Emerging Micro Expert Report On Emerging Microbiological Food Safety Issues Implications For Control
Emerging Micro Expert Report On Emerging Microbiological Food Safety Issues Implications For Control
Emerging Micro Expert Report On Emerging Microbiological Food Safety Issues Implications For Control
policy officials, and other interested groupsof the scientific information on emerging foodborne pathogens (from a broad ecological perspective) relative to public policy issues and strategies
for preventing foodborne illness.
Founded in 1939, the Institute of Food Technologists is a nonprofit scientific society with 28,000 members
working in food science, technology, and related professions in the food industry, academia, and government.
As the society for food science and technology, IFT brings sounds science to the public discussion of food issues.
IFT Staff
Mary Helen Arthur, M.T.S.C.
Lead Editor, Expert Report
Dept. of Science and Communications,
Chicago, IL
2
Table of Contents
Introduction ...................................................................................... 4
Trinity of Factors ..................................................................... 4
Evolution of Controls ............................................................. 4
Fig. 1a-Foodborne Illness ....................................................... 4
Fig. 1b-Reducing One Factor .................................................. 4
Fig. 1c-Reducing Multiple Factors .......................................... 4
Table 1. Evolution of Food Processing .................................... 5
Evolution of Food Safety Policies ........................................... 5
Microbiology 101 .................................................................... 6
Incidence and Prevalence of Foodborne Illness ..................... 8
Emergence of Pathogens ......................................................... 8
Fig. 2. Foodborne Illness Identification .................................. 8
Table 2. Foodborne Disease in the United States ................... 9
Complex Drivers of Change ................................................. 11
Framework for Food Safety Management ............................ 12
EXPERT REPORT
Conclusions .................................................................................... 93
References ..................................................................................... 97
On the Cover
The image on the cover is a scanning electron micrograph of
Listeria monocytogenes, an emerging foodborne pathogen.
2000 S. Lowry/Univ. Ulster/Stone
Introduction
The continued occurrence of
foodborne illness is not evidence of the
failure of our food safety system. In
fact, many of our prevention and
control efforts have beenand
continue to behighly effective. The
U.S. food supply is arguably among the
safest in the world, but significant
foodborne illness continues to occur.
Despite great strides in the area of
microbiological food safety, much
remains to be done.
Foodborne illness is not a simple
problem in need of a solution; it is a complex combination of factors that must be
managed on a continual basis. Aside from
the inherent ability of pathogens themselves to evolve, pathogens victims and
the microbial environment play a role in
the changing nature of foodborne illness,
opening new niches and creating new vulnerabilities. No matter how sophisticated
and complex a system is developed, food
safety management is never finished or
complete, because change is constant.
Recognizing that food safety is a fundamental and continuing issue, the Institute of Food Technologists commissioned an expert panel to review the
available scientific literature related to
emerging microbiological food safety issues. The experts were charged with
identifying the factors that make a microorganism emerge as an important
foodborne pathogen and identifying
mechanisms that use this knowledge to
improve the safety of our food supply.
The objective of this report is to increase
understanding, among IFT members
and other interested parties, of the scien-
Trinity of Factors
At the simplest level, foodborne illness can be reduced to three factors: the
pathogen, the host, and the environment
in which they exist and interact (see Fig.
1a). Complex relationships exist among
these factors, and all three factors are
necessary for foodborne illness to occur.
For example, a susceptible host may consume food that contains a significant
amount of a microorganism, but if the
microorganism does not possess the
traits necessary to cause illness, foodborne disease does not occur. Similarly,
adequately cooking a food to kill the
pathogenic microbes can eliminate the
exposure factor and render the food safe.
When one or more of the three factors changes, new foodborne pathogens
emerge. For example, host susceptibility can increase so much that existing
microorganisms that do not cause illness in the general population achieve
pathogen status in the newly immunocompromised individuals. The change
also can be increased virulence, e.g.,
when a microorganism acquires characteristics that help it invade the human
body. Or it can be new exposure, e.g.,
when fruit from one region carries a
pathogenic microorganism to populations in a different geographic region
that have never before been exposed.
This trinity is also the key to reducing foodborne illness. Prevention and
control efforts often focus on the contribution of one of these factors, such as
washing vegetables to remove surface
Evolution of Controls
New technologies, production practices and food manufacturing processes
are developed to meet the needs of a
changing society. Early food preservation
methodssuch as canning, cheese making, bread making, brewing, fermentation,
pickling, salting, and dryingwere used
to provide sufficient year-round food
availability. Later developments reflected a
new focus on food safety, variety, convenience, and nutritive and sensory quality.
At the beginning of the 20th century,
contaminated milk, meat, and other
foods caused large outbreaks and many
sporadic cases of foodborne disease, often with fatal consequences. The revolution in sanitation and hygiene related to
food and water and the almost universal
adoption of thermal pasteurization for
milk produced tremendous improvements in food safety. New technologies
with increasing sophistication have yielded continued improvements in microbiological food safety while delivering better quality foods with greater nutritional
value and superior sensory characteristics (see Table 1).
Innovations in packaging have been
integral to the developments in food processing and product development. Packages contain and protect their food contents and inform consumers; they also
Table 1. Evolution of Food Processing (Goldblith, 1989; IFT, 2000a, 1989; Lund, 1989)
Early times
Heating
Cooking food kills many foodborne pathogens
1770s-1800s
Canning/Thermal Processing
Significant discoveries in response to industrialization forces and Napoleons armies need for less dependence on local
provisions
1890s
Pasteurization
Thermal treatment of raw milk to prevent milk from transmitting pathogens
1920s - 1930s
1940s
Freezing
Mechanical quick-freezing methods to preserve while maintaining quality
1950s
1960s
Freeze Drying
Rapid deep-freezing followed by sublimation of water by heating the frozen product in a vacuum chamber. Best known applied
to coffee, to preserve delicate aroma compounds and maintain flavor and odor
1940s-1980s
1990s
Carcass Spray Treatments (e.g., water, acid), Steam Vacuuming, Steam Pasteurization
Carcass decontamination interventions to meet microbiological performance criteria
High Pressure Processing
Foods subjected to specified pressures and temperatures to preserve food while maintaining quality
ever-changing challenge both for the industrialized and the developing world.
ways apparent. In these cases, the regulatory response has been to either keep the
hazardous food out of the marketplace
or to forgo regulatory action and rely on
prudent people to protect themselves.
Numerous food safety concerns
have been successfully addressed by this
regulatory paradigm. The promulgation
of regulations for low acid canned food
virtually ended the historic association
of botulism with commercially canned
food. Under the regulations, commercially canned foods undergo a minimum calculated destruction of 12D for
Microbiology 101
The characteristics of the various
microorganisms are part of what
makes microbiological food safety issues so complex. One type of microorganism may thrive under conditions that are fatal to a different microbe. Some microbial pathogens
cause disease by infecting the human
host, while others produce toxins that
cause illness. Some pathogens can
multiply in food during storage while
others cannot. Because most microorganisms can reproduce within a
matter of minutes, these pathogens
can evolve quickly when environmental stresses select for strains with
unique survival characteristics.
Types of Microorganisms
Microorganisms are divided into
three distinct categories: prokaryotes,
eukaryotes, and viruses. Both
prokaryotes and eukaryotes are highly
regulated cells that possess elaborate
sensing systems, enabling them to be
aware of and react to their environment as it changes.
ProkaryotesBacteria
Prokaryotes are single-celled living
microorganisms that have no nuclear
membrane separating their genetic
material from the cytoplasm within
the cell. They are microscopic, in that
they cannot be seen with the unassisted human eye. Bacteria are generally
free-living in the environment, although some have complex nutrient
requirements and can grow only in
special niches.
Bacteria are arguably the worlds
Viruses
Foodborne pathogenic viruses are
comprised of a single type of nucleic
acid surrounded by a protein coat. Viruses are not free-living. In fact, they
are not living beings at all, but are obligate intracellular parasites. They are
smaller than bacteria (10-350nm), and
some viruses prey on bacteria (bacteriophages).
Types of Bacteria
Bacteria come in various shapes,
such as rods, spheres (cocci), and spirals.
As a response to certain adverse environmental conditions, some bacteria can
form spores. The spores start as dense
regions within the cell, but as the cell de-
Roles of Foodborne
Microorganisms
Not all microorganisms in foods are
harmful. In fact, only a small proportion
of foodborne organisms have been asso-
organisms that are associated with disease in humans and/or animals. Pathogenic microbes are capable of causing illness, either by infecting the host or by
producing toxins that cause the host to
become ill. Some microorganisms are
pathogenic in one host species but not in
another. For example, Escherichia coli
O157:H7 causes illness in humans but
not in cattle, its primary host.
Commensal Microbes
Virtually all raw food contains microorganisms. The source of these is the
production environment, where a wide
variety of organisms are environmentally
ubiquitous. Processing and handling of
foods can also contribute to the types
and numbers of commensal organisms
on foods. Most of these commensal organisms are harmless to animals and humans; in fact, they may actually be beneficial in that they provide high levels of
competitive microflora that usually
grow faster than contaminating pathogens. Although the purpose of many
common food processing methods such
as pasteurization and canning is the destruction of pathogens, commensal microorganisms are often destroyed in the
process as well.
Spoilage Microbes
Spoilage may be defined as a condition in which food becomes inedible because of undesirable changes in color,
flavor, odor, appearance, and texture.
This condition occurs primarily because the organisms grow to high levels,
producing enzymes that break down
food components such as fats, proteins,
and sugars. In most instances, spoilage
is caused by commensal organisms that
have been allowed to reach populations
in the range of 105 to 107 CFU/g of
food. Different classifications of foods
(such as red meats, vegetables, fish, etc.)
have different spoilage profiles because
the food environment will dictate which
organisms will grow, dominate, and
EXPERT REPORT
Beneficial Microbes
Perhaps the most widely recognized
group of beneficial foodborne microorganisms are the members of the lactic acid
bacteria (LAB) group. This is a functional
name used to classify fermentative organisms that produce lactic acid as the primary by-product of metabolic activity, although other products, such as alcohol
and carbon dioxide may be produced as
well. These metabolic products are responsible for the characteristic flavor,
odor and texture of fermented food products. The lactic acid bacteria are commonly used in dairy, vegetable and meat
fermentations. Notable members of this
group belong to the Lactococcus, Lactobacillus, Pediococcus, and Leuconostoc genera.
Some species of yeasts and molds can also
be used in the commercial production of
fermented foods, including Saccharomyces
cerevisiae, used frequently for producing
bread, beer and wines, and Aspergillus
oryzae, used in the fermentation of oriental foods such as soy sauce.
Foodborne Pathogens
Foodborne pathogens encompass a
relatively small group of foodborne micro-
Microbiological Indicators
An index or indicator refers to a
single or group of microorganisms, or
alternatively, a metabolic product,
whose presence in a food or the environment at a given level is indicative
of a potential quality or safety problem. Microbiological indicators are
used in place of direct testing for a
pathogen, largely because they are
easier to work with. Indicators may
be a specific microorganism (e.g., E.
coli), a metabolite (e.g., lactic acid titration), or some other indirect measure (e.g., ATP bioluminescence as a
measure of sanitation efficacy). Using
a specific microorganism as an indicator is difficult, because appropriate
indicator organisms are difficult to
identify. An ideal indicator organism: (1) has a history of presence in
foods at any time that the target
pathogen or toxin might be present;
(2) is present at concentrations directly related to that of the target
pathogen or toxin; (3) is absent from
food when the target is not present;
(4) has growth rates equivalent to, or
slightly greater than, the pathogen;
(5) has rapid, simple, and inexpensive
quantitative assays available; (6) has
similar resistance profiles to the target; and (7) is nonpathogenic.
nally, some proportion of foodborne illthan half of all recognized foodborne disness is caused by pathogens that have not
ease outbreaks have unknown causes, inyet been identified and thus cannot be
dicating the real number of disease-causdiagnosed. In fact, many of the pathoing agents is likely much larger than 200.
gens of concern today were not recogThe recognized causes of foodborne illnized as causes of foodborne illness just
ness include viruses, bacteria, parasites,
20 years ago (Mead et al., 1999).
manmade chemicals, biotoxins, heavy
New scientific advances make it posmetals, and prions. The symptoms of
sible to approach foodborne illness from
foodborne illnesses range from mild gasa different, broader perspective. More
troenteritis to life threatening neurologic,
powerful diagnostic procedures and bethepatic, and renal syndromes.
ter communication technology allow imIn the United States, foodborne disproved tracking and surveillance for
eases have been estimated to cause apfoodborne illness. Genetic identification
proximately 76 million illnesses,
methods allow scientists to link geo325,000 hospitalizations, and 5,000
graphically distinct outbreaks of fooddeaths each year (Mead et al., 1999). A
borne illness to a single source. Pathopathogens ability to cause illness can be
gens can appear to emerge simply bevery different from the severity of the
cause scientists develop methods to idenillness it causes. Some pathogens such
tify the presence of certain microorganas Norwalk-like viruses cause a great
isms and link them to foodborne disease.
number of illnesses (9.2 million per
New technologies based on recent
year) but the fatality rate is very small
advances in genomics also give scientists
(0.001%) (see Table 2). Others such as
greater insight into pathogen virulence
Vibrio vulnificus cause few illnesses (47
and evolution, opening the door to better
per year), but many of those illnesses
controls and therapeutics. Future scienare fatal (38.3%). Salmonella, Listeria
tific advances will continue to enhance
monocytogenes, and Toxoplasma gondii
efforts to identify and understand foodare responsible for more that 78% of
borne pathogens, and these insights will
the deaths but only approximately 11%
contribute the data necessary for scienceof total cases of foodborne disease. The
based risk assessment and food safety
issue is further complicated by pathomanagement.
gens, such as L. monocytogenes, that
cause little or no illness in healthy individuals but cause severe illness and
Emergence of Pathogens
death in sensitive populations, including the immunosuppressed, the elderly
The terminology newly emerging
and the developing fetus. Prioritizing
pathogen has become somewhat overfood safety resources can be difficult.
used, or perhaps it is merely ill defined.
Scientists gather data about the inciTrue pathogen emergence could be didence and severity of foodborne illness
rectly linked to evolution, whether that
through surveillance, both passive and
evolution occurs gradually or rapidly.
active. Mild cases of illness often are not
In a broader context, emergence can
captured by surveillance programs because Fig. 2. Foodborne Illness Identification
medical intervention is
not required for recovSick individual seeks medical attention.
ery. Many steps are required for a foodborne
Clinician considers cause to be foodborne,
pathogen to be identirequests
proper tests and collects appropriate specimen.
fied as the cause of illness and for data to be
Proper test is performed correctly.
gathered through surveillance programs (see
Fig. 2). Furthermore,
Results are reported to the health department
because many pathoand ultimately the CDC.
gens transmitted
through food also may
Regulatory agency investigates.
be spread by contaminated water and person-to-person contact,
Identification of the source of the illness
the role of food can be
prompts product recall or other action.
difficult to assess. FiINSTITUTE OF FOOD TECHNOLOGISTS
EXPERT REPORT
Table 2. Foodborne Disease in the United States, Including Estimated Annual Prevalence (FDA/CFSAN, 2002; Mead et al., 1999)
Principal Symptomsa
Onseta
Illnessesb
Deathsb
6-15 hr
0.5-6 hr
27,360
n/a
n/a
0
n/a
n/a
Days to weeksc
777
2-5 days
1,963,141
n/a
99
n/a
Clostridium botulinum
18-36 hr
58
Clostridium perfringens
8-22 hr
248,520
Enterotoxigenic
24 hr
55,594
Enteropathogenic
n/a
n/a
n/a
E. coli O157:H7
1-2 days
62,458
52
Enteroinvasive
12-72 hr
n/a
n/a
2,493
499
Raw meats, poultry, eggs, milk and dairy products, fish, shrimp,
frog legs, yeast, coconut, sauces and salad dressings
n/a
n/a
Microorganism
Bacteria
Bacillus cereus
Brucella spp.
Campylobacter spp.
C. jejuni
Escherichia coli
Listeria monocytogenes
Salmonella spp.
S. Typhi and S. Paratyphi
7-28 daysc
659
6-48 hr
3-4 weeks
1,341,873
553
Shigella spp.
12-50 hr
89,648
14
Staphylococcus aureus
1-7 hr
Meat and meat products, poultry, egg products, salads (egg, tuna,
chicken, potato, macaroni), cream-filled bakery products, milk and
dairy products
185,060
50,920
1-3 days
n/a
n/a
Streptococcus spp.
Group A (S. pyogenes)
a
b
c
10
Table 2. Foodborne Disease in the United States, Including Estimated Annual Prevalence, continued
Principal Symptomsa
Onseta
Illnessesb
Deathsb
2-36 hr
n/a
n/a
49
V. cholerae serogroup O1
6hr-5 days
n/a
n/a
48 hr
n/a
n/a
Vibrio parahaemolyticus
4-96 hr
n/a
n/a
Vibrio vulnificus
16 hr
47
18
Vibrio, other
n/a
n/a
n/a
5,122
13
Yersinia enterocolitica
24-48 hr
86,731
Anisakis simplex
1hr-2 weeks
n/a
n/a
Ascaris lumbricoides
n/a
n/a
n/a
Cryptosporidium parvum
1-12 days
30,000
Cyclospora cayetanensis
1 week
14,638
Giardia lamblia
1 week
200,000
Taenia spp.
n/a
n/a
n/a
Toxoplasma gondii
10-23 daysd
375
112,500
3-14 days
52
10-50 days
Microorganism
Group D (other Streptococcus
spp.)
Vibrio cholerae
Trichinella spiralis
Viruses
Hepatitis A
Norwalk-like viruses
Rotavirus
a
b
c
d
4,170
24-48 hr
9,200,000
124
1-3 days
39,000
been essentially eliminated; however, cases of botulism in the late 1980s and early
1990s led scientists to discover that C.
botulinum could grow and produce toxins in foods such as twice-baked potatoes, grilled onions, and garlic-in-oil.
When these new food associations were
discovered, even C. botulinum was described by some as an emerging pathogen. Large outbreaksalthough recurrences of situations that have happened
in the pastcan propel a pathogen to
emerging status whether truly deserved
or not. In many cases, it is the new food
vehicle that is the surprise, and not so
much the pathogen itself.
Host Factors
Changing demographic characteristics of consumers affect the number of
cases of foodborne illness. As the
worlds population continues to grow,
constant rates of disease will increase the
total number of cases. In addition, the
proportion of the population that is at
high risk of foodborne infections, illness,
and death is rising. Factors that increase
the impact of foodborne diseases include
age, chronic diseases, immunosuppressive conditions, and pregnancy. The immune system functions less effectively in
the elderly, putting them at greater risk. A
growing proportion of our population is
immunocompromised due to HIV infection, cancer chemotherapy, and drugs
used to combat rejection of transplanted
organs. Larger numbers of people with
chronic diseases, like diabetes, now live
longer and also are at increased risk of
foodborne diseases.
Other consumers are at elevated risk
of foodborne illness because of the in11
creased likelihood of exposure. This elevated risk is sometimes due to food preferences based on ethnicity, age, or gender.
Young adult males, for example, are
more likely to eat inadequately cooked
ground beef.
Environmental Factors
Food industry economics and technical factors continue to drive consolidation in primary agricultural production
and food processing. Although this
helps reduce costs and assure uniform
quality, when a large lot of a contaminated food enters distribution, the scope of
the resulting outbreak is increased.
Global sourcing also carries the potential to move pathogens and toxins
from areas in which the pathogen is indigenous to areas in which it has not previously existed. Unfamiliarity complicates diagnosis and containment and can
result in outbreaks that become quite
large before they are recognized. Hazards are truly mobile, and our food safety programs must be very agile to reduce
our risk.
Even slight increases in environmental temperatures can significantly affect
the risk of foodborne illness. The growth
of algae in surface waters, estuaries, and
coastal waters is sensitive to temperature.
About 40 of the approximately 5,000
known species of marine phytoplankton
(algae) can produce biotoxins, which
may reach human consumers through
shellfish. Warmer sea temperatures can
encourage a shift in species composition
of algae toward the more toxic dinoflagellates. Upsurges of toxic phytoplankton blooms in Asia are strongly
correlated with El Nio, and in the United States, paralytic shellfish poisoning
and other marine biotoxin-induced diseases have been associated with shell
stock harvested from beds traditionally
considered safe.
Consumer desires drive food product development. Food manufacturers
respond to desires for fresher food,
low fat products, or ready-to-eat foods
by developing new processes or reformulating existing products. Changes in
the food processing environment or
product formulations can create a new
niche for pathogenic microorganisms.
Producing familiar foods in nontraditional sites also may lead to introduction of new food hazards; such was the
case with the first outbreaks of cyclosporiasis associated with raspberries
12
Pathogen Factors
Stable and accurate transfer of genetic information from parent to offspring
is essential for the preservation of a species. However, keeping pace with an
ever-changing environment also requires
variability. When naturally occurring
bacteria, for example, divide, most of the
offspring look and act just like their parents, but a small proportion of the offspring mutate, increasing the chance that
some might survive in a new, hostile environment. If the environment has not
changed, these new strains may not survive, but this natural occurrence makes it
almost certain that traditional food processes will fail to deliver their predicted
level of safety at some point. This is part
of nature and happens without human
intervention.
