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Instructor's Manual for Jay's

Modem Food Microbiology


Sixth Edition

Peter S. Murano, PhD


Assistant Professor
Department of Animal Science
Texas A & M University
College Station, Texas

Elsa A. Murano, PhD


Associate Professor
Department of Animal Science
Texas A & M University
College Station, Texas

AN ASPEN PUBLICATION
Aspen Publishers, Inc.
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2000
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T ABLE OF CONTENTS

Introduction ........................................................................... v

Chapter 1 - History of Microorganisms in Food .............................................. 1

Chapter 2 - Taxonomy, Role, and Significance of Microorganisms in Foods. . . . . . . . . . . . . . . . . . . . . . .. 3

Chapter 3 - Intrinsic and Extrinsic Parameters of Foods That Affect Microbial Growth . . . . . . . . . . . .. 6

Chapter 4 - Fresh Meats and Poultry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 8

Chapter 5 - Processed Meats ............................................................. 11

Chapter 6 - Seafoods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 13

Chapter 7 - Fermentation and Fermented Dairy Products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 15

Chapter 8 - Fruit and Vegetable Products: Whole, Fresh-Cut, and Fermented ................... 18

Chapter 9 - Miscellaneous Food Products .................................................. 21

Chapter 10-Culture, Microscopic, and Sampling Methods .................................... 24

Chapter ll-Physical, Chemical, Molecular, and Immunological Methods ....................... 27

Chapter 12-Bioassay and Related Methods .................................................. 31

Chapter 13-Food Preservation with Chemicals. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 33

Chapter 14-Food Preservation with Modified Atmospheres ................................... 36

Chapter 1S-Radiation Preservation of Foods and Nature of Microbial Radiation Resistance ......... 38

Chapter 16--Low-Temperature Food Preservation and Characteristics of


Psychrotrophic Microorganisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 41

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Chapter 17-High-Temperature Food Preservation and Characteristics of
Thermophilic Microorganisms .................................................. 43

Chapter 18-Preservation of Foods by Drying. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 47

Chapter 19-0ther Food Preservation Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 50

Chapter 20-Indicators of Food Microbial Quality and Safety. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 52

Chapter 21-The HACCP System and Food Safety .......................................... 55

Chapter 22-Introduction to Foodborne Pathogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 58

Chapter 23-Staphylococcal Gastroenteritis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 62

Chapter 24-Food Poisoning Caused by Gram-positive Sporeforming Bacteria. . . . . . . . . . . . . . . . . .. 65

Chapter 25-Foodborne Listeriosis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 68

Chapter 26-Foodborne Gastroenteritis Caused by Salmonella and Shigella. . . . . . . . . . 72

Chapter 27-Foodborne Gastroenteritis Caused by Escherichia coli . ................... 74

Chapter 28-Foodborne Gastroenteritis Caused by Vibrio, Yersinia, and Campylobacter Species ...... 77

Chapter 29-Foodborne Animal Parasites. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 80


,
Chapter 30-Mycotoxins. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 84

Chapter 31-Viruses and Some Other Proven and Suspected Foodborne Biohazards ............... 87

iv INSTRUCTOR'S MANuAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
Introduction

This Manual is intended to be used as companion terms and definitions provided can help clarify con-
material to the textbook Modern Food Microbiology, cepts covered in the textbook. This section can also
Sixth Edition, by James Jay. Each chapter ofthe Manual serve as a source of questions for quizzes or exams. It
corresponds directly to the chapters in the textbook, can also serve as review material to be used in summary
and is divided into the following sections: discussions.
Finally, the two sections on questions are designed to
1. Learning Objectives. test student-learning ofthe fundamental concepts from
2. Chapter Outline. each chapter. The success of accomplishing the learn-
3. Terms and Definitions. ing objectives is directly related to student performance
4. Basic Knowledge Questions. in answering these questions. In particular, the Basic
5. Critical Thinking Questions. Knowledge Questions section is designed to review
the material covered in the chapter, and serve as a good
The purpose of the Learning Objectives section is indicator of a type of learning known as "lower-level
to help the instructor design the lecture material in a cognition." The Critical Thinking Questions section
way that will ensure a successful outcome, from the is designed to challenge critical thinking skills by pro-
perspective of both the instructor and the student. There viding students with questions that require a more
are two purposes for the Chapter Outline. One is to complex type of learning known as "higher-level cog-
help the instructor organize the material for the lecture, nition." In some instances there is no "correct" answer
aiding in the decision-making regarding which chap- to these questions, but the student is encouraged to
ters and/or sections to include or omit. The second is to develop a reasonable and defensible answer or hypoth-
serve as a summary of the material that is included in esis to explain phenomena for which no definitive
the textbook, enabling the instructor to review it at a answer has been provided by current research. The
glance. instructor is encouraged to use the questions provided
The purpose of the Terms and Definitions section in both of these sections in quizzes or exams, or to
of each chapter is to serve as supplementary material, utilize them to launch discussions on specific topic
providing definitions ofterms that are either only briefly areas in more advanced lecture settings, such as in
defined, or not defined at all in the textbook. Thus, the honors-level or graduate-level courses.

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY v


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 1

History of Microorganisms in Foods

Learning Objectives Terms and Definitions

The reader will be able to: Aflatoxins: Toxins produced by the mold ge-
1. Discuss the events of ancient history related to nus Aspergillus.
the presence and use of microorganisms in food BSE: Condition of cattle, also known as
products. "mad cow disease", which mani-
2. Describe the contributions of notable microbi- fests as tremors and lack of coordi-
ologists to our understanding of the role of mi- nation in the animal, resulting in
croorganisms in food spoilage and food produc- death due to brain damage.
tion. Botulism: Disease caused by ingestion oftoxin
3. List major historical events related to food law, produced by the bacterium
food processing accomplishments, and outbreaks Clostridium botulinum, often re-
offoodbome illness that have shaped the science sulting in death by paralysis of in-
of food microbiology in the last 50 years. voluntary muscles.
Brine: Solution of water and salt, used to
pack foods into cans to promote
Chapter Outline even heat distribution during heat
processmg.
1. Historical account of food microbiology Cyclosporiasis: Condition caused by infection with
a) Food preservation methods in ancient times the parasite Cyclospora cay-
b) The role of microorganisms in food poison- etanensis, usually from consump-
ing and food spoilage tion of contaminated fresh foods
c) Modem food processing such as fruits and vegetables, or
2. Significant dates and events water.
a) Food preservation Ergot: Type of food poisoning caused by
b) Food spoilage ingestion of toxin produced by the
c) Food poisoning mold Claviceps purpurea when
d) Food legislation growing on wheat.
Gastroenteritis: Condition marked by inflamma-
tion of the intestinal tract, usually

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
as a result of infection by micro- changing the flavor and extending
bial pathogens. the shelf-life of the products.
Giardiasis: Condition caused by infection with Spontaneous
the protozoan parasite Giardia Generation: The sudden appearance of living
lamblia. organisms or biological contami-
Hemorragic nants from no apparent source, a
Colitis: Condition caused by infection with rejected theory once used by scien-
verotoxin-producing strains of Es- tists to explain how diseases and
cherichia coli, most notably sero- the spoilage of food occurs.
type 0157:H7, manifesting itself
as profusely bloody diarrhea. Basic Knowledge Questions
Irradiation: Processing treatment based on ex-
posure offoods to ionizing sources 1. What experiment did Spallanzani conduct to dis-
of radiation, such as gamma rays, prove the doctrine of spontaneous generation?
accelerated electrons, or x-rays, for 2. What processing treatment did N. Appert in-
the purpose of reducing or elimi- vent?
nating microbial, parasitic, and in- 3. What two contributions did Pasteur make to the
sect contaminants. field of microbiology?
Listeriosis: Condition caused by infection with 4. Who commercialized the freezing offoods in the
the bacterium Listeria mono- U.S.?
cytogenes, which manifests itself 5. What important food law was enacted in 1906 in
as flu-like symptoms in normal, the U.S.?
healthy adults, but develops into 6. Name three microorganisms known to be re-
complications such as stillbirths, sponsible for outbreaks offoodbome illness only
meningitis, and encephalitis in in the last 20 years.
people with depressed immune sys- 7. What is the Talmadge-Aiken Act?
tems.
Smoking Critical Thinking Questions
(e.g. meats): A processing treatment whereby
foods are exposed to smoke pro- 1. Design an experiment that would show that ozone
duced by burning of wood or other application to foods can kill spoilage bacteria.
fuels, or by addition of extracted 2. How would our lives be different if we did not
smoke products, for the purpose of have canning as a food preservation method?

2 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 2

Taxonomy, Role, and Significance


of Microorganisms in Foods

Learning Objectives d) Gastrointestinal tract


e) Food handlers
The reader will be able to: f) Animal feeds
1. Discuss the various molecular methods used to g) Animal hides
classify bacteria. h) Air and dust
2. List and describe the environments where micro- 4. Synopsis of common foodbome bacteria.
organisms that contaminate food originate. 5. Synopsis of common genera of foodbome molds.
3. List and describe several types or groups of a) Basic mold physiology
foodbome bacteria. b) Common genera found in foods
4. Describe the general physiology of molds. 6. Synopsis of common genera of foodbome yeasts.
5. List and describe several types of foodbome a) Basic yeast physiology
molds. b) Common genera found in foods
6. Describe the general physiology of yeasts.
7. List and describe several types of foodbome
yeasts. Terms and Definitions
8. List ways in which bacteria, yeasts, and molds
differ from each other. Aerobe: An organism that requires oxygen
for optimum growth. If a "strict
Chapter Outline aerobe", presence of oxygen is re-
quired for growth, e.g. Bacillus
1.Introduction to the presence and purpose of mi- species.
croorganisms. Anaerobe: An organism that requires the ab-
2. Bacterial taxonomy. sence of oxygen for optimum
a) rRNA analyses growth. If a "strict anaerobe", ab-
b) Analysis of DNA sence of oxygen is required for
3. Primary sources of microorganisms in foods. growth, e.g. Clostridium species.
a) Soil and water Ascospore: Sexual spores formed within an
b) Plants and plant products enclosed sac, or ascus, of certain
c) Food utensils molds.

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Copyright 2000, Aspen Publishers, Inc.
Cleistothecia: Carp in which ascospores develop Mesophile: An organism that grows best at
within a closed envelope in certain temperatures in the range of 20 to
molds. 40C.
Conidia: Asexual spores produced by myce- Mesotroph: An organism that can grow at tem-
lium of certain molds, used in re- peratures between 20 and 40e.
production. Conidia are resistant Microaerophile: An organism that grows best in a
to UV radiation and to dry condi- low-oxygen atmosphere. Often, the
tions. term is used to refer to organisms
Columella: The neck or tapering base of the that cannot tolerate atmospheric
sporangium of molds. levels of oxygen.
DNA-DNA Microbiota: Microscopic environment, includ-
Hybridization: The joining (annealing) oftwo single ing all of its resident organisms.
strands of DNA into one double MoIG+C: The percent of guanine and cy-
strand, guided by the pairing of to sine bases that contribute to the
bases from one strand to the com- total base content in a DNA mol-
patible bases in the other strand ecule. The DNA of most gram-
(adenine with thymine, and cytosine negative bacteria has a G + C con-
with guanine). If two organisms tent> 55%, while that of most
are related, their base sequences gram-positives has a G + C content
will be very similar, and thus DNA <50%.
from one will anneal to that of the Mycelium: Tangled mass of filaments, or hy-
other. phae, formed by the growth of some
Genus: Classification level of organisms, foodbome molds, visible to the
more specific than "family", but naked eye.
less than "species", based on broad Psychrotroph: An organism that can grow at or
physical, biochemical, genetic and! below 4C and also at tempera-
or other characteristics (plural is tures below 20e.
"genera"). Reverse
Gram- Transcriptase: Enzyme that catalyzes the synthe-
negative: Bacteria that stain red as a result of sis of DNA from RNA. It is used to
gram-staining, due to their thin cell build genetic libraries of DNA
wall structure. molecules from the rRNA of dif-
Gram- ferent organisms, with which stud-
positive: Bacteria that stain blue/purple as a ies can be carried out to determine
result of gram-staining, due to their degree of relatedness.
their thick cell wall structure. Rhizoid: Root-like filament that functions
Hetero- as absorption organs of filamen-
fermentative: An organism that generates vari- tous fungi.
ous organic compounds as a result Ribosomes: Cellular structures, composed of
of carrying out fermentation reac- proteinandrRNA (ribosomal RNA).
tions. RNA: Ribonucleic acid, molecule found
Hyphae: Filamentous structures that make in living organisms, responsible for
up the mycelia of molds. translating the encoded messages
Koji: Cooked rice inoculated with As- in genetic material into proteins.
pergillus oryzae, A. soyae, or A. Sanger method: Biochemically-based method used
japonicus, and allowed to ferment to determine the order, or sequence,
to produce soy sauce. of bases (adenine, cytosine, thymi-

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Copyright 2000, Aspen Publishers, Inc.
dine, and guanine) present in a piece 2. Describe three sources of contamination of foods
of DNA. of animal origin.
Species: Classification of organisms within 3. What are some of the new genera into which
a specific genus, based on unique organisms formerly classified as Micrococcus
physical, biochemical, genetic and! have been placed?
or other characteristics. 4. Describe the general structure of molds.
Stolon: The prolonged extension ofthe body 5. What is a teleomorph?
wall of some molds, from which 6. Name two genera of molds that are commonly
buds develop. responsible for spoilage of fruits.
Taxonomy: Classification of living things ac- 7. Name and describe the genus of yeast used in the
cording to specific characteristics production of breads and beer.
into distinct groupings.
Thermophile: An organism that generally requires
temperatures of 55C or above for
Critical Thinking Questions
growth.
Xerophile: An organism that grows best in
low-humidity conditions. 1. Describe the procedure you would follow to
determine the source of contamination of a par-
Basic Knowledge Questions ticular food.
2. Should molecular-based methods be used as the
1. Why is rRNA used in taxonomical classification definitive way to classify microorganisms? Why
of microorganisms? or why not?

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Copyright 2000, Aspen Publishers, Inc.
CHAPTER 3

Intrinsic and Extrinsic Parameters of


Foods That Affect Microbial Growth

Learning Objectives t) Biological structures


2. Extrinsic Parameters
The reader will be able to: a) Temperature
1. Define pH, water activity, and OIR potential. b) Relative humidity
Describe how these and other intrinsic param- c) Concentration of gases
eters affect microbial growth. d) Presence of other microorganisms
2. Describe how various extrinsic parameters af- 1. General microbial interference
fect the ability of microorganisms to grow in 2. Lactic antagonism
foods. 3. Combined Intrinsic Extrinsic Parameters: The
3. Describe the concept of microbial interference Hurdle Concept
and antagonism between microorganisms.
4. Explain the hurdle concept and describe examples Terms and Definitions
of how it can be applied to inhibit and/or elimi-
nate microorganisms in foods. Adsorption: The process of acquiring moisture
by a system, typically occurring as
Chapter Outline a result of an excess of moisture in
the environment surrounding that
1. Intrinsic parameters. system.
a) pH Arrhenius
1. General facts Equation: Mathematical equation that ex-
2. Effects on microorganisms plains the rate of reactions, such as
b) Moisture content the chemical reaction between an
1. Water activity Caw) acid and a base to form a salt and
2. Effects of low ~ water. In this equation, the rate
c) Oxidation-reduction potential constant, k, is directly and linearly
1. General facts related to the temperature, T, of the
2. Eh effects components.
d) Nutrient content Biological
e) Antimicrobial constituents Acidity: Acidity of a food, caused by the
1. General facts action ofmicroorganisms, typically
2. Lactoperoxidase system through fermentation reactions.

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Copyright 2000, Aspen Publishers, Inc.
~-galactosidase: Enzyme that catalyzes the break- Poising
down of galactose by microbial Capacity: The tendency of a system to resist
cells. changes in O/R potential.
Buffer: A solution or food component that, Shelf-life: Period of time required for a food
because of its biochemical struc- to become spoiled and thus orga-
ture, resists changes in the pH of its noleptically undesirable.
surroundings. Water Activity: The amount of available water in a
Catabolism: Process of breaking down bio- medium for microbial growth. It is
chemical compounds into usable also defined as the ratio of vapor
molecules by a biological system. pressure of a food to the vapor
Desorption: The process oflosing moisture by a pressure of pure water.
system, typically occurring as a
result of a deficit of moisture in the
environment surrounding that sys- Basic Knowledge Questions
tem.
Generation 1. Explain how weak acids (i.e. organic acids) af-
Time: Time required for cell division to fect bacterial cells.
take place. A generation time of20 2. Provide the mathematical definition of water
min. refers to the time it takes for activity (3w), and explain how having a low aw in
one cell to divide, yielding two the medium inhibits cell multiplication.
cells. 3. Name four factors that determine the OIR poten-
Lag Phase: Phase of bacterial growth in which tial of a food system.
synthesis of macromolecules is 4. Name five types of nutrients required by micro-
being carried out, such as for re- organisms.
pairing of damaged structures, and 5. How does the lactoperoxidase system inhibit the
for acclimatization to new envi- growth of microorganisms?
ronmental conditions. In this phase, 6. Name four extrinsic parameters that affect mi-
cell division does not take place. crobial growth.
Osmotic Stress: Condition in which a microorgan- 7. Define "psychrotroph" and "thermophile."
ism is exposed to a high concentra- 8. Define "bacterial interference" and give two ex-
tion of solutes in the surrounding amples of the types of interference that can be
environment, causing diffusion of observed between two organisms.
water from the inside to the outside 9. Give an example of how the hurdle technology
of the cell by osmosis, in order to can be applied to prevent the growth of microor-
equalize the pressure on both sides. ganisms.
Oxidation-
Reduction
Potential: The ease with which a particular
Critical Thinking Questions
substrate gains or loses electrons.
A substrate that loses electrons (be-
comes oxidized), and causes other 1. A new microorganism has been discovered, and
substrates to gain electrons (be- it has been reported that it is able to grow only in
come reduced). A system with a the presence of bacteria used in the fermentation
high OIR potential lacks electrons, of milk. Provide a possible scenario or mecha-
thus it is oxidized. Conversely, a nism to explain this phenomenon.
system with a low O/R potential 2. What is the relationship between OIR potential
has an oversupply ofelectrons, thus and the oxygen requirements of microorganisms
it is reduced. (i.e. aerobes, anaerobes, etc.)?

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 7


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 4

Fresh Meats and Poultry

Learning Objectives 4. Incidence/prevalence of microorganisms in fresh


red meats.
The reader will be able to: a) Traditional ground meats
1. List the sources of contamination of fresh meats. b) Soy-extended ground meats
2. List the steps involved in the slaughter of food c) Mechanically deboned meat, poultry, and fish
animals. d) Hot-boned meats
3. Explain the reasons why ground meats are more e) Effect of electrical stimulation
highly contaminated than whole cuts. f) Organ and variety meats
4. Describe the differences between various types 5. Microbial spoilage of fresh red meats.
of meats (i.e. hot-boned vs. cold-boned meats, a) General considerations
variety meats, etc.) in terms of numbers and b) Mechanism of spoilage
types of microorganisms present. 6. Spoilage of fresh livers.
5. Describe the types of organisms responsible for 7. Incidence/prevalence of microorganisms in fresh
spoilage of fresh red meats. poultry.
6. List examples of methods that can be used to 8. Carcass sanitizing/washing.
detect microbial spoilage of meats.
7. Describe the events that take place during spoil-
age of liver. Terms and Definitions
8. Describe the spoilage of whole, cut-up, and com-
minuted poultry. Actin: Protein found in thin filaments of
9. List and describe carcass washing and sanitation muscle tissue cells. It binds to an-
procedures employed by the industry to decrease other protein, myosin, resulting in
microbial contaminants. muscle contraction.
Bone Taint: Spoilage of fresh meat near the
bone, typically caused by
Chapter Outline Clostridium and Enterococcus spe-
cies.
1. Sources of contamination Cathepsins: Enzymes that break down proteins
2. Biochemical events that lead to rigor mortis. in tissue cells, resulting in the re-
3. The biota of meats and poultry. lease of amino acids.

