PREVALENCE, PERSISTENCE, AND VIRULENCE POTENTIAL OF FOODBORNE PATHOGENS FROM RETAIL FOOD SYSTEMS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: TERMINATED
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1008443
• Project No.: IND011661
• Project Start Date: Dec 1, 2015
• Project End Date: Sep 30, 2020

Project Director
Oliver, HA.

Recipient Organization
PURDUE UNIVERSITY
(N/A)
WEST LAFAYETTE,IN 47907

Performing Department
Food Science

Non Technical Summary
Foodborne pathogens remain concerns due to their ability to cause severe human disease and their common presence in environments from farm to fork. USDA-ERS estimate costs resulting from listeriosis and salmonellosis to exceed $2.8M and $3.7M, respectively (USDA-ERS, 2014); these estimates do not include production costs (e.g., routine testing, recalls) or other cost associated with prevention. There has been no significant reduction in the number of foodborne illness cases caused by major pathogens (e.g. Salmonella) compared to baseline data collected in 2006 (CDC, 2013). This trend indicates that lesser known food vehicles or food handling environments may be responsible for more cases of foodborne disease than currently estimated.The most recent estimates of human foodborne disease indicate that non-typhoidal Salmonella serotypes cause approx. 1 million cases resulting in 378 deaths and 11% of the overall burden of foodborne disease in the United States (Scallan et al., 2011). While consumption of undercooked poultry and poultry products or foods cross-contaminated with raw poultry and/or products remains among the most likely sources of foodborne salmonellosis (Gould et al., 2013), Salmonella is also estimated to cause approximately 50% of produce-associated illnesses and be responsible for the majority of produce-associated outbreaks in the US (Strawn, et al., 2014).

Animal Health Component

0%

Research Effort Categories

Basic

20%

Applied

80%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
712	4010	1100	100%

Knowledge Area
712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins;

Subject Of Investigation
4010 - Bacteria;

Field Of Science
1100 - Bacteriology;

Keywords

listeria

salmonella

retail

produce

food safety

Goals / Objectives
The objectives of this project are to:Objective I. Elucidate foodborne pathogen contamination patterns and persistence in retail food systemsObjective II. Develop, implement, and test practical and feasible control strategies in select establishments to (i) foodborne pathogens in retail food systems and (ii) reduce cross-contamination.Objective III. Assess virulence potential of foodborne pathogens from retail food systems.Our long-term goal is to foundational data that will (i) assist in the identification of food safety hazards in produce at retail and (ii) elucidate effective and practice mitigation strategies to control foodborne pathogens in retail food systems.

Project Methods
Isolation of foodborne pathogens from environmental samples. Versions of the U.S. Food and Drug Association Bacterial Analytical Manual method will be used to detect and isolate foodborne pathogens from environmental samples (www.cfsan.fda.gov/~ebam/bam-10.html). Designated sites will be swabbed with pre-moistened sterile sponges.Whole Genome Sequencing. DNA will be extracted from retail produce environment pathogenic isolates. DNA will be prepared for sequencing using the Nextera XTDNA sample preparation kit and Nextera XT index kit with 96 indices (Illumina, Inc., San Diego, CA). Pooled samples will be sequenced using a HiSeq2500 rapid run with 2-151-bp paired-ends.Pulsed field gel electrophoresis (PFGE). PFGE typing will be performed using the standardized CDC PulseNet protocol (http://www.cdc.gov/pulsenet/protocols/pulsenet_listeria_protocol%20.pdf). DNA from isolates will be digested separately with the restriction enzymes which act as "molecular scissors" to yield DNA fingerprint patterns. PFGE patterns will be analyzed and compared using Bionumerics software (Applied Maths, Austin, TX). Differences and similarities among PFGE patterns will provide insight into transmission patterns of foodborne pathogens isolated from food and processing/handling environments.Single nucleotide polymorphism (SNP) genotyping. A multiplex SNP genotyping assay (SNaPshot Multiplex SNP Genotyping Kit; Applied Biosystems, Foster City, CA) will be used to determine the proportion of L. monocytogenes isolated from produce and other plant-based products and the retail environment that pose reduced human health risk. Briefly, the SNaPshot assay identifies changes (mutations) in DNA sequences that lead to nonfunctional proteins (typically virulence factors).

