ENHANCING MICROBIAL FOOD SAFETY BY RISK ANALYSIS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: TERMINATED
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1003413
• Project No.: WYO-519-14
• Multistate No.: S-1056
• Project Start Date: Jun 24, 2014
• Project End Date: Sep 30, 2018

Project Director
Bisha, BL.

Recipient Organization
UNIVERSITY OF WYOMING
1000 E UNIVERSITY AVE DEPARTMENT 3434
LARAMIE,WY 82071-2000

Performing Department
Animal Science

Non Technical Summary
The long-term goal of this project is the establishment of a multi-disciplinary network of scientists that performs comprehensive and integrated risk-based research and outreach to improve the safety of food from farm to fork. Interested stakeholders, including food producers and/or processors, retailers and consumers, have identified the need for an approach that conducts applied research to determine the prevalence and ecology of foodborne pathogens (including antibiotic resistant bacteria) in fresh and processed foods coupling that to research aimed at establishing effective control methods to decrease pathogen contamination of foods. Several outreach objectives have also been developed in support of this project. These objectives include communication of risk-based management recommendations derived from the research aspects of this proposal to stakeholders as well as to those who interact with stakeholders. Communication strategies will be precisely tailored to the particular audience (processors, distributors, retailers, consumers). Message content will focus on risk-based strategies and microbial control opportunities deemed critical for each target audience to achieve the greatest strides in improving food safety in the U.S. Outreach to those who advise producers and consumers (e.g. educators, extension personnel) who are not part of the project will be achieved through ongoing symposia to disseminate key information concerning lessons learned during the course of this project. This project has been specifically designed to address the critical needs of the fresh and processed food industries by developing a thorough understanding of how these foods become contaminated with foodborne microbial pathogens. It is well established that the heterogeneous distribution of pathogens in food makes studying the ecology of these pathogens difficult. The problems facing the food industry are also sufficiently complex such that solutions to these problems are beyond the scope of any single investigator's programmatic outputs. This means they are most efficiently addressed through multidisciplinary efforts with expertise in risk analysis, microbial ecology, epidemiology of foodborne disease, and food safety microbiology. The research group at the University of Wyoming will focus on the development of rapid detection methods for foodborne pathogens, sample preparation method development for rapid detection, evaluation of currrent and novel processing technologies for enhancement of food safety and quality, and risk assessment of factors resulting in contamination of food including wildlife intrusions.

Animal Health Component

5%

Research Effort Categories

Basic

25%

Applied

45%

Developmental

30%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
501	4010	1100	25%
722	4010	1100	25%
712	4099	1100	25%
723	4099	1100	25%

Knowledge Area
501 - New and Improved Food Processing Technologies; 722 - Zoonotic Diseases and Parasites Affecting Humans; 712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins; 723 - Hazards to Human Health and Safety;

Subject Of Investigation
4099 - Microorganisms, general/other; 4010 - Bacteria;

Field Of Science
1100 - Bacteriology;

Keywords

food safety

sample preparation

microbial detection

rapid detection

rapid characterization

Goals / Objectives
Risk Assessment: Assess food safety risks in agriculture systems Risk Management: Develop science-based interventions to prevent and mitigate food safety threats

Project Methods
We will 1) evaluate current practices and processing technologies used in food production to ensure the safety of food, 2) develop rapid detection technologies and sample preparation methods for rapid detection, and 3) assess risk factors resulting in contamination of foods and develop mitigation approaches. The work will be performed on bacterial isolates and viral particles and not on live animals, thus not requiring IACUC approval.1) Sampling will be performed in commercial processing facilities (including cantaloupe processing facilities) on food products, water and food contact and non-food contact surfaces. Total counts, fecal coliform and E. coli counts will be used to asses the effectiveness of antimicrobial interventions on the quality of the products as well as an indication of safety. Specific pathogen testing will also be performed for Salmonella enterica and Listeria monocytogenes. Culture methods and molecular methods will be emplyed for final identification and typing of isolates.2) We will develop paper-based analytical devices as well as flow cytometry-based assays for detection of foodborne pathogens and indicator microorganisms. We will evaluate sample concentration methods for bacterial and viral pathogens from water, bioaerosols and other matrices to improve detection sensitivity and to streamline detection protocols.3) We will assess the role of wildilife to foodborne disease risk via contamination of leafy green produce, such as spinach and lettuce, when they visit agricultural fields. We will collect data in collaboration with the National Wildlife Research Center on when wildlife enter agricultural production areas, what species enter, where they enter, and how many of them are present at a given point in time, so we can better understand if and where introduction of pathogens causing foodborne diseases might occur. We will also collect fecal samples from wildlife visiting produce fields and test them for major microbial pathogens causing foodborne disease. This information will provide needed insight into the magnitude of the problem and will determine if actions should be taken to limit this risk and what those actions should be.

