ENHANCING MICROBIAL FOOD SAFETY BY RISK ANALYSIS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1018174
• Multistate No.: S-OLD 1077
• Project Start Date: Dec 1, 2018
• Project End Date: Sep 30, 2023
• Program Code: [(N/A)]- (N/A)

Recipient Organization
OHIO STATE UNIVERSITY
1680 MADISON AVENUE
WOOSTER,OH 44691

Performing Department
Food Animal Health Research Program

Non Technical Summary
The Centers for Disease Control and Prevention (CDC) estimates that one in six Americans becomes sick each year from eating contaminated food, with about 48 million cases of foodborne illness, 128,000 hospitalizations, and 3,000 deaths occurring each year from foodborne pathogens in the U.S. (Scallan et al., 2011). The long-term goal of this S1077 project is to perform comprehensive and integrative risk-based research, education, and outreach to improve food safety and advance public health. Objectives are 1) Risk Assessment: Characterize food safety risks in food systems; 2) Risk Management: Develop, validate, and apply science-based interventions to prevent and mitigate food safety threats; and 3) Risk Communication: Convey science-based messages to stakeholders to improve food safety behaviors and practices. At Dr. Gireesh Rajashekara's and Dr. Qiuhong Wang's laboratories, studies focus on the bacterial food-borne pathogens (especially Salmonella and Campylobacter) and human norovirus, respectively. Our efforts will contribute to the effective control of foodborne pathogens across food commodities in the U.S. and the education of next generation of scientists working in food safety field.

Animal Health Component

40%

Research Effort Categories

Basic

50%

Applied

40%

Developmental

10%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
712	5010	1040	100%

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

Subject Of Investigation
5010 - Food;

Field Of Science
1040 - Molecular biology;

Keywords

food safety

risk analysis

microbiology

Goals / Objectives
Risk Assessment: Characterize food safety risks in food systems Risk Management: Develop, validate, and apply science-based interventions to prevent and mitigate food safety threats Risk Communication: Convey science-based messages to stakeholders to improve food safety behaviors and practices

Project Methods
1) Risk Assessment: Assess food safety risks in agricultural systemsHuman noroviruses (HuNoVs) in fresh produceSince human NoVs are still unculturable in routine cell culture, we will use virus-like particles (VLPs) of human NoVs and surrogate viruses (porcine sapovirus and non-human primate Tulane virus) to perform most in vitro experiments. Finally, we will confirm our findings with infectious human NoVs from human diarrhea samples under BSL2 conditions. We will use the VLPs of wild type (WT) human NoV and mutants varying in histo-blood group antigen (HBGA)-binding to investigate binding to lettuce cell wall and identify the viral amino acids involved using ELISA assays. An increase or decrease in binding will indicate that the amino acids confer enhanced or reduced attachment to lettuce cell wall carbohydrates, respectively. Our studies will guide us to look for economic reagents that can bind to the specific amino acid residues on the capsid proteins of HuNoV to prevent contamination in the future.Salmonella in fresh produceWe will study the impact of specific environmental plant growing conditions (temperature and relative humidity), Salmonella-inoculation methods (leaf-spray, cotyledon-clip, or grafting), tomato variety and Salmonella genotype, and co-presence of phytopathogens (Xanthomonas or Clavibacter) on the persistence and dissemination of Salmonella in tomato plant tissues. Inoculated plants will be grown in greenhouses or growth chambers with pre-set growing conditions until the fruit harvesting stage and the Salmonella population will be monitored weekly using direct plating on selective medium. The data generated will enhance our understanding of the biotic and abiotic factors increasing the foodborne pathogens risk in fresh produce production.2) Risk Management: Develop science-based interventions to prevent and mitigate food safety threatsCombination of probiotics and novel small molecules inhibiting the growth of Campylobacter and Salmonella to reduce food safety burden in poultry and fresh produceA high-throughput screening, using library of small molecules (SMs), will be performed to identify novel narrow spectrum growth inhibitors of Salmonella and Campylobacter. The antimicrobial efficacy of these growth inhibitors will be confirmed in animal studies. Further, the antagonistic effect of known biocontrol agents (Bacillus spp. and Enterobacter sp) and probiotics (Bifidobacterium spp., Lactobacillus spp., and Escherichia coli Nissle 1917) will be tested in vitro and in vivo (poultry) against Salmonella and Campylobacter, respectively. The best growth inhibitors and biocontrol/probiotics will be combined together to enhance the reduction in these foodborne pathogens in poultry and/or fresh produces.Study the mechanisms of actions of novel compounds inhibiting the growth of Campylobacter or SalmonellaA random expression library of Salmonella will be generated using fragments of the Salmonella genomic. By over expressing the drug target(s) of the growth inhibitors, we expect to observe reduced antimicrobial efficacy of the SMs with generated mutants overexpressing proteins related to the mode of action of the SMs. The gene encoding the target protein in these mutants will be sequenced to identify the potential drug target(s) of the SMs and/or the mode of action of the SMs. In addition, columns with high affinity for the SMs will be designed to isolate molecules from Salmonella or Campylobacter that bind with the SMs. These results will allow determining the mode of action of each SM in Salmonella.Quantify the impact of several plant health and disease management practices on the populations of antimicrobial resistant (AMR) bacteria in the plant production environmentA model field test-plots that mimic conditions in production agriculture will be develop to study the impact of commercially available pesticides (glyphosate, streptomycin, copper, triazole) on AMR bacterial population from the environment and organic fertilizer. Soil and plant tissues will be collected at strategic time points before and after agricultural practices to monitor the composition of AMR bacteria using direct planting on selective media and the amplification of a selected set of genes linked to AMR. The data generated will enhance our understanding of the impact that antimicrobial use in horticulture has on driving the emergence and survival of AMR bacteria in the environment and will allow to development of guidelines and practices for prudent antimicrobial use in agriculture.3) Risk Communication:Convey science-based messages to stakeholders to improve food safety behaviors and practices

