Frontiers in Food Safety and
Bioscience
Center for Food Safety and Applied Nutrition
Food and Drug Administration
Sufian Alkhaldi, Ph. D.
Overview
7 Food Concerns You Need to Watch
7 Cutting-Edge Technologies You Need to Pay Attention to
Background
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
U. S. Food SupplyU. S. Food Supply
USDA FDA
7 Food Safety Concerns
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
Some of the new foodborne pathogens
found since 1977
1-Shiga toxin-producing E. coli
2- Noroviruses 3- Salmonella 4-Vibrio cholerae O139 5-Listeria
monocytogenes 6- Campylobacter jejuni 7- Spongiform
encephalopathy prions
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%
CDC Estimated % of Total Foodborne Illnesses, Hospitalizations, and Deaths
0
10
20
30
40
50
60
70
%Total FBI %TotalHospital
%TotalDeaths
Salmonella
Listeria
Norwalk-like viruses
Campylobacter
E.coli O157:H7
Data from Mead, et.al., Food Related Illness and Death in the United States,
Emerging Infectious Disease, 1999. Vol.5, No. 5, pp.38
Contributing factors to foodborne outbreaks from 1993-1997
0
100
200
300
400
500
600
700
800
900
Improper holding cold or hot
Inadequate cooking
Cross Cont. Unsafe food source
Poor personal hygiene
Num
bers
of
outb
reak
s
Olsen et al. 2000. MMWR Morb Mortal Wkly Rep, 200; 49:1-62
CDC’s EHSNET OUTBREAK/NONOUTBREAK STUDY
[2002 – 2003]
Bacterial32%
Parasitic5%
Toxin3%
Unknown13%
Viral 47%
Viral
Bacterial
Parasitic
Toxin
Unknown/Missing
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.
Food Purpose Dose
Fresh, non-heated processed pork Control of Trichinella spiralis0.3 kGy min. to 1 kGy max.
Fresh foods Growth and maturation inhibition 1 kGy max.
Foods Arthropod disinfection 1 kGy max.
Dry or dehydrated Enzyme preparations Microbial disinfection 10 kGy max.
Dry or dehydrated spices/seasonings Microbial disinfection 30 kGy max.
Fresh or frozen, uncooked poultry products Pathogen control 3 kGy max.
Frozen packaged meats (solely NASA) Sterilization 44 kGy min.
Refrigerated, uncooked meat products Pathogen control 4.5 kGy max.
Frozen uncooked meat products Pathogen control 7 kGy max.
Fresh shell eggs Control of Salmonella 3.0 kGy max.
Seeds for sprouting Control of microbial pathogens 8.0 kGy max.
Fresh or frozen molluscan shellfish1 Control of Vibrio species and other foodborne pathogens 5.5 kGy max.
Foods Permitted to be Irradiated Under FDA's Regulations (21 CFR 179.26)
The logo, as required by regulation of the US-FDA to show a food has been treated
with ionizing radiation.
Editor: Sufian Alkhaldi Ph.D.
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.
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
Biological Threat Agent Detection: Targeting at
Molecular Level
Bacterial Cells
BacterialSpores
NucleicAcid
Viruses
Antigens
DNA RNA
Toxins
Antigens
ExtracellularAntigens
IntracellularAntigens
Extracellular Antigens
Iqbal et al. 2000. A review of molecular recognition technologies for detection of biological threat agents. Biosensors & Bioelectronics 15: 549-578
Dr. Patrick Brown
Public Database
Spotted DNA Microarray
Target Sequence
Sequence Selection using software (bioinformatic)
Hybridization
DNA Chip Strategy
DNA labeling
DNA Purificatio
n
Hybridization
DNA digestion
DNA Chip Strategy
Vir F Ail Yst blaA 16S
Hybridization Probe of Yersinia enterocolitica
Dr. J. Craig Venter
Crew member of the RV Weatherbird II hauls in water samples containing microbes from
the Sargasso Sea.
Sargasso Sea
Sargasso Sea is one of the best-studied and most well-characterized regions of the global ocean
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.
Metagenomic in Action
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.
Separate oil & water
Add emulsifier Add biopolymer
Primary emulsion Secondary emulsion
Expressed bacterial proteins (Condition 1)
Expressed bacterial proteins (Condition 2)
Tag with Cy5
Tag with Cy3
First dimensional proteinSeparation by IPGphorIEF System
Second dimensional protein Separation by DALT II Vertical System
Protein sequencing and molecular weight determination by MALDI-Reflectron-TOF
Protein picking, digesting, and purification by spot handling workstation
Two dimentional SDS PAGE with two fluorescent tag proteins.
Discovering unknown protein functions.
Drug development. Vaccine development.
Dr. Barry Bochner
DNA RNA PROTEIN PHENOTYPE
Molecular Analyses Cellular Analysiswhich is more
conclusive, kinetic
Biolog, 2001
Wells contain different tests and measure different pathways
Respiration is measured using a Redox Dye
PM Set Up Procedure
Phenotype Microarray Pattern containing different 2000
substrate
Omni Log Phentoype Microarray System
Phenotype kinetic results
Microtiter plate inoculation
Tested bacteria grown in minimal medium
Phenotypic Microarray
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.
+ agarose
+ RestrictionEnzyme
+ lysis buffer
100
90807060 PFGE-AscI PFGE-ApaI
Sandwich
Sandwich
Sandwich
Sandwich
Sandwich
Sandwich
Finished Product
Finished Product
Finished Product
Swab
Swab
Swab
Swab
Swab
Swab
Swab
Finished Product
2000
2000
2000
1999
2000
2001
2005
2005
2005-03-10
2005-03-17
2005-03-10
2005-09-07
2005-09-07
2005-09-07
2005-09-07
% Similarity
Sandwich Producing PlantListeria monocytogenes
Federal Partners(FDA, USDA)
Food and Environmental Isolates
State and Local
PartnersClinical, Food, and
Environmental Isolates
International Partners
Clinical, Food, and Environmental
Isolates
CDCClinical isolates,
State and International Overflow
Questions?And my Thanks!WWW.Suf-Microarray.com
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