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Coliform: A gram-negative, non-spore-form- Putrefactive
ing rod-shaped bacterium, able to Anaerobe: Literally, an organism that breaks
ferment lactose with gas produc- down protein and grows optimally
tion within 48 hrs. at 35C. in the absence of oxygen. An ex-
DFD: Dark, firm, and dry meat, it typi- ample is Clostridium sporogenes,
cally has a pH higher than that of a bacterium that is used often as an
normal meat tissue, causing it to indicator of commercial sterility.
spoil faster. Reticulo-
Fecal endothelial
Coliform: A gram-negative, non-spore-form- System: Portion of the immune system of
ing rod-shaped bacterium, able to humans and animals that is respon-
ferment lactose with gas produc- sible for the production of phago-
tion within 48 hrs. at 44.5C. Since cytic cells (i.e. macrophages, poly-
most microorganisms that are shed morphonuclear leukocytes, etc.).
in the feces of humans and animals These cells scavenge the body
share these characteristics, fecal through the circulatory and lym-
coliforms are used as indicators of phatic systems in order to remove
the presence offecal contamination invading particulates, including
. .
in foods. mlcroorgamsms.
Lymph Nodes: Organs that form part of a network Rigor mortis: Literally "stiffness of death,"
which filters foreign bodies from caused by depletion of ATP, which
the tissue fluid and lymph, collect- results in tension by the interaction
ing microbial and other particulate of actin and myosin in muscle tis-
contaminants. Cells of the immune sue.
system called lymphocytes reside Rumen: The first of four chambers, or
in the nodes, and playa role in di- stomachs, of a ruminant animal
gestion and/or inactivation of in- (i.e. cattle). Upon consumption of
vading organisms. plants by the animal, the unchewed
MPN: Most probable number method, material first passes into the rumen.
used to estimate the number of cells It is then regurgitated and masti-
of a particular class of organisms in cated with saliva in the mouth. The
a food sample, when that number is material is then re-swallowed. It
expected to be too low to be detect- passes through the rumen, as well
able by direct plating methods. as two other chambers (the reticu-
Myofibril: Structure inside a multinucleated lum and the omasum) on its way to
skeletal muscle cell (fiber), com- the third chamber (the abomasum),
posed of thick and thin filaments. where it is digested.
Myosin: Protein found in the thick filaments Sarcoplasm: Fluid surrounding contractile myo-
of myofibrils. It binds to another fibrils in a muscle cell. The term
protein, actin, and catalyzes the "sarcoplasmic fraction" refers to
breakdown of ATP, which drives water-soluble proteins, obtained by
muscle contraction. extraction of the sarcoplasmic
PCR: Polymerase chain reaction, is a fluid.
molecular biology-based method Serovar: Classification within a microbial
used to increase the amount of species, based on antigenic differ-
DNA of a particular organism in a ences (i.e. type of antibodies that
sample, thereby increasing its de- are produced as a result of expo-
tectability by subsequent assays. sure by a human or animal to that

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Copyright 2000, Aspen Publishers, Inc.
particular organism). For example, b) physical methods
Escherichia coli 0157:H7 andEs- c) bacteriological methods
cherichia coli 0104:NM are two d) physicochemical methods
serovars (also termed "serotypes") 4. Describe how fresh meat becomes spoiled, both
of the same organism. in the presence and absence of oxygen. Include
Visceral the names of the types of organisms responsible.
Taint: Microbial spoilage of carcasses in 5. What are the main differences between spoilage
New York-dressed poultry, due to of fresh meats vs. organ (variety) meats?
invasion of inner tissues by micro- 6. Name two products of putrefaction, and describe
organisms in the intestinal tract of the chemical reactions by which they are formed.
the animal. 7. What are some of the decontamination methods
used to remove microorganisms from animal
carcasses?
Basic Knowledge Questions
Critical Thinking Questions
1. Describe how ground meat products become
contaminated. 1. Compare and contrast the role of molds vs. bac-
2. List several types of bacteria and molds that are teria in spoilage of fresh meat and poultry.
responsible for spoilage of fresh meat and poul- 2. Rank the following products according to how
try. quickly you would expect them to spoil, and
3. Name at least three methods that can be used for justify your reasoning:
detecting spoilage of fresh meats for each of the a) chicken cuts (with skin)
following categories: b) beef cuts
a) chemical methods c) vacuum-packaged ground beef

10 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 5

Processed Meats

Learning Objectives Terms and Definitions

The reader will be able to: Choleglobin: Green compound produced by oxi-
1. Define the curing and smoking processes used in dation of myoglobin or oxymyoglo-
the meat industry. bin in meats, often as a result of
2. Describe the biota of cured meats in terms of microbial spoilage.
microbial composition and typical level of each Dry Sausage: Sausage product containing 30-
contaminant. 40% moisture, not smoked or sub-
3. Describe the types of spoilage that occur in cured jected to heat-processing.
and smoked products. Greening: Condition of spoiled cured meat,
4. Describe the various types of fermented meat caused by (1) oxidization of por-
products. phyrin molecule in curing agents
5. Discuss some of the safety concerns associated as a result of reaction with hydro-
with fermented meat products. gen peroxide, the latter being pro-
duced by certain microorganisms,
or by (2) production of hydrogen
sulfide by microorganisms, which
Chapter Outline
reacts with myoglobin to form the
green-colored sulphmyoglobin.
1. Common processing treatments. Hetero-
a) Curing fermenters: Microorganisms that ferment sug-
b) Smoking ars to produce various compounds,
2. Sausage, bacon, bologna, and related products. such as organic acids, diacetyl, car-
a) Processing methods bon dioxide, etc.
b) Spoilage Homo-
3. Bacon and cured ham. fermenters: Microorganisms that ferment sug-
4. Fermented meat products. ars to produce one compound, typi-
a) Processing methods cally lactic acid or other organic
b) Product safety acid.

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Nitrite: Curing agent used in processing Basic Knowledge Questions
some meat products, for the pur-
pose of fixing tissue pigment, re- I. Describe the purpose for the following ingredi-
tain moisture, and prevent germi- ents in curing meats:
nation of Clostridium botulinum a) nitrite
spores. b) sodium ascorbate
Offal: Parts of an animal that are removed c) sugar
during dressing, especially but not d) phosphates
limited to, digestive tract, lungs, 2. Name at least four reasons why meats are smoked.
blood, feet, and head. 3. Define the following terms related to spoilage,
Semidry and give an example of a microorganism that
Sausage: Sausage products containing 50% may cause it:
moisture, heat-treated to achieve a) sliminess
60-68C internal temperature, and b) souring
smoked. c) greening
Sliminess: Condition of spoiled cured meat, 4. Compare the spoilage that takes place in cured
caused by production oflong-chain hams vs. smoked hams.
carbohydrates by certain foodborne 5. Compare dry and semi-dry sausages in terms of
bacteria and yeasts, which impart processing treatments and types of organisms
a slimy texture to the product. used to produce them.
Souring: Condition of spoiled cured meat, 6. Name the types of molds used in production of
caused by production of organic processed meats.
acids through the fermentation of 7. What are some concerns associated with fer-
lactose by some bacteria, such as mented meat products and the organism Escheri-
lactobacilli and enterococci. chia coli 0157:H7?
Sours: Condition of spoiled cured hams,
caused by microbial fermentation Critical Thinking Question
of sugars pumped into the product.
Trichinosis: Health condition caused by infec- 1. You are asked to develop a new meat product
tion with larvae of the worm Tri- that is shelf-stable, yet contains no nitrite. De-
chinella spiralis, typically found in scribe the process and ingredients you would use,
untreated raw pork. and justify your choices.

12 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 6

Seafoods

Learning Objectives 4. Spoilage of fish and shellfish.


a) Fish
The reader will be able to: b) Shellfish
1. Define what is meant by "seafoods." 1) Crustaceans
2. Describe the sources of contamination of fresh 2) Mollusks
seafood.
3. Name some of the organisms that make up the
micro flora of fresh seafood. Terms and Definitions
4. Describe the processes involved in production of
several fermented fish products. Alpha-hemolysis: Type of hemolysis observed on
5. Describe the changes that occur during spoilage blood agar plates as a result ofmi-
of fresh fish, including decarboxylation of spe- crobial enzymatic action on red
cific amino acids by microorganisms. blood cells in the agar. It manifests
6. Discuss the types of spoilage flora that exist in itself by formation of a cloudy
crustaceans and mollusks, and the changes that green-colored zone of hemolysis
take place as a result. surrounding the colonies of spe-
cific types of microorganisms that
may be growing on the surface of
Chapter Outline the agar. It is considered a mild re-
action, when compared with beta-
1. Definition of seafood. hemolysis of red blood cells (ex-
2. Microbiological quality of various fresh and fro- emplified by formation of a com-
zen products. pletely clear zone around the colo-
a) Typical organisms making up the seafood nies).
micro flora Blanching: Processing treatment designed to
b) Levels of various types of microorganisms heat foods at temperatures near
found in seafood boiling for a few seconds. It is con-
3. Fermented fish products. sidered a mild treatment yet it can
a) Fish sauces serve to partially decontaminate the
b) Fish pastes surface of some products, as well

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Copyright 2000, Aspen Publishers, Inc.
as inactivate product enzymes. 2. How are fermented fishery products made? Give
Halophilic: Microorganisms that grow opti- some examples.
mally in the presence of high salt 3. Name the types of microorganisms responsible
concentrations. for spoilage of fresh fish.
Histamine: Volatile compound produced as a 4. What are the components of slime?
result of the break down of the 5. What are the chemical changes that take place
amino acid histidine in fish tissue during spoilage of fresh fish?
by the enzyme histidine decarboxy- 6. Defme the following terms:
lase, which is found in certain mi- a) TVB
croorganisms. Production ofhista- b) TVA
mine is associated with microbial c) TVS
spoilage of these products, as well d) TVN
as scombroid food poisoning. 7. What is the pH scale on which spoilage of oysters
Shucking: Process used to remove oysters is based?
from their shell.
TMAO: Trimethylamine-N-oxide, com-
pound found in fresh fish which is
Critical Thinking Questions
reduced to trimethylamine (TMA)
during microbial spoilage.
Tyramine: Produced from decarboxylation of 1. Describe the shifts that occur regarding the preva-
the amino acid tyrosine found in lence of specific populations of microorganisms
fish tissue, and is thus associated during storage of fresh fish. Speculate on the
with spoilage of fish products. reasons why these shifts occur.
2. What are the physiological differences between
Basic Knowledge Questions crustaceans and mollusks that would affect the
types of microorganisms that would be respon-
1. What are some genera of microorganisms typi- sible for spoilage of each.
cally found in fresh seafood?

14 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 7

Fermentation and Fermented


Dairy Products

Learning Objectives Chapter Outline

The reader will be able to: 1. Fermentation.


1. Define "fermentation." a) Background
2. List the genera of organisms encompassed in the b) Definitions and characteristics of fermenta-
"lactic acid bacteria" group. tion
3. Provide an overview of biochemical pathways c) The lactic acid bacteria
involved in fermentation of glucose by microor- d) Metabolic pathways and molar growth yields
ganisms. 2. Dairy products.
4. Defme "homofermentative" vs. "heterofermenta- a) Milk biota
tive" organisms, and give examples of each. b) Starter cultures, products
5. Describe how microbial growth is measured. c) Cheeses
6. Describe the types of organisms found in raw 3. Apparent health benefits of fermented milks.
mille a) General
7. Define "pasteurization" treatments. b) Lactose intolerance
8. Describe microbial spoilage of milk. c) Cholesterol
9. Discuss the types of starter cultures used in dairy d) Anticancer effects
fermentation and provide examples offermented e) Probiotics
dairy products. 4. Diseases caused by lactic acid bacteria.
10. Describe the steps involved in fermentation of
milk to produce cheese. Terms and Definitions
11. Define "lactose intolerance."
12. Discuss the cholesterol-lowering and anticancer Carcinoma: Malignant growth originating in
effects of fermented products. epithelial tissue.
13. Define "probiotics." Cloning Vector: Any small genetic element that can
14. Discuss the disease-causing ability oflactic acid be used to carry other genes, help-
bacteria. ing to replicate them inside a host.

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Cloning vectors are usually de- yield of cells according to some me-
signed to allow integration of for- dium component that is either used
eign DNA for the purpose of trans- or formed as a result of cell growth.
porting that DNA into whatever Mucopeptide: A class of glycoproteins in which
organism the vector is to be placed. peptides are complexed with an
Crohn's acid, such as muramic acid, as oc-
Disease: Inflammatory bowel disease affect- curs in the cell wall structure of
ing humans, and suspected of be- most bacteria.
ing caused by species of Mycobac- Peptidoglycan: Polymer found in the cell wall of
terium, the agent responsible for a most bacteria, made up of N-
similar condition in cattle and acetylglucosamine and muramic
sheep known as Johne's Disease. acid molecules held together by
Curd: Precipitated casein protein, as a amino acid bridges.
result ofa lowering of the pH dur- Vector Plasmid: Small, circular, double-stranded
ing the fermentation of milk by lac- DNA that serves as a vehicle to in-
tic acid bacteria. sert specific sequences, or genes,
Hetero- into a particular organism. In the
fermentative: Refers to the fermentation that case of starter cultures, a vector
some organisms undergo, in which plasmid is constructed with a gene
glucose is metabolized by a series that encodes for the production of
of reactions, one of which includes specific compounds. This vector
the enzyme phosphoketolase, to plasmid is then introduced into the
produce lactic acid, ethanol, and particular microorganism that is to
other compounds such as acetic carry out the fermentation reac-
acid, and carbon dioxide. tions.
Homo- Probiotic: A food that contains a microorgan-
fermentative: Refers to the fermentation that ism, or consortium of micro organ-
some organisms undergo, in which isms, ingestion of which is thought
glucose is metabolized by a series to promote health and well-being
of reactions, one of which includes of the consumer.
the enzyme hexose isomerase, to Sarcoma: Tumor of fleshy consistency, a
produce lactic acid. malignant growth derived from
Johne's non-epithelial tissue (i.e. connec-
Disease: Inflammatory bowel disease of tive tissue, lymphoid tissue, carti-
sheep and cattle, caused by infec- lage, etc.).
tion with Mycobacterium avium Starter Culture: A culture of various types of mi-
subspecies paratuberculosis. Simi- croorganisms, typically lactic acid
lar to Crohn's Disease in humans, bacteria, which is used for fermen-
for which a causative agent has not tation of milk and other commodi-
been identified. ties for the production of fermented
Molar Yield foods. These are often genetically-
Constant: Can be defined as grams of cells modified in order to promote pro-
formed per mole of nutrient con- duction of specific compounds
sumed (Y x/m)' or grams of cells during fermentation, and to mini-
formed per mole of oxygen con- mize variations in production
sumed (Y x/O2 )' or grams of cells yields.
formed per moles of ATP formed. Titratable
The overall goal is to calculate the Acidity: U sed to monitor the progress of

16 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
fermentation by measuring the 6. Describe production of yogurt, including the or-
amount of base required to neutral- ganisms responsible and the interaction between
ize the amount of acid in a prod- the two that results in the final product.
uct, i.e. lactic acid. It provides a 7. What is lactose intolerance and how can it be
measure of the total acidity rather treated?
than of hydrogen ion concentration.

Basic Knowledge Questions Critical Thinking Questions

1. What is fermentation? Include in your answer a 1. Compare and contrast homo fermentation and
definition for "top" and "bottom" fermentations. heterofermentation. Include differences in terms
2. List the 12 genera of bacteria that constitute the of metabolic pathways, enzymes, and examples
"lactic acid bacteria," and describe them in terms of microorganisms.
of growth requirements and other general char- 2. You work at a cheese production plant. A new
acteristics. strain of Propionibacterium has been purchased
3. What is "molar growth yield"? by the company and you are asked to determine
4. Describe the spoilage of pasteurized milk prod- whether it is as good, or better, than the strain that
ucts. is currently being used in the production of swiss
5. Name some organisms that are commonly used cheese. Design an experiment and the approach
as starter cultures in dairy fermentation. you would follow to answer this question.

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Copyright 2000, Aspen Publishers, Inc.
CHAPTER 8

Fruit and Vegetable Products:


Whole, Fresh-Cut, and Fermented

Learning Objectives Chapter Outline

The reader will be able to: 1. Fresh and frozen vegetables.


1. Describe how fruits and vegetables become con- a) Incidence of microorganisms
taminated with microorganisms. b) Spoilage
2. Describe the characteristics of vegetables that 1) Bacterial agents
make them prone to specific types of spoilage. a. Soft rot
3. Define the various types of spoilage conditions b. Other conditions
of vegetables caused by bacteria and by molds, 2) Fungal agents
and include examples of organisms responsible a. Gray mold rot
for these. b. Sour rot
4. Describe the spoilage of fruit, including a discus- c. Rhizhopus soft rot
sion of the characteristics that enable these com- d. Phytophora rot
modities to resist growth by most microorgan- e. Anthracnose
Isms. 2. Spoilage of fruits.
5. Define fresh-cut produce and discuss the types of 3. Fresh-cut produce.
spoilage and disease-causing microorganisms that a) Examples
may be found. b) Microbialload
6. Describe various plant-based fermented prod- c) Pathogens
ucts, such as breads, olives, pickles, sauerkraut, 4. Fermented products.
wine, and cider within the context of the pro- a) Breads
cesses involved and the types of starter cultures b) Olives, pickles, and sauerkraut
utilized to produce them. 1) Olives
7. Describe fermented products made from beans 2) Pickles
of various plants, such as coffee and cacao. 3) Sauerkraut
8. Describe soy-based and other crop-based fer- c) Beer, ale, wines, cider, and distilled spirits
mented products, including the process involved 1) Beer and ale
and the organisms used for fermentation reac- 2) Wines
tions. 3) Cider

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4) Distilled spirits OTR: Oxygen transmission rate, usually
5) Miscellaneous fermented products refers to the property of a packag-
a) Coffee beans ing film to allow the free passage
b) Cacao beans of oxygen. For example, a low OTR
c) Soybean products film is used for packaging products
1) Soy sauce under vacuum.
2) Tempeh Pathovar: Classification of microorganisms
3) Miso below the species level according
d) Ogi to their ability to cause specific
e) Gari pathogenic effects.
f) Bongkrek Pectin: Complex carbohydrate found in the
g) Ontjom cell wall of most fruits and veg-
etables, composed of galacturonic
acid molecules linked together by
Terms and Defmitions alpha 1-4 linkages.
Superoxide: Radical of oxygen containing an
Auxins: Plant hormones that can desensi- unpaired electron. Highly reactive
tize plants to ethylene and thus de- molecule, capable of interfering
lay ripening. with many biochemical reactions
Blight: Any disease, symptom of disease, of bacteria and other organisms.
or injury ofplants resulting in with- Superoxide
ering, cessation of growth, etc. but Dismutase: Enzyme found in many microor-
without rotting. ganisms, which catalyzes a reaction
Catalase: Enzyme found in many microor- that combines two superoxide mol-
ganisms, which catalyzes a reaction ecules and water to form hydrogen
that breaks down hydrogen perox- peroxide and oxygen.
ide to release oxygen and water. It Wort: Liquid obtained from the mashed
is part of the defense mechanism malt and malt adjuncts after these
of bacteria to prevent interference have been heated and their starches
of hydrogen peroxide in its bio- digested.
chemical reactions.
Cellulose: Complex carbohydrate that forms
the wall of plant cells, composed
Basic Knowledge Questions
of up to 12,000 glucose molecules
linked by beta 1-4 linkages.
Cotyledons: Storehouse of nourishment for the 1. Describe a possible mechanism by which Erwinia
embryo of seed plants, the leaf in a carotovora causes soft rot of vegetables.
seed. 2. Name three types of fruit rot and the organisms
Ethylene: Plant hormone known to trigger responsible.
ripening offruit and promote plant 3. What are the concerns over the safety of fresh-
senescence. cut produce packaged under vacuum? How can
Hops: Flowers used to impart certain fla- these concerns be addressed/minimized?
vors, aromas, and bitterness to wort 4. Compare the production of sour dough bread vs.
during beer production. idli bread, including the organisms used and the
Lenticel: One of the cortical pores in the role that each plays in fermentation.
stems of woody plants by which air 5. Describe the process of brewing beer and ale,
penetrates. including the enzymes involved.
Lye: Strongly alkaline solution such as 6. What is "malo-lactic" fermentation, and why is it
NaOH. important to the wine industry?