Progress 10/01/19 to 09/30/20

Outputs
Target Audience:Food safety researchers, microbiologists, public health state and federal agencies involved in food safety, producers and processors, and consumers. Changes/Problems:The COVID-19 pandemic significantly affected the work of the scientists involved in the projects described above. More precisely, it is only since a few months that the researchers were able to work part time in the lab. We were unable to collect field samples in the small farms of Ohio. Further, greenhouse and chicken rooms are limited due to the high demand for this type of structure. Therefore, several experiments initially scheduled during 2020 were unachieved and postponed for 2021. What opportunities for training and professional development has the project provided?The program is training 3 graduate students, 2 post-docs, 2 Research Associates, and 1 visiting scholar. How have the results been disseminated to communities of interest?We have given oral and poster presentations and published research articles. What do you plan to do during the next reporting period to accomplish the goals? We will improve the delivery method of novel antimicrobial agents (probiotic derived peptides, probiotics and small molecules) effective against Salmonella and Campylobacter in chicken using microencapsulation and by creating derivatives more suitable for large scale applications (higher antimicrobial efficacy, high water solubility, and resistant to gastric acids). We will combine probiotic, probiotic derived peptides, and small molecule treatments to improve the control of foodborne pathogen, Salmonella) in poultry production. We will identify the bacterial targets of the novel small molecules and probiotic derived peptides using genetic and biochemical approaches. We will use metagenomics to assess the impact of specific health (manure and glyphosate) and disease (copper, streptomycin, and triazole) management practices on the microbiome of a tomato field and determine its association with the AMR burden. We will use omics to identify the antimicrobial agent(s) secreted by Salmonella inhibiting the growth of plant pathogens in tomato. We will test the combinatorial approach (i.e. growth inhibitors small molecules, small peptides, biocontrol agents) to enhance the control of plant and foodborne pathogen in tomato production. We will identify the role of specific gene(s) of Salmonella essential for its persistence in tomato plant tissues using directed gene mutagenesis approaches. We will continue assessing the prevalence of foodborne pathogens (Salmonella, Listeria, Campylobacter, and Escherichia) in the small farms of Ohio. Further, we will use metagenomics to identify specific component of the microbiome modulating the persistence of foodborne pathogens. Dr. Wang will collaborate with her former Visiting Scholar Dr. Oka Tomoichiro (now at NIID, Japan) and her former post-doc Dr. Malak Esseili (now Assistant Professor at University of Georgia) to validate various inactivation process for HuSaVs using the newly developed HuSaV in vitro cell culture system.