In addition to this unstimulated hypermutability, the food production and
processing environment can increase the
rate of change in foodborne pathogens.
Bacterial stress responses may increase
pathogen virulence, and other actions
can affect which microorganisms survive
and dominate in a particular environment. For example, use of antimicrobial
agents during livestock production may
select resistant strains from a background of susceptible microorganisms,
increasing the likelihood that the microorganisms in a food are resistant to those
and related antimicrobials. Even if these
microorganisms are not pathogenic, they
can share the genetic material that enables them to resist antimicrobials with
pathogenic microorganisms in the human gut, producing pathogens that cause
infections that may be difficult to treat.
Through improved laboratory techniques, scientists are identifying adverse
health effects associated with ever-smaller levels of exposure to natural and anthropogenic substances. New ELISA and
radioimmunoassays for various mycotoxins are pushing tolerances for common mycotoxins down and are finding
more poorly characterized mycotoxins
in a broad array of commodities. Our
understanding of biology, however, is
not keeping up with our laboratory
skills, and judging the public health significance of positive laboratory results
is becoming more difficult. Unlocking
the human genome and the genomes of
pathogenic microorganisms, however, is
Science of Pathogenicity
Pathogenicity is the ability to cause
illness. Because pathogens are living
organisms that rapidly adapt and
evolve, the methods they use to cause
illness are never static. Pathogen
evolution is continuous and is driven
by a variety of forces, only some of
which relate to human activities. The
continual evolution of foodborne
pathogens forces us to change food
production processes and products to
maintain and improve microbiological
food safety. Control strategies that
were once effective may not remain so
if the pathogens become tolerant.
Fortunately, genomic and improved
molecular and imaging techniques have
vastly expanded scientific understanding
of the organisms that cause foodborne
disease. These tools also have enabled
scientists to attribute foodborne disease
to microorganisms that had not previously been identified as pathogenic or as
foodborne.
However, researchers still have many
questions to answer: What makes one
strain of a microbe pathogenic when
other microorganisms within the same
species are not? How do microorganisms
become pathogenic? Understanding
pathogenicity is not just necessary for
developing methods to treat illness but is
also needed for pathogen control.
Pathogen control includes prevention of food contamination, elimination
from the food, reduction to an acceptable
level, or prevention of multiplication and
toxin formation. In addition, when sciEXPERT REPORT
Nomenclature
Traditionally, the first step in understanding foodborne pathogens has been
to develop a system of nomenclature and
descriptions of microorganisms within
this system. For the purposes of study,
scientists try to classify microorganisms
based on a set of common characteristics
that sometimes include presumed pathogenic attributes. However, as our scientific understanding has improved, the
initial classifications often no longer
present a full and accurate picture. When
nomenclature becomes outdated, questions are raised about the scientific validity of regulatory policies based on classification schemes that predict pathogenic-
Example 1
Example 2
Example 3
Family
Enterobacteriaceae
Enterobacteriaceae
Mycobacteriaceae
Genera/genus
Escherichia
Salmonella
Mycobacterium
Species
coli
enterica
avium
enterica
paratuberculosis
Subspecies
Serovar
O157:H7
Typhimurium
13
Science of Pathogenicity
Pathogenicity is the ability to cause
illness. Because pathogens are living
organisms that rapidly adapt and
evolve, the methods they use to cause
illness are never static. Pathogen
evolution is continuous and is driven
by a variety of forces, only some of
which relate to human activities. The
continual evolution of foodborne
pathogens forces us to change food
production processes and products to
maintain and improve microbiological
food safety. Control strategies that
were once effective may not remain so
if the pathogens become tolerant.
Fortunately, genomic and improved
molecular and imaging techniques have
vastly expanded scientific understanding
of the organisms that cause foodborne
disease. These tools also have enabled
scientists to attribute foodborne disease
to microorganisms that had not previously been identified as pathogenic or as
foodborne.
However, researchers still have many
questions to answer: What makes one
strain of a microbe pathogenic when
other microorganisms within the same
species are not? How do microorganisms
become pathogenic? Understanding
pathogenicity is not just necessary for
developing methods to treat illness but is
also needed for pathogen control.
Pathogen control includes prevention of food contamination, elimination
from the food, reduction to an acceptable
level, or prevention of multiplication and
toxin formation. In addition, when sciEXPERT REPORT
Nomenclature
Traditionally, the first step in understanding foodborne pathogens has been
to develop a system of nomenclature and
descriptions of microorganisms within
this system. For the purposes of study,
scientists try to classify microorganisms
based on a set of common characteristics
that sometimes include presumed pathogenic attributes. However, as our scientific understanding has improved, the
initial classifications often no longer
present a full and accurate picture. When
nomenclature becomes outdated, questions are raised about the scientific validity of regulatory policies based on classification schemes that predict pathogenic-
Example 1
Example 2
Example 3
Family
Enterobacteriaceae
Enterobacteriaceae
Mycobacteriaceae
Genera/genus
Escherichia
Salmonella
Mycobacterium
Species
coli
enterica
avium
enterica
paratuberculosis
Subspecies
Serovar
O157:H7
Typhimurium
13
Nomenclature of
Salmonella
gastroenteritis (caused by S. Typhimuriuim, S. Enteritidis, and others); enteric fever (S. Typhi and S. Paratyphi); and
an invasive systemic disease (S. Choleraesuis). In the United States, nontyphoidal Salmonella account for an estimated 1.3 million illnesses annually,
with several hundred deaths (Mead et
al., 1999). The U.S. incidence of typhoid
fever is relatively lowapproximately
700 cases annually, mainly as a result of
international travel. Worldwide, the in-
14
terial species is in flux. In terms of diversity, genome size may vary by 20%
among subpopulations of a single species (for example, see Bergthorsen and
Ochman, 1998). This variability can
cover as many as 1-2 megabases of DNA
that code for thousands of genes, which
can confer unique characteristicssuch
as virulenceon specific subpopulations. Still, a subset of genes that define
signature characteristics of the species
must be shared among all members of
that species.
These contrasting views of what constitutes a bacterial species challenge the
concepts that underlie microbiological
criteria and testing for the control of
pathogens in food manufacturing. In
some cases, the potential to cause foodborne disease can be characteristic of an
entire species, such as S. enterica, but in
other cases it may be a consequence of
recently evolved virulence characteristics
among specific subpopulations of a species, such as Escherichia coli O157:H7.
The sensitivity of genome-based fingerprinting methods has even called into
question the equality of virulence in genetically distinct subpopulations of E.
coli O157:H7 (Kim et al., 1999). As another example, it is questionable whether
all microorganisms in the Listeria monocytogenes species are capable of causing
disease in humans. Even though each
distinct genetic lineage of the species appears to harbor the known virulence
genes, mounting phylogenetic evidence
indicates that virulence characteristics
differ (Rasmussen et al., 1995; Wiedmann et al., 1997).
New scientific information provides
the opportunity to better target the
pathogenic subpopulations within a species. Present day approaches to control
must be modified to be consistent with
the information presently available. As
improved detection and identification
methods enable scientists to differentiate
between virulent and avirulent organisms, we should be able to allocate our
risk management resources more efficiently, focusing only on pathogenic organisms in our food supply.
Virulence
If pathogenicity is a microorganisms
ability to cause disease, then virulence
can be considered the degree of pathogenicity. Some pathogens are particularly
efficient at causing clinically significant
disease (highly virulent), while others
EXPERT REPORT
Quorum Sensing
Not long ago, bacteria were
thought to lead a solitary existence
and either live or die as a single cell. It
has been established that intercellular
communication is fairly common
among bacteria, and that this intercellular communication can lead to coordinated activities once thought to
be the exclusive domain of multi-cellular organisms. It was not surprising
then that researchers asked whether
intercellular communication was involved in virulence, and it was also
not surprising that genetically well
characterized foodborne pathogens
such as Salmonella and Escherichia
coli would be investigated.
Intercellular communication
among bacteria is carried out by
small molecules called autoinducers.
The theory is that at very low population densities, there is insufficient
autoinducer in the environment to
be detected by those bacteria present,
but that when some threshold
number of bacteria is reached, autoinducer is present in sufficient concentration to trigger some activity in
bacteria capable of detecting the au-
Toxins are perhaps the best understood family of bacterial virulence factors, presumably because they are often
secreted into the area surrounding the
bacteria where they can be isolated and
studied. Toxins have specific mechanisms
to recognize host cell surface receptors
enabling them to transport themselves
into the host cell. Some toxins can modify specific cellular targets to affect fluid
secretion, cytoskeletal structure, or even
nerve functions. Others insert themselves into the host cellular membrane,
resulting in cell disintegration or dissolution (lysis). Despite the vast number
of cellular targets, toxins can be classified
into families based on their structure
and function.
To cause disease, most pathogens require the ability to adhere to host surfaces following ingestion. Mammalian
hosts have several nonspecific defenses
designed to prevent colonization by inhibiting pathogen attachment, such as
peristalsis, the mucocilliary system, and
even cell sloughing (see section on human host, p. 28). Bacterial pathogens
INSTITUTE OF FOOD TECHNOLOGISTS
Virulence of
Salmonella
Recently, scientists have made significant advances toward understanding the molecular mechanisms of the
pathogenicity of Salmonella. Similar
to pathogenic E. coli, Salmonella have
a collection of virulence factors, including factors needed to adhere to
intestinal surfaces, invade host epithelial cells, and survive within phagocytic cells. It has been estimated that
Salmonella contain more than 200
virulence factors, encoded in at least
five pathogenicity islands in addition
to a virulence plasmid and many
pathogenicity islets.
The complex mechanisms used
by Salmonella species to adhere to
intestinal cells continue to be studied
and debated. Many adhesins have
been reported, and because they
appear redundant, defining the role
of each in the disease process has
been difficult.
Significantly more is known
EXPERT REPORT
17
18
ally present in low numbers does not ensure product safety, as the infectious doses
(100-102 infectious units) are presumed to
be low (Iversen et al., 1987; Jaykus, 2000b;
Moe et al., 1998).
Three major routes for viral contamination of foods have been recognized
and include: (1) shellfish contaminated
by fecally polluted marine waters;
(2) human sewage pollution of drinking
and irrigation waters; and (3) ready-toeat and prepared foods contaminated as
a result of poor personal hygiene of infected food handlers (Jaykus, 2000b). In
addition, the NLVs have been shown to
be spread by aerosolization of vomitus
and through fomites (Marks et al., 2000;
Patterson et al., 1997).
Protozoan Parasites
Like viruses, parasitic protozoa replicate in the intestines of infected hosts
and are excreted in the feces. However,
their host range is wider than viruses, being able to replicate in human and nonhuman animal hosts. Since they are
transmitted primarily by the fecal-oral
route, the major source of contamination for foods and water is through contact with human and animal fecal pollution. This contamination may occur directly, through contaminated meat carcasses or poor personal hygiene practices
of infected food handlers, or indirectly,
via contact with fecally contaminated
waters or other cross-contamination
routes. Like the viruses, the parasitic
protozoa (in the cyst or oocyst form) are
environmentally inert, they do not replicate in foods, are extremely environmentally stable, resistant to many of the traditional methods used to control bacterial pathogens, and have notably low infectious doses (DuPont et al., 1995; Haas,
1983). Although transmitted by the fecaloral route, direct person-to-person
transmission of parasitic protozoa is unlikely because excreted oocysts require
days or weeks under favorable condi-
Marine Biotoxins
Marine biotoxins are produced by
several dinoflagellate and diatom spe-
EXPERT REPORT
Mycotoxins
A large group of mycotoxins, known
collectively as trichothecenes, have been
associated with acute human illness
(Pestka and Casale, 1990). The most severe manifestation is alimentary toxic
aleukia. Initial symptoms after a single
contaminated meal include throat inflammation, vomiting, diarrhea and abdominal pain; with continued ingestion,
the illness can progress to oral hemorrhaging, pneumonia, bone marrow depletion, and potentially death. Human
outbreaks of gastrointestinal illness have
been linked to millet contaminated by a
variety of Fusarium that was capable of
producing T-2 toxin and other trichothecenes and also wheat and barley infected with Fusarium that produced the trichothecenes deoxynivalenol (vomitoxin),
nivalenol and fusarenon-x (Bhat et al.,
1989; Luo, 1994).
Other mycotoxins have been associated with acute disease. Aflatoxin ingestion has caused acute liver inflammation in Kenya (Ngindu et al., 1982) and
India (Krishnamachari et al., 1975). In
the early 20th century, cardiac beriberian acute cardiac disease that produces a rapid pulse, abnormal heart
function, and low blood pressure leading to respiratory failure and death
that occurred in Japan and other Asian
countries was linked to contaminated
rice containing the mycotoxin citreoviridin (Ueno, 1974).
Evolution
Pathogen evolution is a continuous
biological process that is influenced by
19
20
Evolution of Salmonella
S. enterica shows significant diversity in its host specificity, but the reasons for host specificity remain undefined. Some serovars, such as S. Typhi,
are very human specific while others
are animal specific (e.g., S. Pullorum
infects chickens). Others, such as S.
Typhimurium and S. Enteritidis, have
a wide host range. S. Typhimurium
typically causes a diarrheal disease in
humans, but a typhoid-like disease in
mice. Because many strains of Salmonella cannot tolerate the low pH of
the gastric environment, the infectious dose for ingested exposure to
Salmonella is usually (but not always)
quite high (greater than 100,000 bacteria required to produce illness).
However, if delivered intravenously, a
mere 10 organisms of strains that require a large oral infectious dose can
kill a mouse.
The incidence of antibiotic resistance among Salmonella and some
occurs rapidly.
Genetic exchange between bacterial
species is obviously frequent and can significantly affect the evolution of pathogens. The acquisition and spread of antibiotic resistance has been an easy way
to follow genetic exchanges, and the efficiency of this spread is demonstrated by
the pervasiveness of resistance in many
bacteria not previously resistant to antibiotics. As discussed above, the genetic
material that encodes virulence factors
can move between bacteria, with the potential to rapidly create a new pathogen,
even from a commensal organism. A
major unanswered question is often
where these virulence factors originated.
Despite the prevalence of Type III secretion systems within several bacterial
pathogens, scientists have not found the
systems ancestor.
As in all organisms, the evolution of
pathogens is continuous. In some cases,
classifying pathogens based on their virulence factors is a more logical method
than classification based on serotype or
some other trait unrelated to virulence.
The evolution of pathogens will become
clearer as scientists sequence and study
additional genomes of related species.
However, even seemingly related organisms can contain significant diversity
EXPERT REPORT
O26:H11/O111:H8 lineages of enterohemorrhagic E. coli demonstrate that acquisition of the same virulence gene elements has occurred on multiple occasions, even through parallel paths (Reid
et al., 2000). This discovery raises a significant question regarding the selective
pressures that cause the abrupt rise to
dominance of particular virulence gene
combinations within a species.
In infectious diseases that are primarily carried by humans and transmitted person-to-person, the appearance of new alleles of virulence genes is
associated with the rise and spread of
more virulent clones (Musser, 1996).
This process likely also occurs among
foodborne pathogens. However, foodborne pathogens must overcome
unique hurdles, including survival in
the pre-harvest environment, as well as
survival during food processing, storage, and preparation.
Many of the hurdles in food production and processing are a different
set of selective pressures than those exerted by the human hosts gastrointestinal tract. Successful foodborne pathogens, such as Salmonella and L. monocytogenes, have acquired not only virulence characteristics, but also physiological and ecological characteristics that
Selection
Evolution and selection are closely
related. Evolution produces microorganisms that are distinctly different
from previous generations; selection
gives these mutant strains an advantage
and causes them to become prominent.
Together, these forces have a profound
impact on the emergence of foodborne
pathogens.
The proposed step-wise model (Feng
et al., 1998) for evolution of E. coli
O157:H7 (as opposed to a gradual evolution) points out some rather interesting features of the process by which new
pathogens emerge. First, mobility of
gene segments appears to be a limiting
factor in the rate at which microorganisms can test new gene combinations.
New high-throughput genome sequencing methods will produce data to develop
a much better estimate of the frequency
of such events and an improved understanding of their underlying mechanisms. Second, the common virulence
genes shared by the O157:H7 and EHEC
21
tage conferred. The combination of genomics and population genetics will provide methods for identifying genetic alterations that correlate with the descent
of specific populations of foodborne
pathogens. However, scientists must begin to devise strategies for identifying
which genetic alterations, among the
many different gene sequences that define a subpopulation, confer selective advantage. Identifying and understanding
these genes may enable scientists to pinpoint the selective forces at work.
Stress
Each microbe prefers a specific set
of environmental conditions. When environmental parameters are significantly different from the desired range, the
microbes undergo stress. To be more
specific, stress is defined as chemical or
physical parameters that impair the
function of the macromolecular machinery of the microorganism. Examples of stress for certain microorganisms might include high and low temperature, acidic pH, low water availability, and presence or absence of oxygen.
Specific genes are activated, producing
proteins that protect the bacterium from
stress. This process, known as an adaptive response, improves the microorganisms ability to survive under the stressful conditions. Although the responses
improve the range of conditions the microorganisms can tolerate, these responses also require energy, so they are
only expressed when needed. Under
normal conditions, bacteria that do not
turn off their stress responses would be
outgrown by those that reroute that energy to other cellular processes. Stress
responses are of particular interest in
foodborne pathogens because they can
render bacteria tolerant to traditional
food processes or the intrinsic parameters of the food.
Bacteria have evolved elaborate networks to protect against or repair damage caused by detrimental conditions.
Bacterial responses to stress are varied
and complex, including both structural
and physiological changes. For most
bacteria, these responses are modulated
by specific sigma (F) factors (Grossman
et al., 1984; Lange and Hengge-Aronis,
1991) or regulators (Christman et al.,
1989) that direct the activation of specific
genes that comprise regulons (large
numbers of coordinately controlled
genes) and encode for the proteins reINSTITUTE OF FOOD TECHNOLOGISTS
Responses to Stress
The cellular responses to stress can
generally be divided into two categoriesgeneral and specific stress responses. In many instances, a certain
stress response gene may be part of
both specific and general stress response pathways. This is usually because the gene has multiple regulatory
elements that are recognized by the distinct machinery that coordinates the
general or specific response. It is therefore difficult to separate the contribution of general and specific stress response pathways to cellular viability under any given stress condition. Rather,
the combination of the two, and specifically the combined fine-tuning of each
pathway, dictates many of the survival
characteristics of the species.
Spore Formation
In addition to general and specific
stress response pathways, some bacteria
and other microorganisms also have
evolved highly sophisticated pathways for
stress adaptation, such as forming
spores. Spores are metabolically inactive
or dormant and are much more resistant
to adverse environmental conditions,
e.g., extremes of temperature, low water
activity, and radiation.
In organisms that form spores, the
adaptive response pathways are somewhat
hierarchical and, depending on the environmental conditions, can be triggered
alone or in combination. Bacillus subtilis,
the model organism for studying spore
formation, relies on the general stress response, several stationary phase and transition state pathways, the development of
competence, and differentiation into the
dormant endospore. The pathways for
spore formation, competence, and normal
growth are mutually exclusive, but each
one can be used in combination with the
general stress response. Before the organism commits to a major step such as normal growth, competence, or spore formation, sophisticated signal transduction
pathways measure environmental nutritional and chemical signals, as well as the
state of cellular processes such as DNA
replication.
Example
Reference
Metabolic changes
Hengge-Aronis, 2000
Protection
Repair
polymerase subunit. F38 accumulates during several different stress conditions and
activates the target genes that produce the
general stress response. The rate of synthesis for the RpoS protein increases little
during stress conditions; it accumulates
rapidly after stress because the degradation rate is slowed. Unlike most response
regulators, which directly modulate gene
transcription by increasing the amount of
signal protein produced, this response
regulator functions by controlling the stability of the protein and thereby changing
its rate of degradation.
In the instance of the GSR of grampositive bacteria, much is known about
B. subtilis. As in the gram-negative bacteria, the GSR in B. subtilis is modulated
primarily by an alternative sigma subunit of RNA polymerase, in this case
known as FB. However, FB is controlled
Emergence of Viruses,
Parasitic Protozoa and
Marine Biotoxins as
Foodborne Pathogens
Although pathogenicity of some
of the marine biotoxins has been
characterized, there is a paucity of
information available regarding the
disease mechanisms or virulence
factors for enteric viruses and parasitic protozoa. The reasons for this
are several-fold; for many of these
agents, in vitro cultivation and/or
animal models are nonexistent. Because they have had relatively less
scientific emphasis over the last few
decades, fewer research dollars have
gone into understanding these
EXPERT REPORT
25
26
Pathogenicity of
E. coli O157:H7
E. coli O157:H7 is typical of what
might be expected in terms of an
emerging pathogen.
