Source: UNIVERSITY OF MISSOURI submitted to
INCREASING THE SAFETY OF FOODS BY RAPID MOLECULAR DETECTION OF FOODBORNE PATHOGENS AND THE USE OF NOVEL ANTIMICROBIALS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
1007829
Grant No.
(N/A)
Project No.
MO-HAFE0001
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Jan 1, 2016
Project End Date
Dec 31, 2020
Grant Year
(N/A)
Project Director
Mustapha, AZ.
Recipient Organization
UNIVERSITY OF MISSOURI
(N/A)
COLUMBIA,MO 65211
Performing Department
Food Systems & Bioengineering
Non Technical Summary
Millions of Americans are stricken with microbial-related foodborne illnesses each year, resulting in thousands of hospitalizations and deaths. Despite much progress in food testing, plant inspections, and manufacturer and consumer education, outbreaks related to contaminated foods continue to occur. This project will utilize molecular techniques to develop rapid, sensitive and selective assays to accurately detect live pathogens in food, and investigate the usefulness and toxicity of novel antimicrobial compounds at inhibiting such pathogens. Results of this research will be published in scientific journal articles, and presented at scientific meetings and invited seminars around the world. The ultimate goal of this project is to increase the safety of foods that we eat by eliminating or reducing the levels of potentially dangerous pathogens by the use of robust detection tools and potent and safe antimicrobial compounds.
Animal Health Component
0%
Research Effort Categories
Basic
5%
Applied
95%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7124010110050%
5014020104025%
7234030115025%
Knowledge Area
712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins; 501 - New and Improved Food Processing Technologies; 723 - Hazards to Human Health and Safety;

Subject Of Investigation
4010 - Bacteria; 4020 - Fungi; 4030 - Viruses;

Field Of Science
1040 - Molecular biology; 1100 - Bacteriology; 1150 - Toxicology;
Goals / Objectives
The goals of this project are to (1) design and develop highly sensitive, selective and rapid molecular detection methods to simultaneously detect various important foodborne pathogens, including STEC, Salmonella and antibiotic resistant strains of these and other pathogens; (2) investigate the antimicrobial effects of inorganic and organic nanoparticles and other novel antimicrobial compounds at inhibiting pathogens in foods; and (3) investigate the toxicity of nanomaterials on mammalian cells.
Project Methods
Real time or q-PCR will be the main platform for molecular detection, but biosensors that use oligonucleotides, aptamers or antibodies may also be explored. In addition to using probe-based q-PCR, other variations, including high resolution melting (HRM) curve q-PCR, isothermal q-PCR and biosensor-based techniques will be developed for simultaneous detection of multiple serotypes of STEC, Salmonella and antibiotic resistant strains of these and other pathogens. Using nucleic acid intercalating dyes or other techniques, the specific detection of viable and viable-but-non-culturable cells will also be developed via these methods. All developed assays will be validated with various types of food samples, including meats and eggs, produce, and juices and compared to culture-based standard methods.Inorganic nanoparticles, such as ZnO, nanoAg, TiO2 and organic nanoparticles, such as graphene oxide and nanocellulose will be studied for their inhibitory actions against foodborne pathogens in vitro and when incorporated into packaging materials. Pathogenic cells will be incubated in the presence of various concentrations of the nanoparticles and their growth monitored over time. Reduction in numbers of cells as compared to a control will be evaluated to determine inhibitory activity.Because not much is known about the toxicity of nanoparticles, this will be evaluated using mammalian cells, such as Caco-2 and HeLa cells and cytotoxicity assays, such as the MTT assay will be conducted. No significant differences in cell viability between treated and control cells would indicate the relative non-toxicity of the nanoparticles if ingested.In addition to training graduate students to conduct these experiments, undergraduate student assistants will also benefit from participating is these research efforts. Some of these techniques will also be covered in various upper level Food Microbiology courses taught in the department to allow for other students to also benefit from the latest findings in these areas.The successes of the projects will be measured by way of outcomes, for example, number of publications/presentations generated and number or graduate students completed.

Progress 01/01/16 to 12/31/20

Outputs
Target Audience: Nothing Reported 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? Nothing Reported 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 goals of this project are to (1) design and develop highly sensitive, selective and rapid molecular detection methods to simultaneously detect various important foodborne pathogens, including STEC, Salmonella and antibiotic resistant strains of these and other pathogens; (2) investigate the antimicrobial effects of inorganic and organic nanoparticles and other novel antimicrobial compounds at inhibiting pathogens in foods; and (3) investigate the toxicity of nanomaterials on mammalian cells.