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Copyright 2000, Aspen Publishers, Inc.
7. Compare production of ogi, gari, bongkrek, and of organisms that would be found, the order in
ontjom in terms of the steps required and organ- which each type would affect the fruit, and ex-
isms of importance., plain your reasoning.
2. You are wanting to develop a new fermented
Critical Thinking Questions beverage from black beans. List the steps you
would follow and include the types of organisms
1. During the spoilage of fruit, explain the sequence you would use and the reasoning behind your
of events that may take place in terms ofthe types decisions.

20 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 9

Miscellaneous Food Products

Learning Objectives Chapter Outline

The reader will be able to: 1. Delicatessen and related foods.


1. Describe the types of microorganisms found in a) Salads and sandwiches.
delicatessen foods and how they cause spoilage b) Cream pies and breadlbiscuitlpie dough
of such products. 2. Eggs.
2. List and define the structural components of a) Structure of eggs
eggs, and describe the types of microorganisms b) Egg contamination and types of microorgan-
that cause spoilage. isms present
3. Describe how mayonnaise is made and discuss 3. Mayonnaise and salad dressing.
the types of microorganisms that cause spoilage a) Formulation
in this product. b) Types of organisms present
4. Describe the types of organisms and the spoilage 4. Cereals, flour, and dough products.
that is seen in bakery products, frozen meat pies, 5. Bakery products.
candy, spices, and nuts. 6. Frozen meat pies.
5. Describe what types of dehydrated foods are 7. Sugars, candies, (' ad spices.
available, the types of microorganisms that cause 8. Nutmeats.
spoilage, and how they do so. 9. Dehydrated foods.
6. Define foods used for medical purposes and 10. Medical foods.
discuss the types of spoilage that can be ob- 11. Single-cell protein.
served, including the types of microorganisms a) Definition
responsible. b) Rationale
7. Define single-cell protein, provide a justification c) Organisms
for its need, and discuss the types of organisms d) Products
and products that are used, and the safety and e) Nutrition and safety
nutritional quality of these.

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Terms and Definitions Mustiness: Spoilage condition of eggs caused
by Pseudomonas graveolens and
Albumen: Multi-layer component of eggs, other bacteria.
consisting mainly of water and pro- Ovoflavoprotein: Molecule that binds the vitamin
teins that surround the yolk. riboflavin in eggs.
Amylase: Enzyme produced by some micro- Ovotransferrin: Protein molecule that chelates
organisms which breaks down metal ions such as iron, located in
starch into individual glucose mol- the albumen of eggs, synonymous
ecules. These can be easily metabo- with conalbumin.
lized by the organism for growth Pink Rot: Spoilage condition of eggs caused
and cell division. by Pseudomonas bacteria.
Avidin: A protein found in the albumen of Pinspots: Spoilage of eggs caused by molds,
eggs, it binds the vitamin biotin. typified by appearance of mycelia
Biotin: Water-soluble vitamin, found in in the inside of the egg, visible by
eggs and many other foods. It acts candling.
as a coenzyme in carboxylation Proteolytic: Quality of some microorganisms
(carboxyl-group-adding) reactions that enable them to break down
in cells. proteins through enzymatic action.
Black Rot: Spoilage condition of eggs caused Red Rot: Spoilage condition of eggs caused
by Proteus, Aeromonas, Pseudo- by some Serratia species.
monas bacteria. Ropiness: Spoilage condition of homemade
Chalazae: A pair of spiral bands of albumen- bread caused by the bacterium Ba-
like substance in the white of eggs. cillus mesentericus.
They extend from the ball of the Rotting: Spoilage of eggs caused by bacte-
yolk and are attached at each end of ria, i.e. "green rot," "pink rot,"
the egg to the vitelline membrane. "colorless rot," and "custard rot."
Colorless Rot: Spoilage condition of eggs caused Shiga-like
by Pseudomonas and Acineto- Toxin: Toxin that is produced by certain
bacter bacteria. bacteria, typically pathogenic
Conalbumin: Synonymous with ovotransferrin, strains of Escherichia coli such as
is one of the proteins found in the 0157 :H7. Toxin is similar in struc-
albumen of eggs. It consists of a ture to Shiga toxin produced by the
peptide chain linked to iron. bacterium Shigella dysenteriae.
Custard Rot: Spoilage condition of eggs caused Vitelline
by Proteus vulgarius and P. Membrane: A membrane enclosing the ball of
intermedium. the yolk of eggs.
*"
Emulsion: An aqueous system that contains
two liquids that are not soluble in Basic Knowledge Questions
each other, one of which is dis-
persed in the other as droplets. 1. Draw a picture of the cross-section of an egg,
Lipolytic: Quality of some microorganisms labeling each structure/layer appropriately.
that enable them to break down lip- 2. How do microorganisms enter eggs and what
ids through enzymatic action. factors affect this?
Lysozyme: Enzyme found in egg white and 3. Describe how mayonnaise spoils.
human tears, destructive to gram- 4. What properties of bakery products make them
positive bacteria. more prone to mold growth than other foods?
Mucin: Glycoproteins found in secretions 5. How does Leuconostoc spoil candy products?
of many organisms, as well as tis- 6. At what stages during processing are contami-
sues of man and animals. nants introduced into nuts?

22 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


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7. List the advantages of the use of single-cell 2. After spending time in the poorest third-world
protein as a source of protein for human con- country in the world, you decide to use your
sumption. knowledge of single-cell protein to design a sys-
tem of food production that will ease the mal-
Critical Thinking Questions nourishment of that country's citizens. Describe
the factors you need to consider and how you
1. Knowing what you now know about some of the could introduce such products into the diet.
components of eggs, discuss how you could use
these to improve the microbial quality of salads.

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 23


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 10

Culture, Microscopic,
and Sampling Methods

Learning Objectives 2. Conventional standard plate count.


a) Factors to consider.
The reader will be able to: b) Homogenization of food samples.
1. Describe the standard plate count method, includ- c) The spiral plater.
ing new technologies for the processing of multiple 3. Membrane filters.
samples. a) Definitions.
2. Discuss membrane filtration techniques. b) Direct epifluorescent filter technique.
3. Describe microscope colony count methods. c) Microcolony-DEFT.
4. Describe the agar droplet method and its applica- d) Hydrophobic grid membrane filter.
tions. 4. Microscope colony counts.
5. Describe and compare three major dry-film-based 5. Agar droplets.
methods. 6. Dry film and related methods.
6. Briefly define the Most Probable Numbers method a) Petrifilm
and how it is applied for enumeration of microor- b) Redigel
ganisms in foods. c) SimPlate
7. Describe dye reduction and roll tube methods. 7. Most probable numbers.
8. Describe direct microscopic count methods, includ- 8. Dye reduction.
ing a discussion on distinguishing viable from non- 9. Roll tubes.
viable cells. 10. Direct microscopic count.
9. List and describe several methods for the evalua- a) Conventional method.
tion of surface contamination. b) Slide method.
10. Define microbial injury and stress, and discuss how c) Howard mold counts.
these processes occur, how the cell may repair the 11. Microbiological examination of surfaces.
damage and recover, and how some cells can re- a) Swab/swab-rinse methods.
main viable but not culturable. b) Contact plate.
c) Agar syringe/"agar sausage" methods.
d) Other surface methods.
Chapter Outline 1) Direct surface.
2) Sticky film.
1. General overview. 3) Swab/agar slant.

24 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
4) Ultrasonic devices. cific for a unique antigenic deter-
5) Spray gun. minant, such as a particular com-
12. Metabolically injured organisms. ponent of the cell membrane of a
a) Definition of injury and stress. given bacterium. They are useful
b) Recover/repair. for detecting the presence of par-
c) Mechanism. ticular microorganisms in food
13. Viable but nonculturable organisms. samples.
Protein A: Protein which binds nonspecifi-
Terms and Definitions cally to antibodies on their Fc por-
tion, serving as an anchor prior to
Acridine Orange: Fluorescent dye which penetrates exposing them to an antigen, such
microbial cells. as a bacterial cell produced by Sta-
Autoclaved: Refers to the condition of a mate- phylococcus aureus.
rial after it has been subjected to Resazurin: Dye used as an indicator of the de-
heating using an autoclave device gree of anaerobiosis of a medium,
to achieve sterility. In this proce- with the dye turning from pink to
dure, the material (i.e. food or me- colorless upon reduction of the
dium) is placed inside a chamber medium when anaerobic condi-
and heated with steam under pres- tions have been achieved. Such
sure for varying times at tempera- conditions can be caused by chemi-
tures of 250F or more. cal means or by depletion of oxy-
Beta- gen due to microbial growth.
Glucuronidase: Enzyme produced by certain mi- Quebec
croorganisms, such as most strains Colony Counter: Device equipped with a lamp, de-
of Escherichia coli, that breaks signed to shine light through a petri
down glucuronic acid. plate on which microbial colonies
Lag Phase: Phase of bacterial growth in which are growing. It allows for low-level
cells are actively undergoing meta- magnification of colonies for ease
bolic reactions, chiefly synthesiz- of counting.
ing macromolecules for cell Stomacher: A blender-like device in which
growth, but no cell division takes food samples in buffer are homog-
place. It is during this phase that enized by gentle pummeling rather
injured cells undergo repair. than high-speed blending.
Methylene TCACycle: Also known as Krebs Cycle, it is
Blue: Dye used as an indicator of the de- the series of reactions that take
gree of anaerobiosis of a medium, place in most microbial cells dur-
with the dye turning from blue to ing the metabolism of sugars,
colorless upon reduction of the yielding approximately 36 mol-
medium when anaerobic condi- ecules ofATP per molecule of glu-
tions have been achieved. Such cose.
conditions can be caused by chemi- Triton-X: Detergent used to remove proteins
cal means or by depletion of oxy- and other organic materials from
gen due to microbial growth. surfaces. Chemical name is
Monoclonal polyoxyethylene p-t-octyl phenoL
Antibody: Antibody molecules produced by Trypan Blue: An acid dye that stains mold and
a single B-cell of an animal or per- other colonies with a blue color.
son. Monoclonal antibodies are Trypsin: Enzyme found in gastric juice that
identical to each other, and are spe- is used to break down proteins.

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Copyright 2000, Aspen Publishers, Inc.
Basic Knowledge Questions 7. Define "culturable but not viable".

1. What factors should be considered in conducting


standard plate counts of microbial contaminants?
Critical Thinking Questions
2. What are the types of fluorescent microscopy
methods that can be used to evaluate microor-
ganisms? How do they work? 1. It is known that the number of pathogenic organ-
3. Compare and contrast the Redigel and SimPlate isms in most foods is very low, provided those
methods to enumerate bacteria. What are the products have not been temperature-abused. What
advantages/disadvantages of each? method(s) would you consider using to enumer-
4. What method would you use to sample the sur- ate such organisms? Why?
face of a food processing plant after it has been 2. You have developed a new isolation medium for
sanitized? Why? the hypothetical bacterium Testina obligata. De-
5. What is microbial injury? sign an experiment that would test the ability of
6. What factors affect the ability of microorgan- this new medium to successfully and accurately
isms to repair after injury? enumerate injured as well as healthy cells.

26 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 11

Physical, Chemical, Molecular,


and Immunological Methods

Learning Objectives 2. .Chemical Methods.


a) Thermostable nuclease
The reader will be able to: b) Limulus lysate for endotoxins
1. Define impedance, microcalorimetry, and flow c) Adenosine triphosphate measurement
cytometry, and be able to cite examples of where d) Radiometry
and how these physical detection methods are e) Fluorogenic and chromogenic substrates
used in the food industry. f) Lux gene luminescence
2. Define and describe chemical methods for detec- g) Ice nucleation assay
tion of microorganisms, comparing those based 3. Methods for Characterizing and Fingerprinting
on enzymatic assays and those based on other F oodbome Organisms
technologies. a) Serotyping
3. Describe serotyping and bacteriophage typing, b) Bacteriophage typing
and compare the two traditional assays for bacte- c) Nucleic acid (DNA) probes
rial classification and identification. d) DNA amplification (Polymerase Chain Re-
4. Describe the basis for various DNA-based meth- action)
ods of fingerprinting microorganisms. e) Multilocus enzyme electrophoresis typing
5. Describe and discuss immunological methods, f) Restriction enzyme analysis
making comparisons of application and limita- g) Random amplification of polymorphic DNA
tions of each. h) Pulsed field gel electrophoresis
6. Explain the principle behind the ELISA method, i) Restriction fragment length polymorphism
and describe its application for the detection of j) Ribotyping
various types of microorganisms and their tox- 4. Immunological Methods
ins. a) Fluorescent antibody
b) Enrichment serology
Chapter Outline c) Salmonella 1-2 test
d) Radioimmunoassay
1. Physical Methods. e) ELISA
a) Impedance and related methods f) Gel diffusion
b) Microcalorimetry g) Immunomagnetic separation
c) Flow cytometry h) Hemagglutination

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 27


Copyright 2000, Aspen Publishers, Inc.
Terms and Definitions Exothermic
HPR: Rate at which microorganisms
ATP: Adenosine triphosphate, it is produce heat during growth,
the primary source of energy which can be correlated to the
for all living cells, microbial as number of organisms present
well as others. It is produced in the medium.
by the metabolism of nutrients Fluorescein
and is used to fuel catabolic Isothiocyanate: Fluorescent stain which binds
and other reactions. to protein.
Antigen: Term used to refer to any mol- Flow Cytometry: The measurement of indivi-
ecule or particulate structure, dual cells as they flow through
considered foreign to a particu- a liquid medium. Cells can be
lar animal or human being. individually measured in terms
Antigens can be part ofthe cell of size, fluorescence, light-ab-
wall of a bacterium, and when sorbing properties, and can be
introduced into an animal, counted as they pass by a laser
elicit the production of anti- beam detector.
bodies by the animal that are Flow Micro-
specific for and bind to that Calorimetry: Microbial identfication
antigen. method in which a flow-
Cation Exchange: Method used to separate pro- through vessel is used contain-
teins in a sample, in which ing a seven-sugar medium.
those with a net positive charge Microorganisms such as yeasts
will bind to a column contain- and lactic acid bacteria are al-
ing a carboxyl group, whereas lowed to grow, producing ther-
proteins with a negative charge mograms unique to each or-
will not. The positively- ganism.
charged proteins are then re- HAntigens: Components of the flagella of
leased from the column by motile bacteria, such as salmo-
adding sodium chloride or an- nellae.
other salt to the column. So- Hybridization: Method in which two single-
dium ions in the buffer will strands of DNA are allowed to
compete with the positively anneal, or bind, at complimen-
charged groups on the protein, tary base pairs along the
and will bind to the column. strands. The degree of hybrid-
Those proteins with a low-den- ization is used as an indicator
sity of net positive charge will of the similarity of the DNA
emerge first out ofthe column, strands. Ifthese come from two
followed by those with a higher separate organisms, hybridiza-
charge density. tion is used as an indicator of
Coagulase: Enzyme produced by some the similarity of the two organ-
microorganisms, such as Sta- isms.
phylococcus aureus, which is IDT: Impedance detection time, re-
used as an indicator of the abil- fers to the time required to de-
ity of the cells to produce en- tect resistance to the flow of
terotoxin. electricity in a medium. IDT
Enthalpy: The amount of heat lost by a can be correlated to the num-
system to the environment. ber of microorganisms present

28 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
in a broth medium, with the known as "restriction endonu-
shorter the IDT the higher the cleases".
number of cells present. Serotyping: The use of antibodies specific
Impedance: The resistance to flow of elec- for particular organisms for the
trical current. It is the opposite purpose of identification and
of conductivity. classification according to dif-
KAntigens: Components of the capsule of ferences on the molecules re-
encapsulated bacteria, such as siding on the surface of cells
streptococci. (i.e. in cell membrane).
Lipopoly- Thermostable
saccharide (LPS): Structural component of the Nuclease: Enzyme produced by Staphy-
bacterial gram-negative cell lococcus aureus, especially
envelope. It is pyrogenic, able those cells that produce entero-
to induce fever in humans who toxin, as well as other organ-
have been infected. One of its isms. It degrades DNA and is
components is Lipid A. Also resistant to boiling for 15 min-
known as endotoxin. utes.
Lysate Protein: Protein found in the amoeb- Vero Toxin: Toxin produced by some
ocytes, or blood cells, of the pathogenic strains of Escheri-
horseshoe crab. It is used to chia coli, such as 0157:H7. It
detect the presence of endot- causes hemorrhagic colitis in
oxin by the formation of a clot humans. Toxin was first iden-
when the protein comes in con- tified as being cytotoxic to
tact with the LPS of gram- Vero tissue culture cells (Afri-
negative cells. can Green Monkey kidney
o Antigens: The 0 polysaccharide side cells), hence the terms "vero
chain that is part of the LPS of toxin" and "verocytotoxin."
gram-negative bacteria. These
are found on the outermost sur-
Basic Knowledge Questions
face of the cell, typically pro-
truding out of the outer cell 1. Compare the use of thermostable nuclease and
membrane. Also known as "so- coagulase as indicators of the presence of entero-
matic antigens". toxin-producing S. aureus.
Phage: A virus that is specific and 2. Discuss the LAL and ATP assays for detection of
generally lethal to bacteria, microorganisms. What do these have in com-
which can be manipulated to mon? What are the limitations of each?
act as a carrier and insert cer- 3. Name several of the chromogenic substrates used
tain desirable genes into the in culture media for detection of microorgan-
cells. isms. What is the difference between fluoro- and
Propidium chromogenic substrates?
Iodide: Fluorescent stain that binds to 4. Describe the various types of antigens found on
DNA. gram-negative bacteria.
Restriction 5. List some applications of the polymerase chain
Enzymes: Enzymes that will bind to spe- reaction assay for detection of microorganisms
cific sites along DNA mol- in food.
ecules (i.e. specific base se- 6. Compare and contrast PFGE and RFLP. Which
quences), lysing or cutting the do you think gives a more accurate fingerprint of
molecule at those sites. Also a particular microbial isolate? Why?