Nomenclature
E. coli O157:H7 belongs to a
group of E. coli (enterohemhorrhagic
E. coli, EHEC) that cause hemorrhagic colitis (severe bloody diarrhea) and,
in a small portion of the cases,
hemolytic uremic syndrome (HUS).
These strains are within a larger
group of Shiga-toxin-producing E.
coli (STEC), but EHEC possess virulence factors that other STEC do not.
EHEC have been the source of many
food- and water-borne outbreaks. It
has been estimated that these pathogens cause about 75,000 cases of diarrhea and several hundred deaths annually in the United States (Mead et
al., 1999). Young children and older
adults are particularly susceptible to
HUS, while people of all ages can get
the diarrhea.
The designation O157:H7 is based
on serological analysis: lipopolysaccharide (O) type 157, and flagellar (H)
type 7. However, scientists have since
discovered that several related E. coli
strains with different O serotypes
cause similar disease, such as
O26:H11. Although O157:H7 is the
most predominant EHEC serotype in
North America, Japan and the United
Kingdom, different serotypes of EHEC
such as O26:H11 and O111:NM dominate in other areas of the world, notably central Europe and Australia. Because of the conservation of virulence
factors, but not serotypes, classification schemes of EHEC strains should
probably be based on virulence factors
rather than the variant serotype. However, O157:H7 (EHEC 1) and
O26:H11/O111:NM (EHEC 2) clearly
comprise two distinct genetic lineages.
Virulence
E. coli O157:H7 and related
strains have the capacity to persist in
cattle without causing disease because
cattle lack a receptor for the illnessproducing Shiga toxin (Pruimboom-
EXPERT REPORT
target certain cells, including the endothelial cells of blood vessels; the
dead cells accumulate and plug the
kidney, causing HUS. Shiga toxin is
encoded within a mobile genetic element, a bacteriophage, that enables it
to move to different strains of bacteria.
As discussed below, it is believed that
the acquisition of this toxin by E. coli is
a relatively recent genetic event.
In addition to producing Shiga
toxin, EHEC adhere to the large bowel,
the pathogens preferred site in the
body, using a variety of virulence factors contained within the LEE pathogenicity island. This 38-kb region of
DNA, inserted near a tRNA gene, contains all the genes necessary for binding to epithelial cells and causing a
pedestal to form on their surface. Pedestals are created when actin in the cytoplasm is accumulated and polymerized beneath the pathogen. In addition, effacement of the microvilli occurs, giving rise to the term attaching
and effacing E. coli. Pedestal formation is a complex process that utilizes a
Type III secretion system, several Type
III E. coli secreted proteins (Esps), and
a key molecule, Tir (Translocated intimin receptor), that is delivered to
host cell membranes. Once in the host
cell membrane, Tir binds to intimin, a
bacterial outer membrane protein, resulting in intimate bacterial adherence.
Because Tir spans the host membrane,
it is also able to recruit host cytoskeletal proteins to cause actin accumulation and pedestal formation.
Emergence
E. coli O157:H7 is believed to have
arisen from a series of fairly recent genetic events. O157:H7 was first reported as a foodborne pathogen following
an outbreak associated with contaminated hamburgers in 1982. Subsequent studies of diarrheal samples
from prior outbreaks and sporadic
cases revealed only a single E. coli
O157:H7 isolate in the Center for Disease Control and Preventions collection. This isolate had been obtained
from an individual in the mid-1970s.
A closely related strain of E. coli called
enteropathogenic E. coli (EPEC) causes
diarrhea in children (but does not
Continued on next page
27
Treatment
Therapies against EHEC infections are extremely limited. Treatment with antibiotics is thought to
worsen the illness, presumably by
breaking up the bacteria, which releases more toxin and increases tox-
pies use an inert substance that mimics the toxins glycolipid receptor. Ingestion of the mimicking substance
should bind excess toxin and thereby
limit disease progression. Experiments indicate it may be effective, but
only when taken very early after infection. Alternate therapies are being
explored.
In addition, significant effort is
focused on developing treatments
such as vaccines and probiotics to
reduce carriage of O157:H7 by cattle. Decreasing the level of O157:H7
in cattle would significantly decrease
the potential for food and water
contamination. Similarly, childhood
vaccines are being developed, but
given the low incidence of disease,
questions remain about whether
universal vaccination should be employed, were an effective vaccine to
be developed. Experimental approaches to block virulence factors
such as the Type III secretion system
also are being studied.
quired by consumption of food containing pathogens or their toxins. The pathogens or their toxins can damage or destroy host cells or processes, or they can
induce a host response to their presence
that is harmful to the human host. Foodborne illness is caused by: viral, bacterial,
or parasitic infections (e.g., Norwalk-like
viral gastroenteritis, Campylobacter enteritis, toxoplasmosis); toxins produced
during microbial growth in food (e.g.,
Clostridium botulinum, Staphylococcus
aureus, Bacillus cereus, and Aspergillus
flavus); and toxins produced by algal and
fungal species (e.g., ciguatera fish poisoning) (see Table 5).
Foodborne infections occur when
pathogenic microorganisms are ingested,
colonize the intestine, and sometimes invade the mucosa or other tissues. Foodborne toxicoinfections arise when a microorganism from ingested food grows
INSTITUTE OF FOOD TECHNOLOGISTS
Treatment
Therapies against EHEC infections are extremely limited. Treatment with antibiotics is thought to
worsen the illness, presumably by
breaking up the bacteria, which releases more toxin and increases tox-
pies use an inert substance that mimics the toxins glycolipid receptor. Ingestion of the mimicking substance
should bind excess toxin and thereby
limit disease progression. Experiments indicate it may be effective, but
only when taken very early after infection. Alternate therapies are being
explored.
In addition, significant effort is
focused on developing treatments
such as vaccines and probiotics to
reduce carriage of O157:H7 by cattle. Decreasing the level of O157:H7
in cattle would significantly decrease
the potential for food and water
contamination. Similarly, childhood
vaccines are being developed, but
given the low incidence of disease,
questions remain about whether
universal vaccination should be employed, were an effective vaccine to
be developed. Experimental approaches to block virulence factors
such as the Type III secretion system
also are being studied.
quired by consumption of food containing pathogens or their toxins. The pathogens or their toxins can damage or destroy host cells or processes, or they can
induce a host response to their presence
that is harmful to the human host. Foodborne illness is caused by: viral, bacterial,
or parasitic infections (e.g., Norwalk-like
viral gastroenteritis, Campylobacter enteritis, toxoplasmosis); toxins produced
during microbial growth in food (e.g.,
Clostridium botulinum, Staphylococcus
aureus, Bacillus cereus, and Aspergillus
flavus); and toxins produced by algal and
fungal species (e.g., ciguatera fish poisoning) (see Table 5).
Foodborne infections occur when
pathogenic microorganisms are ingested,
colonize the intestine, and sometimes invade the mucosa or other tissues. Foodborne toxicoinfections arise when a microorganism from ingested food grows
INSTITUTE OF FOOD TECHNOLOGISTS
Example(s)
Bacterial infection
Campylobacter jejuni
Common
Viral infection
Norwalk-like viruses
Very common
Parasitic infection
Toxoplasma gondii
Relatively common
Bacterial toxin
Clostridium perfringens,
Bacillus cereus
Relatively common
Algal toxin
Less common
Mycotoxin
Aflatoxin
Less common
Prions*
BSE
None
Inorganic contaminants*
Heavy metals
Less common
Organic contaminants*
Pesticide residues
Less common
Pfiesteria piscicida
and Pfiesteria-like
Microbes As Potential
Foodborne Pathogens
An association between seafoodborne illness in humans and the occurrence of Pfiesteria in the marine
environment has not been established, and much remains unknown.
Pfiesteria piscicida and Pfiesteria-like
organisms, discovered in 1988, have
caught the attention of researchers
and funding agencies interested in
characterizing the biological functions and effects of previously unrecognized toxic dinoflagellates (Fleming et al., 1999; Glasgow et al., 1995;
Smith et al., 1988). Initially presenting as the cause of massive fish kills
in North Carolina in the late 1980s,
Pfiesteria-like organisms also have
stimulated public concern over the
potential threat these organisms may
have on human health and seafood
safety. Because of difficulties in identification of the Pfiesteria species,
these organisms are currently characterized on the basis of morphology
and hence referred to as morphologi-
30
term for Crohns disease (CD) and ulcerative colitis (UC), which are both
chronic inflammatory diseases with a
prolonged clinical course. Abdominal
abscesses are a complication of CD
while in UC abdominal perforations
may lead to peritonitis. The cause of
IBD, and the mechanism(s) for spontaneous exacerbations and remissions remain unresolved. Controversial reports
by some investigators about the potential association of Mycobacterium
paratuberculosis, the etiologic agent of
Johnes disease in ruminants, with CD
have appeared in the literature for years.
A conference convened by the National
Institutes of Health Division of Microbiology and Infectious Disease (NIH/
DMID, 1998) and the European Commissions Scientific Committee on Animal Health and Animal Welfare (EC,
2000) concluded that there is insufficient evidence to prove or disprove that
M. paratuberculosis is the cause of CD.
Studies to determine how M. paratuberculosis could be transferred from animals to humans have focused on milk
and results have been conflicting
(CAST, 2001). Both the NIH/DMID
conference and the EC committee recommended further study of this issue.
An association between IBD and the
bacterial L-forms of Pseudomonas, Mycobacterium, Enterococcus faecalis and
E. coli also has been suggested (Herman-Taylor et al., 2000; Korzenik and
Dieckgraefe, 2000).
The preceding examples are representative, but they do not present a comprehensive discussion of foodborne disease. The differing symptoms, duration
and severity make diagnosis and comprehensive tracking of foodborne illness
difficult. The large number of foodborne
pathogenseach with its own virulence
factorsproduce an astonishing array
of illnesses, and the pathogens continue
to evolve. Millions of illnesses and thousands of deaths occur each year as a result of contaminated food in developed
countries, and the situation is much
worse in the developing world.
Resistance to Microbial
Foodborne Disease
Humans are protected from infectious foodborne disease by a variety of
nonspecific (innate) and specific immune system mechanisms. When all of
these systems are functioning optimally,
the chance of foodborne illness is reEXPERT REPORT
Immune Response
Foodborne pathogens and their
toxins enter human tissue via the gastrointestinal (GI) tract. The GI tract is
approximately 30 feet long and includes cells that produce acid, mucus,
antibodies, and other substances that
protect the host from foodborne
pathogens (Burke, 1985). Food entering the mouth is chewed, breaking it
into smaller pieces and mixing it with
saliva. When swallowed, the food travels via the pharynx and esophagus to
the stomach. Upon entering the stomach, the food is broken down by gastric
juices that contain pepsin, lipases and
acids.
From the stomach, the partially digested food enters the small intestine.
The intestine is approximately 20 feet
long and has an irregular, folded lining
that provides a very large surface area
that facilitates digestion and absorption of nutrients. It also facilitates exposure to microorganisms that survived passage through the stomach.
The tissues immediately below the epithelial lining cells contain blood capillaries to absorb monosaccharides and
amino acids and deliver bloodborne
defenses, and lymph capillaries to absorb fatty acids and glycerol. Enzymes
and hormones secreted by the liver and
pancreas assist digestion and absorption in the small intestine. Because of
its length, surface area (equivalent to a
singles tennis court), and digestive capacity, more than 80% of absorption
occurs in the small intestine. Undigested food material enters the large intestine (colon), through which it travels
until it finally exits the anus. Anatomic,
physiologic, and pathologic changes in
the GI tract influence the level of protection from the effects of pathogens.
These GI tract changes also affect the
population of nonpathogenic microorganisms living in the GI tract, which
influences the likelihood of foodborne
infections.
The large surface area, presence of
large amounts of nutrients, and absorptive capacity of the alimentary canal make it particularly prone to pene-
the GI defensive barriers depends on several factors, such as the number of microorganisms and virulence factors (see
virulence, p. 15). Thorne (1986) outlined
five pathogenic mechanisms for bacterial
diarrheal diseases: (1) bacteria produce
toxin but do not generally adhere and
multiply (e.g., B. cereus, S. aureus, C. perfringens, C. botulinum); (2) bacteria adhere to the lining of the intestine and
produce toxin (e.g., enterotoxigenic E.
coli, Vibrio cholerae); (3) bacteria adhere
and damage the villi that make up the
brush border (e.g., enteropathogenic E.
coli); (4) bacteria invade the mucosal layer and initiate intracellular multiplication (e.g., Shigella spp.); and (5) bacteria
penetrate the mucosal layer and spread
to lamina propria and lymph nodes (e.g.,
Yersinia). With each increasing level of
action, the pathogens focus moves from
the mucosal to the systemic compartment, and, hence, the specific immune
response must escalate.
The antigen-sampling process itself
may become a major portal of entry for
pathogens (Owen and Ermak, 1990).
Wells et al. (1988) hypothesized that, in
some instances, a motile phagocyte may
ingest an intestinal bacterium, transport
it to an extraintestinal site, fail to accomplish intracellular killing, and then liberate the bacterium at the extraintestinal
site. This hypothesis was based on the
observation that the intestinal bacteria
that most readily translocate out of the
intestinal tract are facultative intracellular pathogens. Secondly, intestinal particles without inherent motility (e.g., yeast,
ferritin, starch) move out of the intestinal lumen within hours of their ingestion. Thirdly, the rate of translocation of
intestinal bacteria can be altered with
agents that modulate immune functions
such as phagocytosis. Thus, systemic infection by translocating intestinal bacteria could be a result of the antigen-sampling process that evolved to regulate the
immune response to intestinal antigens.
Susceptibility to Microbial
Foodborne Disease
The extent to which the human host is
susceptible to disease influences the likelihood of foodborne illness. Many factors
play a role in the level of susceptibility.
Infectious Disease
Susceptibility to infectious disease is
the inability of the hosts body to prevent
or overcome invasion by pathogenic mi-
Table 6. Factors That Increase Host Susceptibility (adapted from CAST, 1994)
General
Factors
Specific Factors
Reasons
Age
Pregnancy
Hospitalized people
Concomitant infections
Consumption of antibiotics
Sensitive
populations
Underlying
medical
conditions
34
Cryptosporidiosis
Cryptosporidium was first described in the early 1900s but was not
considered to be medically or economically important. During the early 1970s, Cryptosporidium was linked
to diarrhea in calves, and case reports
describing its appearance in a variety
of animal species began to appear in
the literature. In 1976, Cryptosporidium was first associated with severe
watery diarrhea in a patient who had
been receiving immunosuppressive
chemotherapy for 5 weeks. The diarrhea resolved 2 weeks after discontinuation of the therapy. Following this
report, additional case reports of severe, persistent diarrhea in immunosuppressed or immunodeficient individuals appeared in the literature (Pitlik et al., 1983).
During the early 1980s, two series
of observations began to shape the
emerging epidemiology of human
cryptosporidiosis. First, cases of
cryptosporidiosis were reported
among persons who had normally
functioning immune systems and ex-
and Trichinella spiralis. In one study, individuals with chronic heart disease had
an increased risk for listeriosis (Schuchat
et al., 1992). Cellular immunity declines
during pregnancy, which may account
for the severity of certain infections.
Evidence indicates that immune deficiency not only increases the number of
cases but also the severity of infection
from a wide variety of foodborne pathogens. For example, studies conducted in
Los Angeles, San Francisco and New
York City during the mid-1980s demonstrated that patients with acquired immune deficiency syndrome (AIDS) had
rates of Campylobacter and Salmonella
infection that were 19 94 times the general population rates in the same cities
(Celum et al., 1987; Greunewald et al.,
1994; Sorvillo et al., 1994). In addition,
16% of Campylobacter infections and
44% of Salmonella infections resulted in
bacteremia in these compromised patients, much higher rates of severe disease than occurred in the general population. San Francisco residents infected
with the human immunodeficiency virus
(HIV) also have been shown to have inci-
immune function with age, termed immune senescence. The data regarding the
effects of aging are confusing and sometimes conflicting. In general, cell-mediated
immunity declines, including both functional and quantitative cell counts. Superimposed and interrelated with this
generalized impairment are age-related
decreases in organ structure and function.
Nutritional abnormalities in macro- and
micronutrients are common in the elderly
and may compound immune senescence.
The presence of other illnesses and environmental factors also may contribute to
the decline.
EXPERT REPORT
Susceptibility to Biotoxins
The variability of human susceptibility to mycotoxins and other biotoxins can
be attributed to physiologic and environmental factors, host genetics, and the presence of infection and inflammation.
Physiologic Factors
A number of factors can influence a
persons ability to detoxify ingested
35
biotoxins. For example, biotransformation enzyme activity can vary during perinatal and postnatal development (deBethizy and Hayes, 1994).
When the individual is older, environmental forces play a role in host response: the nutritional quality of the
diet, the presence of chemicals in the
diet, and the intake of prescription or
elicit drugs can affect the profile of
biotransformation enzymes. Finally,
environmental toxins in air (e.g., cigarette smoke) and water can increase or
decrease the activity of biotransformation enzymes toward specific mycotoxins or other biotoxins.
Genetic Polymorphisms
Wide differences in the human capacity to biotransform biotoxins appear
to relate to genetic background that influences the presence, amount and activation of various enzyme systems that
metabolize ingested biotoxins into chemical derivatives that are less or sometimes
more toxic than the original mycotoxin
(Daly et al., 1994; Kalow, 1993). For example, the level of expression for
CYP1A2, a cytochrome P-450 (CYP)-dependent monooxygenase that metabolizes aflatoxin B1 varies considerably in the
human liver (Eaton et al., 1995). The activity of microsomal epoxide hydrolase,
which acts coordinately with CYP1A2,
can vary up to 40-fold in human tissue
(Seidegard and Ekstrom, 1997). It has
been suggested that epoxide hydrolase
polymorphisms may alter the risk
of aflatoxin-associated liver cancer
(McGlynn et al., 1995). Specifically, enzymes may vary in both the amount
present and their effectiveness/activity,
resulting in differing risks of a negative
outcome from aflatoxin ingestion.
Other Factors
Other significant host factors, apart
from immune suppression per se, are associated with increased risk of both acute
foodborne disease and chronic sequelae.
Genetic predisposition and underlying
chronic disease have been cited as potential risk factors. For instance, septic
Vibrio vulnificus infections are most
commonly seen in men over 50 years of
age with liver and/or blood disorders
(Desenclos et al., 1991; Tacket et al.,
1984). These underlying conditions frequently result in elevated serum iron levels that play a role in V. vulnificus disease
pathogenesis, although the exact mechanism is not fully understood (Wright et
al., 1981). Similar evidence is available
for yersiniosis in iron-overloaded patients treated with deferioxamine, although again the pathogenic mechanisms are not yet clear (Mandell and
Bennett, 1995). As previously discussed,
genetic predisposition plays a role in the
development of chronic reactive arthritis.
Recent evidence from a quantitative human challenge study for the Norwalklike virus indicates a two-phase dose-response relationship that appears to be
separately associated with prior exposure
(antibody titer) and individual susceptibility, neither of which are associated
with any recognized specific host factors
(Moe et al., 1999). When taken together,
this body of evidence suggests that many
factors apart from immune suppression
influence host susceptibility to foodborne disease agents.
Behavior Changes
The 1990s saw a tremendous increase
in public awareness of food safety issues
in the United States. This awareness
arose in part because of the continuing
interest in personal health and well being, a phenomenon that occurred
throughout the worlds developed countries. Schools, education programs, media communications, and the Internet
have made foodborne and waterborne
diseases important concerns to many
consumers. Outbreaks of foodborne illness that would have gone unnoticed a
decade ago are now the subject of rapid,
in-depth news coverage. The increased
publicity about infectious foodborne
hazards appears to reinforce food safety
messages and to increase motivations to
handle foods safely.
Proper hygiene and sanitation related
to food handling and preparation, appropriate methods of refrigeration and
freezing, and thorough cooking of foods
comprise a very effective approach to
preventing foodborne illness. However,
these behaviors are just one aspect of a
healthy life-style. Additional behavioral
changessuch as consuming probiotics,
eating a balanced diet, and exercising
regularly to maintain a healthy weight
foster proper functioning of the immune
system that may heighten resistance to
occasional pathogens in the food supply.
Consumer awareness of food safety
issues has placed additional pressure on
the food service and food processing industries to improve their efforts to ensure the safety of the products they provide both domestically and internationally. Hazard Analysis and Critical Control Points (HACCP) implementation
has become commonplace in the food
processing and delivery process. Companies have a strong economic incentive
to prevent outbreaks of foodborne illness associated with their product or restaurant.
While many host factors that influence infection, occurrence and severity
of illness are associated with human
physiology, the factors that influence exposure to foodborne pathogens are often
tied to human behavior, specifically consumption, food handling, and preparation behaviors.