Publications


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

    Outputs
    Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Five Ph.D. students and one undergraduate student were trained during this time period. These students conducted the experiments and were involved in writing the research publications that were generated. They also presented the research at the IAFP annual meetings. Three USDA Borlaug Fellows were trained and collaborative food safety research continues with these Fellows. How have the results been disseminated to communities of interest?The results of the research have been disseminated to communities of interest via scientific publications, conference presentations and virtual international meetings. 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 goals of this project are to (1) design and develop highly sensitive, selective and rapid molecular detection methods to simultaneously detect various important foodborne pathogens, including STEC, Salmonella and antibiotic resistant strains of these and other pathogens; (2) investigate the antimicrobial effects of inorganic and organic nanoparticles and other novel antimicrobial compounds at inhibiting pathogens in foods; and (3) investigate the toxicity of nanomaterials on mammalian cells.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2020 Citation: 1. Yu, Z., W. Wang, L. Sun, F. Kong, M. Lin and A. Mustapha. 2020. Preparation of cellulose nanofibril/titanium dioxide nanoparticle nanocomposites as fillers for PVA-based packaging and investigation into their intestinal toxicity. Int. J. Biol. Macromol. 156:1174-1182 https://doi.org/10.1016/j.ijbiomac.2019.11.153
    • Type: Journal Articles Status: Published Year Published: 2020 Citation: 2. Torres Dominguez, E., P. H. Nguyen, A. Hylen, M. R. Maschmann, A. Mustapha, H. K. Hunt. 2020. Design and characterization of mechanically stable, nanoporous TiO2 thin film antimicrobial coatings for food contact surfaces. Mat. Chem. Phys. 251:123001 https://doi.org/10.1016/j.matchemphys.2020.123001
    • Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: 3. Choo, K.-W., L. Mao, M. Lin and A. Mustapha. 2020. Antimicrobial, physical and mechanical properties of polyvinyl alcohol films incorporated with modified bacterial nanocellulose. Presented at the International Association for Food Protection Annual Meeting, October 26-28. P3-50.
    • Type: Conference Papers and Presentations Status: Accepted Year Published: 2020 Citation: 4. Dhital, R., M. Hodel and A. Mustapha. 2020. Detection of virulence and ESBL genes in Salmonella by multiplex high resolution melt-curve real-time PCR assay. Presented at the International Association for Food Protection Annual Meeting, October 26-28. P1-112.


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

    Outputs
    Target Audience:Food science researchers and educators, regulatory agencies, food industry personnel, stakeholders and consumers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?FourPh.D. students were trained. These studens conducted the experiments and were involved in writing the research publications that were generated. They also presented the research at the IFT and IAFP annual meetings. How have the results been disseminated to communities of interest?Via scientific publications, and conference poster and oral presentations. What do you plan to do during the next reporting period to accomplish the goals?Continue to work on all listed objectives.

    Impacts
    What was accomplished under these goals? (1) Risk Assessment: Assess food safety risks in agriculture systems (2) Risk Management: Develop science-based interventions to prevent and mitigate food safety threats (3) Risk Communication: Communicate food safety messages to stakeholders

