Transcript Dr. Sufian Alkhaldi - 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
USDA

The mission of FDA CFSAN is to protect the public health
through



FDA
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
7 Food Safety Concerns
Types of foodborne threats

Microbial




Chemical




1- Bacterial
2- Viral
3- Parasitic
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)




~ 2.4 million
~1.4 million
~ 0.5 million
80%
20%
~ 2 million
10%
~23 million
~4 million
~4 million
40%
1%
1%
Parasitic (2,500,000)


1- Campylobacter
80%
2- Salmonella
3- Shigella
% foodborne
4- Giardia
Viral (31,000,000)



5- Norovirus
6- Rotavirus
7- Astrovirus
CDC Estimated % of Total Foodborne
Illnesses, Hospitalizations, and Deaths
70
60
50
20
Salmonella
Listeria
Norwalk-like viruses
Campylobacter
10
E.coli O157:H7
40
30
0
%Total FBI
%Total
Hospital
%Total
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
Contributing factors to foodborne
outbreaks from 1993-1997
900
Numbers of outbreaks
700
600
500
400
300
200
100
0
ne
s
rc
e
al
h
yg
i
so
u
d
P
oo
rp
er
so
n
fo
o
U
sa
fe
C
on
s
Cross Cont. Unsafe food
source
n
in
at
io
Inadequate
cooking
ta
m
co
C
ro
s
In
ad
eq
u
at
e
co
l
ng
di
H
ol
er
Im
pr
op
ok
in
g
Improper
holding
cold or hot
d
or
ho
t
Numbers of outbreaks
800
Olsen et al. 2000. MMWR Morb Mortal Wkly Rep, 200; 49:1-62
Poor
personal
hygiene
CDC’s EHSNET OUTBREAK/NONOUTBREAK
STUDY
[2002 – 2003]
Unknown
13%
Toxin
3%
Parasitic
5%
Bacterial
32%
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.
Foods Permitted to be Irradiated Under FDA's Regulations
(21 CFR 179.26)
Food
Purpose
Dose
Fresh, non-heated processed pork
Control of Trichinella spiralis
0.3 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.
The logo, as required by regulation of the USFDA 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







1234567-
DNA Microarray
Metagenomics
Food Nanotechnology
Functional Proteomic
Pheneomic (Phenotype Microarray)
Pulse-Field Gel Electrophoresis
Epigenetics
Biological Threat Agent Detection:
Targeting at Molecular Level
Bacterial
Spores
Bacterial
Cells
Nucleic
Acid
DNA
RNA
Viruses
Toxins
Antigens
Antigens
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
Dr. Patrick Brown
DNA Chip
Strategy
Target
Sequence
Sequence Selection using software
(bioinformatic)
Public Database
Spotted DNA
Microarray
Hybridization
DNA Chip Strategy
DNA labeling
DNA digestion
Hybridization
DNA
Purification
Hybridization Probe of
Yersinia enterocolitica
Vir F
Ail
Yst
blaA
16S
Dr. J. Craig Venter
Metagenomic in Action
Sargasso Sea
Sargasso Sea is one of the beststudied 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.


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.
Add emulsifier
Separate oil & water
Add biopolymer
Primary emulsion
Secondary emulsion
Tag with
Cy5
Expressed bacterial
proteins (Condition 1)
First dimensional protein
Separation by IPGphor
IEF System
Second dimensional protein
Separation by DALT II
Vertical System
Tag with
Cy3
Expressed bacterial
proteins (Condition 2)
Protein sequencing and
molecular weight
determination by MALDIReflectron-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
Biolog, 2001
Molecular Analyses
Cellular Analysis
which is more
conclusive, kinetic
Respiration is measured using a Redox Dye
Wells contain different tests and measure
different pathways
PM Set Up Procedure
Phenotypic Microarray
Tested bacteria grown in
minimal medium
Microtiter plate
inoculation
Phenotype Microarray Pattern
containing different 2000
substrate
Omni Log Phentoype
Microarray System
Phenotype kinetic results
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.
+ Restriction
Enzyme
+ lysis buffer
+ agarose
Sandwich Producing Plant
Listeria monocytogenes
100
90
80
70
60
% Similarity
PFGE-AscI
PFGE-ApaI
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
CDC
Clinical isolates,
State and International
Overflow
State and
Local
Partners
Clinical, Food, and
Environmental Isolates
Federal
Partners
(FDA, USDA)
Food and Environmental
Isolates
International
Partners
Clinical, Food, and
Environmental
Isolates
Questions?
And my Thanks!
WWW.Suf-Microarray.com