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 29


Copyright 2000, Aspen Publishers, Inc.
7. Describe the conventional and "sandwich" ELISA gram depicting the steps you would need to
assays. follow in carrying out the assay.
2. Name the factors you would recommend that an
industry person considers when making a deci-
Critical Thinking Questions sion regarding the type of test to use for the
following purposes:
1. You are given antibodies specific for Salmonella a) Verifying that a sanitation procedure on a
species, developed in goats. The only other re- food contact surface has indeed been suc-
agent you have is antibodies developed in hu- cessful.
mans, specific for goat tissue, to which an en- b) Determining whether a bacterial isolate found
zyme is bound. Your job is to design an ELISA on a certain food product is the same as the
assay that will help you decide whether a certain strains found in the food processing plant
food sample contains this organism. Draw a dia- environment.

30 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 12

Bioassay and Related Methods

Learning Objectives c) Human ileal and intestinal cells


d) Guinea pig intestinal cells
The reader will be able to: e) HeLa cells
1. Describe the various types of whole animal as- t) Chinese hamster ovary cells
says, stating the application of each. g) Vero cells
2. Describe bioassays that require surgical inter- h) Y-l adrenal cell assay
vention of animals, such as the ligated ileal loop i) Other assays
test and the RIT ARD test.
3. Describe the various types of tissue culture cells Terms and Definitions
that can be used to detect specific pathogens and!
or their toxins. Buccal Mucosa: The lining of the inside of the
mouth.
Cereulide: Toxin produced by some
Chapter Outline strains ofthe bacterium Bacil-
lus cereus, causes vomiting.
1. Whole-Animal Assays. Depilation: Removal of hair, usually by
a) Mouse lethality shaving, to expose the surface
b) Suncus murinus of the skin prior to intradermal
c) Suckling (infant) mouse injection.
d) Rabbit and mouse diarrhea Erythema: Reddening of the skin, usually
e) Monkey feeding as a result of injection of cer-
t) Kitten (cat) test tain substances intradermally.
g) Rabbit and guinea pig skin tests Emesis: Vomiting.
h) Sereny and Anton tests Hepatocyte: Liver tissue cells.
2. Animal Models Requiring Surgical Procedures. Intracardial (i.e.): One method of injection, liter-
a) Ligated loop techniques ally meaning "through the
b) The RITARD model heart".
3. Cell Culture Systems. Intradermal (i.d.): One method of injection, liter-
a) Human mucosal cells ally meaning "through the
b) Human fetal intestine skin".

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 31


Copyright 2000, Aspen Publishers, Inc.
Intraluminal (i.I.): Method of injection, literally tissue culture cells inoculated
meaning "through the lumen," with 100 uL of medium.
or the inside of the intestines. Scintillation
Intraperitoneal (i.p.): One method ofinjection, liter- Counter: Instrument that measures the
ally meaning "through the wall radioactivity of a sample by
of the peritoneum," the sac-like measuring alpha and beta
material that surrounds the ab- emissions.
dominal cavity of the body. Trypsin: Proteolytic enzyme found in
Kanagawa the gastrointestinal tract that is
Test: U sed to characterize strains of used to activate the toxin of
V. parahaemolyticus. Lysis of some types of Clostridum
human or rabbit red blood cells botulinum.
but not horse on Wagatsuma's
agar denotes a K+ strain ..
Necrosis: Tissue death, caused by the Basic Knowledge Questions
lack of blood circulation.
Orogastric: Through the mouth and into 1. Compare the Anton and Sereny tests.
the stomach. 2. Why is the Suncus murinus test used?
Phagolysosome: Sac-like structure in the cyto- 3. Define the GWIBW ratio as used in the Suckling
plasm of phagocytic immune Infant Mouse assay.
system cells such as macroph- 4. Name and describe the various types of skin tests
ages. It is formed by the merg- used for detection of toxin.
ing of two cytoplasmic struc- 5. Describe the ligated ileal loop test.
tures: a phagosome (contain- 6. Construct a table that lists at least three types of
ing an invading microorganism tissue culture cells and the pathogen or agent that
that has been engulfed by the each test detects.
macrophage) and a lysosome 7. What are Vero cells and what are they used for?
(containing lysozyme and
other lytic enzymes) for the Critical Thinking Question
purpose of digestion of the in-
vading organism. 1. There is a new microorganism that has been
PEU: Plating efficiency unit, or the linked to an outbreak of foodborne illness. De-
amount of enterotoxin that re- scribe the types of tests that you would employ to
sults in a 25% inhibition of200 determine how it causes disease.

32 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 13

Food Preservation with Chemicals

Learning Objectives 2. Sorbic Acid.


3. The Propionates.
The reader will be able to: 4. Sulfur Dioxide and Sulfites.
1. Describe the mode of action of benzoic acid, 5. Nitrites and Nitrates.
parabens, sorbic acid, propionates, sulfur diox- a) Compounds used
ide, and sulfites. b) Organisms affected
2. Explain the uses of nitrites and nitrates, and c) The Perigo factor
describe the types of organisms that are affected d) Interaction with cure ingredients and other
by these compounds. factors
3. Define the "Perigo Factor". e) Nitrosamines
4. Describe how nitrites can be used in combination f) Nitrite-sorbate and other nitrite combinations
with other antimicrobial agents. g) Mode of action
5. Describe the antimicrobial effect provided by h) Summary of nitrite effects
sugars and salts. 6. NaCl and Sugars.
6. Describe how several ingredients, added to food 7. Indirect Antimicrobials.
products for other purposes, act to inhibit micro- a) Antioxidants
organisms. b) Flavoring agents
7. Describe the mechanism of action of lactic and c) Spices and essential oils
acetic acid, and list some of the applications of d) Medium-chain fatty acids and esters
these antimicrobial compounds. 8. Acetic and Lactic Acids.
8. Define the term "bacteriocin," and list and de- 9. Antibiotics and Bacteriocins.
scribe several types of these compounds, and the a) General considerations
organisms that produce them. b) Nisin and other bacteriocins
9. Describe the use of antifungal agents and fumi- c) Monensin
gants for decontamination of plant products. d) Natamycin
e) Tetracyclines
f) Subtilin
Chapter Outline g) Tylosin
10. Antifungal Agents for Fruits.
1. Benzoic Acid and the Parabens. 11. Ethylene and Propylene Oxides.

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 33


Copyright 2000, Aspen Publishers, Inc.
12. Miscellaneous Chemical Preservatives. Germination: Phase in the life cycle of spore-
a) Sodium diacetate forming organisms. It involves
b) Ethanol the shift from the spore-form
c) Diethylpyrocarbonate to the vegetative cell form of
d) Dimethyl dicarbonate the organism. Germination is
e) Acid anionic sanitizers often triggered in the spore by
f) Sucrose fatty acid esters exposure to a short-term stress,
g) Glucose oxidase such as mild heating, or by ex-
posure to environmental con-
ditions suitable for growth,
Terms and Definitions such as the renewed availabil-
ity of nutrients.
Bacteriostatic: An agent or chemical that in- GRAS: Status given to some food in-
hibits growth of bacteria with- gredients, classifying them as
out killing the cell. generally recognized as safe for
Chelate: A compound that sequesters human consumption. Thus, test-
particular ions in solutions, ing and approval by the Food
binding strongly enough to and Drug Administration is not
prevent their interaction with required for these to be used.
other molecules. One example Examples are sugar and salt.
is EDTA, which chelates iron Ionophores: Small hydrophobic molecules
and other multivalent ions. that dissolve within lipid bilay-
Endospore: Physical structure developed ers of membranes, acting as
by spore-forming bacteria, in channels to allow the diffusion
which several layers are added of ions into or out of the cell.
to the cell for protection in Some antibiotics, such as vali-
harsh environments (i.e. dry nomycin, act to transport po-
conditions, lack of nutrients). tassium ions, disrupting trans-
In this stage, the cell is in a port mechanisms of the bacte-
"dormant" state with no rial cell.
growth taking place. The spore pKa: The pH of a weak acid in which
can germinate upon exposure half of the molecules are in the
to proper conditions, at which undissociated state and the
time the cell that results can other half are in the dissociated
resume active growth. state.
Fistulation: A surgical procedure in which Phosphoroclastic: Refers to a chemical reaction
a in-out port is implanted on in which phosphate is removed
the skin of an animal in order from one molecule and used to
to access the contents of par- phosphorylate another, such as
ticular abdominal chambers, in the conversion of ATP from
such as the stomach or rumen. ADP.
With this implant, samples of Plasmolysis: The lysis of microbial cells as
fluid can be drawn from that a result of osmotic imbalance
chamber after specific feeding that causes the cells to lose
regimens are applied to the water to the point of shrinkage
animal, for example. (dehydration). It is the result of
Fungistat: A compound that inhibits having a higher concentration
growth of fungal cells without of solutes in the medium com-
causing death. pared to the inside of the cell.

34 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
PMF: Proton Motive Force, refers to Basic Knowledge Questions
the force that drives the move-
ment of compounds in and out 1. Describe how spore-forming bacteria germinate,
of the cell. It is dependent on and how certain antimicrobial agents can inhibit
the establishment of a charge this process.
differential, or gradient, across 2. Describe the mode of action of sulfites to inhibit
the cell membrane, in which microorganisms.
positively charged ions (pro- 3. What is DNFH and how is it formed?
tons) are on the outside of the 4. Defme the "Perigo Factor", and give an example
cell. of it.
Undissociated: Status of a molecule in which 5. What is the mode of action of nitrite in inhibiting
hydrogen ions are no longer germination of spores?
bound to the rest of the mol- 6. Name some spices and the antimicrobial com-
ecule. The molecule is then pounds found in each.
said to be in the oxidized, 7. What are the classes ofbacteriocins?
charged state, containing one
unpaired electron. Weak acids Critical Thinking Questions
are undissociated when the pH
of the medium is below the 1. Do you think the "Perigo Factor" is a real phe-
pKa of the acid. nomenon? How would you test its existence?
Viristatic: A compound that inhibits the 2. Why do you think bacteriocins are usually in-
proliferation of viruses without hibitory to organisms that are closely related to
necessarily causing death. the one producing them?

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 35


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 14

Food Preservation with


Modified Atmospheres

Learning Objectives 4. The Safety of MAP Foods.


a) Clostridium botulinum
The reader will be able to: b) Listeria monocytogenes
1. Define the various types of modified atmosphere c) Other pathogens
packaging systems. 5. Spoilage of MAP and Vacuum-Packaged Meats.
2. Describe the primary effects of MAP on inhibi- a) Determining factors
tion of microorganisms. b) Volatile components of vacuum-packaged
3. List and describe various food products that can meats and poultry
be packaged with MAP.
4. Discuss the food safety considerations of MAP-
packaged foods. Terms and Definitions
5. Describe the type of spoilage that results in MAP-
packaged foods. CAP: Controlled-atmosphere pack-
aging, refers to the packaging
of foods in a modified atmo-
Chapter Outline sphere within an impermeable
package so tllat the composi-
1. Definitions. tion of the atmosphere inside
a) Hypobaric (low pressure) storage the package does not change
b) Vacuum packaging during storage.
c) Modified atmosphere packaging Captech Process: Packaging system in which
d) Equilibrium-modified atmosphere hygienic conditions are fol-
e) Controlled-atmosphere packaging or storage lowed in the packaging of the
2. Primary Effects of CO2 on Microorganisms. food with either gas-permeable
a) General findings or impermeable film, and stor-
b) Mode of action age at -1.5C.
3. Food Products. EMA: Equilibrium-modified atmo-
a) Fresh and processed meats sphere, refers to the packaging
b) Poultry offoods in gas-permeable film
c) Seafoods or material. The food is either

36 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
gas-flushed prior to sealing of Basic Knowledge Questions
the package, or not flushed at
all. Gas exchange is thus al- 1. List some of the historical events surrounding
lowed to take place between the development of modified atmosphere pack-
the food tissue and the outside aging technology.
environment. 2. What are the two types of modified atmosphere
Hyperbaric: Condition where the pressure packaging?
is higher than atmospheric lev- 3. List some of the primary effects of gas-flushing
els (i.e. 1 atm). on microorganisms.
Hypobaric: Condition where the pressure 4. List the hurdles against Clostridium botulinum
is lower than atmospheric lev- growth that are inherent in modified atmosphere
els (i.e. 1 atm). packaged foods.
OTR: Oxygen transmission rate of a 5. What are the concerns regarding Listeria
packaging material, usually monocytogenes and modified atmosphere-pack-
expressed as volume of oxygen aged foods?
transmitted through the pack- 6. List the determining factors that guide the types
age per a specific area of the of microorganisms that may be found in modi-
surface of the package within fied atmosphere-packaged foods.
a given period of time, at a 7. Describe the products of spoilage of modified
given temperature of storage, atmosphere-packaged meats, including the types
and under a specific relative of organisms responsible.
humidity. For example: 8 mLl
mZ/24 hl37C/700/0 R.H.
Periplasm: Fluid space located between Critical Thinking Question
the inner and outer membranes
of gram-negative bacteria. It 1. What are the possible mechanisms at play in
contains several types of mol- inhibition ofmicroorganisms by COz? How would
ecules, primarily proteins. you test your hypothesis?

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 37


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 15

Radiation Preservation of Foods and


Nature of Microbial Radiation Resistance

Learning Objectives Chapter Outline

The reader will be able to: 1. Definitions and Units of Measure.


1. Define the term "radiation" and describe the 2. Characteristics of Radiations ofInterest in Food
units employed for measuring dose. Preservation.
2. Describe the various types of radiation that could a) Ultraviolet light
be used for decontamination of foods. b) Beta rays
3. Describe the effect of various parameters inher- c) Gamma rays
ent to microorganisms on ability of irradiation to d) X rays
inactivate them. e) Microwaves
4. Describe the effect of various parameters not 3. Principles Underlying the Destruction of Micro-
related to the microorganism (packaging atmo- organisms by Irradiation.
sphere, temperature, food composition) on mi- a) Types of organisms
crobial inactivation by irradiation. b) Numbers of organisms
5. Describe the types of foods most suitable for c) Composition of suspending menstruum
irradiation. d) Presence or absence of oxygen
6. Describe how foods should be processed prior to e) Physical state of food
irradiation. f) Age of organisms
7. Describe the process of irradiation, as it applies 4. Processing of Foods for Irradiation.
to the use of specific sources (i.e. electrons, a) Selection of foods
gamma rays, X rays). b) Cleaning of foods
8. Define the terms "radappertization," c) Packing
"radurization," and "radicidation." d) Blanching or heat treatment
9. Discuss the legal status of food irradiation in the 5. Application of Radiation.
U.S. a) Gamma radiation
10. Discuss the effect of irradiation on the quality of b) Electron beam/accelerated electrons
foods, and how detrimental effects can be mini- 6. Radappertization, Radicidation, and Radurization
mized and!or prevented. of Foods.
11. Discuss the topic of microbial resistance to irra- a) Definitions
diation, citing specific microbial examples. 1) Radappertization

38 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
2) Radicidation Microwaves: Type of energy produced when
3) Radurization electrically neutral foods are
7. Legal Status of Food Irradiation. placed in an electromagnetic
8. Effect ofIrradiation on Food Quality. field of a particular wave-
9. Storage Stability ofIrradiated Foods. length. It causes the molecules
10. Nature of Radiation Resistance of Microorgan- in food to oscillate, creating
isms. friction and thereby heat.
a) Biology of extremely resistant species Rad: Unit of measure for absorbed
b) Apparent mechanisms of resistance dose. It is equivalent to 0.01
joule of energylkg of matter.
Radappertization: Application of ionizing radia-
Terms and Definitions tion to achieve commercial ste-
rility, with typical doses rang-
Beta Radiation: Electrons emitted by a radio- ing from 30 to 50 kGy.
active isotope during its decay, Radiation: Emission of energy and its
such as cobalt-60. propagation through space or
Cold Sterilization: Processing treatment that matter (i.e. food).
causes no noticeable increase Radicidation: Application of ionizing radia-
in temperature of the product. tion to achieve pasteurization,
Term is often used to refer to with typical doses ranging
ionizing radiation. from 2 to 10 kGy.
Electromagnetic Radiolysis: The lysing effect of chemical
Spectrum: Spectrum of all the electro- bonds as caused by exposure
magnetic energy present in of a material to radiation.
nature, ranging from commu- Radiolytic
nication bands to gamma rays, Products: Products of the lysis ofchemi-
and including visible light, cal bonds by application of
microwaves, and ultraviolet various food processing treat-
radiation. ments such as ionizing radia-
Electron Volt: Unit of measure of energy, de- tion, heat, and freezing. For
scribed as the energy gained by example, oxygen radicals de-
an electron as it travels through rived from the lysis of water
a 1 volt potential. molecules.
Gamma Rays: Photons of electromagnetic Radurization: Application of ionizing radia-
energy emitted by a radioactive tion to achieve low-grade pas-
isotope during its decay, such teurization, with typical doses
as cobalt-60. ranging from 0.75 to 2.5 kGy.
Gray: Unit of measure for absorbed X Rays: Photons of energy emitted as
dose. It is equivalent to 100 a result of collision of acceler-
rads, and to 1 joule of energy/ ated electrons with a heavy
kg of matter. metal.
Ionizing
Radiation: Type of radiation in which Basic Knowledge Questions
matter is exposed to high lev-
els of energy within the elec- 1. What are the units used to express dose and
tromagnetic spectrum, causing energy?
ionization of some of its mol- 2. List the types of radiation found in the electro-
ecules. magnetic spectrum of energy.

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 39


Copyright 2000, Aspen Publishers, Inc.
3. How are microorganisms affected by ionizing Critical Thinking Questions
radiation?
4. How do the following parameters affect micro-
1. You need to irradiate a large quantity of a prod-
bial inactivation in foods by ionizing radiation?
uct that is less than one inch thick. What irradia-
a) Oxygen content of the package
tion source would you use, and why?
b) Physical state of the food
2. Under what circumstances or conditions would
c) Age of the microorganisms
you expect a foodbome pathogen to develop
5. List the advantages ofirradiating foods by accel-
resistance to irradiation?
erated electrons vs. gamma rays.
6. What is the legal status of red meat irradiation in
the u.s.?
7. What procedures can be used to reduce or pre-
vent any damage to food quality by irradiation?