Eating outside the home in restaurants and other foodservice venues has
been identified as a risk factor for certain
foodborne diseases (Friedman et al.,
2000), and the number of meals that
Americans eat away from home continues to increase. In the 1990s, food eaten
outside the home accounted for almost
80% of reported foodborne illness outbreaks in the United States (Bean et al.,
1996). Because of the larger number of
people involved, these outbreaks are
more likely to be recognized and, thereINSTITUTE OF FOOD TECHNOLOGISTS
Table 7. Factors That Increase Risk of Foodborne Disease (adapted from CAST, 1994)
General
Factors
Specific Factors
Reasons
Life-style
Stress
Poor hygiene
Geographic location
Consumption of antacids
Diet
Individual Choices
Americans pride themselves on
their individual freedoms. In a culture
based on these individual rights, we allow people to engage in high-risk behavior and offer products that sometimes cater to these risks. Steps are taken to mitigate risk but ultimately certain behaviors are inherently risky.
Sometimes people knowingly engage in
high-risk behavior. If people are aware
of the risks and continue to engage in
risky behavior such as eating raw oysters or eggs, it is appropriate to consider to what lengths our society should
go to protect them.
Surveys conducted from 1998-1999
as part of the FoodNet Active Surveillance Program for foodborne diseases
have documented certain aspects of
consumer behavior. With regard to
consumption of fresh produce that is
known to be at particular risk for microbial contamination, 19% of respondents reported eating a mesclun lettuce
mix in the 7 days before the interview,
and 8% reported eating alfalfa sprouts,
although these eating habits are highly
regional (CDC, 1999a). Among other
potentially risky food exposures, 25%
of the people who had eaten eggs had
chosen to eat eggs that were runny;
11% of persons who consumed hamburgers ate burgers that were still pink
inside; 4.4% drank unpasteurized ap38
Cultural Differences
Diet selection can create subpopulations at greater risk for certain foodborne illnesses. Some reports of foodborne illnesses involve transmission via
foods consumed primarily by immigrant
groups. Outbreaks of trichinosis have
become relatively rare in the United
States because cooking pork thoroughly
has become a widespread cultural practice. An exception occurred in 1990,
when Laotian immigrants in Iowa prepared and ate undercooked pork, a traditional food, as part of a wedding celebration (Stehr-Green and Schantz, 1986).
Other reports involve foods more commonly consumed by ethnic populations.
Y. enterocolitica outbreaks are also rare,
but several outbreaks in African-American communities were associated with
preparation and consumption of pig intestines (Lee et al., 1990). The epidemiology of human brucellosis in California
has shifted from an occupational disease
related to animal husbandry to a foodborne disease most frequently affecting
Hispanics who often consume raw milk
and cheeses made with raw milk while
abroad (Chomel et al., 1994). Consumption of rare hamburgersa risk factor
for E. coli O157 infectionis more common in U.S. Caucasians than in any other racial/ethnic group (CDC, 1998a).
Dietary Recommendations
A host of organizations have issued
dietary recommendations and provided
information to assist in health promotion and chronic disease prevention (U.S.
PSTF, 1997). These recommendations
may advocate increased or decreased
consumption of certain types of foods,
outline specific circumstances for consumption, or caution individuals with
certain medical conditions. Each type of
recommendation has consequences for
foodborne illness. In recognition of the
importance of food safety, the latest edition of the federal governments dietary
guidelines contains a section on food
safety.
Counseling the general population to
limit dietary intake of fat and emphasize
foods containing fiber (i.e., fruits, vegetables, grain products) has increased consumption of foods such as fresh produce
Modification of Susceptibility
Because of the significance of the human host in foodborne illness, it is appropriate to look for host-related opportunities for control or mitigation of illness. Certainly, behavioral and demographic issues influence susceptibility
and exposure. If the host could be rendered immune to infectious agents, illness would cease to occur despite microbial contamination in the food supply.
Although complete protection is currently out of reach for many pathogens, approaches such as immunization and probiotics can decrease human susceptibility. Coupling these approaches with efforts to mitigate exposure would further
boost our ability to control foodborne
illness of infectious origin.
Immunization
Vaccines use the hosts own immune
system to combat disease. Knowing how
the immune system functions enables
scientists to investigate methods to enhance its effectiveness or trigger its protective effects without causing illness.
The medical community has had great
success with vaccination for some infectious diseases, and potential vaccines to
prevent foodborne illness are the subject
of considerable research.
Although no vaccines are currently
available for most enteric bacterial
pathogens, experimental approaches are
under investigation. In general, vaccination strategies to control enteric bacterial
diseases are complicated by many factors,
not the least of which is the complexity
of host immunity as well as a general absence of appropriate animal models for
oral challenge and subsequent disease
presentation for many of these pathogens. For example, an effective broadspectrum vaccine against enterohemorrhagic E. coli (EHEC) will likely need to
target systemic immunity against the
Shiga toxins as well as local intestinal
immunity against intestinal colonization
factors (Nataro and Kaper, 1998). Nonetheless, genetic manipulation has enabled researchers to produce bacterial
EXPERT REPORT
Probiotics
Probiotics represent another opportunity to use our understanding of the
gut microflora and the immune response
to facilitate human health and decrease
susceptibility to illness. Although originally used to describe substances produced by one protozoan that stimulated
another (Fuller, 1989), the definition for
the term probiotic now generally implies a viable microbial supplement that
beneficially affects the host (human or
animal) by improving or maintaining a
desirable microbial balance in the gut.
The reduction of harmful enteric microorganisms is only one of numerous potential health benefits of maintaining a
healthy gut microflora. Probiotic cultures are consumed in foods or capsules
or are facilitated by ingesting prebiotics
(compounds that enhance the proliferation of beneficial indigenous bacteria).
Currently, dairy foods such as yogurt
have been the most popular vehicle of
choice to deliver viable probiotic cultures. Intestinally-derived lactobacilli
and bifidobacteria predominate in this
role (Hughes and Hoover, 1991).
The effect of probiotic cultures varies
based on numerous conditions. The human gut contains 100 trillion viable bacteria and other microorganisms representing anywhere from 100 to 400 different species; the population dynamics are
quite complex. The microflora of the
human intestinal tract is affected by genetic or host factors, the composition of
microbial populations, and the metabolites produced by these microbes; these
factors are in turn influenced by climate,
diet, stress, drugs, age, and disease (Mitsuoka, 1990). One can maintain that
whenever there is a change in the intestinal microflora from its normal state, the
change is detrimental or undesirable. A
foodborne intestinal infection that produces diarrhea can be viewed as a period
of microbial imbalance or instability in
the gastrointestinal tract. However, by establishing themselves in the human GI
tract in proportionally high numbers,
acidulating bacteria, such as lactobacilli
and bifidobacteria, may protect the gut
against invasive pathogenic agents.
39
control methods.
and control are implemented throughout the food production and processing
system. Researchers are continually
searching for a better understanding of
the pathogens and their interaction
with the environment, leading to
improved control technologies. But at
the same time, the pathogens continue
to evolve, and human actions sometimes drive that evolution. Even small
environmental changes can have
unforeseen or even unforeseeable
impact on microbial populations.
40
PRE-HARVEST ENVIRONMENT
Efforts to minimize microbial contamination of food begin in the pre-harvest environment. Raw ingredients are
one way in which pathogens are introduced into the processing environment.
Unfortunately, pathogen control in the
production agriculture environment is
often difficult.
Overarching Issues
In the pre-harvest environment, many
of the significant issues in microbiological
control methods.
and control are implemented throughout the food production and processing
system. Researchers are continually
searching for a better understanding of
the pathogens and their interaction
with the environment, leading to
improved control technologies. But at
the same time, the pathogens continue
to evolve, and human actions sometimes drive that evolution. Even small
environmental changes can have
unforeseen or even unforeseeable
impact on microbial populations.
40
PRE-HARVEST ENVIRONMENT
Efforts to minimize microbial contamination of food begin in the pre-harvest environment. Raw ingredients are
one way in which pathogens are introduced into the processing environment.
Unfortunately, pathogen control in the
production agriculture environment is
often difficult.
Overarching Issues
In the pre-harvest environment, many
of the significant issues in microbiological
Percent
(by $ value)
Mexico
51
Canada
15
Chile
12
Costa Rica
Netherlands
Guatemala
Other
13
Manure
The widespread occurrence and use
of animal manure as fertilizer is a growing environmental concern, because it
contaminates: water for drinking, irrigation, aquaculture and recreation; the
hides, coats, and feathers of farm animals; and farm equipment and buildings. In the United States, cattle, hogs,
chickens and turkeys produce an estimated 1.36 billion tons of manure annually (EPA, 2000), with greater than 90%
attributed to cattle. Each year livestock
create an estimated five tons of animal
manure per person living in the United
States, meaning the amount of animal
manure is 130 times greater than the
amount of human waste produced (U.S.
Senate Agriculture Committee Democratic Staff, 1998).
Many of the most prominent foodborne pathogens in the United States, including Campylobacter jejuni, Salmonella
and Escherichia coli O157:H7, are carried
by livestock and are principally transmitted to foods by fecal contamination. C.
jejuni accounts for an estimated 2 million cases of foodborne illness annually,
with poultry and unpasteurized milk as
its principal vehicles (Mead et al., 1999).
Salmonella causes an estimated 1.3 million cases of foodborne illnesses annually, with eggs, poultry, beef, pork and produce as primary vehicles (Mead et al.,
1999). Both C. jejuni and Salmonella are
carried in the intestinal tract of apparently healthy poultry and livestock. Fecal
contamination of hides, feathers and
skin occurs during poultry and livestock
production and slaughter. This contamination can subsequently carry through
to processing. A case-control study of
patients with Campylobacter infection
identified the following risk factors for
campylobacteriosis: foreign travel; eating undercooked poultry; eating chicken,
turkey or non-poultry meat cooked out-
side the home; eating raw seafood; drinking raw milk; living on or visiting a farm;
and having contact with farm animals or
puppies (Friedman et al., 2000). Fecal
contamination is a common source of C.
jejuni contamination for each risk factor.
For example, poultry feces frequently
contain C. jejuni at populations of 105 to
107 colony forming units of bacteria per
gram (CFU/g), resulting in levels of
greater than 103 CFU C. jejuni per gram
of carcass in 60 to 90% of retail poultry.
E. coli O157:H7 causes an estimated
73,500 cases of infection in the United
States annually. Its principal vehicles of
transmission are beef, produce, water
(both drinking and recreational), and
contact with cattle (Doyle et al., 1997;
Griffin, 1998). Because E. coli O157:H7 is
carried in the intestinal tract of cattle, the
pathogens most frequent origin is direct
or indirect contact with cow manure. Manure can contaminate food when used as
a soil fertilizer, when it pollutes irrigation
water, when cattle defecate near produce
or foods of animal origin, and when intestinal contents or manure-laden hides
contact carcasses during slaughter and
processing. Case-control studies of patients with E. coli O157:H7 infections revealed several major risk factors for illness: eating undercooked ground beef, living on or visiting a farm, and having contact with farm animals, especially cattle
(Kassenborg et al., 1998). Depending on
environmental conditions, E. coli
O157:H7 can survive in manure for many
weeks, and in some instances for more
than one year. Similarly, the pathogen can
survive well in lake water, with as little as a
10- to 100-fold reduction occurring during 13 weeks at 8 C.
Increased proximity and animal density during production contribute to
problems of pathogens in runoff water
because the difficulty of manure management is increased with greater volume. Another issue is composting of manure by farmers, including organic farmers. The conditions that effectively destroy pathogens are not well defined.
Also, manure handling in other countries may be worse than in the United
States, a serious concern for imported
foods. Human feces from field workers
without access to adequate sanitation facilities remains an issue as well.
Import Item
1980
1985
1990
1995
% Change
(1980-95)
45.3
53.8
56.3
55.3
22.1
Fresh fruits
24.2
28.0
30.7
33.3
37.6
Fresh vegetables
7.6
8.9
8.4
11.7
53.9
1.4
7.0
5.7
3.5
150.0
Water
9.1
22.2
57.8
84.9
833.0
EXPERT REPORT
41
Production Practices
and Mycotoxins
A recent example of how changes
in ecology and production practices
have affected mycotoxin incidence in
the United States is the massive increase in Fusarium head scab in Midwestern wheat and barley during the
last decade (McMullen et al., 1997).
Head scab is often accompanied by elevated contamination by deoxynivalenol (vomitoxin) and other trichothecenes. Two factors seem to have driven the head scab epidemic. One is an
increased spring rainfall during early
wheat head formation. The uncharacteristic increase in rainfall may be a
result of the prolonged El Nio dur-
Use of Antibiotics
The widespread use of antibiotics in
animal production and in the treatment
of human illness both facilitate the
emergence of antibiotic resistance. Microorganisms can develop resistance to
antimicrobials through gene mutations
or by acquiring transferable genetic elements, such as plasmids and conjugative
transposons, that harbor resistance
genes. These mobile genetic elements
are important in horizontal transmission of genes from the resident to transient microflora of the intestinal tract
(Levy et al., 1976). In addition to the
mobility of the genetic elements, the antibiotic-resistant bacteria can be transmitted to different animal hosts. A tetracycline-resistant E. coli strain from
cattle was traced to humans, mice, pigs,
and fowl found at the same location
(Levy et al., 1976). The selective pressure
caused by antibiotic administration
causes the microbial populations that
harbor the appropriate resistance
determinant(s) to flourish (Levy, 1992).
These antibiotic-resistant microbes can
make their way to humans through contaminated foods or animal-to-human
transmission (Angulo et al., 2000;
Holmberg et al., 1984), although the
public health impact of the use of veterinary drugs is difficult to measure (Howgate, 1997).
The contribution of sub-therapeutic
levels of antibiotics in animal feed to the
emergence of antibiotic-resistant pathogens has been debated for years (Feinman, 1998). A growing body of evidence
from epidemiological data and traceback studies indicates that agricultural
use of antibiotics plays an important
role in the emergence of some antibiotic-resistant bacteria (Angulo et al.,
2000). A review of Salmonella outbreaks between 1971 and 1983 revealed
that antibiotic-resistant strains were
more likely to originate from animals
than were strains without resistance
(Holmberg et al., 1984). Additionally,
the emergence of Salmonella with decreased susceptibility to fluoroquinolone paralleled approval of the veterinary use of enrofloxacin, a fluoroquinolone antibiotic, even though fluoroquinolones had been used in humans
for the preceding six years with little impact on the development of resistant
Salmonella (Threlfall et al., 1997). Although there is no evidence that the use
of antibiotics in feed is responsible for
the evolution of the multi-drug resistant
strain of Salmonella Typhimurium
DT104, the rapid dissemination of this
strain in animals and humans indicates
there is an advantage for strains with the
antibiotic-resistance phenotype (see
sidebar, p. 44). S. Typhimurium DT104 is
the second leading cause of human salmonellosis in England and Wales
(Anonymous, 1996) and the most common Salmonella species isolated from
cattle (Hollinger et al., 1998). S. Typhimurium DT104 is resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline (known as
R-type ACSSuT). The resistance genes to
the antibiotics are located on the chromosome rather than a plasmid, indicating they are nonmobile and stable
(Threlfall et al., 1995). In the United
States, the ACSSuT resistant pattern was
present in 28% of 976 S. Typhimurium
isolates collected nationally in 1995, a
substantial increase from 7% in 1990
isolates (Hosek et al., 1997). Human infection by S. Typhimurium DT104 has
greater morbidity and mortality than the
other nontyphoid Salmonella infections
(Wall et al., 1994). Because the accumulating data from molecular subtyping
methods and epidemiological investigations suggest that the use of sub-therapeutic levels of antibiotics in animal
feeds plays a role in the emergence of antibiotic-resistant foodborne pathogens,
the prudent and judicious use of antibiotics in both the agricultural and medical sectors is needed.
Feeding Practices
Another animal production practice
with potential ramifications for microbiological food safety is farm animal diet
composition. A change of diet, for example, can change the microbial ecology of
the ruminant digestive system. Many
studies have evaluated the effect of dietary changes on fecal shedding of E. coli
O157:H7 or acid-tolerant E. coli by cattle
or sheep, with conflicting results. Some
investigators have determined that sheep
or cattle fed hay shed E. coli O157:H7 in
their feces considerably longer than animals fed grain (Hovde et al., 1999; Kudva
et al., 1997). In contrast, studies of acidtolerant E. coli, which is a characteristic
of E. coli O157:H7, revealed that a mostly
grain diet promotes shedding of acid-tol43
Development and
Dissemination of
Resistant Organisms
Microorganisms develop resistance to antibiotics encountered in
clinical and environmental settings.
This fact has led to calls for the judicious use of antibiotics in human
medicine and for restrictions on the
use of antibiotics in veterinary medicine and animal production.
There are at least three fundamentally different ways that exposure to antibiotics can promote the development
and/or dissemination of resistant
microorganisms: (1) mutations and selection of mutants capable of surviving
in vivo exposure to the antibiotic (e.g.,
fluoroquinolone resistance in C. jejuni),
(2) mobilization and horizontal transfer of genetic elements containing resistance genes among different species of
bacteria (e.g., vancomycin resistance
among enterococci), and (3) widespread dissemination of strains with
previously developed resistance (e.g., S.
Typhimurium DT 104). The differences
between these mechanisms have important implications for the prevention
and control of antibiotic resistance
among foodborne bacteria.
Resistance of C. jejuni to fluoroquinolones is conferred by a point mutation in the gyr gene (Engberg et al.,
2001). Resistant organisms with the
same molecular subtype characteristics
as sensitive microorganisms have been
isolated from human patients after ap-
parent failure of treatment with ciprofloxicin. During the 1990s, an increased occurrence of resistant C. jejuni isolates in
Minnesota was associated with treatment
with a fluoroquinolone antibiotic and
foreign travel. However, a growing proportion of resistant isolates was not attributable to these sources. Surveys of
chicken at retail markets in Minnesota
demonstrated a 20% prevalence of contamination with resistant C. jejuni strains.
These strains showed considerable diversity based on polymerase chain reaction
(PCR)-restriction fragment length polymorphism (RFLP) (PCR-RFLP) subtyping of the flaA gene, suggesting that mutations and selection of mutants was independently occurring among many C. jejuni strains. The overlap of molecular subtypes obtained from human and chicken
sources suggest that chicken was a primary source of resistant C. jejuni for humans
that were not treated and did not travel
(Smith et al., 1999).
In Europe, a glycopeptide antibiotic,
avoparcin, was used as an antimicrobial
growth promoter. Following its introduction, resistance of enterococci to vancomycin (a similar glycopeptide antibiotic)
was observed in hospitalized patients, exposed animals, and the human population outside of hospitals. Vancomycinresistance genes were transferred horizontally between species of enterococci
(Simonsen et al., 1998; van den Braak et
al., 1998). The potential for vancomycinresistance genes to be acquired by strains
of Staphylococcus aureus represents an
important public health threat
fecal shedding of E. coli O157:H7 by cattle (Buchko et al. 2000). Feces of cattle
fed 85% barley were more frequently E.
coli O157:H7-positive than those from
cattle fed 85% corn, although no major
differences were observed in cell numbers of E. coli O157:H7 in feces throughout most of the study. Before specific
diet and feeding practices can be practically applied to farm production practices for pathogen control, considerably
more research is needed to elucidate the
influences that different dietary practices
contribute to gastrointestinal carriage
and fecal shedding of pathogens.
As with humans, good bacteriain
some cases nonpathogenic normal gut
florain the gastrointestinal tract of animals may be able to prevent colonization by pathogens (Nurmi and Rantala,
1973). In chickens for example, the gut
of the hatchling chick is sterile until it ingests microorganisms from the environment. If the chicks are exposed to adult
bird fecal material, colonization of the
gut occurs rapidly. Todays production
practices remove this route of colonization, and the hatchling may not acquire a
normal gut flora for days or weeks
(Spencer and Garcia, 1995). The lack of
a fully developed normal flora increases the chances of the chick gut becoming
a carrier for microorganisms, such as
Salmonella, that are pathogenic in hu-
44
sewage. For a number of reasons, the fecal coliform index is inadequate for
monitoring the presence of viral contamination in shellfish or their harvesting
waters, and both outbreaks and sporadic
cases of enteric viral disease associated
with the consumption of contaminated
shellfish continue to occur in the United
States (Jaykus et al., 2001). Protozoan
parasites such as Giardia and Cryptosporidium species have recently been identified in feral shellfish (Fayer et al., 1998),
although the significance of this food
with respect to disease transmission has
yet to be determined. Shellfish harvesting beds become contaminated with parasitic protozoa as a result of contamination with animal farm runoff or human
sewage, both treated or untreated.
The indigenous bacterial pathogens
(most notably Vibrio cholerae, Vibrio
parahaemolyticus, and Vibrio vulnificus)
are part of the estuarine microflora and
have excellent survival capabilities in the
marine environment. In addition, V.
cholerae from human sewage may contaminate the marine environment. While
the transmission of V. cholerae by seafood is well documented throughout the
world, the United States has not had a
major outbreak since 1911, although
sporadic cases have occurred, usually
due to the consumption of imported
crustacea such as crabs and shrimp
(Bean et al., 1997). Historically, V. parahaemolyticus outbreaks are rare in the
United States, but two highly publicized
recent outbreaks have challenged this
trend (CDC, 1998b; CDC, 1999b). Shellfish implicated in these outbreaks were
harvested from both Atlantic and Pacific
waters during these outbreaks, but in
both cases, the mean surface water temperatures were significantly higher (1-5
C) than those reported in previous years.