Progress 06/24/14 to 09/30/18

Outputs
Target Audience: Target audiences included undergraduate and graduate students, microbiologists, food safety specialists, extension professionals, mass spectrometrists, biotechnology, animal health industry, and food industry, veterinarians, academic personnel, consumers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? This project has provided opportunities for training of three graduate students and three undergraduate students. How have the results been disseminated to communities of interest? Results have been disseminated via peer-reviewed publications, regional, national, and international conference papers and presentations, extension publications, teaching and symposia.. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Risk Assessment: Assess food safety risks in agricultural systems We assessed whether Salmonella contamination of hydroponic tomato nutrient solution would lead to Salmonella survival and contamination of the tomato fruit and plants, hydroponic nutrient film technique (NFT) troughs, and water receptacles. Although contaminated hydroponic nutrient solution led to surface contamination of roots, such an event may not pose a high risk of contamination of hydroponically grown fruit. Additionally, we sampled produce and fecal samples from mammalian and avian wildlife associated with the produce fields to determine presence and distribution of antibiotic resistance in indicator bacteria, ultimately establishing low levels of antimicrobial resistance, both phenotypically and genotypically. In our studies, we found that wildlife seems to play a minor role in the dissemination of antimicrobial resistance to produce fields in the absence of anthropogenic sources. Conversely, impacted wildlife potentially maintain, reintroduce, and disseminate AMR bacteria across food animal production landscapes. We investigated the genetic context of phenotypically similar AMR Escherichia coli harboring priority AMR phenotypes that were collected from cattle and raccoons. Phenotypic testing was conducted on more than 1,000 bacteria isolated from cattle and wildlife feces collected at six livestock facilities. The main clades of AMR E. coli isolates were identified, with the most highly conserved clade containing isolates from both cattle and raccoons. The variable genomes of isolates were primarily attributable to differential repertoires of Inc-like plasmids and prophage mosaics. The genetic conservation between AMR isolates from cattle and wildlife suggest a complex AMR livestock ecology that has inputs from multiple sources. Risk Management: Develop science-based interventions to prevent and mitigate food safety threats? We have developed a system in which the resin serves as a secondary concentration step for viral particles captured in the impinger liquid. Nucleic acids can then be directly eluted in small volumes, providing a concentrated sample for molecular analyses and improvement in viral detection sensitivity, primarily through reduction in sample volume. The anion exchange resin is expected to capture viruses with net-negative surface charges, including many enteric viruses, influenza viruses and other viruses relevant to public and animal health. We have adapted anion exchange resin-mediated virus concentration methods to liquid impingement-based bioaerosol sampling, improving detection by 6.77×, 3.33×, and 8.26×, for type A and type B influenza viruses, and MS2 bacteriophages, respectively. We have developed paper-based microfluidic devices (µPADs) employing electrochemical detection using stencil-printed carbon electrodes (SPCEs) on inexpensive and disposable transparency film platforms to detect low concentrations of pathogenic or generic E. coli