40 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 16

Low-Temperature Food Preservation and


Characteristics of Psychrotrophic
Microorganisms

Learning Objectives 5: Freezing of Foods and Freezing Effects.


a) Quick freezing
The reader will be able to: b) Slow freezing
1. Define the various classifications for organisms 6. Storage Stability of Frozen Foods.
that can grow or tolerate cold temperatures. 7. Effect of Freezing on Microorganisms.
2. Describe the minimum temperature requirements a) Freezing
for psychrotrophs and other types of organisms. b) Thawing
3. Describe how foods are frozen. 8. Some Characteristics of Psychrotrophs and
4. Understand the difference between slow and quick Psychrophiles.
freezing, and how these affect food quality and a) Increase in unsaturated fatty acid residues
storage stability. b) Synthesis of high levels of polysaccharides
5. Describe the effect of freezing and thawing on c) Pigment production
survival of microorganisms. d) Differential substrate utilization
6. Discuss the various characteristics of 9. The Effect of Low Temperatures on Microbial
psychrotrophic and psychrophilic microorgan- Physiologic Mechanisms.
isms in terms of their physiology and ability to a) Slower metabolic rate
conduct specific functions. b) More efficient transport of solutes
7. Discuss the effect of low temperature on micro- c) Larger cells
bial physiology and growth. d) More efficient synthesis of flagella
8. Explain why psychrotrophic organisms have a e) Aeration effect
low tolerance for temperatures above their opti- t) Increased requirement for organic nutrients
mal range for growth. 10. Nature of the Low Heat Resistance ofPsychro-
trophs.

Chapter Outline
Terms and Definitions
1. General Considerations.
2. Definitions. Blanching: Process by which foods are
3. Temperature Growth Minima. exposed briefly to either boil-
4. Preparation of Foods for Freezing. ing water or steam for the pur-

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Copyright 2000, Aspen Publishers, Inc.
pose of inactivating enzymes, Temperature
among other reasons. Coefficient: Expressed symbolically as
Bound Water: Water that is held by chemical "QIO'" it is defined as the ve-
bonds to other molecules, thus locity of a chemical reaction at
it is not available for microbial a given temperature + 10C,
use. divided by the velocity of the
Cryoprotectant: Chemical compound that low- reaction at that same tempera-
ers the freezing point of water. ture.
Eurypsychrotroph: Psychrotrophic organism that
does not form visible colonies Basic Knowledge Questions
until 6 to 10 days at 7C.
Free Water: Water that is not bound to other 1. List the purposes of blanching.
molecules, thus it is available 2. Compare and contrast quick vs. slow freezing in
for microbial use. terms of their effect on food quality and on
Lipid Solidification microorganisms.
Theory: Theory that seeks to explain 3. Discuss the physiological effects that freezing
the ability of some microor- has on microorganisms (not psychrotrophs or
ganisms to survive and grow at psychrophiles).
low temperatures. It states that 4. Describe the effect of freezing and thawing on
at such temperatures, more food.
unsaturated than saturated fatty 5. List at least three characteristics of psychrophilic
acids are synthesized and in- and psychrotrophic microorganisms that enable
corporated into the cell mem- them to grow at low temperatures.
brane. The presence of these 6. How does fatty acid composition relate to the
fatty acids make the membrane ability of cells to withstand low temperatures?
more fluid, preventing it from 7. Explain the effect oflow temperature on protein
freezing. synthesis.
Psychrophile: Microorganism that can grow
from below zero to 20C, with Critical Thinking Questions
an optimum growth range of
10 to 15C. 1. Describe what happens during "cold shocking"
Psychrotroph: Microorganism that can grow of bacterial cells. Under what circumstances dur-
between zero and 7C, produc- ing food processing could such an event take
ing visible colonies within 7 to place?
10 days. 2. Explain how low temperatures affect the uptake
Stenopsychrotroph: Psychrotrophic organisms that of solutes by psychrotrophic otganisms. What
form visible colonies within 5 would happen to the uptake of solutes if such an
days at 7C. organism were to suddenly be exposed to higher
temperatures?

42 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 17

High-Temperature Food Preservation


and Characteristics of Thermophilic
Microorganisms

Learning Objectives a) Water


b) Fat
The reader will be able to: c) Salts
1. Define the tenns "pasteurization" and "steriliza- d) Carbohydrates
tion." e) pH
2. Discuss how various extrinsic and intrinsic fac- f) Proteins and other substances
tors can affect survival of microorganisms to g) Numbers of organisms
heat treatments. h) Age of organisms
3. Discuss the phenomenon ofheat resistance within i) Growth temperature
the context of specific microorganisms and the j) Inhibitory compounds
mechanisms they employ to survive heating. k) Time and temperature
4. Defme "D value," "z value," "F value," and other 1) Effect of ultrasonics
tenns related to the measurement of heat resis- 3. Relative Heat Resistance of Microorganisms.
tance of microorganisms. a) Resistance of various types
5. Define "thennophiles" and provide examples of b) Spore resistance
such organisms. 4. Thennal Destruction of Microorganisms.
6. Describe how thennophiles can survive high- a) Thennal Death Time
temperature treatments, providing examples of b) D value
the mechanisms and factors that affect this. c) z value
7. Discuss spoilage ofcanned foods, including foods d) F value
that are of various acid content. e) Thennal Death Time curve
f) 12D concept
5. Some Characteristics of Thennophiles.
Chapter Outline a) Growth
b) Enzymes
I. Types of High-Temperature Treatments. c) Ribosomes
a) Pasteurization d) Flagella
b) Sterilization 6. Other Aspects ofThennophilic Microorganisms.
2. Factors Affecting Heat Resistance in Microor- a) Nutrient requirements
ganisms. b) Oxygen tension

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c) Cellular lipids heated at a given temperature.
d) Cellular membranes FValue: The time, usually in minutes,
e) Effect of temperature at 250F, required to destroy all
t) Genetics spores or vegetative cells of a
7. Canned Food Spoilage. particular organism. It is ob-
a) Causes of spoilage tained from a TDT curve by
b) Low acid foods noting the specific temperature
c) Acid foods at which no survivors are ob-
d) High acid foods served. It is used as a way to
measure the ease or difficulty
of sterilizing a particular food
Terms and Definitions product, with the higher the F
value the more difficult it is to
Acid Foods: Foods that have a pH between eliminate all cells of a specific
3.7 and 4.6. organism in a specific food.
"Bot Cook": Term for the heat treatment Flipper: Condition of a can in which
required to destroy 12 log of one end becomes convex due
Clostridium botulinum spores to expansion of the metal as a
in a food product. result of excessive heat, physi-
Commercial cal damage, or pressure at the
Sterilization: Process by which foods are other end.
exposed to levels of heat that Hard Swell: Condition of a can in which
are high enough to destroy all both ends appear bulged, and
vegetative cells, as well as neither can be dented by apply-
spores of Clostridium botuli- ing pressure. This is usually
num, such that no microbial due to release of CO2 and H2S
growth is observed during gas by spoilage microorgan-
room-temperature storage. isms, or to formation ofH2 gas
DValue: The time of heating, usually by a non-microbial chemical
expressed in minutes, required reaction between iron in the
to decrease a microbial popu- can and acidic compounds in
lation by 90%, or by 1.0 10gIO the food.
in a specific medium and at a High-Acid Foods: Foods that have a pH less than
specific temperature. It is an 3.7.
indirect measure of how heat Leakage-Type: Type of spoilage of canned
resistant a microorganism is. It foods caused by nonspore-
is also used to compare the heat forming microorganisms.
resistance of two different or- These organisms would not be
ganisms, with the higher the D able to survive the canning pro-
value the higher the resistance. cess, thus their presence is due
The D value can be calculated to faulty seams and small holes
by taking the reciprocal of the in the can.
absolute value of the slope of Low-Acid Foods: Foods that have a pH higher
a "survivor curve." A survivor than 4.6.
curve is generated by plotting Pasteurization: Process by which foods are
the relationship between the exposed to levels of heat that
number of cells in a medium are high enough to destroy all
vs. the time that the cells were disease-producing vegetative

44 INSTRUCTOR'S MANuAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
cells, or to reduce the number ZValue: The temperature, in degrees
of spoilage organisms. Farenheit, necessary to de-
Protoplast crease the TDT by 90%, or 1.0
Dehydration: Reduction of the water content 10glO" It is a constant that can
of the inside of a bacterial be used, along with the D value
spore as a way of increasing the of an organism at a particular
spore's resistance to heating. temperature, to determine what
Diminution of water lowers the D value would be at a dif-
heat conductivity thus protect- ferent temperature. It can be
ing the spore from damage. calculated by taking the recip-
Soft Swell: Condition of a can in which rocal of the absolute value of
both ends appear bulged, but the slope of a TDT curve. A
which can be dented if pressed TDT curve is generated by
with moderate force. plotting the relationship be-
Springer: Condition of a can in which tween the loglo of D values
one or both ends appear con- obtained at various tempera-
cave and spring out upon push- tures vs. the temperatures
ing, resulting in bulging. themselves.
Sterilization: Process by which foods are
exposed to levels of heat that
are high enough to destroy all Basic Knowledge Questions
viable microbial cells.
Thermal 1. How does the concentration of salt, fat, carbohy-
Death Time: Time, usually in minutes, nec- drates, and proteins affect microbial resistance to
essary for a heat treatment to heat?
kill a specific number of cells 2. How does pH affect microbial resistance to heat?
at a specific temperature. Ex- 3. How does the number and age of cells affect their
periments are conducted to survival to heat?
determine the D value at vari- 4. What physiological characteristics make some
ous temperatures for a particu- microbial cells more heat-resistant than others?
lar organism, and the relation- 5. Define "D" and "z" values, both in mathematical
ship between the 10gIO of each and in practical terms.
D value vs. temperature is plot- 6. What are the three types of enzymes found in
ted. From this curve one can thermophilic organisms?
obtain the "z value" of an or- 7. Describe low -vs. high-acid foods, giving ex-
ganism. amples of organisms that can spoil each type as
Thermophile: Microorganism that grows at a well as the condition of the product due to spoil-
minimum temperature of45C, age.
with an optimum range of 50
to 60C, and a maximum at
70C or above. Critical Thinking Questions
UHT: Ultra-high temperature pas-
teurization, refers to a process 1. Discuss the various theories explaining why bac-
whereby milk and milk prod- terial spores are more resistant to heating than
ucts are exposed to very high vegetative cells.
temperatures (140 to 150C) 2. Assume that you obtained the following data
for a few seconds, rendering after heating a bacterial spore suspension in cream-
them commercially sterile. style corn:

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 45


Copyright 2000, Aspen Publishers, Inc.
Minutes Number of survivors b) Calculate the time you would have to heat the
o 108 product in order to achieve a l2D process if
20 106 you started with 105 spores.
40 l~
a) Plot a survivor curve, properly label each
axis, and calculate the D value.

46 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 18

Preservation of Foods by Drying

Learning Objectives e) Freezing as a drying treatment


3. Effect of Drying on Microorganisms.
The reader will be able to: a) Bacteria
1. Define the various categories of dried foods, b) Yeasts and molds
according to moisture content. c) Death and injury
2. Describe how low-moisture foods are prepared, 4. Storage Stability of Dried Foods.
citing examples. a) Browning
3. Describe how various types of organisms are b) Relative humidity
affected by drying, and what causes microbial 5. Intermediate-Moisture Foods.
death. a) General definitions and history
4. Explain browning and how certain parameters b) Preparation of IMF
can be manipulated to minimize this effect dur- c) Microbial aspects of IMF
ing drying. d) Storage stability of IMF
5. Define intermediate-moisture foods (lMF) and e) IMF and glass transition
describe how these are processed.
6. Discuss spoilage ofIMF during storage, includ-
ing microbial and nonmicrobial factors. Terms and Definitions
7. Define glass transition and explain how it can be
used as an alternative to water activity measure- Alarm Water
ments. Content: The amount of water of a food,
above which mold growth will
take place.
Chapter Outline Case-Hardening: A condition of vacuum-dried
food in which a hard layer
1. Categories of Foods. forms at the surface of the
2. Preparation and Drying of Low-Moisture Foods. product, causing a slowing-
a) Fruits and vegetables down of the drying process,
b) Meat and of reconstitution at a later
c) Milk time.
d) Eggs Drum-Drying: A process in which milk-based

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Copyright 2000, Aspen Publishers, Inc.
and other products, such as causing moisture to escape. A
mashed potatoes, are dried by vacuum is applied, helping to
allowing it to coat the outside draw the moisture out more
of stainless steel drums, which efficiently, exhausting it from
have been heated to 100-150C the chamber to prevent it from
with steam. Product dries into condensing on the surface of
sheets, which peel off the sur- the product.
face of the drums and are cut Humectant: A compound that binds and
into flakes as they drop into an retains water.
auger at the bottom of the Hysteresis: The condition resulting from
drum-dryer. an excess in adsorbed moisture
ERR: Equilibrium Relative Humid- by a system above its equilib-
ity, refers to the water that is rium level.
present in a food product, Intermediate-
which can be lost due to des- Moisture Foods: Also known as IMF, refers to
orption. It can be defined foods that have a moisture con-
mathematically as the ratio of tent between 15 and 50% and
the partial pressure of water a water activity between 0.60
vapor generated by a food and 0.85. Examples are cakes,
sample, to the partial pressure jams, fruit juice concentrates,
of water vapor generated by and sweetened condensed
pure water. milk.
Freeze-Drying: A process in which egg-based Low-Moisture
products, coffee, juices, and Foods: Also known as LM, refers to
other products that are deemed foods that have a moisture con-
to be sensitive to heat are dried. tent of less than 25% and a
Food is frozen inside a cham- water activity between 0 and
ber. A vacuum is then applied, 0.60. Examples are freeze-
causing moisture in the food to dried coffee and crackers.
sublime (i.e. goes directly from Maillard Reaction: Also known as "non-enzy-
a solid to a gas). This results in matic browning," it refers to
drying of the product from the the development of dark pig-
surface to the inside until the ments as a result of reaction
moisture content is below 5%. between aldehydes and amino
The food develops a sponge- groups of sugar and protein
like structure due to space-gaps molecules. It is favored by high
caused by the escaping water temperature ana low moisture,
molecules. since these increase the con-
High-Temperature centration of reactants.
Vacuum-Drying: Method of drying particulate Maillard browning is seen to
foods such as citrus juices and occur fastest when the mois-
instant tea. The temperature of ture content of foods decreases
the food and the rate at which to about 15-20% during dry-
water is removed from it are ing, and is inhibited below 2%
controlled by heating it under moisture.
a vacuum. The food is placed Sorption Isotherm: The relationship between the
in a chamber on top of plates amount of water either
that are heated by circulating adsorbed or lost (desorbed) by
a heated fluid through them, a system (i.e. food), as a func-

48 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
tion of its relative humidity or 2. Explain the relationship between water activity
water activity. An example of and spoilage of foods. Provide examples of mi-
a food undergoing adsorption: croorganisms responsible for spoilage at various
crackers inside a humid cham- water activity levels.
ber. An example of a food un- 3. List at least two strategies that can be used to
dergoing desorption: fruit in- minimize quality changes during drying.
side a dry chamber. 4. Present the mathematical equation for calculat-
Sun Drying: The process of drying foods by ing water activity, based on Raoult's law of mole
exposing them to sunlight for fractions.
long periods of time. Raisins 5. List the four techniques that can be used to
(dried grapes) are an example. change the water activity during production of
Water Dynamics: Food components that respond intermediate-moisture foods.
to changes in the moisture and 6. Define Equilibrium Relative Humidity, and ex-
temperature of the food by plain its relationship with the amount of dis-
changing their physical state, solved solids in a food.
from a viscous glassy state to 7. What is "glass transition"?
a rubbery structure. This tran-
sition occurs at a specific tem-
perature, termed "T g". The Critical Thinking Questions
higher the moisture content,
the lower the Tg' thus this value 1. Research has shown that microorganisms are
could be used as an alternative more susceptible to the lowering of water activ-
to water activity for the pur- ity in foods that have been prepared by adsorp-
pose of predicting microbial tion, compared with foods that have been pre-
activity in foods. pared by desorption. Provide a theory that would
explain this phenomenon.
Basic Knowledge Questions 2. Compare freezing and drying offoods relative to
the effect of these treatments on survival and
1. Name some advantages of freeze-drying over growth of microorganisms, and on overall prod-
high-temperature vacuum-drying. uct quality.

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Copyright 2000, Aspen Publishers, Inc.
CHAPTER 19

Other Food Preservation Methods

Learning Objectives c) Effects ofHHP on specific foodborne organ-


isms
The reader will be able to: 2. Pulsed Electric Fields.
1. Discuss alternative, or unconventional methods a) Mode of action
of food preservation. b) Effects on microorganisms
2. Describe the principle behind high-hydrostatic 3. Aseptic Packaging.
pressure (HHP) processing of foods. a) Description of the process
3. Explain how HHP affects food components. b) Applications
4. Explain the mechanism of action ofHHP against 4. Manothermosonication(Thermoultrasonification).
microorganisms, citing specific examples of how a) Definitions
certain pathogens are affected. b) Effect on microbial contaminants in foods
5. Describe the process of applying pulsed electric
fields in food processing.
6. Discuss the principle behind the antimicrobial Terms and Definitions
effects of pulsed electric fields.
7. Discuss aseptic packaging as a food preservation Aseptic
technology, and how it is used in conjunction Packaging: Refers to the packaging of ster-
with conventional food processing treatments ile product'> into sterile con-
such as heat pasteurization. tainers. Sterility is maintained
8. Discuss manothermosonication, describing the by conducting all activities in
principles behind the process and how it affects a "clean room" environment in
microbial cells. order to prevent introduction of
microbial contaminants into
the food.
Chapter Outline Baroresistant: Describes organisms that are
relatively resistant to high pres-
1. High-Pressure Processing. sures (above atmospheric lev-
a) Description of the process els).
b) Some principles and effects ofHHP on foods Hydrostatic: Refers to the uniform distribu-
and organisms tion of a treatment in a men-

50 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
struum. When applied as part Basic Knowledge Questions
of the high-hydrostatic pressur-
ization of foods, the pressure 1. Cite some examples of other processes that have
increases instantaneously and been used in a hurdle approach in conjunction
is distributed evenly through- with HHP to reduce microbial contaminants in
out the product. foods.
Nisin: A bacteriocin produced by 2. How does microbial resistance to HHP relate to
some strains of Lactococcus resistance to other treatments?
iactis, which is inhibitory to 3. Describe the effect of water activity on effective-
certain microorganisms. ness ofHHP in reducing microorganisms.
PediocinAcH: A bacteriocin produced by the 4. Describe some examples of how HHP can affect
bacterium Pediococcus acidi- the quality of vegetables.
iactici, and which is inhibitory 5. What causes cell death by PEF?
to certain organisms. 6. List 3 advantages of aseptic packaging over other
Plasmin: Enzyme found in milk which processing methods.
hydrolyzes casein proteins. It 7. What is manothermosonication?
is inactivated by pasteuriza-
tion. Critical Thinking Question
Sonication: Treatment by high-frequency
sound waves. When applied 1. Experiments have shown that bacteria differ in
simultaneously with heat, the terms of their ability to survive HHP treatment.
combined treatment is known Refer to the list of organisms on page 378 and
as "manothermosonication". provide a theory that may help explain this phe-
nomenon.

