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Frontiers in Food Safety and Bioscience

Frontiers in Food Safety and Bioscience. Sufian Alkhaldi, Ph. D. Center for Food Safety and Applied Nutrition Food and Drug Administration. Overview. 7 Food Concerns You Need to Watch 7 Cutting-Edge Technologies You Need to Pay Attention to. Background. U. S. Food Supply.

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Frontiers in Food Safety and Bioscience

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  1. Frontiers in Food Safety and Bioscience Sufian Alkhaldi, Ph. D. Center for Food Safety and Applied Nutrition Food and Drug Administration

  2. Overview • 7 Food Concerns You Need to Watch • 7 Cutting-Edge Technologies You Need to Pay Attention to

  3. Background U. S. Food Supply • The mission of FDA CFSAN is to protect the public health through • Safe and secure food supply • Nutrition information through the food label • FDA has a variety of tools to achieve this mission • Regulations • Guidance • Risk communication FDA USDA

  4. 7 Food Safety Concerns

  5. Types of foodborne threats • Microbial • 1- Bacterial • 2- Viral • 3- Parasitic • Chemical • 4- Antibiotics • 5- Carcinogens (e.g. dioxins) • 6- Heavy metals • Other • 7- Prions

  6. 1-Shiga toxin-producing E. coli 2- Noroviruses 3- Salmonella 4-Vibrio cholerae O139 5-Listeria monocytogenes 6- Campylobacter jejuni 7- Spongiform encephalopathy prions Some of the new foodborne pathogens found since 1977

  7. Major causes of foodborne disease (Mead et al 1999) • Bacterial (5,200,000) % foodborne • 1- Campylobacter ~ 2.4 million 80% • 2- Salmonella ~1.4 million 80% • 3- Shigella ~ 0.5 million 20% • Parasitic (2,500,000) • 4- Giardia ~ 2 million 10% • Viral (31,000,000) • 5- Norovirus ~23 million 40% • 6- Rotavirus ~4 million 1% • 7- Astrovirus ~4 million 1%

  8. CDC Estimated % of Total Foodborne Illnesses, Hospitalizations, and Deaths Data from Mead, et.al., Food Related Illness and Death in the United States, Emerging Infectious Disease, 1999. Vol.5, No. 5, pp.38

  9. Contributing factors to foodborne outbreaks from 1993-1997 Numbers of outbreaks Poor personal hygiene Improper holding cold or hot Unsafe food source Inadequate cooking Cross Cont. Olsen et al. 2000. MMWR Morb Mortal Wkly Rep, 200; 49:1-62

  10. CDC’s EHSNET OUTBREAK/NONOUTBREAK STUDY [2002 – 2003]

  11. Food and Radiation • The FDA took an additional step in December 1997 when it approved irradiation to control pathogens in fresh and frozen red meat. Gamma rays from a solid radioactive source (cesium-137 or cobalt-60) penetrate a food product and kill any present pathogens or parasites by disrupting bacterial DNA. • The USDA is currently writing regulations for its use.

  12. Foods Permitted to be Irradiated Under FDA's Regulations (21 CFR 179.26)

  13. The logo, as required by regulation of the US-FDA to show a food has been treated with ionizing radiation.

  14. Bad Bug Book Editor: Sufian Alkhaldi Ph.D.

  15. Bad Bug Book • The provide basic facts regarding foodborne pathogenic microorganisms and natural toxins. • It brings together in one place information from the Food & Drug Administration, the Centers for Disease Control & Prevention, the USDA Food Safety Inspection Service, and the National Institutes of Health. • http://www.cfsan.fda.gov/~mow/intro.html • Editor: Sufian Alkhaldi Ph.D.