This phenomenon suggests the potential
for emergence of this pathogen is associated with natural changes in the temperature of the Gulf Stream (El Nio) or
global warming from human activities.
V. vulnificus has been of greater concern in the United States in recent years,
since this organism results in a syndrome characterized by gastrointestinal
disease followed by primary septicemia,
with mortality rates approaching 50%.
Individuals with underlying liver dysfunction, circulatory problems particularly related to diabetes, or those who are
immunocompromised are especially at
risk (Hlady and Klontz, 1996). Evidence
exists that other Vibrio species also can
45
the fecal coliforms and important human pathogens such as enteric viruses,
the pathogenic vibrios, and perhaps the
parasitic protozoa. In addition, shellfish
harvested from other countries with less
stringent standards than those of the
United States may be contaminated, and
some countries have much higher domestic rates of shellfish-associated disease. Pre-harvest control methods such
as relaying (movement of shellfish from
one harvest site to an alternative, pristine
site to allow the animals to purge them-
46
Aquaculture
One area where food production
has undergone dramatic change is
HARVEST ENVIRONMENT
The harvest environment is relatively
commodity specific, because the methods used and harvest locations depend
on the commodity in question. The harvest environment is especially important
for foods such as fruits and vegetables
that undergo minimal additional processing prior to consumption.
Produce
Many different approaches are used
today to harvest produce depending on a
variety of determinants, including the
type of produce, site of growing operation, and labor availability. However,
from a microbiological safety perspective, there are many common hazards for
harvest operations. Water quality, field
worker hygiene, field sanitation, truck
sanitation, and temperature control are
all food safety issues related to the harvest environment (IFT, 2002).
Water as ice or in the liquid form can
readily transmit microorganisms to produce if contaminated. Most fruits and
vegetables are washed with water at least
once, and many types of produce are
treated with water several times during
processing. In addition to being used for
washing, water is used for cooling, conveying produce (flume water), and for
applying disinfectants and fungicides.
Disinfectants are added to about 50% to
60% of water used in packing facilities.
Care must be taken to control the sanitary quality of water.
Field worker hygiene is a major consideration because of the widespread use
of human hands in cutting or picking
vegetables and fruits in fields or orchards. Approximately 90% of farms
that grow fruit or vegetables harvest produce exclusively by hand (USDA/NASS,
2001). The amount of human hand contact that occurs during harvesting varies
depending on the type of produce. Melons are handled at most steps of the operation, whereas apples receive considerably less frequent contact. Vegetables
such as leaf lettuce may be harvested,
trimmed, sorted and bagged by hand. It
is very difficult to uniformly enforce
proper hand washing and glove use
48
among workers.
An interesting practice in recent
years has been the production of freshcut produce in the field. Presumably
done to maintain product freshness, this
practice essentially brings food processing into the harvest environment. For example, many fresh-cut processors are
now removing the outer leaves and coring lettuce in the field. Some are even
cutting lettuce where it is grown. Unfortunately, it is much more difficult to control contamination in the harvest environment, and this practice may increase
the chances for contamination of these
products. Certainly contamination of
wash and rinse water would be of concern under these circumstances, as
would be issues associated with poor hygiene of food handlers and inadequate
sanitation of equipment.
Fruits and vegetables frequently
come into contact with harvesting
equipment (such as knives, machetes,
clippers and scissors) and containers
(such as bins, boxes, buckets, pans,
trailers, and truck beds). Such equipment and containers should be properly
washed and disinfected, although studies indicate that washing and sanitizing
is done only about 75% and 30% of the
time, respectively (USDA/NASS, 2001).
Packing equipment such as tables, conveyor belts, flumes, and washing and
cooling bins are washed and sanitized
about 75% and 50% of the time, respectively.
Equipment and containers used during harvest operations are frequently
made of materials, such as wood, that are
difficult to clean. Soil from the field,
which may contain pathogens, often encrusts equipment and containers. If not
removed, soil adhering to equipment
used for washing and disinfecting produce will reduce the sanitizing capacity
of the disinfectant. Produce operations
must prevent accumulation of soil on
equipment and containers to enable effective disinfection of food contact surfaces.
Proper temperature control of fruits
and vegetables is critical for both safety
and quality purposes. Optimal temperatures vary according to the commodity; however, the temperature range for
storing fruits and vegetables is usually
quite narrow. Most pathogens, but not
all, are inhibited by the cool temperatures at which produce is stored. In addition, cool temperatures tend to prolong the survival of viruses and para-
Food Animals
Production livestock are naturally
contaminated with a variety of potential
human pathogens, both externally and
internally. These microorganisms originate from the environment in which the
animals are produced, as well as from
feedstuffs and the co-mingling of animals from various sources. The animals
also can readily transmit these microorganisms among themselves, once they
are moved from the production environment to the transportation system. Because of centralized slaughter establishments, production livestock may be
transported considerable distances before slaughter.
During transportation, livestock are
confined to prevent excessive movement. As the animals are in close physical contact, microorganisms may be
transferred from one animal to another
either by contact with each other or
their excreta. Aerosol transmission of
salmonella also has been demonstrated
in chickens and mice (Clemmer et al.,
1960; Darlow et al., 1961). The upper
respiratory tract may be important in
transmission, and the tonsils and lungs
may be important sites for the invasion
and dissemination of Salmonella in pigs
(Fedorka-Cray et al., 1995; FedorkaCray et al., 2000; Gray et al., 1996).
Even under the best of conditions,
transportation produces measurable
stress upon live animals. Transport
stress may have several physiological effects on the live animal that may ultimately impact food safety. Transportation of animals may increase fecal shedding of potential human pathogens, such
as salmonella. This increase in shedding
can contaminate the trucks or trailers
used for transportation, and potentially
increase the population of foodborne
pathogens in and on many of the animals within the truck or trailer.
Once animals arrive at a slaughter
establishment, they may be unloaded
into holding pens prior to slaughter, depending on the animal species in question. Animals from several trucks or
INSTITUTE OF FOOD TECHNOLOGISTS
POST-HARVEST ENVIRONMENT
In the post-harvest environment,
food safety becomes less commodity oriented, as the food moves through processing into the distribution and retail
sectors. Microbiological food safety issues in the processing environment have
the potential to affect many different
foods. Also, at the processing stage, ingredients from many different commodity sectors may be combined into a single
product. As ingredients are combined,
the physical characteristics of the food
change, and its microbiological profile is
altered.
The microbiological controls applied in the post-harvest environment
are often designed to intentionally stress
the microorganisms present in the food.
These stresses may be designed to be lethal on their own or in combination.
Environmental conditions also may be
modified to limit microbial growth,
through such techniques as drying or
Role of Inspection
An important aspect of meat inspection in the United States and other
developed countries is antemortem inspection. All animals presented for
slaughter are inspected prior to slaughter. At present, this inspection focuses
on observable clinical illness, and not
on general hygiene of the animals.
However, because of the recognition of
the impact of animal hygiene on meat
contamination, there have been some
attempts to regulate this on the live animal. For example, part of the antemortem inspection in the United Kingdom
is an evaluation of the overall hygiene
of the animal, and the animals may be
rejected for slaughter if they are too
dirty. Efforts to reduce the level of external carcass contamination of live animals have typically focused on either
washing the entire animal or trimming
the hair from the most heavily contaminated areas of the animal (typically the
rump and midline of a steer or cow).
These procedures must be balanced
with animal welfare issues, which prevent the introduction of excessive, needless or unnecessary stresses on live animals.
Source of Contamination
Contamination of animal carcasses
during slaughtering procedures is undesirable but unavoidable in the conversion of live animals to meat for consumption. It is assumed that the muscle
tissue of healthy animals entering the
49
slaughter establishment is free of microorganisms (Ayres, 1955). However, intrinsic bacteria, occurring in the deep
muscle tissue of healthy animals, have
been reported for many animal species
(Ingram, 1964; Ingram and Dainty,
1971). The most frequently characterized intrinsic bacteria are Clostridium
spp. (Canada and Strong, 1964; Jensen
and Hess, 1941; Narayan, 1966; Zagaevskii, 1973). Other potential human
pathogens, such as salmonella, have
not been reported as intrinsic bacteria
in the muscle tissue of healthy animals.
The assumption is bacteria that contaminate the muscle tissue are from extrinsic sources (gastrointestinal tract,
lymph nodes, external carcass surfaces,
and environmental sources). The majority of the microflora transferred to
the tissue surfaces, while aesthetically
undesirable, is nonpathogenic.
Carcass processing may be divided
into processes that impact the external
surfaces of the carcass and those that
open the body cavity (evisceration).
Examples of surface processes include
the scalding of poultry and the removal of beef carcass hides. These processes in fact remove significant populations of bacteria from the carcass, but
also expose the edible tissue to contamination. While the process of hide
removal may be thought of as a primary source of contamination of beef carcasses, it also removes significant contamination from the carcass. In a similar fashion, scalding and de-feathering
of chickens removes significant contamination from the chicken carcass,
although some still remains on the carcass. The microflora deposited by
these processes is primarily from environmental sources, that which is on the
carcass from the livestock production
environment and transportation. This
microflora may include enteric pathogens, such as salmonella and E. coli
O157:H7, and microorganisms such as
Listeria and Clostridium. From a microbiological perspective, contamination of edible tissue by external surface
processing is a relatively common occurrence.
Processes that open the body cavity
expose edible tissue to bacteria from
the gastrointestinal tract. The primary
bacteria of potential public health concern from this source are the enteric
pathogens. In the case of red meat processing, this source of contamination is
infrequent, and usually confined to
50
Impact of Interventions
Over the last twenty years, a number
of interventions have evolved specifically
to address microbial contamination of
animal carcasses. These may be divided
into physical methods and chemical
methods. In practice, these methods are
used in combination, resulting in a series
of process interventions or hurdles to
improve the microbiological quality of
meats. Among the physical methods,
prevention of contamination has received considerable attention. From the
perspective of microbial adaptation, it is
better to prevent contamination than to
address it through processing methods.
However, there are practical limitations
to this, given the nature of the process
(i.e., converting a live animal to food for
human consumption). After prevention
of contamination to the extent possible,
physical interventions involve either removal of contamination (trimming),
heat, or chemical treatments.
Effective physical removal of contamination is dependent on first identifying the area of contamination and then
removing the affected area without
transferring the contamination to other
areas. There are practical limits to the
identification of affected areas, as microorganisms cannot be seen. Therefore,
identification of an affected area requires
that there be sufficient contamination
(mud, manure, etc.) to become visible to
the operator. Once the area has been
identified, the operator must then remove the area in an aseptic manner. The
probable outcome of this operation, under controlled conditions, is the complete
removal of the contamination with a virtually sterile surface remaining around
the affected area. However, while this is
easily accomplished under controlled
conditions, it becomes much more unlikely in a processing environment. The
operator must clean and sanitize the
equipment (knife and hook) prior to the
operation, remove the affected area from
a carcass which is frequently moving on
a processing line, and then re-sanitize the
equipment prior to trimming the next
affected area. In practice, trimming is
substantially less effective in processing
environments than in laboratory environments (compare Gorman et al.
(1995) with Reagan et al. (1996)). Trimming as a process is limited to a specific
area of a carcass.
In contrast to trimming, heat may
be applied as either a localized treatment or as a whole carcass treatment.
The most common form of localized
treatment currently in use is the steam
vacuum. Steam vacuuming, as the name
implies, applies a steam treatment to
both loosen contamination and kill
bacteria, along with a vacuum process
to physically remove contamination.
Steam vacuuming may be highly effective in reducing microbial populations
under controlled conditions, but as with
trimming, becomes less effective under
processing conditions (Castillo et al.,
1999; Dorsa et al, 1996). As an alternative to a localized treatment, heat may
be applied as a whole carcass treatment.
Common examples of this are singeing
of hog carcasses, hot water washing and
steam pasteurization. Heat in singeing
processes is commonly applied as open
flame from gas jets, and while there is a
reduction in microbial populations on
the carcass surface, the primary function of this process is to remove residual
hair from the carcass. In contrast, hot
water washing and steam pasteurization
were specifically developed as antimicrobial processes, applied as a final operation before chilling. Hot water washing applies hot water (>80 C) as a whole
carcass rinse, while steam pasteurization places the carcasses in a chamber
and applies steam to briefly raise the
temperature of the carcass surface.
Both hot water and steam pasteurization have been demonstrated to be effective in controlling microbial populations on animal carcasses (Barkate et
al., 1993; Gill et al., 1995; Phebus et al.,
1997). However, surviving pathogens
will have undergone stress that may either increase or decrease the likelihood
of their survival during the remaining
time before consumption.
Chemical interventions involve the
application of food grade chemicals to
the carcass surfaces to inhibit or kill microorganisms (Dickson and Anderson,
(1992; Siragusa, 1996). Typically, the
mode of action of these antimicrobials is
pH, with organic acids, such as lactic or
acetic (low pH) and trisodium phosphate (high pH), the most commonly
used. The concerns with the use of any
chemical intervention process are both
the potential to induce resistance in potential human pathogens and the potential to select for resistant organisms out
of the overall microbial population. If reINSTITUTE OF FOOD TECHNOLOGISTS
Chilling
Rapid chilling of hot (body temperature) animal carcasses is essential to reduce the outgrowth of contaminating
EXPERT REPORT
microbes. The common methods of carcass chilling involve either forced air,
forced air and water, or water chilling.
While the primary intention of chilling is
to limit microbial growth, some chilling
methods do contribute to a reduction in
microflora. For example, forced air chilling dries the carcass surface and may injure or kill some microorganisms by dehydration. The initial combination chilling process using forced air and water
was the Chlor-Chil process (Swift and
Company, 1973), which used chilled
chlorinated water to simultaneously reduce microbial populations and chill the
carcasses more rapidly. Although the use
of chlorine in the water is no longer
widely practiced, the process now commonly known as spray chilling is almost
universally used in the beef industry. Although it does not have the benefit of
drying the carcass, spray chilling reduces
the surface temperatures of carcasses
more rapidly than air chilling, and, since
virtually all of the bacteria are on the
carcass surfaces, it effectively reduces microbial growth to a greater extent than
air chilling. Water chilling, widely used in
the poultry industry, has evolved from a
significant source of contamination between poultry carcasses to a potential
microbial intervention process, with the
use of counter-flow chillers and the addition of processing aids.
Fabrication
The disassembly of chilled carcasses is
referred to as fabrication. The outputs of
fabrication are fresh meat to go to the
consumer, and meat intended for further
processing. During fabrication, microorganisms may be transferred from carcass
surfaces to other parts of the carcass, and
may be transferred from one carcass to
another by common contact surfaces. Because of the latter phenomenon, one carcass has the potential to contaminate several other carcasses. As the fabrication
process proceeds, the carcass identity is
lost, and the potential for trace back is seriously diminished. Other than the physical intervention of preventing or reducing
contamination, there are essentially no
commercially viable interventions in fabrication at this time. This certainly provides opportunities for research and development, but it must be kept in mind
that the objective of the slaughter operation is to place the least contaminated carcass into the chilling and fabrication processes. Assuming that the microbiological
Ground Meats
Ground meats are a special category
of fresh meats. Ground meats are made
from the less desirable cuts of fresh meat
and from the trimmings of the more desirable cuts of meats. Unlike a steak or a
chicken breast, which originate from a
single carcass, ground meat may contain
meat from many different carcasses. The
nature of the process is to grind and mix
the meat, which increases the likelihood
that a single contaminated carcass may
contaminate a larger quantity of meat. An
additional factor is that the trimmings
from the intact cuts of meat are often
from the surface of the cut. Because the
most consistent source of contamination
on animal carcasses are the processes
which affect the external surfaces of the
carcass, the raw materials used for grinding have a higher probability of being
contaminated with bacteria of potential
human health significance. A consequence of these factors is that the microbial populations in ground meat are consistently higher than those on intact meat.
Other than irradiation, there are essentially no commercially viable interventions
for raw ground meats at this time.
A process that has also been used to
recover edible tissue from bones is mechanical de-boning. This process was
first developed in the poultry industry,
but is also used in the red meat industry.
Meat produced from mechanical deboning processes resembles ground
meat, but is used almost exclusively as a
processing ingredient for cooked meats.
This meat does not generally reach the
consumer as fresh meat. Because of the
nature of the process, mechanically deboned meat commonly has a higher microbial population than other meats, including ground meats.
Processing
Fresh meats are frequently further
processed for specific flavor characteristics or for increased shelf life. The processes involved with meats are the processes commonly used with other foods,
including thermal processing (cooking,
canning), dehydration, and fermentation. These processes do not result in any
51
Future Challenges
The net result of processing changes
has been improved microbiological control and reduced levels of enteric pathogens. Since the adoption of these more
comprehensive systems for pathogen
control, the prevalence of salmonella
has decreased as determined by the
USDA/FSIS monitoring program. But
52
into fillets and other value-added products. Fishery products may be consumed
raw, or they may be processed into
ready-to-eat products. Salmon, trout
and other fish species are often processed
into cold smoked and hot smoked fishery
products. Fishery products also are used
in a variety of salads and spreads, and
they are cured, fermented, pickled, dried,
and canned. Rapid harvesting, cooling
and processing can reduce or prevent
the occurrence of certain biological and
chemical food safety hazards associated
with some wild caught and cultured fishery species (e.g., molluscan shellfish and
scombrotoxin susceptible species) (NAS,
1991). In other instances, the use of food
additives or other post processing interventions (e.g., high hydrostatic pressure,
irradiation, or thermal pasteurization)
may be required to control food safety
hazards.
Regardless of the commodity, processing hazards can include pathogens
that may be naturally present on the
food, that may be introduced during
processing and handling, or that may increase to hazardous levels during distribution and storage. Strict attention to
good manufacturing practices, sanitation
control procedures and hygienic practices of plant employees are effective in
controlling many of these hazards.
Water
Water is used extensively in the postharvest processing environment, making
water quality a significant concern. Advances in water treatment over the last
100 years have resulted in dramatic improvements to the microbiological safety
of the public water supplies of developed
countries (Dawson and Sartory, 2000).
Perhaps of greatest present-day concern
in these countries are large communitywide waterborne outbreaks of parasitic
protozoa that are associated with either
unfiltered or inadequately flocculated or
filtered water, such as the Cryptosporidium outbreak that occurred in Milwaukee
in the early 1990s (MacKenzie et al.,
1994; Moe, 1996). It is important to note
that, although drinking water is quite
safe in the United States, it remains a significant cause of morbidity and mortality in developing countries, where water
remains a common source of bacteria,
viruses, and parasitic protozoa that impact human health. If this water is used
in food processing, waterborne diseases
of developing countries can be passed on
INSTITUTE OF FOOD TECHNOLOGISTS
to the consuming public in more developed regions of the world through importation of contaminated foods.
From the perspective of post-harvest
issues for produce, wash water, rinse water, and ice can serve as a potential
source for contamination of fresh produce (Beuchat and Ryu, 1997). Perhaps
predictably, recent evidence suggests that
puncture wounds on fruit usually harbor greater numbers of pathogens to
greater depths than seen at other locations of the intact fruit (Burnett et al.,
2000). More significant, Burnett et al.
(2000) found that a negative temperature
differential (application of cold rinse water to warm fruit) may increase the relative attachment and infiltration of bacterial pathogens in intact fruit. This observation has significance for the use of hydrocooling technology, which has been
applied to produce items such as strawberries, cherries, and field crops. Although hydrocooling water is frequently
decontaminated by chlorination, inadequate control of pathogens in water recirculated through hydrocoolers could
provide a source of pathogens that then
infiltrate produce items because of the
temperature differentials.
Historically, water for food processing in the United States has originated
from municipal systems. However, with
increased volume, as well as stricter regulations, water usage in the processing
environment has increased dramatically
over the last decade. Water reclamation
and reuse has received considerable attention, with guidelines provided by the
Environmental Protection Agency (EPA)
(EPA/AID, 1992), although actual standards remain the responsibility of state
agencies. These EPA guidelines address
water reclamation for nonpotable urban,
industrial and agricultural reuse; direct
potable reuse is not practiced in the
United States. Food processors have expressed some interest in water reuse as
well, particularly in animal slaughter facilities and the USDA/FSIS has issued
guidance for this application. In general,
the reuse of water from any one location
in the slaughter process is restricted to
certain other point locations in the
slaughter process, as provided by specific
recommendations of the agency. In almost every instance, decontamination
steps such as addition of chlorine, as well
as microbiological monitoring, must be
done on the reconditioned water, with
very specific standards recommended.
