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ARS Home » Southeast Area » Athens, Georgia » U.S. National Poultry Research Center » Egg and Poultry Production Safety Research Unit » Research » Publications at this Location » Publication #354755
Research Project: Reduction of Invasive Salmonella enterica in Poultry through Genomics, Phenomics and Field Investigations of Small Multi-Species Farm Environments

Location: Egg and Poultry Production Safety Research Unit

Title: Semi-quantification of total Campylobacter and Salmonella during egg incubations using a combination of 16S rDNA and specific pathogen primers for qPCR

Author
item Rothrock, Michael
item FEYE, KRISTINA - University Of Arkansas
item KIN, SUN AE - University Of Arkansas
item PARK, SI HONG - Oregon State University
item LOCATELLI, AUDE - Oak Ridge Institute For Science And Education (ORISE)
item HIETT, KELLI - Food And Drug Administration(FDA)
item GAMBLE, JOHN - Oregon State University
item SELLERS, HOLLY - University Of Georgia
item RICKE, STEVEN - University Of Arkansas

Submitted to: Frontiers in Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/25/2018
Publication Date: 11/2/2018
Citation: Rothrock Jr, M.J., Feye, K.M., Kin, S., Park, S., Locatelli, A., Hiett, K., Gamble, J., Sellers, H., Ricke, S.C. 2018. Quantification of total Campylobacter and Salmonella during egg incubations using a combination of 16S rDNA and specific pathogen primers for qPCR. Frontiers in Microbiology. 9:2454.Doi: 10.3389/fmicb.2018.02454

Interpretive Summary: Rapid molecular techniques that evaluate eggs for the presence of food-borne pathogens is an essential component to poultry food safety monitoring. Interestingly, it is not just table eggs that contribute to outbreaks of food-borne disease. Broiler layer production actively contributes to the sustainment of food-borne pathogens within a flock. Mainly, the surface contamination of production eggs with invasive pathogens such as Salmonella enterica, Campylobacter jejuni, and Listeria monocytogenes during embryogenesis results in gastrointestinal tract (GIT) colonization. Pathogens that are able to secure a niche within the GIT during the embryonic stage of chick development are nearly impossible to eradicate. Therefore, the current monitoring paradigms are not comprehensive as they fail to capture the presence of invasive pathogens within the embryonic GIT. By doing so, producers are not able to truly spot evaluate eggs for their potential risk as carriers for food-borne pathogens. In this study a novel qPCR assay was developed that quantifies pathogen load relative to total bacterial burden. Eggs sampled from three independent production broiler flocks of different ages were assayed for Salmonella enterica, Campylobacter jejuni, and Listeria monocytogenes. Within flock, eggs were sampled at one-day post-set, two weeks post-set, after vaccination (at 2.5 weeks) and one-day post-hatch. The eggs were washed, and the yolk and embryonic chick gastrointestinal tract were collected. The DNA was extracted and subjected to a qPCR assay. The results confirm a novel technique for pathogen monitoring relative to total bacterial load and a unique method for monitoring the dynamics of food-borne pathogen invasion throughout broiler egg production

Technical Abstract: Rapid molecular techniques that evaluate eggs for the presence of food-borne pathogens is an essential component to poultry food safety monitoring. Interestingly, it is not just table eggs that contribute to outbreaks of food-borne disease. Broiler layer production actively contributes to the sustainment of food-borne pathogens within a flock. Mainly, the surface contamination of production eggs with invasive pathogens such as Salmonella enterica, Campylobacter jejuni, and Listeria monocytogenes during embryogenesis results in gastrointestinal tract (GIT) colonization. Pathogens that are able to secure a niche within the GIT during the embryonic stage of chick development are nearly impossible to eradicate. Therefore, the current monitoring paradigms are not comprehensive as they fail to capture the presence of invasive pathogens within the embryonic GIT. By doing so, producers are not able to truly spot evaluate eggs for their potential risk as carriers for food-borne pathogens. In this study a novel qPCR assay was developed that quantifies pathogen load relative to total bacterial burden. Eggs sampled from three independent production broiler flocks of different ages were assayed for Salmonella enterica, Campylobacter jejuni, and Listeria monocytogenes. Within flock, eggs were sampled at one-day post-set, two weeks post-set, after vaccination (at 2.5 weeks) and one-day post-hatch. The eggs were washed, and the yolk and embryonic chick gastrointestinal tract were collected. The DNA was extracted and subjected to a qPCR assay. The results confirm a novel technique for pathogen monitoring relative to total bacterial load and a unique method for monitoring the dynamics of food-borne pathogen invasion throughout broiler egg production.