and E. faecalis or E. faecium within 4 and 8 hours of enrichment, with the method also applied to detection in food (alfalfa sprouts) and water (lagoon water samples). µPAD-based were also employed for detection of C. sakazakii in powdered infant formula allowing for unambiguous detection of C. sakazakii from spiked PIF at ~ 101 CFU/g after 18 hours enrichment. Risk Communication: Communicate food safety messages to stakeholders? Food safety messages have been communicated to stakeholders (extension educators, food safety and nutrition trainers) at the Connecting Research with Extension Conference, produce safety symposia, and livestock (sheep) producer symposia. Additionally, food safety messages were communicated through material published in the Reflections magazine, which is a publication that reaches producers across the state of Wyoming.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Kraft TB, Keith J, Bisha B, Larson-Meyer E, Griebel A. 2019. The impact of daily kimchi: a pilot study. Scholarly Journal of Food and Nutrition, 1, (4). doi: 10.32474/sjfn.2019.01.000120.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Cadieux B, Colavecchio A, Jeukens J, Freschi L, Edmond-Rheault JG, Kukavica-Ibrulj I, Levesque RC, Bekal S, Chandler JC, Coleman SM, Bisha B, Goodridge LD. 2018. Prophage induction reduces Shiga toxin producing Escherichia coli (STEC) and Salmonella enterica on tomatoes and spinach: a model study. Food Control. 246:38-41. https://doi.org/10.1016/j.foodcont.2018.02.001.
• Type: Other Status: Published Year Published: 2018 Citation: Bisha B. Microbial Diagnostics and Characterization for Produce Safety. Department of Horticulture & Landscape Architecture Seminar Series, Colorado State University. February 12, 2019, Ft. Collins, CO (invited talk).
• Type: Other Status: Published Year Published: 2018 Citation: Bisha B. Source Tracking, Typing, and Field-Based Diagnostics. Symposium: What Does the Future Hold for Produce  The Innovation Frontier. November 30, 2018, Ft. Collins, CO (invited talk).
• Type: Other Status: Published Year Published: 2018 Citation: Bisha B. Food Safety Interventions. Wyoming Wool Growers association  Summer Meeting. August 7, 2018, Laramie, WY.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: 4. Aljasir S, Chandler J, Franklin A, Bevins S, Bentler K, Ellis J, Broten CJ, Bisha, Bisha B. Low Levels of Antimicrobial Resistance Among Indicator Bacteria Isolated from Wildlife Associated with Produce Fields. International Association for Food Protection Annual Meeting. July 8 - July 11, 2018, Salt Lake City, UT.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: 6. Broten CJ, Wydallis JB, Reilly T III, Bisha B. Colorimetric Detection of Cronobacter sakazakii in Artificially Contaminated Powdered Infant Formula Using Microfluidic Paper-Based Analytical Devices. International Association for Food Protection Annual Meeting. July 8 - July 11, 2018, Salt Lake City, UT.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: 8. Colavecchio A, Joseph S, Zhong Z, Zahirovich-Jovich Y, Coleman S, Chandler J, Bisha B, Perez-Mendez A, McEgan R, Danyluk M, Probasco K, Marshall D, Jeukens J, Freschi L, Emond Rheault J, Hamel J, Kukavica-Ibrulj I, Levesque R, Goodridge LD. A Comparison of In Silico Methods to Serotype Salmonella enterica Isolates from Food and Agricultural Environments. International Association for Food Protection Annual Meeting. July 8 - July 11, 2018, Salt Lake City, UT.
• Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: 9. Bisha B. Colorimetric and Electrochemical Bacteria Detection Using Printed Paper- and Transparency-Based Analytic Devices. Fifth Annual Rapid Detection for Food Safety Conference, June 27, 2018, Bethesda, MD (invited talk).