Impacts
What was accomplished under these goals? 1) Risk Assessment: Characterize food safety risks in food systems Whole genome sequencing data identified specific genes in Salmonella enterica that may be involved in its persistence of in tomato plant tissues. We are creating knockout mutants using directed mutagenesis approaches (Lambda red recombination plus CRISPR-Cas9 systems) to assess the biological role of these genes in the persistence on Salmonella enterica in tomato plant tissues. We demonstrated that foodborne pathogens (Listeria, Salmonella, and Escherichia coli O157 ) can be frequently detected in field samples collected from small farms (n=18) of Ohio since 2016 (up to 18.3% in manure, 24% in irrigation water, and 12% in fresh produce samples). Further, the application of animal manure (especially dairy manure) increased the prevalence of foodborne pathogens in the soil and associated fresh produce. 2) Risk Management: Develop, validate, and apply science-based interventions to prevent and mitigate food safety threats We identified three probiotic derived peptides that completely inhibit the growth of multiple Salmonella serotypes by disrupting their cell membrane integrity. One of these peptides inhibited the growth of Salmonella Typhimurium in broiler chickens (approx. 2-log reduction). One recombinant attenuated Salmonella vaccines (RASVs) significantly inhibited the colonization of Campylobacter jejuni in chicken ceca at 17 days post challenged (up to 2.4-log reduction), even when high dosage of Campylobacter (104 CFU/chicken) were used for the inoculum. An indirect ELISA approach was optimized to test the Campylobacter jejuni specific IgY and IgA antibodies in chicken blood serum. We identified that specific management practices (manure, glyphosate, and antimicrobials [copper, streptomycin, and triazole]) increased the antimicrobial resistant burden (extended spectrum beta-lactamase and Aspergillus fumigatus) in the soil and plant tissues of a tomato field. Human sapoviruses (HuSaV) are an important cause of diarrhea and foodborne infections worldwide. Despite the discovery of HuSaVs over 40 years ago, no in vitro cell culture system has been established, limiting research on this important pathogen. Collaborated with scientists in Japan, we successfully propagated HuSaV in a human duodenum cell line in the presence of bile acids. mpacts: Our studies will identify management practices contributing to the AMR and foodborne pathogens (Listeria, Escherichia coli, Campylobacter and Salmonella) burdens in plant agricultural; and thus, will provide new guidance to improve the safety of the fresh produce and to reduce the emergence and dissemination of AMR in the environment. Our studies will highlight gene(s) essential for the persistence of foodborne pathogens such as Salmonella enterica in plant tissues. These genes could represent novel drug target(s) to control foodborne pathogens in plant agriculture. Our novel therapeutics (small molecules, probiotic derived peptides, RASVs) will enhance food safety by mitigating the colonization of poultry by foodborne pathogens such as Campylobacter and Salmonella. The inexpensive, reproducible and scalable in vitro cell culture system for HuSaV provides a fundamental scientific tool for HuSaV research and development of infection control strategies in the future.

