Source: UNIVERSITY OF MISSOURI submitted to
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.
1008946
Grant No.
(N/A)
Project No.
MO-MSFE0009
Proposal No.
(N/A)
Multistate No.
S-1056
Program Code
(N/A)
Project Start Date
Dec 7, 2015
Project End Date
Sep 30, 2018
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
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%
7234030105025%
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; 1050 - Developmental biology;
Goals / Objectives
Risk Assessment: Assess food safety risks in agriculture systems Risk Management: Develop science-based interventions to prevent and mitigate food safety threats Risk Communication: Communicate food safety messages to stakeholders
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 10/01/17 to 09/30/18

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?Three Ph.D. and one M.S. students were trained. These students 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 conferences. How have the results been disseminated to communities of interest?Via scientific publications, 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? Risk assessment, management and communication were addressed by development ofnovel PCR assays to detect foodborne pathogens in food, development of novel nanocellulose packaging films containing nanomaterials to increase food safety, and investigation ofnovel antimicrobials to mitigatefoodnorne pathogens. Data fromour research were published in high ranking scientific journals and presented at international conferences.

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.
  • 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 12/07/15 to 09/30/18

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. andtwo M.S. students were trained. These students 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 conferences. How have the results been disseminated to communities of interest?Via scientific publications, conference poster and oral presentations. 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, management and communication were addressed by development of novel PCR assays to detect foodborne pathogens in food, development of novel nanocellulose packaging films containing nanomaterials to increase food safety, and investigation of novel antimicrobials to mitigate foodnorne pathogens. Data from our research were published in high ranking scientific journals and presented at international conferences.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: 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.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: 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.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: 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.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: 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.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: 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.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: 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.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: 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.
  • Type: Conference Papers and Presentations 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. Presented at the International Association for Food Protection Annual Meeting, Tampa, FL, July 9-12. P2-199.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: 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.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: 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.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Mustapha, A. 2016. Engineered nanoparticles: Antimicrobial properties and toxicity (Plenary presentation). Asia-Pacific International Food Safety Conference, Penang, Malaysia, October 11-13.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Liu, Y., P. Singh and A. Mustapha. 2016. Multiplex real-time PCR assay for reliable detection of Salmonella. Presented at the International Association for Food Protection Annual Meeting, St. Louis, MO, July 31-August 3. P1-108.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Yuan, Y., G. Zheng and A. Mustapha. 2016. Detection of viable Escherichia coli in environmental water using a combined propidium monoazide staining-real-time PCR. Presented at the International Association for Food Protection Annual Meeting, St. Louis, MO, July 31-August 3. P3-65.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: 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.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: 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.


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

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?Training of graduate and undergraduate students in Food Science and Engineering. How have the results been disseminated to communities of interest?Via scientific publications, 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? Risk assessment, management and communication were addressed by scientific experiments to detect foodborne pathogens in food and investigate novel antimicrobials to mitigate these pathogens. Data from the studies outlined below were published in high ranking scientific journals and presented at international conferences. Development of multiplex melt-curve q-PCR assays for detection of antibiotic resistant pathogens, Shiga toxin producing Escherichia coli and Salmonella. Study and analyses ofplasma coated food contact surfaces at preventing biofilms of mixed strain food pathogens. Investigation into the antimicrobial properties and toxicity of nanomaterials. Green synthesis of silver nanoparticles and investigations into their antimicrobial properties. Development of food packaging films using nanocellulose polymers and investigations into their antimicrobial properties.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 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.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: 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.


Progress 12/07/15 to 09/30/16

Outputs
Target Audience:Educators, researchers, consumers, regulatory agencies Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Graduate and undergraduate students were trained while conducting the experiments for the research. How have the results been disseminated to communities of interest?By way of scientific publications, presentations at IAFP and MO Milk, Food and Environmental Health Associationmeeting and international presentations at the Asia-Pacific International Food Safety Conference in Malaysia and at a Food Safety Symposium at the National Research Center, Cairo, Egypt. What do you plan to do during the next reporting period to accomplish the goals?Continue working on novel research in nanomaterials and rapid molecular pathogen detection.

Impacts
What was accomplished under these goals? We havegenerated important data on novel interventions for preventing and mitigating food safety threats via the use of nanomaterials and rapid molecular pathogen detection techniques.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: 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. 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. 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. 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. 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. 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. 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. 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.