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Copyright 2000, Aspen Publishers, Inc.
CHAPTER 20

Indicators of Food Microbial


Quality and Safety

Learning Objectives a) Criteria for indicator organisms


b) Examples of indicators
The reader will be able to: 2. Indicators of Food Safety.
1. Discuss the types of organisms and assays that a) Criteria for indicator organisms,
can be used to indicate product quality, listing b) Coliforms
the criteria that should be used in selecting such 1) Strains
orgamsms. 2) Growth
2. List the criteria used for selecting indicator or- 3) Detection and enumeration
ganisms for food safety. 4) Distribution
3. Define the term "coliform" and describe the 5) Coliform criteria and standards
general characteristics of this class of organisms 6) Some limitations for food safety use
(i.e. growth, distribution, etc). c) Enterococci
4. Discuss the use of coliforms as indicator organ- 1) Historical background
isms of food safety, including limitations that 2) Classification and growth requirements
should be considered. 3) Distribution
5. Describe enterococci and discuss their use as 4) Relationship to sanitary quality of foods
indicator organisms. d) Bifidobacteria
6. Discuss the use ofbifidobacteria and coliphages e) Coliphages
as indicators of food safety. 1) Assay
7. Discuss the correct use of fecal coliforms as 2) Utility for water
indicators of food safety, including the potential 3) Utility for foods
for their overuse. 3. Possible Overuse of Fecal Indicator Organisms.
8. Define predictive microbiology and discuss its 4. Predictive MicrobiologylMicrobial Modeling.
use as a method to predict spoilage and/or growth
of pathogenic microorganisms in foods.
Terms and Definitions

Chapter Outline Citrate Test: Assay used to indicate whether


a particular organism can uti-
1. Indicators of Product Quality. lize citrate as a sole carbon

52 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
source for growth. Most Es- Fecal Coliform: A gram-negative, nonspore-
cherichia coli strains are un- forming rod that can ferment
able to do so, thus yield a nega- lactose, producing gas within
tive reaction with this test. 48 hours at 44.5C. Such or-
Coliform: A gram-negative, nonspore- ganisms are used as indicators
forming rod that can ferment of the potential for contamina-
lactose, producing gas within tion of food and water with fe-
48 hours at 35C. Such organ- ces, and thus of the possible
isms are used as indicators of presence of intestinal patho-
the potential for contamination gens. It is considered by some
of food and water with feces, as a more accurate indicator of
and thus of the presence of in- fecal contamination than
testinal pathogens. coliforms.
Coliphage: A virus that infects coliform HACCP: Hazard analysis critical control
bacteria. Coliphages are found point, refers to a management
in close proximity with system that seeks to control
coliforms, thus have been used foodborne hazards through
as indicators of the presence of prevention. It can be applied
these bacteria. during production, processing,
Diacetyl: A compound produced from distribution, and preparation of
fermentation of milk by food, with the goal of minimiz-
heterofermentative lactic acid ing or eliminating health haz-
bacteria. It imparts specific fla- ards to the consumer.
vor notes and buttermilk aroma Hfr Strain: A cell that has integrated the F
to fermented products. It can factor into its chromosomal
also be used as an indicator of DNA. Such a strain is able to
spoilage of foods where fer- initiate conjugation, and thus
mentation by bacteria is con- transfer other plasmids it may
sidered undesirable (i.e. fruit have to other cells.
juices). Indole Test: Assay used to indicate the pro-
F+ Strains: Cells that have the F factor, duction of indole by a micro-
piece of DNA containing genes organism during metabolism.
that enable the cell to transfer Typically, Escherichia coli will
genetic information to other provide positive results while
cells through conjugation. This other coliforms such as
is accomplished by the synthe- Enterobacter will not.
sis of a structural appendage Methyl Red Test: Assay used to indicate whether
known as a pilus, which serves acidic compounds have been
as a conduit to physically ex- produced by a microorganism,
change genetic material (in the resulting in the lowering of pH
form of plasmids) with cells of to below 4.5. Most strains of
the same or similar species. F+ Escherichia coli are positive
strains are also referred to as for this reaction.
"male" cells. If the F factor Murein: Molecule that forms part of the
becomes integrated into the bacterial cell wall.
chromosome of the F+ cell, the PFU: Plaque-forming unit, it refers
cell becomes an Hfr strain, en- to the empty space left on the
abling it to begin preparations surface of an agar plate by a
for the conjugation event. colony of bacteria after being

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 53


Copyright 2000, Aspen Publishers, Inc.
destroyed by phages. The pres- during metabolism. Escheri-
ence of PFUs is an indication chia coli strains are unable to
that the bacteria on the plate produce this compound.
were destroyed by phages, thus
it serves to identify those bac-
teria to group or species. Basic Knowledge Questions
Scombroid
Poisoning: A foodbome illness caused by 1. List all the criteria that should be used for select-
consumption of seafood con- ing a good indicator organism for food quality,
taining high levels of hista- and one for food safety.
mine. This compound is pro- 2. Explain how spoilage offood can be determined
duced by microorganisms dur- without microbial testing.
ing metabolism of the amino 3. List the four genera of coliform bacteria.
acid histidine. 4. What are the two basic questions that should be
Selective Culturing: Culturing method that utilizes considered in establishing enumeration standards
a medium that contains certain for E. coli and/or coliforms in food?
ingredients that will inhibit 5. List the main characteristics of enterococci that
specific organisms, allowing make them useful indicator organisms.
(selecting for) the growth of 6. What are coliphages most useful for?
others. An example is Brilliant 7. Discuss the relationship between P+ bacterial
Green Lactose Broth, which strains and coliphages.
contains the dye brilliant green
that inhibits most gram-posi- Critical Thinking Questions
tive bacteria, thus selecting for
gram-negative organisms, 1. Is it a good idea to set limits on the number of
mostly of intestinal origin. coliforms that can be found in meat? Why or why
Voges-Proskauer not?
Test: Assay used to indicate whether 2. What indicator organisms would you choose to
a certain microorganism pro- determine whether fecal contamination of fresh-
duces acetic methylcarbinol water catfish had occurred? Defend your choices.

54 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


Copyright 2000, Aspen Publishers, Inc.
CHAPTER 21

The HACCP System and Food Safety

Learning Objectives 3) Principle 3: establish critical limits


4) Principle 4: establish procedures to monitor
The reader will be able to: CCPs
1. Discuss the concept ofthe Hazard Analysis Critical 5) Principle 5: establish corrective actions
Control Point system (HACCP). 6) Principle 6: establish procedures forveri-
2. Describe the prerequisite programs that must be fication
in place before HACCP can be implemented. 7) Principle 7: establish effective record-
3. Define various terms related to the HACCP sys- keeping systems
tem, such as "hazard", "CCP", etc. e) Flow diagrams
4. List the seven principles ofHACCP. f) Application ofHACCP principles
5. Describe the seven principles ofHACCP. 1) Principle 1: hazards and risks
6. Discuss how the seven principles would be ap- 2) Principle 2: CCPs
plied in the manufacture of a particular example, 3) Principle 3: critical limits
cooked beef patties. 4) Principle 4: monitoring CCPs
7. Discuss some limitations of the HACCP system. 5) Principle 5: corrective actions
8. Defme "microbiological criteria" for foods. 6) Principle 6: verification
9. Discuss the various classes of sampling plans. 7) Principle 7: recordkeeping
10. List some of the criteria that have been estab- g) Some limitations of HACCP
lished for various food products. 2. Microbiological Criteria.
a) Definitions
Chapter Outline b) Sampling plans
c) Microbiological criteria and food safety
1. Hazard Analysis and Critical Control Point Sys- d) Microbiological criteria for various products
tem. e) Other criteria!guidelines
a) Introduction and history
b) Prerequisite programs Terms and Definitions
c) Definitions
d) HACCP principles Advisory
1) Principle 1: assess hazards and risks Criterion: A microbiological specifica-
2) Principle 2: determine CCPs tion for an end product which

INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY 55


Copyright 2000, Aspen Publishers, Inc.
is intended to increase assur- public health significance only.
ance that hygienic significance However, limits could also be
has been achieved. It may also set for nonpathogenic organ-
refer to a microbiological isms.
guideline applied in a food es- Monitoring: A planned sequence of obser-
tablishment at a point during vations or measurements de-
or after processing in order to signed to produce an accurate
monitor hygiene. record and to determine if a
Control Point: Any point in a specific food critical limit is being violated.
system where loss of control Standard: Part of a law or regulation that
does not lead to an unaccept- is enforceable by a particular
able health risk. regulatory agency. In the case
Critical Control of microbial standards, it is the
Point: CCp, refers to any point or pro- limit of the number of organ-
cedure in a food system where isms, or of positive samples,
control can be exercised and a allowable by that agency.
hazard can be minimized or Validation: The collection and evaluation
prevented. of scientific and technical in-
Critical Limit: One or more prescribed toler- formation to determine
ances that must be met to en- whether the HACCP plan,
sure that a CCP effectively when properly implemented,
controls a microbiological will effectively control the haz-
health hazard. ards.
CCP Decision Tree: A sequence of questions to as- Verification: Methods,procedures,andtests
sist in determining whether a used to determine whether the
control point is a CCP. HACCP plan is being properly
Corrective Action: Procedures that must be fol- implemented.
lowed when a deviation of the
critical limit occurs.
HACCPPlan: A written document that delin- Basic Knowledge Questions
eates all the procedures to be
followed in implementing the 1. What are the three categories of hazards one
HACCP system. could find in food?
Hazard: Any biological, chemical, or 2. List the seven principles ofHACCP.
physical property of a food that 3. What are some examples of verification docu-
may cause an unacceptable risk ments?
to the health of the consumer. 4. List some of the limitations ofHACCP.
Examples are: bacteria (bio- 5. List the 5 components of the defmition of a
logical), pesticides (chemical), microbiological criterion, according to the Co-
glass (physical). dex Alimentarius Commission.
Mandatory 6. What are the main differences between a 2-c1ass
Criterion: A microbiological standard and a 3-c1ass sampling plan?
that normally contains limits 7. List the types of organisms for which standards
for pathogenic organisms of have been established.

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Critical Thinking Questions 2. Which of the following sampling plans is more
conservative in terms of not allowing hazardous
1. Why is recording the time/temperature of pro- product from reaching the consumer? Why?
cessing of a food product a better monitoring Plan A: n= 10, c=5 and m= 103
method than enumerating the organisms that may Plan B: n=1000, c=100 and m=104
have survived the process?

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CHAPTER 22

Introduction to Foodborne Pathogens

Learning Objectives 2) Listeria monocytogenes


b) Gram-negative bacteria
The reader will be able to: 1) Salmonella
1. Explain how foodbome pathogens are transmit- 2) Escherichia coli
ted via the "fecal-oral" route. 3) Yersiniae
2. List and describe the various hurdles that foodbome 4) Shigellae
pathogens must overcome to cause disease in 5) Vibrios
humans. 3. Summary.
3. Discuss the acid tolerance response.
4. Discuss how gram-positive bacteria cause dis-
ease, listing examples of significant foodbome Terms and Definitions
pathogens.
5. Describe the general mechanisms by which List- (note that the following do not include terms compiled
eria monocytogenes causes illness. in the textbook in Table 22-3)
6. Discuss the virulence factors of salmonellae,
Escherichia coli, yersiniae, shigellae, and vibrios, Adenylate Cyclase: Enzyme that catalyzes a reac-
explaining how these serve to cause disease. tion in which ATP is broken
down to cAMP. Certain bacte-
rial toxins, such as cholera
Chapter Outline toxin, activate this reaction,
resulting in accumulation of
1. Introduction. cAMP by the intestinal tissue
a) The fecal-oral transmission offoodbome patho- cell. This disrupts the transport
gens. of ions across the cell mem-
b) "Universal" requirements brane, causing eftlux of water
c) Attachment sites to the outside of the cell. The
d) Sigma factors and the acid tolerance response resulting effect is profuse wa-
2. Pathogenesis. tery diarrhea.
a) Gram-positive bacteria ATR: Acid tolerance response,
1) Examples mechanism of microbial cells

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that enables them to survive back into the abdominal cav-
low-pH conditions. It is trig- ity of the animal. After some
gered by exposure of the cells time, the animal is sacrificed
to pH levels below 7.0 for spe- and the intestines examined to
cific periods of time. see if the ligated loop has in-
Competitive creased in size. This is an indi-
Exclusion: Mechanism whereby harmless cation that water diffused into
bacteria residing in the intesti- the loop as a result of toxin
nal tract of humans and ani- being present in the sample.
mals serve to exclude patho- Kupffer Cells: Macrophages residing in the
gens by taking up all available liver.
attachment sites on the intesti- LLO: Listeriolysin 0, a compound
nal mucosa. produced by Listeria monocy-
Cytokines: Compounds produced by im- togenes, which lyses red blood
mune system cells such as cells. It also enables the patho-
PMNs. They serve a multitude gen to survive inside of mac-
of functions, including the sig- rophages, and helps remove
naling for specific T cells to mucus from the surface of the
help eliminate infected body intestinal mucosa. This enables
cells. Examples are the the pathogen to adhere to the
interleukins and Tumor Necro- surface better. It is considered
sis Factor. one of the main virulence fac-
Endotoxins: Components ofthe outer mem- tors of this organism.
brane of gram-negative bacte- Lysogen: A virus that integrates its ge-
ria, which elicit a toxic effect netic material into the chromo-
on the host. some of the host cell. The lat-
Exotoxic: Toxic substances produced by ter is not lysed by the virus, but
some pathogenic organisms, rather continues to grow and
which are secreted by the cell. divide, replicating the viral
Fecal-Oral Route: Route of transmission of genes as it replicates its own
foodborne pathogens, where chromosome.
organisms residing in feces of Macropinocytosis: The uptake of foreign bodies
humans or animals contami- (i.e. bacteria) by mammalian
nate the fingers, or drinking tissue cells, such as intestinal
water, or utensils used for food epithelial cells.
preparation, or food itself. Microvilli: Epithelial cells lining the small
These then serve as vehicles to intestine, which have protrud-
carry pathogens into the body ing, "finger-like" appendages
through contact with the on the side facing the intesti-
mouth. nallumen. This helps facilitate
Ileal Loop Test: Bioassay used to determine the nutrient uptake by increasing
presence oftoxigenic bacterial the surface area of the cell.
cells. A rabbit is anesthetized, Multilocus
and its intestines are pulled out Enzyme
and ligated at various points to Electrophoresis: Technique used to examine dif-
create sacs, or loops. The ferences among bacteria in a
sample homogenate is injected population. Since each indi-
into one of the ligated loops vidual enzyme produced by an
and the intestines are placed organism represents a discrete

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and independent characteristic Septicemia: Infection of the circulatory
of that particular isolate, this system by virulent microor-
technique can be used to dif- ganism, which can then reach
. ferentiate among isolates of the body tissues to begin new in-
same bacterial species. The fections.
method consists of subjecting Sigma Factor: Protein subunit of the enzyme
cell extracts to electrophoresis RNA polymerase which plays
in non-denaturing starch gels. a role in translation of genetic
Enzymes travel down the gel material during synthesis of
according to their electric proteins. An organism can have
charge, and are then reacted several sigma subunits, with
with their substrate. The posi- each one being responsible for
tion of the enzyme on the gel translation of specific genes.
is thus determined by the ap- Stx: Shiga-like toxin, produced by
pearance of a visible reaction specific strains of Escherichia
product. Variations in the mo- coli, such as OI57:H7.
bility of the enzyme down the Suckling Mouse
gel reflects amino acid substi- Assay: Mice are inoculated intragas-
tutions that change the charge trically with a preparation of
of the protein. These changes the suspected organism, and
are due to variations in the the animals are sacrificed af-
chromosomal genes encoding ter 4 hrs. The ratio of intesti-
each enzyme. Thus, the pattern nal tract weight to body weight
for a specific isolate represents is measured. A ratio >0.83 is a
a particular "multilocus geno- positive test for the presence of
type". toxin. This assay is carried out
nvCJD: New variant Creutzfeldt-Jakob to detect heat-stable toxin pro-
Disease, it is suspected to be duced by Enterotoxigenic E.
caused by the prion that causes coli strains, as well as other
"mad cow" disease. pathogens.
PMNs: Polymorphonuclear leuko- T Cell-Mediated
cytes, are white blood cells Immunity: Mechanism of the immune sys-
whose main function is to tern in which T cells are in-
phagocytize foreign bodies. volved. Examples are T helper
They circulate through the cells, T suppressor cells, Natu-
body via the lymphatic system. ral Killer (NK) cells, and Cyto-
PMN s produce enzymes and toxic T Lymphocytes (CTLs).
oxygen radicals that help inac- In general, T cells are involved
tivate and kill the ingested in- in inducing the destruction of
truder, and release compounds tissue cells that have been in-
known as cytokines. vaded (by pathogens such as
RpoS: Sigma factor that controls the bacteria and viruses) by NK and
induction of specific genes that CTLs, and help regulate pro-
help the cell survive environ- duction of antibodies by B cells.
mental stressors, such as heat, Virulon: Set of genes that control and
starvation, and low pH condi- code for specific virulence fac-
tions. tors in bacteria.

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Basic Knowledge Questions 6. How do scientists think Enterohemorrhagic
(EHEC) E. coli strains developed?
1. List the five hurdles that a microorganism must 7. List and explain the three types of secretory
overcome to cause disease. systems of gram-negative bacteria.
2. What is the Sigma38 factor?
3. List the sites of infection of the following patho- Critical Thinking Questions
genic organisms:
Helicobacter 1. If you were to construct the ultimate foodbome
Hepatitis A pathogen, what virulence mechanisms would it
Vibrio cholerae have, and why?
Trichinella spiralis 2. Present at least two examples of how immune
4. What is LLO and how does it help Listeria system cells are used by some pathogenic bacte-
monocytogenes cause disease? ria to further their ability to cause disease in the
5. What are pathogenicity islands? host.

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CHAPTER 23

Staphylococcal Gastroenteritis

Learning Objectives 6. Temperature Growth Range.


7. Effect of Salts and Other Chemicals.
The reader will be able to: 8. Effect of pH, Water Activity, and Other Param-
1. Discuss coagulase-positive and coagulase-nega- eters.
tive strains ofS. aureus and their role in foodborne a) NaCI and pH
illness. b) pH, aw ' and temperature
2. Discuss the distribution of S. aureus in nature c) NaN0 2,Eh, pH, and temperature of growth
and the types of foods where it can be found. 9. Staphylococcal Enterotoxins: Types and Inci-
3. Discuss the growth requirements of this organ- dence.
Ism. a) Toxin types
4. List and discuss the various toxin types of S. b) Chemical and physical properties
aureus, including their chemical and physical c) Production
characteristics. d) Mode of action
5. Discuss the conditions in which toxin production 10. The Gastroenteritis Syndrome.
is optimal. 11. Incidence and Vehicle Foods.
6. Discuss the disease and symptoms caused by 12. Ecology of S. aureus Growth.
intoxication with S. aureus. 13. Prevention of Staphylococcal and Other Food-
7. Discuss strategies to prevent staphylococcal Poisoning Syndromes.
foodborne illness.

