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Campylobacter jejuni is the leading cause of bacterial gastroenteritis in the United States with an estimated cost of treatment and loss of productivity at $1 billion, annually. The clinical symptoms include fever, severe abdominal cramps, and diarrhea that can be bloody. Complications include arthritis and Guillain-Barre Syndrome, a neurological disorder leading to paralysis. The major risk factor in acquiring campylobacteriosis is the handling and consumption of contaminated poultry. Recent work has demonstrated the presence of C. jejuni in biofilms colonizing the nipple drinkers of poultry houses. Biofilms are communities of microorganisms held within a slimy matrix attached to a surface such as metal or plastic and therefore, offer a continuous source of infection.
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There are four objectives to this research with the overall objective being to define how the biofilm formation of C. jejuni in poultry water systems affect the colonization of commercial broiler chickens. The first objective relates to determining the relationship of C. jejuni found in poultry house water systems to the isolates that colonize commercial broilers. The second objective aims at determining the type of bacterial populations that comprise the biofilms formed in the watering systems of broiler houses and how they contribute to the colonization of poultry. The third objective aims at testing various surfaces and chemical sanitizers and determining which ones minimize biofilm formation and the final objective relates to offering extension services to integrate research personnel and poultry producers and management to concur on practical intervention strategies to decrease the load of C.jejuni in poultry.
NON-TECHNICAL SUMMARY: Campylobacter jejuni is the leading cause of bacterial gastroenteritis in the United States with an estimated cost of treatment and loss of productivity at $1 billion, annually. Symptoms include fever, severe abdominal cramps, and diarrhea and complications include arthritis and Guillain-Barré Syndrome, a neurological disorder leading to paralysis. The major risk factor in acquiring campylobacteriosis is the handling and consumption of contaminated poultry and recent work has demonstrated the presence of C. jejuni in biofilms colonizing the nipple drinkers of poultry houses. Biofilms are communities of microorganisms held within a slimy matrix attached to a surface and therefore, offer a continuous source of infection. The objective of this study is to define the role of C. jejuni in the formation of biofilms and to identify the various bacterial species in biofilms from poultry houses. Expected results will determine the contribution of C. jejuni as well as the contribution of other pathogens of concern, such as Salmonella spp. to biofilms and the subsequent colonization of poultry. Various surfaces such as polyvinyl chloride and sanitizers such as surfactants will be tested to determine methods to prevent biofilm development. We will work with industry at the corporate and association levels to develop intervention strategies. Ultimately, effective control strategies will be implemented that will reduce the C. jejuni load in poultry products and therefore, lead to a considerable decline in human illness.
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APPROACH: Cecal droppings from the center isle of poultry houses will be sampled to identify 4 positive flocks for C. jejuni from 3 different states. From these flocks, biofilm samples will be collected on day 42 of the grow-out period via scraping of the drinking nipples (n=5) and cultured for biofilm producers. Ceca from the broilers will be swabbed at evisceration to isolate C. jejuni. C. jejuni isolated from the biofilms and the cecas will be subjected to MRP-PFGE via SmaI and MLST and if necessary, to short variable region flagellin (SVR flaA) gene sequencing to determine their genetic relatedness. Along with culture analysis of the bacterial biofilms from broiler houses that had broilers colonized with C. jejuni, culture analysis of bacterial biofilms will also be performed from broiler houses not colonized with C. jejuni to determine if certain biofilm formers can exclude C. jejuni from biofilms. R2A agar will be incubated at various temperatures and atmospheres and the DNA will be extracted from the bacteria and the organisms identified (e.g., genus and species) by 16S rRNA gene amplification and DNA sequencing. Surfaces and chemical sanitizers will be identified that minimize biofilm formation. Abiotic surfaces of PVC, ABS, high density polyethylene (HDPE), galvanized steel, stainless steel, and copper will be examined for C. jejuni biofilm formation using a virulent poultry isolate, an avirulent poultry isolate and a clinical control. Chemical sanitizers to be tested include surfactants, acidic and alkaline solutions, sodium hypochlorite and iodophors using these same isolates and PVC as the abiotic surface. These studies should help define surfaces and sanitizers that are effective in the reduction of the C. jejuni load in biofilms. Intervention steps are essential in the reduction of colonization of C. jejuni and other pathogens of concern such as Salmonella spp. However, due to economic concerns, practical intervention strategies that would be accepted by the poultry industry need to be developed. In order to accomplish this goal, once the initial results including the make up of the bacterial communities of biofilms are available, sequential live interactive webcasts will be scheduled using the University of Illinois Meeting Place web conferencing capabilities to set up meetings between the participating producers, the company that owns the poultry that the producers are contracted to (as the chicken industry is vertically integrated), representatives from the US Poultry and Egg Association, and the researchers to collaborate on what can or cannot be implemented. These results will be used to create factsheets and lesson plans and publicized to state/national meat and poultry regulators/inspectors, extension agents and specialists, and the poultry industry.