  16. 7 Cutting Edge Technologies You Need to Pay Attention to • 1- DNA Microarray • 2- Metagenomics • 3- Food Nanotechnology • 4- Functional Proteomic • 5- Pheneomic (Phenotype Microarray) • 6- Pulse-Field Gel Electrophoresis • 7- Epigenetics

  17. Biological Threat Agent Detection: Targeting at Molecular Level Bacterial Spores Viruses Toxins Bacterial Cells Antigens Nucleic Acid Antigens DNA RNA Intracellular Antigens Extracellular Antigens Extracellular Antigens Iqbal et al. 2000. A review of molecular recognition technologies for detection of biological threat agents. Biosensors & Bioelectronics 15: 549-578

  18. Dr. Patrick Brown 1- Microarray

  19. Target Sequence Hybridization DNA Chip Strategy Sequence Selection using software (bioinformatic) Public Database Spotted DNA Microarray

  20. DNA Chip Strategy DNA labeling DNA digestion Hybridization DNA Purification

  21. Hybridization Probe of Yersinia enterocolitica Vir F blaA 16S Yst Ail

  22. 2- Metagenomic Dr. J. Craig Venter

  23. Metagenomic in Action Sargasso Sea Sargasso Sea is one of the best-studied and most well-characterized regions of the global ocean Crew member of the RV Weatherbird II hauls in water samples containing microbes from the Sargasso Sea. The Sargasso Sea has been intensively studied as part of 50-year time series of ocean physics and biogeochemistry and provides an opportunity for interpretation of environmental genomic data in oceanographic contest.

  24. 3-Food Nanotechnology • Nano emulsions can encapsulate functional ingredients with their droplets, which can facilitate a reduction in chemical degradation. • Functional component encased within one component of a multiple emulsion system could be released in response to specific environmental trigger.

  25. Food Nanotechnology Addemulsifier Addbiopolymer Separate oil & water Primary emulsion Secondary emulsion

  26. 4- Functional Proteomics Tag with Cy5 First dimensional protein Separation by IPGphor IEF System Second dimensional protein Separation by DALT II Vertical System Expressed bacterial proteins (Condition 1) Tag with Cy3 Expressed bacterial proteins (Condition 2) Two dimentional SDS PAGE with two fluorescent tag proteins. Protein sequencing and molecular weight determination by MALDI-Reflectron-TOF Protein picking, digesting, and purification by spot handling workstation

  27. Application of Functional Proteomic • Discovering unknown protein functions. • Drug development. • Vaccine development.

  28. Biolog, 2001 Cellular Analysis which is more conclusive, kinetic Dr. Barry Bochner 5- Phenotypic Microarray DNA RNA PROTEIN PHENOTYPE Molecular Analyses

  29. Respiration is measured using a Redox Dye Wells contain different tests and measure different pathways

  30. PM Set Up Procedure

  31. Tested bacteria grown in minimal medium Microtiter plate inoculation Phenotype Microarray Pattern containing different 2000 substrate Phenotype kinetic results Omni Log Phentoype Microarray System Phenotypic Microarray

  32. Application of Phenotypic Microarray • Studying cell growth in presence of 700-2000 substrates. • Animal cloning. • Improving bacterial mutant to produce enzymes. • Identifying unknown sequences and mutant effect on cells.

  33. + Restriction Enzyme + lysis buffer + agarose 6- PFGE Pulsed-Field Gel Electrophoresis

  34. % Similarity PFGE-AscI PFGE-ApaI 100 80 90 70 60 Sandwich 2000 Sandwich 2000 Sandwich 2000 Sandwich 1999 Sandwich 2000 Sandwich 2001 Finished Product Finished Product 2005 Finished Product 2005 Swab 2005-03-10 Swab 2005-03-17 Swab 2005-03-10 Swab 2005-09-07 Swab 2005-09-07 Swab 2005-09-07 Swab 2005-09-07 Finished Product Sandwich Producing PlantListeria monocytogenes

  35. Federal Partners (FDA, USDA) Food and Environmental Isolates CDC Clinical isolates, State and International Overflow State and Local Partners Clinical, Food, and Environmental Isolates International Partners Clinical, Food, and Environmental Isolates

  36. Questions?And my Thanks!WWW.Suf-Microarray.com

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