Progress 12/01/15 to 09/30/20

Outputs
Target Audience:Food industry food safety professionals, academics, and regulatory agencies. Changes/Problems:Covid-19 significantly impacted research progress in the lab/on campus beginning in March 2020. Graduate students continued to make progress drafting and completing manuscripts, thesis, and dissertations. What opportunities for training and professional development has the project provided?One MS student completed her degree. Three PhD students are currently working on the stated Objectives of this project in addition to two undergraduate research assistants. How have the results been disseminated to communities of interest?Results of this project were presented at the International Association for Food Protection annual meeting (online) and in peer reviewed publications. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? In 2020, we continued to explore solutions to foodborne pathogens in retail. Data were collected to enhance a model sampling plan the retail grocery stores can use to identify stores among their operations that require additional food safety interventions (Obj I, II). No research was conducted toward Obj III. Manuscripts were submitted for peer review during research delays due toCOVID-19.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: S. T. Wu, J. L. Burnett, J. Wang, S. R. Hammons, D. R. Veenhuizen, and H. F. Oliver*. 2020. Infrastructure, sanitation, and management practices impact Listeria monocytogenes prevalence in retail grocery produce environments. Food Control. 109. doi.org/10.1016/j.foodcont.2019.106911
• Type: Journal Articles Status: Published Year Published: 2020 Citation: A. Boonmee, H. F. Oliver+, and S. Chaturongakul. Submitted. Listeria monocytogenes ?A is sufficient to survive gallbladder bile exposure. Front Microbiol
• Type: Journal Articles Status: Published Year Published: 2019 Citation: 3. A. J. Etter, A. M. West, J. L. Burnett, S. T. Wu, D. R. Veenhuizen, R. A. Ogas, and H. F. Oliver*. 2019. Salmonella enterica ser. Heidelberg food isolates associated with a salmonellosis outbreak have enhanced stress tolerance capabilities. Appl Environ Microbiol. 85(16).

Progress 10/01/18 to 09/30/19

Outputs
Target Audience:Food retailers, food microbiologists, and government agencies interested in retail food safety. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two graduate students were train in the lab working directly on the objectives of this project as well as two undergraduates. How have the results been disseminated to communities of interest?Presentations were made at IAFP by Dr. Oliver and students; peer-reviewed manuscripts were published in high impact journals. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The Oliver lab continued to address each of the objectives stated in this project as evidenced by publishedpeer-reviewed manuscripts targeting each aim.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: A. Boonmee, H. F. Oliver, and S. Chaturongakul. 2019. Listeria monocytogenes ?A is sufficient to survive gallbladder bile exposure. Front Microbiol.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: H. Jo, A. M. West, P. J. Teska, H. F. Oliver+, and J. Howarter. 2019. Assessment of early onset surface damage from accelerated disinfection protocol. Antimicro Resis Infect Control. 8(1): 24. doi: 10.1186/s13756-019-0467-9
• Type: Journal Articles Status: Published Year Published: 2019 Citation: S. T. Wu, S. R. Hammons, J. Wang, C. Assisi, B. DiPietro, and H. F. Oliver*. 2019. Predictive risk models combined with employee- and management-implemented interventions identified and reduced Listeria monocytogenes prevalence in retail delis. Food Control.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: S. T. Wu, S. R. Hammons, R. Silver, J. A. Neal, and H. F. Oliver*. Submitted. Retail deli managers and associates have better food safety culture in stores with lower L. monocytogenes contamination.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: A. J. Etter, A. M. West, J. L. Burnett, S. T. Wu, D. R. Veenhuizen, R. A. Ogas, and H. F. Oliver*. 2019. Salmonella enterica ser. Heidelberg food isolates associated with a salmonellosis outbreak have enhanced stress tolerance capabilities. Appl Environ Microbiol.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: A. M. West, P. J. Teska, and H. F. Oliver*. 2019. There is no additional bactericidal efficacy of EPA-registered disinfectant towelettes post-surface drying or beyond label contact time. Amer J Infect Control. 47(1), 27-32. doi: 10.1016/j.ajic.2018.07.005.