Publications

• Type: Journal Articles Status: Published Year Published: 2020 Citation: Deblais, L., S. A. Miller, and G. Rajashekara. Impact of plant pathogens infection on Salmonella enterica subsp. enterica serotype Typhimurium persistence in tomato plants. Journal of food Protection. 2020 Nov 12. doi: 10.4315/JFP-20-291.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Hirotaka Takagi *, Tomoichiro Oka *, Hiroyuki Saito, Takayuki Kobayashi, Tomoko Takahashi, Chika Tatsumi, Takashi Shimoike, Michiyo Kataoka, Qiuhong Wang*, Linda J. Saif*, Mamoru Noda. 2020. Human sapovirus propagation in human cell lines supplemented with bile acids. PNAS first published November 30, 2020; https://doi.org/10.1073/pnas.2007310117
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Tomoichiro Oka, Hiroyuki Saito, Takayuki Kobayashi, Tomoko Takahashi, Takashi Shimoike, Michiyo Kataoka, Qiuhong Wang, Linda J. Saif, Mamoru Noda, Hirotaka Takagi. Cell culture trials for human sapoviruses. The 7th International Calicivirus Conference in Sydney, Australia from October 13  October 16, 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Malak A. Esseili1, Xiang Gao, Patricia Boley, Yixuan Hou, Linda J. Saif, Paul Brewer-Jensen, Lisa Lindesmith, Ralph S. Baric, Robert L. Atmar, Mary K. Estes and Qiuhong Wang. Human norovirus HBGA binding pocket mediates the virus specific interactions with lettuce carbohydratesThe 7th International Calicivirus Conference in Sydney, Australia from October 13  October 16, 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Closs, Jr G., Y. A. Helmy, A. Howell, D. Kathayat, V. Srivastava, L. Deblais, and G. Rajashekara. 2020. Antimicrobial efficacy of probiotic Lactobacillus rhamnosus GG in Salmonella infected chickens. International Association for Food Protection Annual Meeting, Cleveland, OH
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Closs, Jr G., Y. A. Helmy, D. Kathayat, S.Y. Wanda, R. Curtiss, and G. Rajashekara.2020. Recombinant attenuated Salmonella vaccines reduce Campylobacter colonization and induce IgY antibodies in chickens. CRWAD, Chicago, IL.
• Type: Conference Papers and Presentations Status: Published Year Published: 2020 Citation: Closs, Jr G., Y. A. Helmy, D. Kathayat, V. Srivastava, L. Deblais, R. Curtiss, and G. Rajashekara. 2020. Recombinant Attenuated Salmonella Vaccines to Control Campylobacter in Poultry. Edward Hayes Graduate Research Forum, OSU, Columbus, OH.

Progress 12/01/18 to 09/30/19

Outputs
Target Audience:Food safety researchers, microbiologists, public health state and federal agencies involved in food safety,producers and processors, and consumers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The program is training 5 graduate students, 2 post-docs, and 1 visiting scholar. How have the results been disseminated to communities of interest?Yes. We have given oral and poster presentations and published research results and extension articles.? What do you plan to do during the next reporting period to accomplish the goals?Next reporting period's plans are listed here We will identify the antimicrobial agent(s) secreted by Salmonella inhibiting the growth of plant pathogens in tomato. We will test the combinatorial approach (i.e. growth inhibitors small molecules, small peptides, biocontrol agents) to enhance the control of plant and foodborne pathogen in tomato production. We will improve the delivery method of novel antimicrobial agents (probiotics and small molecules) effective against Salmonella and Campylobacter in chicken using microencapsulation and by creating derivatives more suitable for large scale applications (higher antimicrobial efficacy, high water solubility, and resistant to gastric acids). We will combine probiotic and small molecule treatments to improve the control of foodborne pathogens (Salmonella and Campylobacter) in poultry production. We will identify the drug targets of the novel small molecules using genetic and biochemical approaches. We will identify the role of specific gene(s) of Salmonella essential for its persistence in tomato plant tissues. We will continue to adapt human sapoviruses in routine cell culture. We will look for nontoxic and economic inhibitors to block human norovirus binding to lettuce to reduce the transmission of human norovirus via leafy greens.