Terms and Definitions


Chapter Outline
Coagulase: Enzyme produced by some sta-
1. History. phylococci which causes
2. Species of Concern in Foods. clumping or clotting of blood
a) Coagulase-positive strains plasma. Production of this en-
b) Coagulase-negative strains zyme was considered as an in-
3. Habitat and Distribution. dicator of the ability of strains
4. Incidence in Foods. to produce enterotoxin. How-
5. Nutritional Requirements for Growth. ever, several coagulase-nega-

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tive strains have been found to well and the plate is incubated.
produce toxin. As the antigen diffuses into the
De novo Protein gel, it forms a precipitate when
Synthesis: Synthesis of proteins by syn- it reacts with antibodies in the
thesizing amino acids "from gel. The size of the precipitate
scratch," as compared with ring around the well is indica-
synthesis of proteins utilizing tive of the antigen concentra-
amino acids already present. tion.
ELISA: Enzyme-linked immunosor- MHC: Major histocompatibility com-
bent assay, refers to a method plex, refers to protein mol-
used to detect the presence of ecules present on the surface
specific organisms or toxins in of animal cells. T cells of the
a food or clinical sample. It is immune system have receptors
based on the use of antibodies that recognize either foreign
specific for a particular anti- cells (containing MHC Class I
gen (microorganism or toxin). molecules) or body cells (con-
If the sample contains the an- taining MHC Class II mol-
tigen, the antibodies will bind ecules). MHC class II mol-
to it, anchoring it to the surface ecules not only help T cells
of the microtiter plate. After identify other body cells, but
washing any unbound antigen, they also serve to aid in the
a second set of antibodies spe- communication between im-
cific for the antigen is then mune system cells, such as be-
added, which are themselves tween macrophages and T
linked to an enzyme. These cells.
will bind to the antigen, if TNAse: Thermonuclease (also known
present on the plate. Addition as heat-stable nuclease), refers
of substrate will result in an to an enzyme produced by
enzyme-catalyzed reaction, some staphylococci which di-
producing a color compound. gests DNA. It is relatively re-
Emesis: Vomiting. sistant to heating.
Enterotoxin: Toxin produced by a microor-
ganism that affects the intesti-
nal tract.
Basic Knowledge Questions
Gel Diffusion
Assay: Also known as "Ouchterlony
test," is a method used to con- 1.Explain what is meant by coagulase-positive or
firm the presence of a toxin or coagulase-negative, and list several species of
other antigen in a sample. A staphylococci under each of these categories.
layer of agar is poured into a 2. What effect does water activity, pH, and tem-
dish or a glass plate. The gel is perature have on growth and toxin-production of
usually pre-mixed with a solu- S. aureus?
tion of antibodies specific for 3. Based on the types of conditions in which the
the antigen to be examined. organism grows, what types of food products are
After solidifying, holes 1-2 prone to harbor this organism?
mm in diameter are punched 4. List the toxin types produced by S. aureus, and
into the gel slab to provide describe the general properties of these, compar-
wells for the antigen samples. ing them according to heat resistance and quanti-
Antigen solution is added to a ties produced.

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5. List the symptoms and state the onset of illness Critical Thinking Questions
caused by S. aureus.
6. What food products are more frequently associ- 1. How does pH and water activity affect production
ated with staphylococcal food poisoning? of specific toxin types by S. aureus?
7. List, in order, the five most common practices 2. What is the mode of action of staphylococcal tox-
that yield to staphylococcal illness. ins? What experiment(s) would you conduct to de-
termine whether this is correct?

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CHAPTER 24

Food Poisoning Caused by Gram-positive


Sporeforming Bacteria

Learning Objectives b) Characteristics of the organism


c) The enterotoxin
The reader will be able to: 1) Toxin types
1. Discuss the general characteristics of Clostridium 2) Mode of action
perfringens and where it is found in nature. d) Vehicle foods and symptoms
2. Describe the toxins produced by C. perfringens e) Prevention
and how they operate to cause disease. 2. Botulism.
3. Discuss the symptoms of the foodborne disease a) History
caused by C. perfringens and how this organism b) Distribution of C. botulinum
can be transmitted via foods. c) Growth of C. botulinum strains
4. Discuss how to prevent foodborne illness by this d) Ecology of C. botulinum growth
organism. e) Concerns for Sous Vide and related food
5. Describe the general characteristics of Clostridium products
botulinum and where it is found and distributed f) Nature of the botulinal neurotoxins
in nature. g) The adult botulism syndrome: incidence and
6. Discuss the toxin types of C. botulinum and their vehicle foods
mechanism of action. h) Infant botulism
7. Describe how certain food products and process- 3. Bacillus cereus Gastroenteritis.
ing methods (i.e. sous vide) could increase the a) General characteristics
risk of botulism. b) B. cereus toxins
8. Define and describe infant botulism. c) Diarrheal syndrome
9. Discuss the characteristics of Bacillus cereus. d) Emetic syndrome
10. Describe the types oftoxin produced by B. cereus
and the disease syndromes they cause.
Terms and Definitions

Chapter Outline Boticin E: Bacteriocin produced by cer-


tain spoilage organisms that
1. Clostridium perfringens Food Poisoning. inhibits type E Clostridium
a) Distribution of C. perfringens botulinum cells.

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Cereolysin: Hemolysin produced by Bacil- plays a role as a virulence fac-
lus cereus. It is similar to tor in causing disease.
perfringolysin but does not Sous Vide: Processing treatment whereby
, playa virulence role. foods are cooked at relatively
Cereulide: A water-insoluble peptide that low temperatures under
is structurally similar to the vacuum in order to retain fla-
antibiotic valinomycin. It is vors and improve the overall
relatively heat-stable, and is organoleptic quality. It is be-
produced by emetic strains of lieved that this process may
B. cereus. provide the right conditions
Dialysis Sac: A method used to produce and such that growth of C. botuli-
separate spores of C. num cells would take place
perfringens from vegetative preferentially, since their com-
cells of the organism. Gener- petitors are destroyed during
ally, cells are inoculated into cooking.
liquid medium lacking certain Tenesmus: Urge to defecate with inability
nutrients, and placed inside a to empty the bowels at the
dialysis sac with a pore size same time.
smaller than that of spores. The Terminal/Sub-
sac is placed inside a container Terminal Spores: Terminal spores are those that
with the same growth medium. develop at one end of a veg-
During incubation, some ofthe etative cell, making the sporu-
cells will sporulate due to the lated organism appear as a
lack of certain nutrients. The club-shaped body under the
cells that do not sporulate will microscope. Sub-terminal
migrate across the dialysis spores are those that develop
membrane into the medium on between the center and the end
the outside, leaving the spores of the vegetative cell.
inside the sac. Trypsin: Enzyme that digests some pro-
Hemolysin BL: Hemolytic peptide consisting teins at specific amino acid
of three parts (components B, sites. It causes the breakage of
Ll' and L), which causes lysis certain chemical bonds in
of red blood cells, cellular ly- botulinal progenitor toxin, ac-
sis, and necrosis of epithelial tivating it.
cells, among other effects.
Lactic Antagonism: The inhibition of certain patho- Basic Knowledge Questions
genic bacteria by lactic acid
bacteria, such as Lactobacillus 1. List the toxins produced by C. perfringens.
and Lactococcus. 2. What are the factors that affect heat-resistance of
Mouse LD: Mouse lethal dose, it refers to spores?
the amount of a toxin required 3. What is the relationship between enterotoxin
to kill a particular number of production and sporulation of C. perfringens?
mice. It can be expressed as 4. List at least 3 recommendations that should be
mouse LD 50' which refers to followed to avoid foodbome illness outbreaks
the concentration of toxin re- due to C. perfringens.
quired to kill 50% of a certain 5. What are the various groupings oftoxin-produc-
number of mice. ing C. botulinum strains, and what are some of
Perfringolysin: Hemolytic compound pro- their characteristics? Include in your answer relative
duced by C. perfringens which heat resistance, growth temperature and habitat.

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6. Discuss the mechanism of action of botulinal in which mice are injected intraperitoneally to
toxin and the symptoms it elicits. induce symptoms. You are asked to use this
7. Describe the diarrheal and emetic syndromes assay to determine whether a particular food
caused by B. cereus, including the toxins respon- contains type A toxin. Design an experiment,
sible and symptoms experienced. including the controls that you would use, to
test the sample for the presence of this type of
Critical Thinking Questions toxin.
2. Speculate as to why the diarrheal syndrome caused
1. In testing for the presence of botulinal toxin in by B. cereus does not require as many cells to be
a food sample, the mouse bioassay is employed, present as with the emetic syndrome.

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CHAPTER 25

Foodborne Listeriosis

Learning Objectives Chapter Outline

The reader will be able to: 1. Taxonomy of Listeria.


1. Discuss the taxonomical classification of List- a) General characteristics
eria species. b) Serotypes
2. Describe the nutritional requirements of Listeria c) Subspecies typing
species, including the effect that various param- 2. Growth.
eters have on growth of these organisms (i.e. a) Nutritional requirements
temperature, pH, ~). b) Effect of pH
3. Discuss how Listeria species are distributed in c) Combined effect of pH and NaCI
the environment, and the types offoods that have d) Effect of temperature
been found to harbor these organisms, especially e) Effect of aw
L. monocytogenes. 3. Distribution.
4. Describe the thermal properties of L. a) The environment
monocytogenes, particularly within the context b) Foods and humans
of milk pasteurization, and including the effect c) Prevalence
of heat shock on acquired thermotolerance. 4. Thermal Properties.
5. Describe the mechanisms of virulence of L. a) Dairy products
monocytogenes, including how it invades and b) Nondairy products
survives within phagocytic cells. c) Effect of sublethal heating on thermotolerance
6. Discuss the methods used to determine infec- 5. Virulence Properties.
tious dose of this pathogen. a) Listeriolysin 0 and Ivanolysin 0
7. Describe the symptoms of listeriosis and the b) Intracellular invasion
disease syndromes it causes. c) Monocytosis-producing activity
8. Discuss the mechanisms of resistance to listerio- d) Sphingomyelinase
sis in humans, and the regulatory limits that have 6. Animal Models and Infectious Dose.
been imposed on ready-to-eat foods in various 7. Incidence and Nature of the Listeriosis Syn-
countries to prevent outbreaks of this disease. dromes.

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a) Incidence added by streaking the culture
b) Source of pathogens perpendicularly to the listerial
c) Syndromes inoculum. After incubation, L.
8. Resistance to Listeriosis. monocytogenes will cause
9. Persistence of L. monocytogenes in Foods. hemolysis of the red blood
10. Regulatory Status of L. monocytogenes in Foods. cells in the agar, with the reac-
tion being magnified at the
points where the two organ-
Terms and Definitions isms intersect.
Filopodium: Structure similar to pseudo-
ActA: Surface protein of L. mono- pods which helps in the trans-
cytogenes, which induces the fer of L. monocytogenes from
production of actin filaments one phagocytic cell to another
that help propel the cell across the membrane.
through the cytoplasm of ph- FIM: Factor-increasing monocyto-
agocytes en route to the mem- poiesis, refers to a compound
brane. Once there, the bacte- produced by macrophages af-
rium can be transmitted to an- ter ingestion of L. monocyto-
other phagocyte through the genes cells. This factor is se-
formation of vacuoles that creted and stimulates produc-
transport the organism across tion of more macrophages by
the membrane of one cell to the immune system.
that of another. Gamma-
Anton Test: Assay used to confirm the Interferon: Cytokine compound produced
presence of invasion-type of by T cells, it elicits the produc-
microorganisms, such as L. tion of other cytokines (i.e.
monocytogenes. A culture of Interleukin-l). This induces
the suspected organism is Natural Killer cells (a type of
added to the eye of a rabbit or T cell) to destroy macrophages
guinea pig. A positive reaction infected with organisms like L.
is depicted by the development monocytogenes.
of conjunctivitis. Granulocytes: Polymorphonuclear leuko-
Bacteremia: The presence of bacteria in the cytes, or white blood cells of
bloodstream. the immune system. Examples
Beta Hemolysis: Lysis of red blood cells in agar are neutrophils, basophils, and
medium, seen as a complete eosinophils. They contain
zone of clearing surrounding granules within the cytoplasm,
the colonies of bacteria respon- which are used to digest invad-
sible for it. L. monocytogenes ing microorganisms during
is an example. phagocytosis.
CAMP Test: Christie, Atkins, Munch- HTST: High-temperature, short-time
Petersen, refers to a biochemi- pasteurization, typically car-
cal test that is used to confirm ried out in milk at 71.7C for
the identity of Listeria mono- 15 seconds.
cytogenes. The suspected cul- ILO: Ivanolysin 0, refers to a com-
ture is streaked on an agar plate pound produced by Listeria
containing sheep blood. An in- ivanovii. It is similar to
oculum of Rhodococcus equi listeriolysin 0, and causes a
or Staphylococcus aureus is complete zone of hemolysis

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immediately around red blood Nude Mice: Mice that are deficient in their
cells. ability to produce T cells. They
Internalins: Proteins found on the surface are born without hair.
of L. monocytogenes, required Phagolysosomes: Membrane-bound cavities in-
for the organism to invade epi- side the cytoplasm of phago-
thelial cells (Internalin A) and cytes. They are formed by the
mouse hepatocytes in culture fusion of a phagosome (the
(Internalin B). cavity formed when a phago-
LALAssay: Limulus amoebocyte lysate cyte engulfs and internalizes
assay is a method used to de- an invading microorganism
tect LPS of gram-negative bac- into its cytoplasm) and a lyso-
teria. Aliquots of food some (cytoplasmic vacuole
homogenates are added to containing lytic enzymes that
LAL reagent, which is a com- help in digestion of invading
ponent found in amoebocytes bacteria). Most engulfed mi-
of the horseshoe crab. It reacts croorganisms are destroyed in-
with LPS of the bacteria. side phagolysosomes, with the
LLO: Listeriolysin 0, refers to the notable exception of L.
main virulence factor of L. monocytogenes.
monocytogenes. It has a mo- Phospholipases: Enzymes produced by L.
lecular weight of 60,000 monocytogenes that serve to
daltons and is responsible for partially digest the membrane
the hemolytic activity of the of phagocytic cells to achieve
organism. It causes death of vacuole formation in order to
phagocytes that have ingested transport the organism to an-
the bacteria. other cell.
LTLT: Low-temperature, long-time Septicemia: Infection of the bloodstream,
pasteurization, also known as such as caused by bacteria.
vat-pasteurization, it is typi- Sphingomyelinase: Enzyme produced by some
cally carried out at 62.8C for Listeria species, responsible
30 minutes. for incomplete hemolysis ob-
Lipoteichoic served near the Rhodococcus
Acid: Component of the cell wall equi colonies in the CAMP
peptidoglycan of some bacte- test.
rial cells. "Zero Tolerance": Policy that allows no cells of a
Meningitis: Inflammation of the meninges, particular pathogen to be de-
or the membrane surrounding tected in 25- or 50-g samples
the brain and spinal chord. offoods. For example, the U.S.
Monocytes: Phagocytic cells of the im- has a "zero tolerance" regard-
mune system, also knows as ing L. monocytogenes in ready-
macrophages. In general, the to-eat foods.
term "monocyte" is used to
refer to a specific type of mac-
rophage: those that are in the Basic Knowledge Questions
circulatory system. The term
"macrophages" refers to those 1. List the five species of Listeria.
that are fixed to specific tissues 2. Describe the habitats of Listeria species, and
(i.e. liver). their prevalence in foods.

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3. How is Listeria disseminated in the environ- Critical Thinking Questions
ment?
4. What are LLO and ILO? 1. What is the role of T cells in the initiation of
5. Describe the effects of infection with this patho- infection by L. monocytogenes, as well as in
gen on both healthy and immuno-compromised resisting infection by this organism?
patients. 2. Various studies have been conducted to prove or
6. List some of the sources of infection of this disprove the notion that L. monocytogenes can
organism in nature, and in the food processing survive pasteurization. Review the section of this
plant. chapter in which this topic is discussed, and
7. What is the current policy regarding L. present a theory as to why there is such a dispar-
monocytogenes in the U.S. and other countries? ity. What type of experiment would you conduct
to settle the issue one way or another?

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CHAPTER 26

Foodborne Gastroenteritis Caused by


Salmonella and Shigella

Learning Objectives a) Taxonomy


b) Serotyping
The reader will be able to: c) Distribution
1. Describe the taxonomy of salmonellae and how 1) Animals
it has changed over the years. 2) Animal feeds
2. Describe the methods used to serotype the organ- 3) Food products
ism, and list the major types that are involved in d) Growth and destruction of salmonellae
foodbome illness. e) The Salmonella food-poisoning syndrome
3. Discuss the distribution of salmonellae in nature, f) Incidence and vehicle in foods
including food products. g) Prevention and control of salmonellosis
4. Describe the general characteristics of salmonel- h) Competitive exclusion to reduce salmonellae
lae, including growth and their susceptibility to carriage in poultry
processing treatments. 2. Shigellosis.
5. Discuss the virulence factors of salmonellae and a) General characteristics of Shigella species
their role in causing disease. b) Foodbome cases
6. Discuss the types of foods in which salmonellae c) Virulence properties
are found and which have been linked to infec-
tion by these organisms, as well as what can be
done to prevent it. Terms and Definitions
7. Define "competitive exclusion" and discuss how
this concept is used to prevent colonization of Alpha Amylase Test: Assay used to determine
salmonellae in live animals. whether liquid egg has been
8. Describe the main characteristics of shigellae treated at the correct tempera-
and their role in foodbome illness. ture to achieve pasteurization.
9. Discuss the symptoms of shigellosis and the With such a treatment, alpha-
virulence factors that lead to this infection. amylase is inactivated.
Nurmi Concept: This concept was first devel-
Chapter Outline oped to explain how coloniza-
tion of animal tissue by some
1. Salmonellosis. bacteria could be prevented as

72 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


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a result of competition by other 2. Describe the system used to serotype salmonel-
microorganisms living in the lae, based on 0 and H antigens.
same environment. It is also 3. List the types of products where salmonellae are
known as "competitive exclu- typically found.
sion." 4. Name the top 3 Salmonella serotypes in terms of
S. senftenberg 77 5W: Serotype of salmonellae recog- their being isolated most frequently from clinical
nized as the most heat resistant. samples.
It requires longer heat treat- 5. Describe "competitive exclusion" as applied to
ment than other serotypes to poultry flocks.
eliminate it from foods. 6. List the three species of shigellae responsible for
Transovarial: Traveling from the ovaries (of outbreaks of foodbome illness.
the hen) into the developing 7. What is the most common factor involved in the
embryo inside an egg. It is con- incidence of shigellosis worldwide?
sidered one way in which eggs
could be contaminated with
salmonellae.
Critical Thinking Question

Basic Knowledge Questions


1. Provide a theory as to why Salmonella Enteriti-
1. List and describe the three groups into which dis infections are on the rise. What routes of
salmonellae are placed for epidemiological pur- transmission could account for its prevalence in
poses. eggs and in poultry?