    Publications

    • Type: Journal Articles Status: Other Year Published: 2019 Citation: Yu, Z., W. Wang, L. Sun, F. Kong, M. Lin and A. Mustapha!. 2019. Preparation of cellulose nanofibril/titanium dioxide nanoparticle nanocomposites as fillers for PVA-based packaging and investigation into their intestinal toxicity. Int. J. Biol. Macromol. In Press https://doi.org/10.1016/j.ijbiomac.2019.11.153
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Torres Dominguez, E., Ph. H. Nguyen, H. K. Hunt! and A. Mustapha!. 2019. Antimicrobial coatings for food contact surfaces: Mechanical properties, potential applications and legal framework. Comprehensive Rev. of Food Science and Food Safety 18:1825-1858 https://doi.org/10.1111/1541-4337.12502
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Wang, W., Z. Yu, F. K. Alsammarraie, F. Kong, M. Lin and A. Mustapha!. 2019. Properties and antimicrobial activity of polyvinyl alcohol-modified bacterial nanocellulose packaging films incorporated with silver nanoparticles. Food Hydrocolloids 100(2020) 105411 https://doi.org/10.1016/j.foodhyd.2019.105411
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Lee, J.-H., A. Mustapha and J. Hwang. 2019. Enhancing ethnic restaurant visits and reducing risk perception: The effect of information and protection motivation. J. Hosp. and Tourism Insights 2(4):341-357. https://doi.org/10.1108/JHTI-10-2018-0068
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Yu, S., W. Wang, R. Dhital, F. Kong, M. Lin! and A. Mustapha!. 2019. Antimicrobial effect and toxicity of cellulose nanofibril/silver nanoparticle nanocomposites prepared by an ultraviolet irradiation method. Colloids Surfaces B: Biointerfaces. 180:212-220. https://doi.org/10.1016/j.colsurfb.2019.04.054
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Yu, Z., R. Dhital, W. Wang, L. Sun, W-C. Zeng, A. Mustapha! and M. Lin!. 2019. Development of multifunctional nanocomposites containing cellulose nanofibrils and soy proteins as food packaging material. Food Packaging and Shelf Life 21 (2019) 100366. https://doi.org/10.1016/j.fpsl.2019.100366
    • Type: Journal Articles Status: Published Year Published: 2019 Citation: Yu, Z., W. Wang, F. Kong, M. Lin! and A. Mustapha!. 2019. Cellulose nanofibril/silver nanoparticle composite as an active food packaging system and its toxicity to human colon cells. Int. J. Biological Macromol. 129:887-894. https://doi.org/10.1016/j.ijbiomac. 2019.02.084
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Choo, K.-W., W. Wang and A. Mustapha. 2019. Antimicrobial and physical properties of chitosan/acetylated starch edible films containing cinnamon and clove essential oils. Presented at the International Association for Food Protection Annual Meeting, Louisville, KY, July 21-24. P2-29.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Alsammarraie, F., A. Mustapha and M. Lin. 2019. Rapid determination of thiabendazole in juice by SERS coupled with novel gold nano-substrates. Presented at the Institute of Food Technologists Annual Meeting, New Orleans, LA, June 2-5. P02-107.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Yu, Z., M. Lin, A. Mustapha and W. Wang. 2019. Synthesis of cellulose nanofibril/titanium dioxide nanoparticle composites and investigation into their toxicity. Presented at the Institute of Food Technologists Annual Meeting, New Orleans, LA, June 2-5. P02-123.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Yu, Z., M. Lin, A. Mustapha and W. Wang. 2019. Antimicrobial effect and toxicity of cellulose nanofibril/silver nanoparticle nanocomposite prepared by the ultraviolet irradiation method. Presented at the Institute of Food Technologists Annual Meeting, New Orleans, LA, June 2-5. P04-093.


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

    Outputs
    Target Audience:Food Science and food safety researchers, educators, food industry, consumers, regulatory agencies Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Three Ph.D. students and one M.S. student in Food Science were trained and conducted the experiments to accomplish the goals of this project. The students presented their research at the IFT and IAFP annual meetings and were involved in writing the manuscripts for publications that were generated from the research. How have the results been disseminated to communities of interest?Via conference presentations (International Association for Food Protection and Institute of Food Technologists annual meetings), scientific journal article publications and Food Microbiology courses at the University of MO. What do you plan to do during the next reporting period to accomplish the goals?Continue working on our goals by designing more novel molecular pathogen detection assays and studying the antimicrobial and toxic effects of new nanomaterials and natural antimicrobial compounds.