Impacts
What was accomplished under these goals? 1) Risk Assessment: Characterize food safety risks in food systems Lettuce is often implicated in human norovirus (HuNoV) foodborne outbreaks. We previously identified H-like histo-blood group antigens (HBGAs) on lettuce leaves as specific binding moieties for HuNoV GII.4/HS194/2009 strain. We determined that HuNoV-lettuce binding is mediated through the viral HBGA binding sites, which are essential for viral infection of human cells. Salmonella reduced Clavibacter michiganensis and Xanthomonas gardneri population in planta when inoculated together at the same time. The antagonistic effect of Salmonella on these two phytopathogens seems to be caused by the production of antimicrobial agents secreted in the supernatant. Tomato grafting using Salmonella contaminated blades allowed a rapid, systemic, and long-term infestation of the plant tissues by Salmonella, especially in the roots where Salmonella could survive for over 242 days post inoculation and resist to drought conditions. However, Salmonella was not detected in the fruits. Internalization of Salmonella in tomato fruits during post-harvest cleaning procedures principally occurred through the stem abscission zone. Once internalized in the fruits, Salmonella was protected against sanitation procedures (bleach and antimicrobial agents) Salmonella spp. and Listeria monocytogenes can survive in the hydroponic growing systems during the lettuce production. The pathogen species survived and increased in the concentration in the recycled fertilizer, lettuce roots and rock wool cubes, and were detectible in edible portions of the crop during the lifecycle of hydroponic lettuce. We identified behaviors specific to food insecure population receiving cancer treatment that should be targeted in future interventions. We assessed food safety knowledge and readiness to deliver food safety message among dietetics students in Ohio and Internationally. 2) Risk Management: Develop, validate, and apply science-based interventions to prevent and mitigate food safety threats We have grown human sapoviruses (HuSaVs) in human cell lines, which can be used to evaluate anti-virals and inactivation methods for HuSaVs. The combination of biocontrol agents (Bacillus spp. or Enterobacter sp.; as preventive control method) with our novel small compounds inhibiting the growth of Salmonella (as curative control method) and cleared Salmonella in up to 87% of infested tomato plants, depending on the combination tested. Using high-throughput chemical screens, we identified two novel small compounds inhibiting the growth of Campylobacter in broiler chickens (up to 2.7-log reduction), with low impact on the chicken microbiota in ceca. We have identified probiotic derived peptides that completely inhibit Salmonella. Probiotic and microencapsulated probiotic bacteria significantly reduced the amount of Salmonella Typhimurium present in chicken ceca. Recombinant attenuated Salmonella vaccines (RASVs) were tested in SPF layers and two RASVs significantly inhibited the colonization of Campylobacter jejuni in chicken ceca during a 28-day trial. We are currently assessing the impact of specific health (manure and glyphosate) and disease (copper, streptomycin, and triazole) management practices on antimicrobial resistant populations (extended spectrum beta-lactamase and Aspergillus fumigatus) and their genes in tomato field. We have been working on the experiments to show effectiveness of various sanitizers in hydroponic system in eliminating human pathogens (Salmonella spp. ad Listeria monocytogenes) while supporting optimal plant health. 3) Risk Communication: Convey science-based messages to stakeholders to improve food safety behaviors and practices 9/16/2019 - 9/19/2019. Research members service on a booth at the Farm Science Review (fsr.osu.edu), in London Ohio, for outreach on Zoonosis, Animal Disease, and Food Safety. During the 2018 training season we delivered 5 GAP courses reaching 263 produce growers in Ohio. 178 of these were Amish and Old Order Mennonite, primarily from the Shiloh, Ashland, Homerville, Holmes/Wayne, and Geauga communities. We developed Hydroponic Food Safety Farm to Fork, urban youth grower specialized training and a food safety plan writing workshop.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Esseili MA, Gao X, Boley P, Hou Y, Saif LJ, Brewer-Jensen P, Lindesmith LC, Baric RS, Atmar RL and Wang Q. 2019. Human Norovirus HBGA Binding Sites Mediate the Virus Specific Interactions with Lettuce Carbohydrates. Viruses. 11:833.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Deblais, L., Y. A. Helmy, A. Kumar, J. Antwi, D. Kathayat, U. M. Acuna, H. Huang, E. C. de Blanco, J. R. Fuchs, and G. Rajashekara. 2019. Novel narrow spectrum benzyl thiophene sulfonamide derivatives to control Campylobacter. The Journal of Antibiotics. 72(7):555-565.