.org/10.1016/j.psj.2020.11.058.
Pineda, M., Kogut, M.H., Genovese, K.J., Farnell, Y.Z., Zhao, D., Wang, X., Milby, A., Farnell, M. 2021. Competitive exclusion of intra-genus Salmonella in neonatal broilers. Microorganisms. 9(2). Article 446. https://doi.org/10.3390/microorganisms9020446.