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

Outputs
Target Audience:Graduate students, undergraduate students, food scientists, producers, federal and state government employees, extension educators, scientists. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided opportunities for training of two graduate students and three undergraduate students. How have the results been disseminated to communities of interest?Results have been disseminated via peer-reviewed publications, regional, national, and international conference papers and presentations, extension publications and teaching. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? 1) Risk Assessment: Assess food safety risks in agricultural systems We assessed whether Salmonella contamination of hydroponic tomato nutrient solution would lead to Salmonella survival and contamination of the tomato fruit and plants, hydroponic nutrient film technique (NFT) troughs, and water receptacles. Although contaminated hydroponic nutrient solution led to surface contamination of roots, such an event may not pose a high risk of contamination of hydroponically grown fruit. Additionally, we sampled produce and fecal samples from mammalian and avian wildlife associated with the produce fields for presence and distribution of antibiotic resistance indicator bacteria, finding low levels of these indicator bacteria in all samples. We found that in our study, wildlife seem to play a minor role in the dissemination of antimicrobial resistance to produce fields in the absence of anthropogenic sources. 2) Risk Management: Develop science-based interventions to prevent and mitigate food safety threats We adapted an anion exchange resin-based method, initially developed to concentrate negatively charged viruses from water, to liquid impingement-based bioaerosol sampling, improving detection by 6.77×, 3.33×, and 8.26×, for type A and type B influenza viruses, and MS2 bacteriophages, respectively. We developed paper-based microfluidic devices employing electrochemical detection using stencil-printed carbon electrodes (SPCEs) on inexpensive and disposable transparency film platforms. Low concentrations (101 CFU/mL) of pathogenic or generic E. coli and (100 CFU/mL) E. faecalis and E. faecium strains were detected within 4 and 8 hours of pre-enrichment, with the method also applied to detection in food (alfalfa sprouts) and water (lagoon water samples). 3) Risk Communication: Communicate food safety messages to stakeholders Food safety messages have been communicated to stakeholders (extension educators, food safety and nutrition trainers) at the 'Connecting Research with Extension Conference which took place December 12-14, 2017 in Laramie, WY. Additionally, food safety messages were communicated through a publication in 'Reflections' magazine which is a publication that reaches producers across the state of Wyoming.

Publications

• Type: Journal Articles Status: Published Year Published: 2017 Citation: Chandler, J. C., J. W. Schaeffer, M. Davidson, S. L. Magzamen, A. P�rez-M�ndez, S. J. Reynolds, L. D. Goodridge, J. Volckens, A. B. Franklin, S. A. Shriner, and B. Bisha. 2017. A Method for the improved detection of aerosolized influenza viruses and the male-specific (F+) RNA coliphage MS2. J Virol Methods. 246:38-41. doi: 10.1016/j.jviromet.2017.04.004.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Coleman, S. M., B. Bisha, M. Bunning, S. E. Newman, and L. D. Goodridge. 2017. Transmission and persistence of Salmonella spp. in nutrient solution of hydroponic grown tomatoes. HortScience. 52(5):713718. 2017. doi: 10.21273/HORTSCI11200-16.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Adkins J, K. Boehle, C. Friend C, B. Chamberlain, B. Bisha, and C. Henry. 2017. Colorimetric and electrochemical bacteria detection using printed paper- and transparency-based analytic devices. Anal Chem. 21;89(6):3613-3621. doi: 10.1021/acs.analchem.6b05009.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Chandler JC, P�rez-M�ndez A, Paar J, Doolittle MM, Bisha B, Goodridge LD. 2017. Field-based evaluation of a male-specific (F+) RNA coliphage concentration method. J Virol Methods. 239:9-16. doi: 10.1016/j.jviromet.2016.10.007.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Chandler, J., J. Schaeffer, M. Davidson, S. Magzamen, A. Perez-Mendez, S. Reynolds, L. Goodridge, J. Volckens, A. Franklin, S. Shriner, and B. Bisha. 2017. A Method for the Improved Detection of Aerosolized Influenza Viruses Using Impingers that Incorporate Anion Exchange Resin. International Association for Food Protection Annual Meeting Abstract P3-212.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Aljasir, S., J. Chandler, A. Hamidi, D. Sylejmani, B. Wang, K. Schwam, and B. Bisha. A Survey of Antimicrobial Resistance among Dairy Cattle in Kosovo. International Association for Food Protection Annual Meeting Abstract P1-220.
• Type: Conference Papers and Presentations Status: Other Year Published: 2017 Citation: Bisha, B. 2017. Exploring alternative tools for diagnostics and characterization of foodborne pathogens. Colorado State University, Department of Food Science and Human Nutrition Seminar Series, November 9, 2017 (invited talk).
• Type: Conference Papers and Presentations Status: Other Year Published: 2017 Citation: Bisha, B. 2017. Small Things Considered: From Bioaerosols to Microfluidics. Iowa State University, Department of Food Science and Human Nutrition Seminar Series, March 29, 2017 (invited talk).

Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Students, producers, academia, food safety professionals, goverment, food producer associations. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Two graduate students, one undergraduate student, and one research scientist have been trained under this ptroject. How have the results been disseminated to communities of interest?Conference presentations and abstracts and peer reviwed 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? (1) Risk Assessment: Assess food safety risks in agriculture systems We have conducted research in the San Luis Valley, Colorado to determine the risk of contamination of leafy greens with select foodborne pathogens from wildlife. We collected over 361 fecal samples from wildlife (174 from mammals, 184 from birds, and 3 from unidentified species from 10 fields of leafy greens. Moreover, we analyzed 168 spinach and 9 lettuce samples from 6 of the 10 fields for the target pathogens (with another 800 additional lettuce samples tested indipendently). Most samples from mammals were from rabbits and rodents. The majority of avian samples were from Horned Larks and unidentified passerine species. We did not confirm the presence of Salmonella or E. coli STEC in wildlife feces or crop samples (12 samples were positive for E. coli, none were confirmed as STEC by PCR). None of the 110 fecal samples analyzed for the presence of norovirus GI and GII tested positive. In conclusion, while we documented multiple wildlife intrusions in produce fields, we determined that the risk of contamination of produce fields in this specific location was low. This may be due to a number of facts, including the low number of intensive animal operations nearby, current mitigation practices in place by farmers, the composition of wildlife and other reasons. (2) Risk Management: Develop science-based interventions to prevent and mitigate food safety threats Our work has focused on the development of rapid diagnostics. We are currently optimizing protocols and technologies for the detection of foodborne bacterial pathogens Salmonella enterica and Listeria monocytogenes using paper-based analytical devices (µPADs). We have successfully screened several enrichment media and determined optimized one-step enrichment protocols for the sensitive and inexpensive overnight detection of these pathogens.

Publications

• Type: Journal Articles Status: Submitted Year Published: 2016 Citation: Chandler J, Franklin A, Shriner S, Root J, Mooers N, Bentler K, Ellis J, LeJeune J, Goodridge L, Stahl R, Russell M, Robeson M, and Bisha B. Oceanobacillus spp., Antimicrobial Resistant Bacteria at the Wildlife-Livestock Interface. Scientific Reports (submitted.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Chandler J, P�rez-M�ndez A, Paar J, Doolittle M, Bisha B, Goodridge L. Field-based evaluation of a male-specific (F+) RNA coliphage concentration method.J Virol Methods. 2016 Oct 21;239:9-16. doi: 10.1016/j.jviromet.2016.10.007.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Harris L, Lieberman V, Mashiana R, Atwill E, Yang M, Chandler J, Bisha B, Jones T. Prevalence and Amounts of Salmonella Found on Raw California Inshell Pistachios.J Food Prot. 2016 Aug;79(8):1304-15. doi: 10.4315/0362- 028X.JFP-16-054.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Maus A, Anders J, Bisha B, and Basile F. Differentiation of Bacteria at the Strain Level by MALDI-MS of Proteins >15kDa. 64th American society for Mass Spectrometry Annual Conference. June 5-9, San Antonio, TX
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bisha B. Paper-Based Analytical Devices for Detection of Foodborne Bacteria. Third Annual Rapid Detection for Food Safety Conference, June 27-28, Baltimore, MD.
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Chandler C, Franklin A, Shriner S, Root J, Anders J, Wang B, and Bisha B. Synanthropic Wildlife Associated with Livestock Production as Carriers of High Priority Antimicrobial Resistances. International Association for Food Protection Annual Meeting. July 31- August 3, 2016, St. Louis, MO
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Anders J, Wang B, Chandler C, Prenni J, Franklin A, Carlson J, LeJeune J, and Bisha B. MALDI-TOF MS Biotyping in the Characterization of Antimicrobial-resistant Enterococcus spp. from Wildlife Associated with Concentrated Animal Feeding Operations. International Association for Food Protection Annual Meeting. July 31- August 3, 2016, St. Louis, MO.

Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Food safety professionals, researchers, students, and producers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided training and professional opportunities for one graduate student (MS), one postdoctoral fellow, and one research scientist. How have the results been disseminated to communities of interest?Results have been disseminated via peer-reviewed publications and conference presentations and abstracts. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? 1) Risk Assessment: Assess food safety risks in agricultural systems We have been focused on determining the role of wildlife on the contamination of leafy green crops with foodborne pathogens with a special emphasis on Salmonella enterica and E. coli STEC. 10 lettuce and spinach fields in the San Luis Valley, Colorado were sampled for wildlife visitation rates using remote cameras, with approximately 1500 fecal and produce samples taken for downstream microbiological analyses. Results for presence of Salmonella enterica and E. coli STEC as well as microbial indicators are pending. 2) Risk Management: Develop science-based interventions to prevent and mitigate food safety threats a) We have developed Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) methods for rapid and specific identification and typing of antimicrobial resistant (AMR) bacteria isolated from wild mammals, cattle, and environment on livestock facilities. PCA-type analyses which consider the entirety of the data set were able to distinguish phenotypes below the species level, as indicated by the clustering patterns which show that isolates can be differentiated below the taxonomic level of species. That differentiation did not extend to specific antimicrobial susceptibility. Subsets of the isolates were tested phenotypically for antimicrobial susceptibility via phenotypic methods; then through peak alignment and correlative analyses, specific biomarkers were predictive of antimicrobial susceptibility within this dataset. b) We have improved current bioaerosol sampling methods and endpoint molecular detection of viral indicators through enhanced sample preparation. Modification of SKC BioSamplers for capture of viruses from improved detection sensitivity by 8.5x and 2x for the virus surrogates MS2 and Φ6 bacteriophages, respectively. The modifications to the SKC BioSampler are straight forward and add minimum cost, allowing for simplification of sampling procedures and addition of Sample buffer incorporated protectants. 3) Risk Communication: communicate food safety messages to stakeholders We have communicated our findings regarding the current status of antimicrobial resistance and its importance to food safety in several meetings and symposia, including the International Association for Food Protection Annual Meeting and the Annual Public Health Symposium at Colorado State University.

Publications

• Type: Journal Articles Status: Submitted Year Published: 2015 Citation: Chandler JC, Manley WA, P�rez-M�ndez A, Bisha B, Adkins JA, Henry CS, Prenni JE, Goodridge LD. 2015. Molecular and phenotypic properties of cantaloupe-associated Listeria monocytogenes. Food Microbiology (submitted).
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Johnson DC, Bzdek JP, Fahrenbruck CR, Chandler JC, Bisha B, Goodridge LD, Hybertson BM. 2015. An innovative non-thermal plasma reactor to eliminate microorganisms in water. Desalination and Water Treatment. DOI: 10.1080/19443994.2015.1024752
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Bisha B, Brehm-Stecher BF. 2015. Flow cytometry for rapid detection of Salmonella spp. in seed sprouts. ScienceOpen Research. doi: 10.14293/S2199-1006.1.SOR-LIFE.AJ19WR.v1.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: J Anders, B Wang, J Chandler, J Prenni, A Franklin, J Carlson, J LeJeune, B Bisha. MALDI-TOF MS Biotyping for Characterization of Antimicrobial-resistant Escherichia coli from Concentrated Animal Feeding Operations and Associated Wildlife. International Association for Food Protection Annual Meeting. July 25- July 28, 2015, Portland, OR.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: S Coleman, H Kessler, X He, J Avens, J Chandler, B Bisha, L Goodridge, M Bunning. Evaluation of Consumer Washing Techniques to Reduce Natural Microbiota on the Surface of Whole Cantaloupes. International Association for Food Protection Annual Meeting. July 25- July 28, 2015, Portland, OR.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: J Chandler, M Davidson, J Schaeffer, A Perez-Mendez, J Volckens, S Magzamen, L Goodridge, S Reynolds, B Bisha. Development of an Improved Sampling Method for Concentrating Viruses from Bioaerosols. International Association for Food Protection Annual Meeting. July 25- July 28, 2015, Portland, OR.
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: A Franklin, S Shriner, J Root, B Bisha, J Chandler. The Role of Wildlife in Disseminating Antibiotic-resistant Bacteria to and from Livestock Facilities. 3d International Symposium on the Environmental Dimension of Antibiotic Resistance. May 17 - May 21, 2015, Wernigerode, Germany
• Type: Conference Papers and Presentations Status: Other Year Published: 2015 Citation: B Bisha. Antimicrobial Resistance: Food Safety. Microbial Ecology in a Changing World: Emerging Issues in Antimicrobial Resistance. The 6th Annual Public Health Symposium at Colorado State University. April 15, 2015, Fort Collins, CO.