Reuse water from advanced wastewater
EXPERT REPORT
Alternative Processing
Technologies
Preservation techniques currently
act in one of three ways: (1) preventing
pathogen access to foods, (2) inactivating them should they gain access, or (3)
preventing or slowing their growth
should the previous two methods fail
(Gould, 2000a). Traditional food processing has relied on thermal treatments
to kill/inactivate microbiological contaminants. Unfortunately, thermal processing induces physical and chemical
changes in the food. Canned green
beans do not have the same taste and
texture as fresh, despite having similar
nutritional profiles. Chemical preservatives and naturally occurring antimicrobial compounds also have been used
extensively for food preservation
(Davidson, 1997). Again, a pickled or
acidified food such as cauliflower does
not have the same role on the menu as a
fresh stalk of cauliflower, despite their
identical origin. Beyond the use of singular food preservation techniques,
many strategies employ a combination
of preservation techniques, e.g., refrigerated storage under modified atmosphere, reduced heat treatment with
some acidification, and mild heat with
reduced water activity (Gould, 2000b).
Leistner (2000) states that the important preservation approaches often use
combinations of several factors to assure microbiological safety. These factors, often called hurdles, include heat,
acidity, water activity, redox potential,
preservatives, competitive flora, low
temperatures, and more than 40 other
possible factors. Increasingly, the American public has sought fresh products,
fueling efforts to develop many alternative processing technologies that result
in products that have minimal processinduced changes in sensory and nutritional characteristics. It is expected that
these technologies will play an increasing role in food processing in the future.
Any discussion of emerging food
safety issues must consider the impact of
these alternative food processing technologies. This analysis must consider
both the immediate impact and the potential ramifications further down the
farm-to-table chain. Altering any parameter along the entire food chain can have
consequences beyond the immediate
change, be they intended or unintended.
For example, modifying the feed of a
food animal may alter the microbial contamination of the final product that may
in turn contaminate the kitchen of the
unaware consumer. A new method of
heating food (e.g., microwave) may speed
up the heating and change the types of
microorganisms that survive the heating
process, whether the heating takes place
in a food processing plant or the consumers kitchen.
Overview
A recent report generated by IFT for
FDA (IFT, 2000b) defined alternative
technologies, identified the pathogens of
public health concern that are the most
resistant to various technologies, described the mechanisms of pathogen inactivation including the inactivation kinetics, identified ways to validate the effectiveness of microbial inactivation,
identified critical process factors, and
described process deviations and ways
to handle them. The report also described synergistic effects among technologies, when data were available, and
articulated future research needs for
each technology.
Kinetic parameters and models are
frequently used to develop food preservation processes that ensure safety. They
also allow scientists to compare the ability of different process technologies to reduce microbial populations. Kinetic parameters, with their recognized limitations, use empirical coefficients experimentally determined from microbial reduction measurements to document the
relationship between microbial population decreases and different process conditions. Kinetic parameters for microbial
populations exposed to thermal treatments have been assembled over a significant period of time. Published literature
has included kinetic parameters needed
to control most process, product and microbial situations (Pflug and Gould,
2000).
IFT (2000b) used the models and kinetic parameters to present and compare
microbial inactivation data from thermal, pressure and electromagnetic processes. Thermal parameters apply to microwave energy, electrical resistance
53
Table 10. Limitations to Alternative Processing Technologies Currently Under Development (IFT, 2000b)
Limitations applicable to all or most of the technologies:
Linear first-order survivor curve model
may be inadequate. Appropriate model(s)
would be beneficial to all preservation
technologies.
Standardized experimental protocol(s) for
obtaining statistically reliable kinetic
parameters to describe survivor curves for
microbial populations exposed to various
alternative technologies.
Electrothermal:
54
Others:
Some of the technologies present greater
limitations than others or are at a
development stage that requires extensive
further scientific research before they can
be commercially used. For these
technologies, data are insufficient to
calculate kinetic parameters. For
example, high voltage arc discharge
(application of discharge voltages through
an electrode gap below an aqueous
medium) causes electrolysis and highly
reactive chemicals. Although microorganisms are inactivated, more recent designs
care should be taken to compare the resistance of different microbial populations or to identify the appropriate target
microorganisms. When a new alternative
processing technology is under review, it
is essential to determine the pathogens of
greatest public health concern.
When exploring the new preservation technologies, their preservation level
should be compared to that of classical
thermal pasteurization or commercial
sterilization technologies. Thermal pasteurization focuses on inactivating vegetative cells of pathogenic microorganisms, i.e., microbial cells that are not dormant, highly resistant spores. However,
to have a commercially sterile product,
the process must control or inactivate
any microbial life (usually targeting
spores of C. botulinum) capable of germinating and growing in the food under
normal storage conditions. Commercially sterile products generally require more
EXPERT REPORT
To preserve food using high hydrostatic pressure, the food (normally packaged) is submerged in a liquid (usually
water) contained in a vessel that generates pressure by pumping more liquid
into the pressure vessel or reducing the
volume of the pressure chamber.
In the 19th century, scientists realized
55
Irradiation
Food irradiation, first commercially
introduced in the early 1960s, uses ionizing radiation to decontaminate and disinfect food and inhibit sprouting and
ripening. The ionizing radiation is usually in the form of gamma rays produced
by radionuclides such as 60Co (cobalt) or
137
Cs (cesium). Newer irradiation technologies include e-beam, where ordinary
electricity is used to produce a stream of
electrons, or x-ray irradiation, where the
electron beam is bounced off metal to
create x-rays (Farkas, 1998). To get away
from using the term irradiation, food
INSTITUTE OF FOOD TECHNOLOGISTS
accepted by FDA for food safety purposes include the more recent addition of
microbial control in fresh and frozen
meat and poultry.
To establish the safety of a proposed
food irradiation application, FDA requires data on the radiological, toxicological, and microbiological safety, as
well as the nutritional wholesomeness of
the irradiated product (Pauli and Tarantino, 1995). From the perspective of radiological safety, the energy produced by
approved radiation sources is too low to
induce radioactivity in foods. The issue
of toxicological safety is more complex
and has been thoroughly studied in the
past. Recent petitions in the meat and
poultry area have indicated that FDAs
principal current interest lies in specification of the conditions for food irradiation (such as temperature and packaging
atmosphere) and their impact on microbiological safety and nutritional adequacy (Olson, 1998). The two most important concerns related to the microbiological safety of irradiated foods are (1) the
potential to create highly virulent mutant
pathogens, and (2) the potential that reducing the harmless background microflora could eliminate competitive microbial forces and allow uncontrolled
pathogen growth.
The relative radiation resistance of
microorganisms can be summarized as
follows (in order of most resistant to
least resistant): viruses>spores>grampositive bacteria>gram-negative
bacteria>yeasts and molds>parasites
(Monk et al., 1995). However, there are
some nonsporeforming bacteria, such as
Deinococcus radiodurans and Acinetobacter radioresistens, that are extremely
radiation resistant by virtue of their genetic make-up. Although the exact cellular mechanism(s) responsible for such
resistance is unknown, scientists believe
that these radiation-resistant bacteria
possess particularly effective nucleic acid
repair mechanisms. Although these organisms are not known to be pathogens,
they may provide a genetic pool from
which pathogens could theoretically pick
up resistance genes through mechanisms
of genetic exchange. Furthermore, some
researchers have proposed concerns that
irradiation may produce mutant strains
and/or radiation-resistant pathogens by
means of natural selection. To date, various studies and independent reviews by
researchers and international organizations have found no indication of specific bacteriological hazards associated with
sults from the effect of the modified environment on microbial growth and, for
respiring food products such as fruits
and vegetables, on the product (Molin,
2000). Low oxygen levels negatively affect aerobic microorganisms. Carbon dioxide inhibits the growth of some microorganisms, however, it is not inhibitory
for all microorganisms, and it is important to understand that under modified
atmospheres, the growth rates of some
microorganisms will be reduced, while
others may increase or stay the same.
MAP is not without safety concerns.
Beneficial microorganisms may be inhibited, potentially allowing certain
pathogens to proliferate and cause
foodborne illness. C. botulinum, L.
monocytogenes and potentially pathogenic psychrotrophs are of primary
concern. Psychrotrophs grow well at or
below 7 C and have their optimum
growth temperature between 20 and 30
C, meaning they can multiply under refrigeration conditions. If the oxygen level of the modified atmosphere is too
low, the anaerobic environment could
facilitate C. botulinum growth. Although studies indicate that many foods
become inedible before any toxin is produced (Molin, 2000), foods that remain
organoleptically acceptable after toxin
production are a significant public
health concern. For example, recent
studies have shown that C. botulinum
can grow and produce toxin in products
such as MAP pizza and English-style
crumpets, while the products remain
organoleptically acceptable (Daifas et
al., 1999a,b). With respect to meat and
fish products, Molin (2000) concluded
that most products are judged inedible
well before any toxin is produced. However, there have been studies where toxin
production in MAP meats preceded or
coincided with the development of unacceptable sensory characteristics
(Lawlor et al., 2000), as well as reports
of botulism due to consumption of vacuum-packaged smoked fish products
(Korkeala et al., 1998). L. monocytogenes is a problem with products, such as
ready-to-eat products, fruits and vegetables, that are not heated adequately
before consumption. Low concentrations of CO2 (less than 10%) used in
MAP of some products may inhibit the
natural microflora and increase the
growth rate of L. monocytogenes. Combined with storage at refrigeration temperatures, which can select for L. monocytogenes, a low-CO2 MAP environment
EXPERT REPORT
Key applications for intelligent technology are in the areas of tamper evidence, quality monitoring (i.e., temperature abuse), counterfeiting (i.e., holograms), theft protection and supply
chain management and traceability (i.e.,
automatic data capture coupled to the
Internet). Traceability in the food chain
is a high profile issue, because of its importance in determining which foods are
part of a potentially contaminated lot.
Tracing technology is already being used
in many parts of the food chain, however,
a number of gaps exist.
An example of quality management
packaging is a newly developed method
of preparing packaging material for food
and other products that contain diagnostic properties. The technology involves
immobilizing and protecting antibodies
(or other ligands) on the polymer film
surface. These substances react with targetssuch as food pathogens and toxins, pesticide residues or proteins in the
foodand create a visual sign on the
film surface, alerting the consumer or retailer that the food may be contaminated.
However, this technology would likely
only be applicable for surface contamination. Other examples of quality management technologies are temperature
indicators, time indicators (aging strip),
time/temperature indicators, and microwave cook indicators.
normally transported to holding, shipping, or processing facilities. Even processed foods that have received microbicidal treatment are frequently transported to other locations for bottling, packaging, and/or shipping. Transport conveyances are thus a part of the food
chain where contamination can occur.
In 1994, an estimated 224,000 persons developed salmonellosis from a nationally distributed brand of ice cream
(Hennessy et al., 1996). S. Enteritidis was
the cause, and the most likely scenario
was that pasteurized ice cream premix
was transported by a tanker trailer that
had carried non-pasteurized eggs just
before being loaded with the premix.
Eggs are a known source of S. Enteritidis.
The authors concluded that to prevent
more such occurrences, food products
not destined to be re-pasteurized before
use should be transported in dedicated
containers.
As more and more food commodities
are grown and harvested in the United
States and abroad, transportation will
continue to be a factor in foodborne illness and, in fact, may grow in importance. This is not an easy issue to deal
with, as the types of conveyances are as
varied as the types of commodities they
transport.
The storage of foodstuffs also can be
an entry point for pathogenic microorganisms or permit the growth of pathogens if present. It is generally accepted
that most foods need to be maintained at
cold temperatures from harvest to consumption. This cold chain is subject to
abuse at several steps, and temperature
abuse can contribute to the growth of
pathogens that can increase the likelihood of foodborne illness.
Storage of foodstuffs is carried out in
warehouses and specialized storage facilities, as well as in virtually all institutions
that serve food, including hospitals,
nursing homes, schools, restaurants, retail stores and the home. FDA has determined that improper cold holding of
food is the most frequent temperature
violation for nearly all facility types. For
example, in a survey of fast food restaurants, 31% were out of compliance in
that potentially hazardous foods were
being stored at temperatures above 41 F
(FDA, 2000).
The situation is no better, and is
probably worse, in the home setting. In a
recent survey of homes, 16% were holding refrigerated ingredients at too high a
temperature, and 55% of the participants
INSTITUTE OF FOOD TECHNOLOGISTS
that were improperly holding cold ingredients did not know at what temperature
a refrigerator should hold product. The
remaining 45% of participants were unaware that their refrigerators were not
holding products at the proper temperatures (Daniels et al., 2000). A previous
survey showed that 23% of the 106
households had refrigerators that held
food at too high a temperature (Daniels,
1998).
While improper transportation and
storage are hardly new or emerging
food safety issues, they appear to be
problems that do not go away. As such,
they will continue to contribute to the
burden of foodborne illness. As the
Outbreaks of Shigella
sonnei Infection Associated with Fresh Parsley
In August 1998, the Minnesota Department of Health (MDH) received
multiple independent complaints of illness and reports of confirmed Shigella
infections among persons who had eaten at two restaurants (CDC, 1999c).
The restaurants were in different cities,
had separate water supplies, and had no
employees in common. Preliminary results of interviews with patrons and
food handlers at both restaurants suggested that ill food handlers likely
played a role in contaminating ice and
fresh produce items.
S. sonnei isolates from ill patrons
and food handlers at the two restaurants were submitted to MDH for molecular subtyping by pulsed-field gel
electrophoresis (PFGE). Results of
PFGE demonstrated that both restaurant outbreaks were caused by the same
strain of S. sonnei and that it was a
strain that had not previously been isolated in Minnesota. Strains with similar PFGE patterns had been isolated
from travelers returning from Mexico.
Because the outbreaks at the restaurants appeared to have a common
source, food histories of restaurant patrons were re-evaluated by food ingredients rather than by menu item. In
one restaurant, uncooked chopped
parsley was associated with illness
(odds ratio 4.3, 95% confidence interval 2.4, 8.0). In the other restaurant,
parsley was not associated with illness,
EXPERT REPORT
food chain becomes even more complex, perhaps their effect will become
even greater.
The retail and food service environments are part of the post-harvest environment. As more meals are eaten away
from home, this environment becomes
increasingly significant. No matter how
well or how poorly food safety measures
are applied prior to consumption, avoiding illness often depends on how well
food is handled immediately prior to
consumption.
While the role of human handling
in foodborne disease has been recognized for years, recent food safety initiatives have increased our awareness of
particular risks. For instance, strong
epidemiological evidence supports the
transmission of Salmonella and Campylobacter to ready-to-eat (RTE) food
products via cross-contamination with
uncooked poultry (DAoust, 1989;
Deming et al., 1987; Harris et al., 1986;
Hopkins et al., 1984). Equally strong
evidence exists for the transmission of
viral foodborne disease by poor personal hygiene of infected food handlers,
with recent data suggesting that 50-95%
of confirmed viral foodborne disease
outbreaks are attributable to human
61
Resistance to Controls
The efficacy of a preservation technology is influenced by a number of microorganism-related factors that are generally independent of the technology itself. These factors include the type and
form of the target microorganism; the
genus, species and strain of microorganism; its growth stage; selection by environmental stresses; and sublethal injury.
Among the foodborne microbiological hazards, bacteria are generally the
primary targets for most preservation
processes, and bacterial susceptibility to
sublethal cellular injury is of special
concern. Processes designed to inactivate pathogens also must address the resistance properties in foods of other microorganismssuch as viruses, yeasts,
ous exposure or through cross protection triggered by environmental or processing factors including stresses such as
heat or acid (Davidson, 1999). Some researchers have attempted to differentiate
various multiple stresses, e.g., lethal pH
and water activity. Shadbolt et al. (2001)
hypothesized that pH-induced stress
causes an energy drain that sensitizes
the cell to other environmental constraints. Similarly, the effect of habitual
exposure to reduced water activity increased the heat tolerance of Salmonella
spp. (Mattick et al., 2000). Another example of cross protection is the increased radiation resistance caused by
the induction of acid resistance in enterohemorrhagic E. coli (Buchanan et al.,
1999).
Because environmental stress can
increase a pathogens resistance to control technologies, the potential for increased resistance must be factored into
the process. The analysis should consider: (1) if the food environments are likely to induce stress conditions in the microorganisms; (2) whether stress-induced resistance could possibly occur at
any point in the food processing operation; and (3) if it did, whether it would
significantly impact the inactivation
process and lead to possible underprocessing. Considering that most food
processing systems are designed to expose microorganisms only once to any
given stress-inducing factor (for example, heat, acid, or antimicrobials), a resistant population is unlikely to develop.
However, it is possible that sublethal injury to E. coli O157:H7 as it passes
through the low pH of a cows gastrointestinal tract and then through an
acid rinse at slaughter may change its resistance to heat inactivation during
preparation. Likewise, L. monocytogenes
cells in the production environment that
are sublethally injured by repeated exposure to sanitizers may have altered
survival or virulence characteristics. An
additional exception is previously processed material that is reintroduced into
the process stream. In this case, indepth studies of the impact of processing-induced stress are needed.
Sublethal Injury
The microbial ecology of food is influenced dramatically by food processing and preservation technologies.
Whether or not the actions are directly
or indirectly aimed at microorganisms,
64
low for the recovery of many of the injured cells (see sublethal injury, p. 63).
In direct plating, the sample is placed
directly on or in selective media, which
may inhibit the growth of the injured
cells. The MPN technique, however, is
extremely labor intensive. Molecular
techniques, such as the coupling of immunochemical or nucleic acid-based
assays to the MPN, or nucleic acid
probe hybridizations of colony blots (de
Blackburn and McCarthy, 2000; FDA,
1998; Miwa et al., 1998), are increasingly being applied to traditional methods
for obtaining results more quickly.
PCR, originally developed for screening/detection applications, has been
modified for use as a quantitative assay
(Nogva et al., 2000). Recombinant bioluminescent strains of food pathogens
have been used to quantitatively assess
survival in foods and the food processing environment by measurement of
light emission (Ramsaran, et al., 1998;
Siragusa et al., 1999). These techniques
and others need to be further developed,
refined and validated as our need for
quantitative information on pathogens
increases.
The development of genetic fingerprinting techniques has provided the
ability to finely discriminate strains
within a species of pathogen. Several
variations of fingerprinting techniques
are available (Farber et al., 2001), and
new ones continue to be developed. Genetic fingerprinting has been extremely
useful in epidemiological work and
identification of foodborne illness outbreaks (e.g., PulseNet) (CDC, 1999d).
The technology has begun to be applied
to the study of pathogens in the food
plant environment (Norton et al., 2001).
This novel approach allows questions
about pathogen evolution, persistence
and routes of contamination in the food
plant to be examined.
New tools also need to be developed
and applied for assessing stress and injury of pathogens in the food production environment. Sublethally injured
cells may have different survival characteristics during food storage or after
consumption, and may show greater resistance to control measures and
heightened virulence than the original
population (Abee and Wouters, 1999;
Bower and Daeschel, 1999; Gahan and
Hill, 1999). If the injured cells are not
detectable by the method chosen, the
safety of a food process may be assumed, when, in fact, a food safety risk
Ability of Pathogens
To Survive in the
Environment
The Vaudois University Hospital Medical Center in Lausanne,
Switzerland, generally diagnoses a
mean of three listeriosis cases per
year. However, a cluster of 25 listeriosis cases (14 adults and 11 maternal/fetal) was observed at the
same medical facility between January 1983 and March 1984, with 15
additional cases diagnosed at surrounding hospitals (Bille, 1988; Malinverni et al., 1986).
Epidemiology
This epidemic appeared atypical because of the high number of
healthy, immunocompetent individuals affected, the high rate of
brain-stem encephalitis, and a
mortality rate of 45%. The organisms isolated from clinical specimens from thirty-eight of the 40
patients involved in the outbreak
were serotype 4b. A high number
(92%) of the L. monocytogenes serotype 4b cultures were of two
unique phage types, compared
with only 44% of the serotype 4b
cultures obtained during the previous 6 years. This evidence indicated the outbreak might be traced
to a single source. A thorough
case study did not identify the
source or mode of Listeria transmission, however, public health officials initiated a prospective casecontrol study, assuming that a
similar listeriosis outbreak was
likely the following winter. Overall, 16 additional cases were identified between November 1984 and
April 1985.
After a 1985 listeriosis outbreak
in California was linked to consumption of Mexican-style cheese,
Swiss officials initiated a baseline
study to detect Listeria spp. in a variety of dairy products. While surveying soft, semi-hard, and hard
cheeses, L. monocytogenes was iso-
66
Management
Immediately following the recall,
Swiss officials began investigating how
the cheese could have become contaminated. The cheese implicated in this
outbreak was produced at 40 different
factories located in western Switzerland,
and all contaminated cheese was reportedly prepared from Listeria-free bovine
milk. After coagulating the milk, the resulting curd was dipped into wooden
hoops and allowed to drain for 1-2
days. When drained, the cheese was
transported to one of 12 cellars located
throughout the area and ripened for 3
weeks on wooden shelves, during which
Application of Science to
Food Safety Management
Effective application of our current
scientific knowledge is a vital part of
continuing efforts to improve food
safety. Our food safety policies are
based on the best scientific information available at the time they were
created, but our knowledge continues
to improve. Flexible, science-based
policies should incorporate new
information as it becomes available.