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Deblais, L., Y. A. Helmy, A. Testen, C. Vrisman, A. M. J. Madrid, D. Kathayat, S. A. Miller, and G. Rajashekara. 2019. Specific Environmental Temperature and Relative Humidity Conditions and Grafting Affect the Persistence and Dissemination of Salmonella enterica subsp. enterica Serotype Typhimurium in Tomato Plant Tissues. Appl. Environ. Microbiol. 85:e00403-19.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Deblais, L., C. Vrisman, D. Kathayat, Y. A. Helmy, S. A. Miller, and G. Rajashekara. 2019. Imidazole and Methoxybenzylamine Growth Inhibitors Reduce Salmonella Persistence in Tomato Plant Tissues. Journal of Food Protection 82:9971006.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Paden, H, Hatsu, I., Kane, K., Lustberg, M., Grenade, C., Bhatt, A., Diaz, D., Beery, A. and Ilic, S. (2019) Assessment of Food Safety Knowledge and Behaviors of Cancer Patients Receiving Treatment. Nutrients, 11, 1897; http://doi:10.3390/nu1108189
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Her, E., Seo, S., *Choi, J., *Pool, V. & Ilic, S., (2019). Assessment of Food Safety at University Food Courts Using Surveys, Observations, and Microbial Testing. Food Control, 103 (9), pp.167-174. https://doi.org/10.1016/j.foodcont.2019.04.002
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Gould, V.J., Evans, E.W., Redmond, E.C., Marklinder, I.M., Quinlan, J.J., & Ilic, S. (2019). Exploring the Role of Dietitians in the Delivery of Food Safety Information. Food Protection Trends, (39) 3, pp. 277-282.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Deblais, L., J. Scaria, L. Antony, C.M. Vrisman, V. Srivastava, Y.A. Helmy, S.A. Miller, and G. Rajashekara. 2019. Salmonella Typhimurium strain and tomato genotypes affect the persistence of Salmonella in tomato plants. American Phytopathology Society Meeting Abstract 689-P2.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Closs Jr, G., Yosra A. Helmy, Dipak Kathayat, Allison Howell, Gireesh Rajashekara (2019). Efficacy and Antimicrobial Characterization of Known Probiotics LA, LGG, and BB12 against Salmonella Infections in Vitro. ASM Microbe Abstract AAR-680.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Closs Jr, G., Yosra A. Helmy, Allison Howell, Dipak Kathayat, Gireesh Rajashekara (Dec. 2018). Efficacy and antimicrobial characterization of different probiotic bacteria against Salmonella infections in-vitro. The 99th annual conference of research workers in animal Diseases (CRWAD).
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Gould, V.J., Evans, E.W., Radmond, E.C., Ilic, S., Alwan, N. Hjeij, L. 2019 IAFP European Symposium on Food Safety 2019. Food Safety Attitudes of Undergraduate Dietitians.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Evans, E., Gould, V., Redmond E.C,, Alwan, N., Hjeji, L., Ilic S. 2019 International Association of Food Protection 2019. How Does the Food Safety Knowledge of Student Dietitians Compare at a University in Wales, Lebanon and Ohio?
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Gomez, C. Bradley, M., Ilic, S., Paden, H.*, Ryser, E., Mitchel, J. 2019 International Association of Food Protection 2019. Creating a Risk Model for Nosocomial Listeriosis in Cancer Patients Who Consume Ready-to-Eat Salad.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Moodispaw, M., Saint-Preux, C., Srivastava, V., Lewis Ivey, M., Ilic, S. 2019. International Association of Food Protection 2019. Survival of Listeria monocytogenes in Hydroponic Lettuce Systems.
• Type: Other Status: Other Year Published: 2019 Citation: Ilic, S. (2019, Jun). Food safety and behavior change. Food and Agriculture Organization of the United Nations (FAO), Rome, Italy
• Type: Other Status: Other Year Published: 2019 Citation: Ilic, S. (2019, Jun). Biocides and antimicrobial resistance in food processing. Food and Agriculture Organization of the United Nations (FAO), Rome, Italy
• Type: Other Status: Other Year Published: 2019 Citation: Lewis Ivey, M.L. (2019, Jun). Antimicrobial Use on Crops and Effects on Antimicrobial Resistant Microorganisms and Gene Frequency in the Plant Production Environment. Food and Agriculture Organization of the United Nations (FAO), Rome, Italy
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Tomoichiro Oka, Hiroyuki Saito, Takayuki Kobayashi, Tomoko Takahashi, Takashi Shimoike, Michiyo Kataoka, Qiuhong Wang, Linda J. Saif, Mamoru Noda, Hirotaka Takagi. Cell culture trials for human sapoviruses. The 7th International Calicivirus Conference in Sydney, Australia from October 13  October 16, 2019.
• Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Malak A. Esseili1, Xiang Gao, Patricia Boley, Yixuan Hou, Linda J. Saif, Paul Brewer-Jensen, Lisa Lindesmith, Ralph S. Baric, Robert L. Atmar, Mary K. Estes and Qiuhong Wang. Human norovirus HBGA binding pocket mediates the virus specific interactions with lettuce carbohydratesThe 7th International Calicivirus Conference in Sydney, Australia from October 13  October 16, 2019.