Frontiers in Food Safety and Bioscience

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