    Impacts
    What was accomplished under these goals? Under objective 1, we have developed a high resolution melt curve real time PCR methods for detecting majority of Salmonella in foods. Under objective 2, we have developed and have been testing various nanocellulose films incorporated with nanosilver and essential oils at inhibiting pathogens in foods;as well assynthesized nanosilver using green chemistry and tested its antimicrobial activity against foodborne pathogens. Under objective 3, we have tested our developed nanocellulose filmsand green-synthesized nanosilver against Caco2 and HFC mammmalian cells.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2018 Citation: Yu, Z., S. Lin, W. Wang, W. Zeng, A. Mustapha, and M. Lin!. 2018. Soy protein-based films incorporated with cellulose nanocrystals and pine needle extract for active packaging. Ind. Crops. Prod. 112:412-419. https://doi.org/10.1016/j.indcrop.2017.12.031
    • Type: Journal Articles Status: Published Year Published: 2018 Citation: Alsammarraie, F. Y., M. Lin, A. Mustapha, H. Lin, X. Chen, Y. Chen, H. Wang and M. Huang. 2018. Rapid determination of thiabendazole in juice by SERS coupled with novel gold nanosubstrates. Food Chem. 259:219-225. https://doi.org/10.1016/j.foodchem. 2018.03.105
    • Type: Journal Articles Status: Published Year Published: 2018 Citation: Liu, Y., P. Singh and A. Mustapha!. 2018. Multiplex high resolution melt-curve real-time PCR assay for reliable detection of Salmonella. Food Control 91:225-230. https://doi.org/10.1016/j.foodcont.2018.03.043
    • Type: Journal Articles Status: Published Year Published: 2018 Citation: Alsammarraie, F., A. Mustapha! and M. Lin!. 2018. Green synthesis of silver nanoparticles using turmeric extracts and their antibacterial activities. Colloids and Surfaces B: Biointerfaces 171:398-405. https://doi.org/10.1016/j.colsurfb.2018.07.059
    • Type: Journal Articles Status: Published Year Published: 2018 Citation: Yuan Y., M. Lin, G. Zheng and A. Mustapha!. 2018. Detection of viable Escherichia coli in environmental water using a combined propidium monoazide staining-qPCR. Water Research 145:398-407. (https://doi.org/10.1016/j.watres.2018.08.044)
    • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Wang, W. Z. Yu, M. Lin and A. Mustapha. 2018. Antimicrobial ability of modified bacterial cellulose film against spoilage microorganisms. Presented at the International Association for Food Protection Annual Meeting, Salt Lake City, UT, July 8-11. P1-06.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2018 Citation: Alsammarraie, F., A. Mustapha and M. Lin. 2018. Green synthesis of silver nanoparticles using turmeric extracts and investigation of their antibacterial activities. Presented at the Institute of Food Technologists Annual Meeting, Chicago, IL, July 15-18. S406.


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

    Outputs
    Target Audience:Food Science and food safety esearchers, educators, food industry, consumers, regulatory agencies Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Undergraduate and graduate students were trained while they conducted the research experiments for this project. How have the results been disseminated to communities of interest?Via conference presentations (International Association for Food Protection and Institute of Food Technologists annual meetings), scientific journal article publications and Food Microbiologycourses at the University of MO. What do you plan to do during the next reporting period to accomplish the goals?Continue working on our goals by designing more novel molecular pathogen detection assays and studying the antimicrobial and toxic effects of new nanomaterials and natural antimicrobial compounds.

    Impacts
    What was accomplished under these goals? Under objective 1, we have developed two new high resolution melt curve real time PCR methods for detecting only E. coli O157:H7 and majority ofSalmonella, respectively. Under objective 2, we have developed and have been testing various nanocellulose films incorporated with nanosilver and essential oils at inhibiting pathogens in foods. Under objective 3, we have tested one of our developed nanocellulose films against Caco2 cells and are continuing experiments in this objective using HFC cells.

    Publications

    • Type: Journal Articles Status: Under Review Year Published: 2017 Citation: Yuan Y., G. Zheng and A. Mustapha. 2017. Detection of viable Escherichia coli in environmental water using a combined propidium monoazide staining-qPCR. Water Research. Submitted. Liu, Y., P. Singh and A. Mustapha. 2017. Multiplex high resolution melt-curve real-time PCR assay for reliable detection of Salmonella. Food Control. Submitted.
    • Type: Journal Articles Status: Published Year Published: 2017 Citation: Liu, Y., P. Singh and A. Mustapha. 2017. High-resolution melt curve PCR assay for specific detection of E. coli O157:H7 in beef. Food Control 86:275-282. Ouf, J., Y. Yuan, P. Singh and A. Mustapha. 2017. Detection of viable but nonculturable Escherichia coli O157:H7 in ground beef by propidium monoazide real-time PCR. Int. J. Agric. Sci. Food Technol. 3(2):026-031. Yu, Z., F. Alsammarraie, F. X. Nayigiziki, W. Wang, B. Vardhanabhuti, A. Mustapha and M. Lin. 2017. Effect and mechanism of cellulose nanofibrils on the active functions of biopolymer-based nanocomposite films. Food Res. Int. 99(1):166-172. Nguyen, T. H.-D., B. Vardhanabhuti, M. Lin and A. Mustapha. 2017. Antibacterial properties of selenium nanoparticles and their toxicity on Caco-2 cells. Food Control 77:17-24. Shen, Z., A. Mustapha, M. Lin and G. Zheng. 2017. Biocontrol of the internalization of Salmonella enterica and enterohaemorrhagic Escherichia coli in mung bean sprouts with an endophytic Bacillus subtilis. Int. J. Food Microbiol. 250:37-44. Liou, P., F. X. Nayigiziki, F. Kong, A. Mustapha and M. Lin. 2017. Cellulose nanofibers coated with silver nanoparticles as a SERS platform for detection of pesticides in apples. Carb. Pol. 157:643-650.
    • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Chenggeer, J. Jones, M. Chen, Q. Yu, and A. Mustapha. 2017. Prevention of mixed-species biofilm formations on nanoscale plasma-coated surfaces. Presented at the International Association for Food Protection Annual Meeting, Tampa, FL, July 9-12. P3-131. Liu, Y., P. Singh and A. Mustapha. 2017. High-resolution melt curve PCR assay for specific detection of E. coli O157:H7 in beef. Presented at the International Association for Food Protection Annual Meeting, Tampa, FL, July 9-12. P2-199. Wang, W., M. Lin and A. Mustapha. 2017. Antimicrobial ability of modified bacterial cellulose film Presented at the International Association for Food Protection Annual Meeting, Tampa, FL, July 9-12. P3-145. Yu, Z, W. Wang, F. Nayigiziki, B. Vardhanabhuti, A. Mustapha, and M. Lin. 2017. Effect of cellulose nanofibers on active functions of polysaccharide-based edible films. Institute of Food Technologists Annual Meeting, Las Vegas, NV, June 25-28. Mustapha, A. 2016. Engineered nanoparticles: Antimicrobial properties and toxicity (Plenary presentation). Asia-Pacific International Food Safety Conference, Penang, Malaysia, October 11-13.


