Location: Poultry Microbiological Safety and Processing Research Unit
2019 Annual Report
Objectives
Objective 1. Develop reliable and reproducible challenge models with Salmonella and Campylobacter for use in accurately developing, evaluating, and validating processes for reducing pathogen load using various chemical sanitizers.
Objective 2. Develop, evaluate, and validate current and novel chemicals, operational protocols, and sampling methodologies used during poultry production and processing of broilers for the reduction and/or control of foodborne pathogens.
Sub-objective 2.1. Assess the ability of commercial and novel chemical sanitizers to reduce or eliminate Salmonella, Campylobacter, and Pseudomonas species from inoculated broiler carcasses and parts.
Sub-objective 2.2. Examine the effectiveness of chemical sanitizers applied to carcasses before defeathering or before chilling to reduce contamination by Salmonella and Campylobacter carcasses in postchill carcasses.
Sub-objective 2.3. Formulate novel microbicidal surfactants from mixtures of medium-chain fatty acids (MCFA) and organic acids (OA) to be used as sanitizers to significantly reduce microbial contamination during poultry processing.
Objective 3. Identify and evaluate risk factors in the production, management, transportation, or processing that impact bird/egg contamination with foodborne pathogens and develop intervention strategies to control pathogens in the absence of antibiotics.
Sub-objective 3.1. Assess the ability of chemical sanitizers to reduce contamination of inoculated, fertile eggs by Salmonella.
Sub-objective 3.2. Identify and evaluate risk factors in the production, management, transportation, or processing that impact broiler contamination with foodborne pathogens and develop intervention strategies to control pathogens in the absence of antibiotics.
Objective 4. Determine the extrinsic factors that impact the survival and attachment of microorganisms including evaluating media and growth factors. Develop and validate new improved technologies to isolate and propagate foodborne pathogens.
Sub-objective 4.1. Evaluate media and growth factors and use the findings to develop new, improved technologies for the isolation and propagation of Campylobacter.
Sub-objective 4.2. Assess accuracy of current laboratory methods in recovering Salmonella from poultry, animal feeds, and dry environmental samples with fermentable substrates available and development of a more efficient pre-enrichment media.
Approach
Poultry products contaminated by Salmonella and Campylobacter continue to be major sources of human foodborne illnesses. Live poultry are sporadically colonized by these pathogens, and the birds may serve as reservoirs for the bacteria without displaying any signs of illness or declines in performance. Cross contamination of carcasses during processing may spread the bacteria to poultry meat which may cause foodborne infections if the meat is not properly handled. Therefore, the primary goal of our research will be to develop novel interventions that may be used by commercial poultry producers and processors to reduce contamination of poultry by Salmonella, Campylobacter, and indicator microorganisms. Novel chemical sanitizers that may be used during processing to reduce carcass contamination by foodborne pathogens will be formulated or identified. Factors that enhance survival of these pathogens will be identified and used to formulate a novel bacteriological medium that will be utilized in research projects to determine the efficacy of currently available and newly developed interventions. The project outcomes will result in additional control measures that will reduce the levels of Salmonella and Campylobacter in broiler flocks and reduce contamination of processed carcasses by these pathogens. These outcomes will enable the poultry industry to achieve Food Safety Inspection Service (FSIS) performance standard goals and to reduce the number of cases of human foodborne illness associated with contaminated poultry products. Research goals will be achieved by utilizing an interdisciplinary approach that incorporates knowledge and skills of the scientists and other scientists who possess skills and resources required to successfully compete this project.
Progress Report
ARS researchers in Athens, Georgia, continued to conduct research on new methods to reduce the contamination of processed poultry meat by human, bacterial pathogens. Studies were conducted on evaluating vegetable oils, such as olive, canola and corn oils as carriers to inoculate Salmonella onto feathered carcasses. The oils may aid in the attachment of pathogens to carcasses during challenge studies. Research was conducted on a new procedure utilizing antimicrobial broiler carcass washes to reduce Campylobacter contamination of processed poultry meat. Findings demonstrated that dipping poultry meat in antimicrobial chemical solutions in a specific, sequential order for 6 seconds each, produced significant reduction in contamination of the meat by this pathogen. Additional studies were conducted on the ability of carcass washing to reduce bacterial contamination of broiler carcasses between slaughter and evisceration operations in the pilot-plant poultry processing facility. These, studies indicated that washing the carcasses with water under high pressure could produce 1 to 2 log reductions in contamination by Campylobacter and other bacteria. Furthermore, in vitro studies were conducted to examine the antibacterial activity of medium-chained-fatty acids (MCFA) dissolved in amino acid solutions containing other compounds. Disc diffusion assays were conducted with the MCFA-amino acids and indicated that some of the mixtures could inhibit the growth of pathogenic bacteria. Studies were conducted to determine methods for increasing the recovery of Salmonella from eggs shells 1, 6, and 24 hours after inoculating. Recovery of the bacterium from eggs shells inoculated with the pathogen was enhanced by the partial removal of the eggshell cuticle (prior to inoculation), by complete removal of albumen and yolk (prior to inoculation), or by inoculation through the shell into the air cell (avoiding direct contact with the cuticle). Other studies were conducted on methods to reduce the contamination of broiler litter by Salmonella by adding alum powder to recycled litter and providing broilers pro-biotics in the drinking water. Findings indicated that both methods could reduce the number of Salmonella recovered from the litter and from the ceca of broiler chickens. Research was conducted on using a newly formulated medium containing pyruvate and carrageenan to grow Campylobacter in containers without the creating an artificial, microaerobic atmosphere. Progress was also made on the use of Clustered-Regularly-Interspaced-Short-Palindromic-Repeats (CRISPR) technology to detect broiler contamination by Salmonella. CRISPR detected significantly more types of Salmonella on the carcasses.