Progress 10/01/17 to 09/30/18

Outputs
Target Audience:Retail food safety experts, government regulators, ris assessors, consumers, other food safety experts. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?The results from a major retail produce study will be submitted in early 2019 What do you plan to do during the next reporting period to accomplish the goals?1. Submit 3 manuscripts on retail produce food safety 2. Graduate one PhD student supporting this work 3. Communicate major research finding to retail stakeholders and offer direction interaction sessions with food safety management personnel.

Impacts
What was accomplished under these goals? Objective I.We have elucidatedListeria andSalmonellacontamination patterns and persistence in retail produce systems in 30 stores in 6 states. Objective II. We have developed, implemented, and tested practical and feasible control strategies in retail produce departmentsto reduce foodborne pathogens and cross-contamination in collaboration with retail food safety experts. Further, we have conducted numerous studies on disinfectant efficacy that have ramifications for food and healthcare systems. Objective III. We sequenced Listeria genomes from retail produce systems isolates

Publications

• Type: Journal Articles Status: Accepted Year Published: 2018 Citation: A. M. West, P. J. Teska, and H. F. Oliver. 2018. There is no additional bactericidal efficacy of EPA-registered disinfectant towelettes post-surface drying or beyond label contact time Infect Control Hosp Epidem. doi: 10.1016/j.ajic.2018.07.005.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: R. E. Novoa, M. Bailey, D. R. Jones, R. K. Gast, K. Anderson, J. Brar, R. Taylor, H. F. Oliver, M. Singh. 2018. Prevalence, persistence, and antimicrobial resistance of Campylobacter spp. from eggs and laying hens housed in five commercial housing systems. Foodborne Pathog Dis. 15 (8):506-516. doi: 10.1089/fpd.2017.2404.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: A. M. West, P. J. Teska, C. B. Lineback, and H. F. Oliver. 2018. Strain, disinfectant type, concentration, and contact time quantitatively impact disinfection efficacy. Antimicro Resis Infect Control. 7:49. doi: 10.1186/s13756-018-0340-2.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Lineback CB, Nkemngong CA, Wu ST, Li X, Teska PJ, Oliver HF. Hydrogen peroxide and sodium hypochlorite disinfectants are more effective against Staphylococcus aureus and Pseudomonas aeruginosa biofilms than quaternary ammonium compounds. Antimicrob Resist Infect Control. 2018 Dec 17;7:154. doi: 10.1186/s13756-018-0447-5.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Bailey MA, Taylor RM, Brar JS, Corkran SC, Vel�squez C, Novoa Rama E, Oliver HF, Singh M. Prevalence and antimicrobial resistance of Campylobacter from antibiotic-free broilers during organic and conventional processing. Poult Sci. 2018 Oct 15. doi: 10.3382/ps/pey486
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Ebner PD, Deering A, Mojadady M, Rahimi Z, Amini R, Popal M, Eshaqzai N, Barak S, Amini S, Azizi E, Morshid E, Mohammadi N, Rahimi M, McNamara K, Oliver HF. Capacity Building through Water Quality and Safety Analyses in Herat, Afghanistan. J Food Prot. 2018 Sep;81(9):1467-1471. doi: 10.4315/0362-028X.JFP-18-051
• Type: Journal Articles Status: Published Year Published: 2018 Citation: West AM, Nkemngong CA, Voorn MG, Wu T, Li X, Teska PJ, Oliver HF. Surface area wiped, product type, and target strain impact bactericidal efficacy of ready-to-use disinfectant Towelettes. Antimicrob Resist Infect Control. 2018 Oct 11;7:122. doi: 10.1186/s13756-018-0416-z.

Progress 10/01/16 to 09/30/17

Outputs
Target Audience:Retail food safety experts, government regulators, risk assessors, consumers, other food safety experts Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Two gradaute students compelted degrees under these objectives in 2017 Retail food safety managers for produce departments were re-trainined in cleaning, sanitation, and cross-contamination in stores enrolled in on-going studies How have the results been disseminated to communities of interest? Four abstracts have been submitted to IAFP 2018 to desiminate our recent findings Six abstracts were presented at IAFP in 2017 which encompasses regulator, industry, and academic stakeholders Two peer reviewed papers were published in the Journal of Food Protection, which is the primary journal for communittees interested in these topics What do you plan to do during the next reporting period to accomplish the goals? Publish recent studies in produce Publish remaining data in retail delis Sequence >250 Listeria genomes to differentiate persistent and transient Listeria in retail produce systems Graduate 2 MS students