    Progress 01/01/16 to 09/30/16

    Outputs
    Target Audience:Researchers, educators, consumers, regulatory agencies Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Undergraduate and graduate students were trained while they conducted the research experiments for this project. How have the results been disseminated to communities of interest?Via journal articles, and presentations at the International Association for Food Protection and MO Milk, Food and Environmental Health Association Annual conferences, as well as at the Asia-Pacific International Food Safety Conference in Kuala Lumpur, Malaysia and at a Food Safety Symposium at the National Research Centre in Cairo, Egypt. What do you plan to do during the next reporting period to accomplish the goals?Continue working on our goals by designing more novel molecular pathogen detection assays and studying the antimicrobial and toxic effects of new nanomaterials and natural antimicrobial compounds.

    Impacts
    What was accomplished under these goals? All goals weremet and more research iscontinuing to obtain even more data to support these goals.

    Publications

    • Type: Journal Articles Status: Published Year Published: 2016 Citation: 1) Liou, P., F. X. Nayigiziki, F. Kong, A. Mustapha, and M. Lin. 2016. Cellulose nanofibers coated with silver nanoparticles as a SERS platform for detection of pesticides in apples. Carbohydrate Polymers 157:643-650. 2) Mao, X., T. H.-D. Nguyen, M. Lin, and A. Mustapha. 2016. Engineered nanoparticles as potential food contaminants and their toxicity to Caco-2 cells. J. Food Sci. 18(8):T2107-T2113. 3) Nguyen, T. H.-D., P. Zhou, A. Mustapha, and M. Lin. 2016. Use of aminothiophenol as an indicator for the analysis of silver nanoparticles in consumer products by surface-enhanced Raman spectroscopy. Analyst 141:5382-5389. 4) Singh, Y. Pfeiffer, and A. Mustapha. 2016. Multiplex real-time PCR assay for the detection of extended-spectrum ?-lactamase and carbapenemase genes using melting curve analysis. J. Microbiol. Methods 124:72-78. 5) Shen, Z., N. Zhang, A. Mustapha, M. Lin, D. Xu, D. Deng, M. Reed, and G. Zheng. 2016. Identification of host-specific genetic markers within 16S rDNA intervening sequences of 73 genera of fecal bacteria. J. Data Mining Genomics & Proteomics 7(1):1-9. 6) Singh, P. and A. Mustapha. 2015. Multiplex real-time PCR assays for detection of eight Shiga toxin-producing Escherichia coli in food samples by melting curve analysis. Int. J. Food Microbiol. 215:101-108. 7) Nguyen, D.-T., M. Lin and, A. Mustapha. 2015. Toxicity of graphene oxide on intestinal bacteria and Caco-2 cells. J. Food Prot. 78(5):9961002. 8) Zhang, Z., Q. Yu, H. Li, A. Mustapha, and M. Lin. 2015. Standing gold nanorod arrays as reproducible SERS substrates for measurement of pesticides in apple juice and vegetables. J. Food Sci. 80(2):N450-458.