Progress 06/24/14 to 09/30/14

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Training was provided for one graduate student and one postdoctoral fellow. How have the results been disseminated to communities of interest? Results have been disseminated in the form of peer-reviewed journal publications and conference presentations and abstracts. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? 1) Risk Assessment: Assess food safety risks in agricultural systems We have worked with commercial cantaloupe producers to survey presence of select foodborne pathogens in cantaloupe and to assess the effectiveness of intervention technologies for enhancement of microbial safety and quality of the product. We have assessed the shedding patterns of antimicrobial resistant (AMR) bacteria by raccoons and the risk they pose for transfer of AMR bacteria to livestock operations. 2) Risk Management: Develop science-based interventions to prevent and mitigate food safety threats We have developed, optimized and validated microfluidic paper-based devices for colorimetric detection of foodborne pathogens and bacterial indicators from agricultural water. We have greatly improved sensitivity of molecular detection of enteric viruses and viral indicators through enhanced sample preparation. 3) Risk Communication: Communicate food safety messages to stakeholders We have communicated our findings regarding effectiveness of processing treatments for improvement of microbial safety of cantaloupe directly to producers during sampling in the processing plant, as well as through talks and symposium proceedings in a meeting aimed at informing cantaloupe growers.

Publications

• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: S Coleman, B Bisha, K Blume, V Sandoval, N Drury, H Kesseler, A Perez-Mendez, J Chandler, B Franz, Ryan Jaebker, Donald Schaffner, Jeffrey LeJeune, Marisa Bunning, Steven Newman, Lawrence Goodridge. Evaluation of Risk Factors Affecting Transmission and Persistence of Salmonella spp. in Hydroponically Grown Tomatoes. P1-118. International Association for Food Protection Annual Meeting. July 31- August 3, 2014, Indianapolis, IN.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: J Chandler, A Perez-Mendez, S Coleman, W Manley, M Danyluk, M Bunning, L Goodridge, B Bisha. Control of the Microbial Quality of Cantaloupes via Employment of a Processing Line Incorporating Chlorine Dioxide as an Intervention Step. P1-109. International Association for Food Protection Annual Meeting. July 31- August 3, 2014, Indianapolis, IN.
• Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: B Bisha. Food Safety: Rapid Diagnostics and Typing. Student American Veterinary Association
• Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: B Bisha. Food Safety: Rapid Diagnostics and Typing. University of Prishtina International Summer University. July 7 - July 18, 2014, Pristina, Kosovo.
• Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: B Bisha. Control of the Microbial Quality of Cantaloupes via Employment of a Processing Line Incorporating Chlorine Dioxide as an Intervention Step. Food Safety Update for Cantaloupe Growers: What do the new developments mean to me? April 17, 2014, Gulf Coast Research & Education Center, Wimauma, FL.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Bisha B, Adkins JA, Jokerst JC, Chandler JC, P�rez-M�ndez A, Coleman SM, Sbodio AO, Suslow TV, Danyluk MD, Henry CS, Goodridge LD. 2014. Colorimetric paper-based detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from large volumes of agricultural water. J Vis Exp. 9;(88). doi: 10.3791/51414.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: P�rez-M�ndez A, Chandler JC, Bisha B, Goodridge LD. 2014. Concentration of enteric viruses from tap water using an anion exchange resin-based method. J Virol Methods. 206:95-8. doi: 10.1016/j.jviromet.2014.05.025.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: P�rez-M�ndez A, Chandler JC, Bisha B, Goodridge LD. 2014. Evaluation of an anion exchange resin-based method for concentration of F-RNA coliphages (enteric virus indicators) from water samples. J Virol Methods. 204:109-15. doi: 10.1016/j.jviromet.2014.03.024.