Impacts
What was accomplished under these goals? We completed a > 10,000 sample study on Listeria and Salmonella in retail produce systems (Obj 1) Two key manuscripts were published on preavlence in retail delis (Obj 1, 2) Six abstracts were presented at IAFP 2017 on related work (Obj 1, 2, 3) Studies continued to eliminate Listeria from retail produce handling environments in grocery stores (Obj 2)

Publications

• Type: Journal Articles Status: Published Year Published: 2017 Citation: A. J. Etter, S. R. Hammons, S. Roof, C. Simmons, T. Wu, P.W. Cook, A. Katubig, M. J. Stasiewicz, E. Wright, S. Warchocki., J. Hollingsworth, H. S. Thesmar, M. Wiedmann, and H. F. Oliver. 2017. Enhanced standard SSOPs have limited impact on Listeria monocytogenes prevalence in retail delis. J Food Prot. 80 (11): 1903-1912.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: S. Hammons, A. J. Etter, J. Wang, T. Wu, T. Ford, M. T. Howard, and H. F. Oliver. 2017. Evaluation of third-party deep cleaning as a Listeria monocytogenes control strategies in retail delis. J Food Prot. 80 (11): 1913-1923.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Y. Hong, P. J. Teska, H. F. Oliver. 2017. The effects of contact time and concentration on bactericidal efficacy of three disinfectants on hard non-porous surfaces. Infect Dis Health. 22(1):17.
• Type: Book Chapters Status: Awaiting Publication Year Published: 2018 Citation: C. Assisi, A. Etter, and H. F. Oliver*. 2018. The future of whole genome sequencing in retail food safety in Food and Feed Safety Systems and Analysis. Editors S. Ricke, G. Atungulu, S. Hong Park, and C. Rainwater. Elsevier, San Diego, California.

Progress 12/01/15 to 09/30/16

Outputs
Target Audience:The target audience is any stakeholderinvolved in retail food systems that are producing, processing, or distributing foods. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?My laboratory has 6 graduate students under my leadership that are being trained in food safety How have the results been disseminated to communities of interest?Peer reviewed journal articles, professional meeting symposia, invited talks. What do you plan to do during the next reporting period to accomplish the goals?Obj 1: Continue our investigations into retail produce, hospitals, and long-term care Obj 2: Implement control strategies, food safety behavior, and facilities design changes in retail systems Obj 3: Publish the transcriptome forSalmonellaHeidelberg outbreak strains exposed to heat stress.

Impacts
What was accomplished under these goals? Obj 1: We have broadened our investigations into retail produce, hospitals, and long-term care Obj 2: We are exploring new control strategies, food safety behavior, and facilities design as contributers to foodborne disease transmission Obj 3: We have defined the transcriptome for Salmonella Heidelberg outbreak strains exposed to heat stress.

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: M. J. Stasiewicz, H. F. Oliver, M. Wiedmann, H. C. den Bakker. 2015. Whole-genome sequencing allows for improved identification of persistent Listeria monocytogenes in food-associated environments. Appl Environ Microbiol 2015, 81(17):6024-6037.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: A. M Pleitner, T. K. Chapin, S. R. Hammons, A V Stelten, K. K. Nightingale , M. Wiedmann, L. M. Johnston, H. F. Oliver. 2015. Development and evaluation of a multi?Institutional case studies?based course in food safety. J Food Sci Edu 14(3):76-85.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: S. McCoy, J. W. Chang, K. McNamara, H. F. Oliver, and A. Deering. 2015. Quality and safety attributes of Afghan raisins before and after processing in Afghanistan. J Food Process Preserve. doi: 10.1002/fsn3.190
• Type: Journal Articles Status: Published Year Published: 2015 Citation: S. R. Hammons, M. J. Stasiewicz, S. Roof, and H. F. Oliver*. 2015. Aerobic plate counts and ATP levels correlate with�Listeria�monocytogenes detection in retail delis. J Food Prot doi: 10.4315/0362-028X.JFP-14-500.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: J. Wang, A. Ray, S. R. Hammons, and H. F. Oliver. 2015. Persistent and transient Listeria monocytogenes strains from retail deli environments vary in their ability to adhere and form biofilms and rarely have inlA premature stop codons. Foodborne Pathogen Dis doi: 10.1089/fpd.2014.1837