Accomplishments
1. Exsanguination during euthanasia does not accelerate the time to death in broiler chickens. ARS Researchers in Athens, Georgia conducted research to determine if exsanguination during poultry processing would assist a more humane form of euthanasia. Results demonstrated that immediately disrupting blood flow in both carotid arteries in the neck did not accelerate the time to death, and thus would not improve animal welfare. These findings will impact slaughter procedure used by commercial poultry processors.
Review Publications
Jacobs, L., Bourassa, D.V., Harris, C.E., Buhr, R.J. 2019. Euthanasia: Manual versus mechanical cervical dislocation for broilers. Animals. 9:47. https://doi.org/10.3390/ani9020047.
Berrang, M.E., Meinersmann, R.J., Cox Jr, N.A., Thompson, T.M. 2018. Multi-locus sequence subtypes of Campylobacter detected on the surface and from internal tissue of retail chicken livers. Journal of Food Protection. 81(9):1535-1539. https://doi.org/10.4315/0362-028X.JFP-18-131.
Berrang, M.E., Meinersmann, R.J., Cox Jr, N.A. 2017. Passage of Campylobacter jejuni and Campylobacter coli subtypes through 0.45 and 0.65 µm pore size nitro-cellulose filters. Journal of Food Protection. 80(12):2029-2032. https://doi.org/10.4315/0362-028X.JFP-17-211.
Cox Jr, N.A., Cosby, D.E., Thippareddi, H., Ritz, C.W., Berrang, M.E., Jackson, J.S., Mize, S.C., Kumar, S., Howard, A.K., Rincon, A.M., Ukidwe, M., Landrum, M., Frye, J.G., Plumblee Lawrence, J.R., Hiott, L.M., Jackson, C.R., Hinton Jr, A., Cook, K.L. 2018. Incidence, species and antimicrobial resistance of naturally occurring Campylobacter isolates from quail carcasses sampled in a commercial processing facility. Journal of Food Safety. 2018:38:e12438. https://doi.org/10.1111/jfs.12438.
Adhikari, P., Cosby, D.E., Cox Jr, N.A., Franca, M.S., Williams, S.M., Gogal Jr, R.M., Ritz, C.W., Kim, W.K. 2018. Effect of dietary fructooligosaccharide supplementation on internal organs Salmonella colonization, immune response, ileal morphology and ileal immunohistochemistry in laying hens challenged with Salmonella Enteritidis. Poultry Science. 97:2525-2533. https://doi.org/10.3382/ps/pey101.
Landrum, M.A., Cox Jr, N.A., Cosby, D.E., Berrang, M.E., Hinton Jr, A., Mize, S.C., Jackson, J.S. 2018. Reduction of Campylobacter on chicken livers using a low acid processing aid. Advanced Food and Nutritional Sciences. 3:1-6.
Oladeinde, A., Cook, K.L., Orlek, A., Zock, G.S., Herrington, K., Plumblee Lawrence, J.R., Hall, M.C., Cox Jr, N.A. 2018. Hotspot mutations and ColE1 plasmids contribute to the fitness of Salmonella Heidelberg in poultry litter. PLoS One. https://doi.org/10.1371/journal.pone.0202286.
Adhikari, P., Lee, C.H., Cosby, D.E., Cox Jr, N.A., Kim, W.K. 2018. Effect of probiotics on fecal excretion, colonization of internal organs and immune gene expression in the ileum of laying hens challenged with Salmonella Enteritidis. Poultry Science. 97(1):2525-2533. https://doi.org/10.3382/ps/pey443.
Oakley, B., Calloway, K., Richardson, E., Meinersmann, R.J., Cox Jr, N.A., Berrang, M.E. 2018. The cecal microbiome of commercial broiler chickens varies significantly by season. Poultry Science. 97:3635-3644. https://doi.org/10.3382/ps/pey214.
Cox Jr, N.A., Cosby, D.E., McLendon, B.L., Wilson, J.L., Berrang, M.E., Hinton Jr, A. 2018. Persistence of Campylobacter and Salmonella in the ceca, spleen and liver/gallbladder of inoculated broilers. International Journal of Poultry Science. 17(8):374-377.
Richardson, K.E., Cox Jr, N.A., Cosby, D.E., Berrang, M.E., Holcombe, N.L., Weller, C.E. 2019. Dry and heat stress affects H2S production of Salmonella on selective plating media. Journal of Environmental Science and Health. 54:313-316.
Hinton Jr, A., Gamble, G.R., Berrang, M.E., Buhr, R.J., Johnston, J.J. 2019. Development of neutralizing buffered peptone water for salmonella verification testing in commercial poultry processing facilities. Journal of Food: Microbiology, Safety, and Hygiene. 10:359.