Everyone benefits from science-based
policies, although factors such as
economic impact and other trade-offs
Risk Assessment
are part of the policy-making environment as well. Ideally, food policy will
draw on a variety of science-based tools
and be structured to provide the
flexibility to apply these tools as
science dictates.
The final decades of the twentieth
century saw tremendous change in our
knowledge of pathogenic microorganisms, their toxins and their metabolites.
Scientists identified a wide array of microbial hazards, and foods previously
thought to be safe were found to be important vehicles of foodborne disease.
This new knowledge resulted in new
policies to protect consumers. This scenario will continue into the new century.
Unfortunately, current systems cannot deliver a risk-free food supply. The
scientific knowledge, technology and
equipment are not available to eliminate
all microbial hazards from all foods.
This vulnerability will continue to influence regulatory policy as regulatory
agencies and industry seek solutions to
these hazards. The primary weakness in
todays control system is the presence of
enteric pathogens on raw agricultural
commodities (e.g., meat, poultry, milk,
eggs, fruits, vegetables, and seafood).
As a result, an essential part of sciEXPERT REPORT
frequently done to model the many circumstances that may surround exposure. And since foodborne pathogen
growth, inactivation and survival are
highly dependent upon intrinsic (foodrelated) and extrinsic (environmental)
factors such as relative humidity and
storage temperature, mathematical
models such as USDAs Pathogen Modeling Program can be used to estimate
the effect of these factors on microbial
persistence and levels in foods.
The relationship between the ingestion of pathogenic microorganisms and
possible health effects may be described
as the quantitative relationship between
the intensity of exposure (dose) and the
frequency of the occurrence of illness
within the exposed population of hosts
(response). For pathogenic microorganisms, this is dependent upon the
number of units of infectious agent ingested in the food, the infectivity and
pathogenicity of the infectious agent,
and the vulnerability of the host. The
purpose of the hazard characterization
step is to quantify or statistically describe the relationship between the risk
agent and the magnitude of the adverse
effect. Included in this step is a full description of the severity and duration of
adverse effects that may result from the
ingestion of a microorganism or toxin
in a food. The source of data used for
hazard characterization is usually human challenge studies, whereby a defined population of consenting adults is
given various doses of the infectious
agent, and their response is measured as
infection or disease. For pathogens
causing mild disease, this is feasible; for
those associated with more severe diseases, or affecting particularly susceptible populations, risk assessors must rely
on animal models of disease or other
data sources. The raw input data from
these types of studies is used in conjunction with statistical methods that
describe the dose-response relationship
in mathematical terms. Some frequently used dose-response models include
the Beta-Poisson, Exponential, and
Gamma-Weibull models.
Risk characterization is the final step
of risk assessment and represents the integration of the exposure assessment and
hazard characterization to obtain a risk
estimate of the likelihood and severity of
the adverse effects that would occur in a
given population. The final risk estimate
should incorporate information about the
variability, uncertainty and assumptions
INSTITUTE OF FOOD TECHNOLOGISTS
FSO Approach
Risk Management
Consider options, including a Food Safety Objective
Hazard Control and Monitoring
Agricultural interventions implemented
GMPs/GHPs implemented
Baseline level of microbial hazard measured
Processing safety objective calculated
(FSO minus growth during storage and distribution)
HACCP
Approach
70
Table 12. FSOs in the Food Safety Management Framework (Derived from ICMSF, 2002)
Managing Microbiological Food Safety
Public health concern
(often based on epidemiological data)
Risk Assessment
Hazard identification
Exposure assessment
Hazard characterization
Risk characterization
Risk Management
Food safety objective
Performance criteria
essary, to further verify that the processing safety objective has been met.
Pasteurization
Fewer than 100 cells can cause disease, especially among young
children
One estimate found 26 x 104 patties per year nationwide may contain
viable E. coli O157:H7 after cooking
Thermal processing
Parameters for sanitation and control measures (prevention of contamination during slaughter and decontamination) may be defined
cover sanitation issues, such as equipment design and cleaning, and pest control.
Performance Criteria
FSOs are met using performance
criteria, which are the required outcome of a control step or a combination of steps. At certain points in food
processing, control measures can be
applied to either prevent an unacceptable increase in a microbiological hazard or reduce the hazard to an acceptable level. Chilling cooked meats and
stews prevents the growth of Clostridium perfringens (i.e., increase of a hazard), and pasteurization of milk or fruit
juices eliminates enteric pathogens (i.e.,
decrease of a hazard). The performance criteria are the reduction necessary (e.g., a 5-log reduction) to achieve
the processing safety objective; they are
calculated from the baseline level of the
microbial hazard (see Fig. 8). Performance criteria also may address the
prevention of pathogen growth (e.g.,
less than 1-log growth).
Under the FSO approach, it is important not to confuse performance
criteria and performance standards.
Baseline
Processing
safety objective
Food
processing
facility
106
Agricultural
intervention
Baseline
Processing
safety objective
Food
processing
facility
74
101
Baseline
Processing
safety objective
105
Food
processing
facility
no reduction required;
HACCP
HACCP is a management tool used
by the food industry to enhance food
safety by implementing preventive measures at certain steps of a process.
When HACCP principles are properly
implemented, microbiological hazards
that have the potential to cause foodborne illness are controlled, i.e., prevented, eliminated or reduced to an acceptable level.
The pathway to HACCP began in
1959 because existing quality control
techniques could not provide the desired level of safety for food produced
for the space foods program (Bauman,
1992). Traditional microbiological testing of finished product was impractical
and ineffective because of the small
quantities of food produced, and the
high product cost limited the amount
available for sampling. In addition, the
food industry had no uniform approach
to managing food safety. HACCP was
developed to meet this need.
Over the years, the fundamental
concepts that comprise a HACCP program have been refined, and their application has become more practical. The
U.S. National Advisory Committee on
Microbiological Criteria for Foods
(NACMCF)a federal advisory committee assembled to provide impartial,
scientific advice to federal food safety
agencies for use in policy developmenthas revised and expanded the
original principles based on industry
experience (NACMCF, 1992; 1998). The
committee consists of experts in microbiology, risk assessment, epidemiology,
public health, food science, and other
relevant disciplines. At the same time as
the NACMCF activities, Codex adopted
a similar HACCP document (CAC,
1997c). NACMCF adopted seven
INSTITUTE OF FOOD TECHNOLOGISTS
108
Food
processing
facility
Pathogen
level
Food
processing
facility
5 log reduction
103
HACCP principles:
Principle 1: Conduct a hazard
analysis.
Principle 2: Determine the critical
control points.
Principle 3: Establish critical limits.
Principle 4: Establish monitoring
procedures.
Principle 5: Establish corrective
actions.
Principle 6: Establish verification
procedures.
Principle 7: Establish recordkeeping and documentation procedures.
The fundamental HACCP principles,
associated definitions, and principles of
application were intended to help the food
industry implement food safety management systems. First, companies identify
hazards that have the potential to cause
illness or injury to the consumer and determine whether or not these hazards are
significant. For significant hazards, the
company identifies critical control points
(CCPs) in the food system, where possible, and establishes critical limits for their
control. Critical limits are based on evidence that the control is effective in practice. Next, CCPs must be monitored to
assure critical limits are not violated. This
monitoring must be a systematic procedure that verifies that the HACCP system
is functioning as intended and that it is effective. Finally, the company must maintain comprehensive records associated
with the HACCP system.
HACCP is an essential part of implementing an FSO. Through the identification of CCPs, the process and/or product
criteria can be translated into processspecific critical limits. Monitoring of
these critical limits ensures that the performance criteria are met.
Using the principles identified in the
NACMCF and Codex documents as a
standard, HACCP was widely adopted by
EXPERT REPORT
Baseline
106
101
Pathogen
level
Number of Samples
10
20
30
50
100
200
300
0.90
0.82
0.74
0.61
0.37
0.13
0.05
0.82
0.67
0.55
0.39
0.13
0.02
<
0.74
0.54
0.40
0.22
0.05
<
0.66
0.44
0.29
0.13
0.02
0.60
0.36
0.21
0.08
0.01
0.54
0.29
0.16
0.05
<
0.48
0.23
0.11
0.03
0.43
0.19
0.08
0.02
0.39
0.15
0.06
0.01
10
0.35
0.12
0.04
0.01
Samples
Positive
Broilers
22,484
10.2%
Market hogs
8,483
7.0%
Cows/bulls
3,695
2.1%
Steers/heifers
2,088
0.3%
Ground beef
50,515
3.7%
Ground chicken
735
14.5%
Ground turkey
3,192
29.2%
EXPERT REPORT
PV(+) =
(prevalence) (sensitivity)
(prevalence) (sensitivity) + (1-prevalence) (1-specificity)
PV(+) =
(0.60) (0.95)
(0.60) (0.95) + (0.40) (0.05)
PV(+) = 0.966, or 96.6% of the time the test is positive, the sample is truly contaminated
PV(+) =
(0.01) (0.95)
(0.01) (0.95) + (0.99) (0.05)
PV(+) = 0.161, or only 16.1% of the time the test is positive, the sample is truly contaminated
77
Microbiological Criteria
Historically, attempts have been
made to apply microbiological criteria
for the purpose of classifying foods as
either microbiologically acceptable, or
microbiologically unacceptable. In
1985, the Food and Nutrition Board of
the National Research Council (FNB/
NRC) addressed the subject of microbiological criteria, and found that such
criteria were of limited use, particularly if safety assurance is the goal, and
that HACCP should be applied wherever possible for safety assurance
(FNB/NRC, 1985). The HACCP systems envisioned by the FNB/NRC did
not rely on end-product testing for
pathogens, but rather, if analyzed microbiologically at all, analyses were
performed at points along the foods
production chain, particularly to verify
that CCPs were under control.
Microbiological criteria considered
by FDA generally include standard
plate count, coliform counts, yeast and
mold counts, and E. coli (generic)
counts. Coliforms and E. coli were believed to be indicators of possible fecal
contamination, and therefore, it is pos78
sible that food containing either bacteria at a prescribed level was unsafe.
The zero tolerance for L. monocytogenes in RTE foods was established as
a safety-related criterion. The zero
tolerance actually means that L.
monocytogenes must be absent from
two 25g samples of foods under FDA
inspection. The total absence requirement was derived at a time when there
were no effective methods for finding
L. monocytogenes in food (or any environment outside of the human or animal). There was no understanding of
the very widespread existence of L.
monocytogenes throughout the environment, including food processing
environments, nor an appreciation of
the number of foods in which the bacteria historically had been present.
Thus, contemporary knowledge about
human exposure suggests that many
humans are routinely exposed to the
bacteria with no consequence to health,
although L. monocytogenes does cause
illness in sensitive subpopulations.
The zero tolerance has acted as a disincentive for the application of quantitative (enumerative) methods, and
thus, the body of human exposure data
is incomplete.
Today, there is growing acceptance
of the need for management systems
based on GMPs and HACCP to control
food safety hazards. Microbiological
testing is sometimes valuable in verifying the effectiveness of GMP and
HACCP systems and validating CCPs
within HACCP systems. There may be
a role for testing certain ingredients
when they can influence the safety of a
finished product, but because the testing of ingredients faces the same weakness as end-product testing, auditing
of suppliers control programs has increased to provide greater assurance.
In addition to product testing, environmental testing may be necessary.
Salmonella and L. monocytogenes have
the ability to become established as
residents in food processing establishments. Environmental sampling programs assess the degree of control and
indicate when corrective actions are
needed. They may or may not indicate
a safety problem in the finished product.
The optimal regulatory approach to
environmental testing would use our
understanding of basic human nature
to encourage and reward diligence. Depending on the type of food and the
processing conditions, it should be expected that these pathogens will be periodically introduced into the food
processing environment by various
pathways. To prevent the pathogens
from becoming established and multiplying, the sampling program should
aggressively look for these pathogens.
Finding a positive sample should be
treated as a success, because corrective
actions can then be applied and consumer protection assured. Treating a
positive sample as a failure and applying a penalty decreases the desire to detect the pathogens, discouraging the
aggressive nature of the environmental
sampling program.
Testing Methods
Disease surveillance and control efforts will benefit greatly from new
pathogen detection methods that offer
greater precision, rapid results, and decreased cost. However, efforts to adapt
these new technologies to the challenges in the food environment are ongoing.
Many methods of detection are currently available for foodborne pathogens, but food microbiologists must often choose between enumeration and
identification without the option of
both. Enumerative methods are usually
based on the ability of the normal
healthy bacterial cells to multiply in a
nutrient-rich medium. Although selective agents are sometimes added to favor
the growth of a specific group of organisms, most of these methods are still
reasonably nonspecific. With respect to
pathogen identification, methods have
historically relied on cultural enrichment to increase the numbers of the target microorganism and allow resuscitation of injured cells. When followed by
selective and differential plating, these
methods provide discrimination of the
target organism from the background
microflora, but are non-enumerative.
For both enumerative and non-enumerative methods, the combined effect
of low levels of contamination and the
need for cultural growth results in
lengthy assays, frequently extending beyond four days for even preliminary results.
Most rapid method developments
have sought to shorten detection time
by replacing the selective and differential plating steps with more rapid technologies such as ELISA and DNA hyINSTITUTE OF FOOD TECHNOLOGISTS
and reports of illnesses among food handlers led to early concern that the restaurant was experiencing a large outbreak of
viral gastroenteritis. However, as the
clinical and epidemiologic features of the
outbreak emerged from interviews with
patrons, it appeared typical of previously
described outbreaks caused by enterotoxigenic E. coli (ETEC).
The recognition that the outbreak
may have been caused by an uncommon
foodborne pathogen led to extensive efforts to obtain stool from ill patrons to
confirm the etiology. A lactose-negative
non-motile E. coli O39 was isolated from
10 of 22 cases. No Salmonella, Shigella,
Campylobacter, Yersinia, Vibrio, or Plesiomonas species were isolated from ill patrons. Although the clinical and epidemiologic features of the outbreak suggested ETEC, the outbreak-associated
O39 strain did not possess ETEC heatlabile (LT) or heat-stable (ST) enterotoxins. Extensive testing of the outbreak-associated strain by a battery of gene
probes detected the presence of intimin
(eae), an adherence factor associated
clined 48% during 1996 -1999, suggesting that these control strategies are beginning to work (CDC, 2000a, b).
Unknown Agents
Surveillance for foodborne diseases
is based on detection of specific pathogens or the occurrence of illnesses, such
as diarrhea, in defined groups. An outbreak may be recognized because a state
public health laboratory detected the increased occurrence of a specific subtype
of E. coli O157:H7, or because half of the
people who attended a specific event developed vomiting and diarrhea shortly
after the event. In the first case, surveillance is limited by what clinical laboratories routinely identify when processing
human stool samples, whether by direct
examination, culture, or use of non-culture diagnostic tests. In the second case,
surveillance has a better chance of identifying foodborne agents that are not part
of routine clinical microbiology, but it is
still limited by the epidemiologic and
laboratory resources available to public
health departments.
Published estimates of foodborne disease occurrence highlight the limitations
of our current surveillance system. Of the
76 million cases of foodborne illness estimated to occur each year in the United
Integrated Surveillance
The modern concept of public health
surveillance, first articulated by Alexander Langmuir, views surveillance as a
process (Foege, 1996). As it relates to
food safety, the process is concerned not
only with outcomes in the human population but also with the occurrence of
foodborne hazards in all types of foods,
their sources, and the various stages in
their conversion to consumable food.
Operationally, surveillance involves the
systematic monitoring of disease and
hazard reportsin animal and plant
populations, food production and processing environments, foods and ingredients, and in human populations
through the systematic collection, analysis, and interpretation of outcome-specific data, closely integrated with the
timely dissemination of these data to
those responsible for preventing and
controlling disease or injury (Thacker
and Berkelman, 1988).
Foodborne disease surveillance has
traditionally been viewed as a subset of
public health surveillance. The links between surveillance for foodborne diseases in humans and surveillance for foodborne hazards in foods have only recently received increased attention. Foodborne hazard surveillance monitors the
conditions that can lead to foodborne illnesses (Guzewich et al., 1997). For example, hazard surveillance systems can detect microbial pathogens at various facilities that handle food (e.g., farms, meat
and poultry processors, and restaurants). Hazard surveillance typically involves the collection of data on foodborne hazards in food products and
food sources, follow-up data when hazards are present at unusual levels, and
information that helps define the sources
of hazards in foods.
Animal health surveillance as it relates to food safety is a component of
foodborne hazard surveillance. Comprehensive animal health surveillance systems were nonexistent until the National
Animal Health Monitoring System
(NAHMS) was implemented in 1983
(King, 1990). Current resources limit
these surveillance programs; the NAHMS
on-farm monitoring system does nar-
82
Pathogen identification
New pathogen identification technologies are faster, cheaper, more powerful
and increasingly automated. No single
Genotyping Methods
Genotyping has many advantages
over traditional typing procedures (Olive
and Bean, 1999; Spratt, 1999; Tompkins,
1992; Versalovic et al., 1993). The major
advantage lies in its ability to distinguish
between two closely related strains. Oth83
84
Future Issues
Although genome-based typing
methods are increasingly powerful tools
in molecular epidemiology, several issues
need to be addressed if these methods are
to be incorporated more routinely. Firstly, one should not forget about the advantages of classifying organisms to the
genus and species level, as well as doing
serotyping and phage-typing for some
bacteria, before interpreting banding
patterns resulting from molecular typing. A good example of this is a 2001 salmonellosis outbreak associated with raw
almonds that was detected only because
both serotyping and phage-typing of salmonella isolates were done. As well, in
the absence of a gold standard by
which to judge a typing method (van
Embden et al., 1993), careful standardization of and adherence to laboratory
protocols is essential, if individual methods are to be accepted for classification
of strains. The lack of reproducibility of
certain techniques is another contentious
issue. Consistent reproducibility is essential, if these methods are to be of value in the long-term analysis and categorizing of bacterial strains. Another extremely important issue is in the interpretation of minor (ca. 1 to 3) banding
differences between strains. Some scientists argue that a single difference in the
production of an enzyme or the shift of a
single band on a gel is not enough to say
that two isolates are different, and that
clonality should be considered as a relative concept (Arbeit, 1995). In addition,
strain relatedness should only be judged
in the presence of other data, especially
epidemiologic data.
The ultimate goal is an ideal molecular typing method; one that is easy to
perform, cost-effective, relatively rapid,
amenable to statistical analysis and automation, able to type all possible strains,
reproducible, and properly balanced between increased discriminatory power
and applicability. Rapid advances in typing methods based on whole organism
DNA sequencing are helping us to approach such an ideal method. In the future, the use of molecular typing in foodborne disease investigations will assist us
in identifying the source of many more
outbreaks, will lead to the earlier detection
of outbreaks, and will be beneficial in
identifying and eliminating areas of persistent contamination in food plants.
Resource Priorities
The Centers for Disease Control and
Prevention (CDC) estimates that 76 million cases of foodborne illness occur annually in the United States. A large number of known pathogens are responsible
for 14 million cases; for the other 62 million cases, the pathogen causing the illness is not known. Of the known pathogens, Norwalk-like viruses (9,280,000
cases), Campylobacter species (1,963,000
cases), and nontyphoid Salmonella
(1,332,000 cases) are responsible for
most illness, while Trichinella spiralis,
Vibrio cholerae, and Vibrio vulnificus are
the known pathogens responsible for the
proach to develop critical food safety information regarding the control of pathogens in specific foods. For example, risk
assessments of Salmonella Enteritidis in
eggs, Escherichia coli O157:H7 in ground
beef, V. parahaemolyticus in raw molluscan shellfish, and Listeria monocytogenes
in ready-to-eat (RTE) foods have been
drafted. When sufficient data are available, quantitative risk assessments can:
identify what foods are of greatest risk
and contribute most to specific foodborne
illnesses, estimate the levels of pathogens
in foods that are unsafe, and identify what
points within the food continuum have
the greatest influence on exacerbating or
preventing foodborne illnesses.
Examples of the use of case-control
studies to identify risk factors for sporadic illnesses include E. coli O157:H7,
Campylobacter, and Cryptosporidium.
Major risk factors associated with sporadic cases of E. coli O157:H7 infection
in the United States are eating undercooked ground beef and visiting a farm.
Risk factors for E. coli O157:H7 infection
in Scotland are handling/preparing raw
food (40%), being involved in gardening/garden play (36%), living on or visiting a farm (20%), having direct/indirect
contact with animal manure (17%), having private water supplies (12%), and recent failures with high coliform counts
of water supplies (12%) (Coia et al.,
1998). Risk factors for sporadic Campylobacter infections in the United States,
identified in a case-control study of six
FoodNet sites from January 1998
through March 1999 involving 1,463 patients with Campylobacter infection and
1,317 controls included: foreign travel,
eating undercooked poultry, eating
chicken or turkey cooked outside the
home, eating non-poultry meat cooked
outside the home, eating raw seafood,
drinking raw milk, living on or visiting a
farm, contact with farm animals, and
contact with puppies (Friedman et al.,
2000). Risk factors associated with
cryptosporidiosis cases in Minnesota
from July 1-December 31, 1998, were
swimming in public pools (e.g., hotel or
school pools), drinking well water, visiting a farm, living on a farm for those less
than age 6, and exposure to cattle and to
manure for those not living on a farm
(Soderlund et al., 2000). The underlying
vehicle largely responsible for transmitting these pathogens to humans is contaminated manure.
The vast quantities of manure produced each year as a by-product of ani-
mal agriculture present a challenge. Cattle, hogs, chickens and turkey produced
an estimated 1.37 billion tons of manure in 1997 (U.S. Senate Agriculture
Committee, 1998). Because many of the
most prominent foodborne pathogens
in the United States, including C. jejuni,
Salmonella, and E. coli O157:H7, are
carried by livestock and are principally
transmitted to foods by fecal contamination, the amount of manure created
in the United States is a growing environmental threat.
Manure-related food safety issues
on the near term horizon include issues
related to fresh produce and organic
produce in particular. For example, recent outbreaks of E. coli O157:H7 infection and salmonellosis have been associated with organically produced alfalfa
and clover sprouts and mesclun lettuce.
Use of contaminated cow manure is a
major concern. The lack of an established, proven composting protocol to
assure elimination of pathogens and
prevent recontamination contributes to
this concern. Another issue on the horizon is the importation of fruits and vegetables from countries with poor agricultural practices, i.e., use of contaminated irrigation water, improper preparation and application of manure as fertilizer, and harvesting and washing produce under unsanitary conditions.
Food irradiation has received considerable attention as a means to address food safety issues. The suggestion,
however, that irradiation is a single solution to eliminating most pathogens
associated with fresh or RTE foods lacks
foundation. For some foods, irradiation
results in foods with unacceptable sensory characteristics (Olson, 1998). Irradiation is a tool with broad applicability,
but it is not a comprehensive solution
for all infectious foodborne hazards in
all foods.
Emerging Pathogens
Unfortunately, pathogens can be addressed only after they evolve. Consider,
for example, the relatively recent identification of E. coli O157:H7. E. coli
O157:H7 received relatively little attention from food safety scientists and the
medical community for more than a decade after its discovery. It was not until
1993, following a large outbreak involving more than 700 patients infected by
eating undercooked fast-food hamburgers, that this pathogen rose to promiINSTITUTE OF FOOD TECHNOLOGISTS
Outbreak Investigation
A comprehensive system of foodborne disease surveillance must include
a system for detecting and rapidly responding to potential outbreaks. Outbreaks caused by specific foodborne
pathogens, such as Salmonella and E. coli
O157:H7, may be identified by detecting
unusual case clusters or increased occurrence of cases by routine surveillance of
cases reported by medical clinics and
clinical laboratories. Recently, molecular
subtyping of isolates by pulsed-field gel
electrophoresis (PFGE) has increased
both the sensitivity and specificity of
pathogen-specific surveillance for detecting outbreaks caused by relatively comEXPERT REPORT
mon pathogens, such as Salmonella Typhimurium (Bender et al., 2001). Routine subtyping and transmission of subtype patterns through electronic communication networks, such as PulseNet,
creates the potential to detect widely dispersed outbreaks that might not be recognized in any individual state (Swaminathan et al., 2001). Investigation of
these outbreaks is required to determine
the source and mode of transmission of
the outbreak-associated strain.
In addition to detecting outbreaks
through laboratory-based surveillance,
outbreaks may also be recognized because of the occurrence of similar illnesses among persons who attended an
event or establishment together. Many of
these outbreaks are recognized before a
causative agent has been diagnosed.
Thus, investigation of these outbreaks
must be conducted to identify the agent
as well as the source and mode of transmission.
Outbreak investigations require the
close collaboration of epidemiologists,
environmental health specialists and public health laboratories. Collection of stool
samples and interviews of ill persons and
healthy comparison groups must be conducted rapidly. Epidemiologists must coordinate their activities with the public
health laboratories to maximize the potential to isolate the agent. Information
collected by epidemiologists can help
guide environmental health evaluation of
an establishment and interviews of food
service workers. Results of environmental
health evaluations can further guide epidemiologic investigations. Epidemiologic
data need to be analyzed and interpreted
in light of the results of laboratory tests
and environmental investigations. Strategies for outbreak control and prevention
need to be identified and implemented as
soon as can be justified by the results of
the investigation. Depending on the scope
of the outbreak and nature of the response, coordination with other state and
local agencies, FDA, USDA, and CDC
may be needed.
Most outbreak investigations are initiated by local or state health departments. Because outbreak investigations
are complex activities that need to be
rapid, thorough, and well-coordinated,
CDC issued a report (CDC/NCID/
DBMC, 2000) intended to assist state
and local health departments assess their
outbreak response capacities and to help
guide them in developing and strengthening their foodborne disease surveil-
National Initiatives
Foodborne illness has no easy solutions. However, major strides can be
made by developing and implementing a
well-conceived strategic approach that
prioritizes the hazards and defines the
strategies that will most effectively reduce
hazards. This approach must include a
strategy to address emerging hazards.
This strategic approach should be a national initiative that includes state, local,
and international involvement, and perhaps reorganizes existing federal food
safety agencies and programs.
The importance of an expanded surveillance system that covers animal
health and the environment cannot be
overstated. The additional information
from an expanded and coordinated surveillance system would enable a broader
vision of the flow of pathogens and potential pathogens throughout the food
chain, and it would fill some important
data gaps in risk assessment. Coupled
with the new genetic tools, potential
foodborne pathogens may be detected
before they cause confirmed human illness. While it may be politically or economically impractical to respond vigorously to a likely pathogen before cases
are identified and linked to the food/
pathogen combination, prior knowledge
of the potential pathogen will decrease
response time and enable a more appropriate first response. Expanded surveillance will require state and local participation, coupled with leadership and coordination at the national level. Thus, expanded surveillance should be part of
any national initiative. These activities
also are consistent with international efforts being planned by the World Health
Organization (WHO) towards establishing a coordinated, expanded, worldwide
surveillance system (Archer, 2001).
found to access data from food manufacturers. This is far from simple under
current conditions. From their own
quality assurance (in-line, and environmental) and/or finished product monitoring programs, manufacturers gather
huge amounts of data. If they were available, these data could provide valuable
exposure information to risk assessors
and information on the prevalence of
pathogens in various food processing environments. Food manufacturers currently do not often share such data or
even collect potentially useful information, because of potential regulatory
ramifications or for product liability reasons. Ways must be found to collect and
share this information in a penalty-free
manner.
For example, food producers concerned with L. monocytogenes are hesitant to test below the genus level, such
that data show only Listeria spp. While
somewhat useful, further speciation and
subtyping could yield even more useful
data, but the finding of L. monocytogenes
in a finished, RTE food would result in
regulatory action under current policy of
both FDA and USDA. Knowing how the
presence of other Listeria species in food
or the processing environment relates to
the possible presence of L. monocytogenes
is thus not achievable.
Recent studies question whether all
subspecies of L. monocytogenes are virulent, or of equal virulence (Wiedmann et
al., 1997). Perhaps this finding, when
further developed, will help define more
appropriate policies that foster collection
of good and meaningful data. Additionally, knowing that the presence of L.
monocytogenes alone may not necessarily
mean the food is potentially harmful
may be an incentive to manufacturers to
speciate further, and to apply methods
that determine or indicate virulence or
lack of virulence.
Interdisciplinary Research. A growing
area in federal research funding has been
the formation of interdisciplinary teams
to examine complex problems. This shift
from more traditional projects with a single researcher has reached all the major
federal funding agencies. Some of these
new interdisciplinary programs have been
highly successful, most notably in the areas of vaccine development, epidemiologic surveillance, and genome sequencing.
These different program structures have
created new paradigms for the generation
and sharing of data that provide added
benefit for detecting the emergence of
A Cooperative
Approach to the
Safety of Sprouts
Outbreaks of foodborne illness
associated with the consumption of
raw vegetable sprouts are a recently
emerged food safety issue addressed
using a combination of industry and
regulatory action.
Sprouts can harbor large populations of microorganisms, because the
conditions used for sprouting seeds
also promote rapid microbial growth.
If present on the seeds, pathogens
grow to high levels. In the 1980s and
1990s, consumption of fresh, uncooked vegetable sprouts became
popular, and commercial sprout suppliers developed broad distribution
systems. The number of reported illnesses increased significantly. In
1997-1998 alone, at least 7 documented outbreaks of Salmonella and
E. coli O157:H7 infections were
caused by consumption of various
types of raw vegetable sprouts. One
of these outbreaks, which occurred in
Japan, was the largest outbreak of E.
coli O157:H7 ever recorded (Taormina et al., 1999).
Consumer Behavior
Research indicates that people consider themselves fairly knowledgeable
about food safety guidelines, and for the
most part they are. However, as in other
areas of health and safety, knowledge and
awareness does not always translate into
behavioral changes. Between 1988 and
1993, indicators of concern about food
increased significantly, suggesting an
emerging public awareness and interest
in food safety problems. At the same
time, data suggest that unsafe food consumption and preparation behavior actually increased (Levy, 1997).
Behavioral surveillance systems can
provide data identifying people or
groups in which behaviors associated
with foodborne diseases are more common and who are at higher risk for foodborne illness, thus assisting in the development of food safety education programs (Yang et al., 1998). Further, surveillance data can be used to evaluate the
90
progress of education programs (Altekruse et al., 1999; Yang et al, 1998). Data
collected through the Behavioral Risk
Factor Surveillance Systems during
1995-1996, which included 19,356 survey participants, showed that several high
risk food handling, preparation, and
consumption behaviors were common,
and some varied by gender, age, race/ethnicity, education and income. For example, 50.2% of respondents reported eating undercooked eggs, and 19.7% reported eating undercooked hamburgers. All
high risk food handling, preparation,
and consumption behaviors were more
prevalent in men than in women. The
prevalence of reported consumption of
undercooked hamburgers decreased with
age, increased with education, and increased with income.
Decisions about behavior frequently
are guided by risk perception rather than
risk awareness (Frewer et al., 1994). Defining risk as hazard + outrage, Sandman (1997) stated that when people misperceive hazards it is often because they
are outraged. Sandman noted that generally, even when the hazard is serious,
the public is apathetic and the least dangerous hazard often generates the greatest outrage. He said, Too often, experts
focus on the hazard and ignore the outrage while the public focuses on the outrage and ignores the hazard. Under
these circumstances, the hazard cannot
be mitigated without addressing their
outrage. Sandman suggested that experts
determine why the outrage is high and
what can be done to lower it so that people want to hear or acknowledge the extent of the hazard. As an example, Sandman stated that consumers know how
to cook and generally will get angry if
you tell them how to do something they
already know about.
If people do not recognize and accept
their role in food safety problems, behavior change is unlikely. One way to break
through public misconceptions is to describe the magnitude of food safety
problems and challenge peoples understanding of themselves as experts. New
data from the FoodNet surveillance system may be the best way to challenge
peoples understanding of themselves as
experts (Levy, 1997).
Consumer Education
Recent federal initiatives have sought
to improve the safety of the U.S. food
supply using a farm-to-table approach,
For example, CFSAN developed materials (press kits, consumer brochure, video news release, and public service announcement) explaining the risk that
unpasteurized or untreated juices may
pose to vulnerable populations. The
materials were targeted to a variety of
audiences (senior citizen groups, day
care centers, elementary schools, and
PTA offices). Similarly, FSIS launched a
Thermy the Thermometer campaign
in 2000 to encourage use of thermometers to ensure sufficient cooking of meat
and poultry.
Consumer Trust in Information.
Consumers place different levels of
trust in information from different
sources. A survey of more than 1000
Americans conducted by the University
of Kentucky found that more consumers completely trusted the accuracy of
the information from government publications and food labels than from any
other source (Buzby and Ready, 1996).
Of those respondents who trusted food
safety information from government
publications, 10.8% trusted the accuracy completely. Of those who trusted
food safety information on food packaging and labels, 10.2 percent did so
completely. Consumers complete trust
of store brochures and advertisements
was lower than other sources, 3% and
1.4% respectively. The authors stated
that it is not surprising that these were
the least trusted of the seven sources of
food-safety information, because people
may feel that advertisers have incentives
to make positive claims about their
products.
Addressing the data on how competing motivations, risk perception, and
taste preferences affect hamburger preparation, Ralston et al. (2000) reported
that several information channels appear to be effective for communicating
the risks of unsafe food preparation.
Respondents who said they get their information from magazines, television,
cookbooks, or government hotlines had
15-17% higher risk motivation, i.e.,
were more risk averse, than those who
did not cite these sources of information. Respondents who said that they
get information from labels did not
have a higher risk motivation index after accounting for other factors that also
increase awareness. Consumers who
cited brochures as their information
source had lower risk motivation than
respondents who did not. Ralston et al.
(2000) noted that it is difficult to sepaEXPERT REPORT
rate the effects of different forms of information because consumers are exposed to several information sources simultaneously and information sources
may interact in their effect on perceptions. The authors concluded that the
results show that consumers who are
more aware of risk from undercooked
hamburger are more likely to adopt safe
behavior and thus contribute to a reduction of foodborne disease.
Risk Communication
Risk communication is a necessary
and critical tool to appropriately define
issues and to produce the best risk management decisions (FAO/WHO, 1998).
Risk communication has been defined as
the interactive exchange of information
and opinions concerning risk and risk
management among risk assessors, risk
managers, consumers and other interested parties (CAC, 1997b). Others have
added risk-related factors to the definition, reflecting a wider risk communication concept (FAO/WHO, 1998).
Several factors play a role in understanding and communicating risk:
whether a risk is voluntary or involuntary, whether the distribution of risk
and benefit is equitable, the degree of
personal control, the individual dread
of the adverse event, the catastrophic
potential of the event, and the extent of
trust in the risk managers (Covello et
al., 1988; NRC, 1989; Sandman, 1987).
A Consultation of the Food and Agricultural Organization (FAO) of the
United Nations and WHO (FAO/WHO,
1998) described these principles of risk
communication: (1) know the audience,
(2) involve the scientific experts,
(3) establish expertise in communication, (4) be a credible source of information, (5) share responsibility, (6) differentiate between science and value
judgment, (7) assure transparency, and
(8) put the risk in perspective. Barriers
to effective risk communication can occur within the risk analysis process (e.g.,
inadequate access to vital information
and inadequate participation of interested parties) or in a broader context
(e.g., differing perceptions among participants, limited understanding of the
scientific process, lack of credibility of
the information source, and societal
characteristics). Elements of effective
risk communication include:
the nature of the risk (e.g., its magnitude and severity);
Microbial evolution has always happened and will always happen. Bacteria,
for example, have an enormous capacity
for mutation, integration of new genetic
material, and recombination of genetic
material in order to assure survival. Bacteria can sense and react to their environment and genetically change themselves in response. Unfortunately, newly
evolved pathogens are first recognized
when they cause an outbreak of illness.
Using new technologies and genomics,
perhaps surveillance of food animals
and the environment for newly emerging
microorganisms with pathogenic potential will become a reality in the future,
and there will be no need to wait for human illness. Another hope for the future
is a better understanding of how human
activities affect foodborne pathogens.
For example, does the cross protection
afforded a pathogen by exposure to an
environmental insult have a negative impact on further processing? Genomics
may also provide a better snapshot of
how a microorganism manifests viru-
At-Risk Subpopulations
It is likely that the number of persons
at higher risk for foodborne disease
agents will continue to increase with time.
The population of the United States is aging, and clearly aging is a risk factor for
more serious outcomes from agents such
Pathogen Evolution
Consumer Understanding
Although consumers are only a small
part of the food safety chain, as consumers we all need to take responsibility for
our contribution to food safety. Those
that have not already done so must accept that zero risk is not a reality. These
two concepts may be difficult for some
consumers to accept. Education and risk
communication will be necessary to provide consumers with a more accurate
perception of food safety risks and to encourage behavior modification, where
needed.
Conclusions
History has demonstrated that science,
food safety.
This report articulates the science behind microbiological food safety, especially as it relates to emerging microbiological hazards. Interpretation and analysis of this scientific information pro-
EXPERT REPORT
93
Microbial evolution has always happened and will always happen. Bacteria,
for example, have an enormous capacity
for mutation, integration of new genetic
material, and recombination of genetic
material in order to assure survival. Bacteria can sense and react to their environment and genetically change themselves in response. Unfortunately, newly
evolved pathogens are first recognized
when they cause an outbreak of illness.
Using new technologies and genomics,
perhaps surveillance of food animals
and the environment for newly emerging
microorganisms with pathogenic potential will become a reality in the future,
and there will be no need to wait for human illness. Another hope for the future
is a better understanding of how human
activities affect foodborne pathogens.
For example, does the cross protection
afforded a pathogen by exposure to an
environmental insult have a negative impact on further processing? Genomics
may also provide a better snapshot of
how a microorganism manifests viru-
At-Risk Subpopulations
It is likely that the number of persons
at higher risk for foodborne disease
agents will continue to increase with time.
The population of the United States is aging, and clearly aging is a risk factor for
more serious outcomes from agents such
Pathogen Evolution
Consumer Understanding
Although consumers are only a small
part of the food safety chain, as consumers we all need to take responsibility for
our contribution to food safety. Those
that have not already done so must accept that zero risk is not a reality. These
two concepts may be difficult for some
consumers to accept. Education and risk
communication will be necessary to provide consumers with a more accurate
perception of food safety risks and to encourage behavior modification, where
needed.
Conclusions
History has demonstrated that science,
food safety.
This report articulates the science behind microbiological food safety, especially as it relates to emerging microbiological hazards. Interpretation and analysis of this scientific information pro-
EXPERT REPORT
93
critically important that regulatory policies be based on the best science currently available. Regulatory policies
based on sampling and testing may incorrectly imply an absence of pathogens, causing some individuals to assume that it is unnecessary to engage in
proper food selection and handling
practices. Given the characteristics of
some foods, available technologies, and
our desire for year-round availability of
a diverse array of foods, it is unlikely
that the marketplace can be made free
from the presence of pathogenic microorganisms at all times.
The large-scale production of some
ready-to-eat (RTE) foods consistently
free of Listeria monocytogenes appears
practically impossible. A great deal of
progress has been made during the past
quarter century to reduce the levels and
frequency of contamination of readyto-eat foods during their manufacture,
but consistently assuring the absence of
Listeria has remained out of reach. This
bacterium is commonly present in the
environment and is constantly reintroduced into the processing environment
on raw ingredients and via other means.
Listeria survives well in the manufacturing and retail environment, and it grows
at refrigerator temperatures. At present,
a limit-of-detection standard exists
for L. monocytogenes in an RTE food
such that its mere detection is grounds
for legal action against the company
distributing the food and is the legal basis for removing the food from the marketplace by regulatory agencies. A true
farm-to-table food safety system would
consider downstream handling and
consumption patterns and epidemiologic characteristics of cases; such a system would not destroy foods that are
unlikely to cause illness in the general
population. In addition, some subtypes
of L. monocytogenes found in or on
foods have not been associated with illness, and additional research may demonstrate that some subtypes are not
pathogenic.
Current technologies are also unlikely to consistently satisfy the demand
for large volumes of fresh fruits and
vegetables that are free of harmful microorganisms. Because these raw agricultural commodities are often consumed without cooking, effective interventions are needed that will diminish
or prevent the presence of pathogens.
Policies that result in minimizing contamination and preventing illness from
EXPERT REPORT
out the food. As the amount of contamination in the food decreases, the food
safety emphasis should focus on further controlling processing conditions
through the application of sciencebased HACCP plans.
Because fresh produce undergoes
very little processing, preventing contamination is the primary emphasis for
ensuring the microbiological safety of
fresh fruits and vegetables. Thus, careful consideration must be given to
growing conditionsincluding soil,
water, manure, livestock, wildlife, pets,
environmental pollution and effluent/
sewage, and humansand their effect
on food safety throughout the food
chain.
Many of the most prominent foodborne pathogens in the United States
are carried by livestock and are principally transmitted to food by fecal contamination. Manure, a significant vehicle for pathogens, is a growing source
of fertilizer. Use of manure fertilizer is
an increasing environmental concern
because it may contaminate water for
drinking, irrigation, and recreation.
Manure also is applied with or without
composting to the soil used to grow
food crops. Manure used in the production of food crops is of special concern because the available scientific information is insufficient to ensure that
foodborne pathogens are killed by current agricultural practices. Intensive
farming practices can contribute to the
rapid spread of human and animal
pathogens by creating more concentrated environments for pathogens to
multiply and evolve and by generating
larger quantities of subsequently contaminated food.
An examination of the science reveals that foodborne illness is caused
by a complex combination of factors
that must be managed on a continual
basis. To achieve our public health
goals, everyone along the farm-to-table
continuum must take responsibility for
their role in food safety management.
Foodborne disease is widely recognized for acute effects on the gastrointestinal tract but also includes
other effects throughout the body. In
addition, foodborne pathogens may
cause chronic disease, which may occur
independently or accompany an acute
illness. Many of these chronic diseases
have only recently been linked to foodborne pathogens. In addition, a growing proportion of foodborne illnesses
96
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