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CHAPTER 27

Foodborne Gastroenteritis Caused


by Escherichia coli

Learning Objectives 2. The Recognized Virulence Groups.


a) Enteroaggregative E. coli
The reader will be able to: b) Enterohemorrhagic E. coli
1. Describe the classification scheme used for Es- 1) The toxins
cherichia coli serotypes. 2) Growth and Stx production
2. Describe the general characteristics of 3) Effect of environmental and physical
Enteroaggregative E. coli, including its preva- agents
lence and its role in disease. 4) Prevalence in foods
3. Describe the general characteristics of 5) Prevalence in dairy cattle
Enterohemorrhagic E. coli. 6) Human disease syndromes/prevalence
4. Discuss the toxins produced by enterohemorrhagic c) Enteroinvasive E. coli
E. coli, including the circumstances in which it is d) Enteropathogenic E. coli
optimally produced and how certain conditions e) Enterotoxigenic E. coli
can affect it. 1) The enterotoxins
5. Describe the prevalence of Enterohemorrhagic 3. Prevention.
E. coli in foods and its origin in animals. 4. Traveler's Diarrhea.
6. Describe the symptoms of disease caused by
Enterohemorrhagic E. coli.
7. Describe Enteroinvasive and Enteropathogenic Terms and Definitions
E. coli serotypes, including the disease syndromes
they cause. AlE: Attachment/effacement, refers
8. Describe the general characteristics of to the mechanism by which
Enterotoxigenic E. coli, and the toxins produced several types of pathogenic E.
by this organism. coli, such as Enterohemor-
9. Discuss the disease syndrome caused by rhagic strains, colonize the in-
Enterotoxigenic E. coli. testinal tract. The bacteria at-
tach to tissue cells, damaging
Chapter Outline them and causing an efface-
ment of the lining of the intes-
1. Serological Classification. tinal tract. This mechanism has

74 INSTRUCTOR'S MANUAL FOR MODERN FOOD MICROBIOLOGY


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been linked to the possession in production of cAMP which
of a particular gene by the bac- disrupts the transport of ions
terium, designated "eaeA." by the intestinal cells, causing
CFA: Colonization factor antigen, efflux of water into the intesti-
refers to fimbrial elements pro- nallumen.
duced by Enterotoxigenic E. HUS: Hemolytic uremic syndrome,
coli strains, which help them refers to the syndrome devel-
colonize intestinal tissue cells. oped in children due to infec-
EAST1: Heat-stable enterotoxin pro- tion generally with Enterohe-
duced by some Enteroaggre- morrhagic E. coli strains. It
gative E. coli strains. results in kidney damage
Enterobacteriaceae: Family of bacteria to which which often necessitates dialy-
several genera of important sis and can ultimately end in
foodborne pathogens belong, death of the patient.
such as Salmonella, Escheri- Intimin: Protein produced by several
chia, Shigella, Yersinia, and pathogenic strains of E. coli,
others. such as Enteropathogenic and
Enteroinvasive: Refers to the property of a Enterohemorrhagic strains. It
pathogenic organism in terms allows for the intimate binding
of its ability to invade intesti- of the bacteria to the microvilli
nal tissue cells. of intestinal tissue cells, setting
Esps: Enteropathogenic-E. coli-se- the stage for either their inva-
creted-proteins, refers to pro- sion or effacement.
teins produced by these organ- LT: Heat-labile toxin, refers to a
isms, which block phagocyto- toxin produced by Enterotoxi-
sis and cause phosphorylation genic E. coli strains, made up
of certain surface receptors of of an A and a B subunit. The A
intestinal tissue cells. The lat- subunit stimulates the adeny-
ter facilitates the intimate at- late cyclase system while sub-
tachment of the E. coli cells to unit B helps the toxin bind to
the microvilli of the intestinal intestinal tissue cells. The toxin
cells. is destroyed by heating at 60C
Hemolytic for 30 min.
Anemia: One of the manifestations of OMP: Outer membrane protein, such
hemolytic uremic syndrome, in as that in the outer membrane
which red blood cells lyse, re- ofEnteroaggregative strains of
sulting in a decrease in their E. coli, believed to be respon-
number. This develops in some sible for adherence to tissue
children as a result of infection cells.
with Enterohemorrhagic E. ST: Heat-stable toxins produced by
coli strains. Enterotoxigenic strains of E.
G Protein: Protein found inside intestinal coli. They withstand heating at
tissue cells. Some bacterial 100C for 15 minutes. These
toxins, such as that produced toxins stimulate the guanylate
by Enterotoxigenic E. coli cyclase system.
strains, cause ribosylation of Sereny Test: Assay performed to determine
the G protein, which itself ac- whether a bacterial strain is
tivates the adenylate cyclase able to invade tissue cells.
system. This activation results Drops of a culture are added

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Copyright 2000, Aspen Publishers, Inc.
to the eye of a guinea pig, with Basic Knowledge Questions
development of conjunctivitis
deemed as a positive for infec- 1. Define the following: EaggEC, EHEC, EIEC,
tivity. andETEC.
Stx Toxins: Also known as Shiga-like tox- 2. What two toxins are produced by EHEC, and
ins, verotoxins, and SLT-l and what is their mode of action?
SLT-2 toxins. There are two 3. How do pH and temperature affect production of
major Stx toxins, Stxl and toxins by EHEC?
Stx2, as well as some variants. 4. Describe the disease conditions caused by EHEC.
The toxins bind to ganglio- 5. How do Esps help EIEC cause disease?
sides, and become internalized 6. What are the modes of action of the two toxins
into the cell. The mechanism produced by ETEC?
of action of the toxins is to in- 7. What organisms are responsible for causing
hibit protein synthesis by caus- traveler's diarrhea?
ing the release of an adenine
residue from the rRNA of ri- Critical Thinking Questions
bosomes.
Thrombo- 1. What could be some reasons why the rate of
Cytopenia: Damage to thrombocytes re- incidence of EHEC in Northern Ireland, En-
sulting in clot formation due to gland, Wales, and Scotland are so much lower
infection with Enterohemor- than in the U.S.?
rhagic E. coli strains. This oc- 2. WhydoyouthinkthatcalvesshedE. coli0157:H7
curs in the elderly, and can re- longer and are usually infected with this organ-
sult in embolism, or stroke. ism more frequently than adult cattle?

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CHAPTER 28

Foodborne Gastroenteritis Caused by Vibrio,


Yersinia, and Campylobacter Species

Learning Objectives c} Virulence properties


d} Gastroenteritis syndrome and vehicle foods
The reader will be able to: 2. Other Vibrios.
1. Describe the various Vibrio species that are re- a} Vibrio cholerae
sponsible for foodbome illness, in terms ofgrowth b} Vibrio vulnificus
conditions and prevalence in nature. c} Vibrio alginolyticus and V. hollisae
2. Describe the virulence factors of Vibrio 3. Yersiniosis (Yersinia enterocolitica).
parahaemolyticus and the disease syndrome it a} Growth requirements
causes. b} Distribution
3. List and describe other Vibrio species and ex- c} Serovars and biovars
plain how they cause foodbome illness. d} Virulence factors
4. Describe Yersinia enterocolitica in terms of its e} Incidence of Y. enterocolitica in foods
growth requirements and distribution in nature. f) Gastroenteritis syndrome and incidence
5. List the serovars and biovars associated with 4. Campylobacteriosis (Campylobacter jejuni).
yersiniosis and their incidence in foods. a} General characteristics
6. Describe the virulence factors of Y. enterocolitica b} Distribution
and the disease. it causes. c} Virulence properties
7. Describe the general characteristics of d} Enteritis syndrome and prevalence
Campylobacter jejuni and its distribution in na- 5. Prevention.
ture.
8. Explain the virulence factors of C. jejuni, spe-
cifically the toxins it produces, and describe the Terms and Definitions
disease syndrome it causes.
Cholecystitis: Inflammation of the gall blad-
der, usually due to infection
Chapter Outline with a microorganism, such as
Y. enterocolitica.
1. Vibriosis (Vibrio parahaemolyticus). CIT: Campylobacter jejuni toxin,
a} Species refers to a heat-labile endot-
b} Growth conditions oxin produced by this organ-

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ism. It is composed of one A NAG-ST: Toxin produced by non-agglu-
subunit surrounded by five B tinating vibrios. It is similar to
subunits. Its mode of action is toxins produced by Citrobacter
through activation of the ade- freundii and Yersinia entero-
nylate cyclase system, much colitica.
like V. cholerae toxin and Ogawa: Specific serotype within the El
Enterotoxigenic E. coli LT Tor biotype of V. cholerae. It is
toxin. serologically distinct from the
Guillain-Barre Inaba serotype.
Syndrome: Chronic condition caused by OmpU: Outer membrane protein which
prior infection with some or- has been shown to playa role
ganisms, most commonly in adherence of V. cholerae
Campylobacter jejuni. It mani- strains to intestinal tract cells,
fests itself as a temporary pa- a necessary step for invasion.
ralysis of the facial muscles. Penner: Serotyping scheme used to dif-
El Tor: Biotype of serovar 0 group 1 ferentiate Campylobacter
V. cholerae. It was first recog- jejuni isolates, based on differ-
nized as unique due to its abil- ences due to heat-stable anti-
ity to agglutinate chicken red gens on the cell surface. Ap-
blood cells. proximately 60 serotypes have
Erythema been identified according to
Nodosum: Chronic skin disease, marked this system.
by the appearance of tender Peritonitis: Inflammation of the peritoneal
pink nodules, usually caused by cavity due to fluid accumula-
infection with a microorganism tion as a result of infection with
such as Y. enterocolitica. certain bacteria, such as Y.
Inaba: Specific serotype within the El enterocolitica.
Tor biotype of V. cholerae. It is Pseudo-
serologically distinct from the Appendicitis: Condition attributed to infec-
Ogawa serotype. tion with Y. enterocolitica in
Isoelectric Point: pH of a molecule at which the which the patient suffers from
net molecular charge is zero. similar symptoms to appendi-
At this point, migration citis. Cases ofyersiniosis have
through a pH gradient stops. sometimes led to appendecto-
Kanagawa mies by physicians unfamiliar
Reaction: Assay used to determine the with this pathogen.
presence of virulent strains of R Plasmids: Plasmid DNA that contains
V. parahaemolyticus. It is based gene( s) for resistance to spe-
on the development of a beta- cific antimicrobial agents. It is
hemolysis reaction on transferred between species of
Wagatsuma's agar. The hemol- the same genus, but can be
ysin responsible for this is ther- transferred across genera as
mostable. well.
NAGs: Non-agglutinating vibrios, re- Reactive
fers to strains of V. cholerae Arthritis: Condition caused by infection
that do not react with antibod- with some foodbome patho-
ies specific for serovar 0 group gens, notably Y. enterocolitica
I vibrios, and thus have been and some salmonellae sero-
placed into their own. types. It can develop 7 days

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after infection, and is marked Basic Knowledge Questions
by inflammation of the joints,
especially wrists and knees. It 1. Define the Kanagawa reaction and explain its
is believed to result from role in the virulence of Vibrio species.
stimulation ofT helper cells to 2. List the various biovars and serotypes of V.
induce production of cytotoxic cholerae.
T cells by the engulfing ofbac- 3. List the minor species of Vibrio that have been
teria by macrophages. The connected with foodborne illness (other than V.
CTLs then produce certain parahaemolyticus and V. cholerae), and provide
cytokines which continue a short description of the types of outbreaks they
stimulation of the T helper have caused.
cells, resulting in fluid accu- 4. Describe the major characteristics of Y.
mulation. enterocolitica and list the serotypes that have
TDH: Thermostable direct hemol- been identified.
ysin, responsible for the 5. What are the virulence factors of Y. enterocolitica?
Kanagawa reaction. It is pro- 6. Describe the major characteristics of C. jejuni
duced by most virulent strains and how it differs from most other foodborne
of V. parahaemolyticus. pathogens.
TRH: Thermostable-related hemol- 7. What is the principal virulence factor of C. jejuni
ysin, hemolysin present in and what is its mechanism of action?
some strains of V. parahae-
molyticus. In addition to TDH, Critical Thinking Questions
it is an indicator of virulence
for these organisms. 1. Provide an explanation for the fact that despite
YOPs: Yersinia outer-membrane pro- the major differences among these organisms, V.
teins, refers to a class of about cholerae, C. jejuni, and Enterotoxigenic E. coli
20 proteins produced by strains all possess a toxin with the same mode of action.
of Y. enterocolitica, which may Consider environmental as well as other factors
help the bacteria adhere to in- 10 your answer.

testinal tissue cells, and even 2. In spite of the fact that C. jejuni is a fastidious
escape phagocytosis. Proteins organism, able to grow only under very strict
are encoded by genes carried conditions in the laboratory, it is considered the
in plasmids. leading cause of diarrhea in developing coun-
tries. How can this be explained?

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CHAPTER 29

Foodborne Animal Parasites

Learning Objectives 2. Protozoa.


a) Taxonomy
The reader will be able to: b) Giardiasis
1. List the main differences between bacteria and 1) Environmental distribution
animal parasites. 2) Syndrome, diagnosis, and treatment
2. Describe the protozoa, and provide a list of or- 3) Incidence in foods and foodborne cases
ganisms as examples. c) Amebiasis
3. Describe the protozoan diseases giardiasis, ame- 1) General characteristics
biasis, toxoplasmosis, sarcocystosis, cryptospor- 2) Syndrome, diagnosis, and treatment
iodiosis, and cyclosporiasis, in terms of organ- d) Toxoplasmosis
isms responsible, foods associated that serve as 1) General characteristics
vehicles of infection, disease symptoms, and the 2) Symptoms, diagnosis, and treatment
treatment that should be applied. 3) Distribution of T. gondii
4. Describe the main characteristics of flatworms 4) Food-associated cases
and roundworms. 5) Control
5. Describe the diseases caused by flatworms, such e) Sarcocystosis
as fascioliasis, fasciolopsiasis, paragonimiasis, f) Cryptosporiodiosis
clonorchiasis, diphyllobothriasis, and taeniasis, 1) General characteristies
including the organisms responsible, the foods 2) Symptoms, diagnosis, and treatment
associated with these infections, disease symp- 3) Waterborne and foodborne outbreaks
toms, and treatment and prevention measures. g) Cyclosporiasis
6. Describe the causative agent of trichinosis, the 1) General characteristics
disease symptoms it causes, the mode of infec- 2) Prevalence and outbreaks
tion, and how this can be prevented. 3. Flatworms.
7. Describe the causative agent of anisakiasis, the a) Classification
disease symptoms it causes, the mode of infec- b) Fascioliasis
tion, and how this can be prevented. 1) General characteristics
2) Symptoms, diagnosis, and treatment
Chapter Outline c) Fasciolopsiasis
1. Differences from Other Microorganisms. d) Paragonimiasis

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1) General characteristics as within 6 hours after inges-
2) Symptoms, diagnosis, and treatment tion. Stomach pain, nausea,
e) Clonorchiasis vomiting, with fever and
1) General characteristics bloody stool possible in some
2) Symptoms, diagnosis, treatment, and pre- cases. Peritonitis can result if
vention the worms are able to penetrate
f) Diphyllobothriasis the gut wall.
1) General characteristics Bradyzoites: Dormant forms oftachyzoites,
2) Prevalence develop after cluster of
3) Symptoms, diagnosis, and treatment tachyzoites become encircled
4) Prevention by a protective wall inside
g) Cysticercosis/Taeniasis muscle and other tissues. They
1) General characteristics do not multiply but can persist
2) Symptoms, diagnosis, and treatment in an infected person for life.
3) Prevention When the immune system of
4. Roundworms. the victim is compromised,
a) Classification bradyzoites will break, result-
b) Trichinosis ing in the emergence of
1) General characteristics tachyzoites.
2) Prevalence Cercariae: Larval stage of some trema-
3) Symptoms and treatment todes such as F hepatica, hav-
4) Diagnosis ing the shape of a tadpole with
5) Prevention and control a tail-like appendage. This
6) Microwave cooking form is produced by a redia,
c) Anisakiasis and are able to swim and at-
1) General characteristics tach to grasses.
2) Symptoms, diagnosis, and treatment Clonorchiasis: Disease caused by infection
3) Prevalence and distribution with the trematode Clonorchis
4) Prevention sinensis. Onset of symptoms is
similar to that of other fluke-
associated illness (approxi-
Terms and Definitions mately 1 month). An infected
person may be asymptomatic,
Amebiasis: Disease caused by infection but in some cases liver dam-
with Entamoeba histolytica, age takes place, which can lead
also knows as amoebic dysen- to liver cancer.
tery. Onset of symptoms is 2- Cryptosporiodiosis: Disease caused by infection
4 weeks, with loose stools con- with Cryptosporidium parvum.
taining mucus and blood last- Onset of symptoms is approxi-
ing for several months, or even mately 2 weeks, and these in-
years. These develop into se- clude voluminous watery diar-
vere abdominal pain, fever, rhea, impaired respiration, de-
vomiting, and back pain, with hydration, abdominal discom-
weight loss a typical outcome. fort, which develop into weight
Anisakiasis: Disease caused by infection loss. These can last about 4
with the roundworms Anisakis days.
simplex and Pseudoterranova Cyclosporiasis: Disease caused by infection
decipiens. Onset of symptoms with Cyclospora cayetanensis.
is very fast, detected as early Onset and symptoms are simi-

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lar to cryptosporiodiosis, but sea, severe diarrhea, vomiting,
can last 6 weeks or more. and flatulence that result in
Cysticercosis: Disease caused by infection weight loss. Can last up to 3
with the flatworms Taenia months and is considered
saginata or Taenia solium. highly contagious.
Symptoms are usually not ex- Intermediate Host: The animal in which a juvenile
perienced at all. However, in or larval form of a parasite de-
60% of infected person, cys- velops and matures.
ticerci develop in the central Metacercaria: Encysted form of cercariae of
nervous system resulting in trematodes such as F. hepatica.
symptoms like epileptic sei- This is the form that is typi-
zures. In 5% of infected per- cally ingested by a person.
sons, cysticerci form in the Once inside the body, it
muscle yielding to muscle en- excysts, traveling to the liver
largement. In 3% of cases they where it matures in the bile
form in the eye. ducts.
Definitive Host: The animal in which an adult Nosocomial: Refers to an infection acquired
parasite carries out its sexual in hospital.
cycle and multiplies. Oocysts: Term used to describe the en-
Diphyllobothriasis: Disease caused by infection cysted, inactive, form of most
with the parasite Diphyllobo- protozoans. Oocysts will often
thrium latum. Onset of disease develop or excysts to active
is approximately one month, forms, which cause disease.
with symptoms including ab- Paragonimiasis: Disease caused by the parasites
dominal cramps, vomiting, Paragonimus westermani and
dizziness, followed by loss of P. kellicotti. Onset is 2-3
appetite yielding weight loss. months, with symptoms in-
Fascioliasis: Disease caused by infection cluding severe chronic cough-
with the trematode Fasciola ing and sharp chest pain.
hepatica. It is also known as Proglottids: Segments of the body of a tape-
parasitic biliary cirrhosis, or worm, such as T. saginata, con-
liver rot. Onset is approxi- taining both male and female
mately 30 days, consisting of reproductive organs.
fever, fatigue, loss of appetite, Rediae: Larvae produced within the
and pain in the liver region of sporocysts of certain trema-
the abdomen. todes, such as F. hepatica. They
*"
Fasciolopsiasis: Disease caused by the parasite develop into cercariae.
Fasciolopsis bush. Onset is 1- Sarcosystosis: Disease caused by Sarcocystis
2 months, with symptoms in- hominis or S. suihominis. On-
cluding violent diarrhea, ab- set of symptoms is relatively
dominal pain, weakness, and short (3-48 hours), and include
weight loss. nausea, stomach cramps, and
Giardiasis: Disease caused by infection diarrhea.
with the protozoan Giardia Scolex: Head of a flatworm, such as T.
lamblia, carried through meat saginata, either in the larval or
or water contaminated with adult stage.
animal feces harboring this or- Sporozoites: Life forms that result from ger-
ganism. Onset of symptoms is mination of T. gondii oocysts.
7-13 days, with cramps, nau- These pass through the intes-

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tinal walls and are able to reach place. They penetrate the intes-
the circulatory system. From tines through a falling-leaf
there, they are transported to type of motility. The same term
muscle tissue and other parts is used to describe the ameba
of the body, where they de- stage of E. histolytica, which
velop into tachyzoites. are also motile.
Strobila: The chain of segment (proglot- Zoonotic: Refers to an infection acquired
tids) forming the body of a through direct contact with a
tapeworm, such as T. saginata. vertebrate animal.
Tachyzoites: Term used to refer to the rap-
idly-multiplying form of
sporozoites of T. gondii and
Basic Knowledge Questions
similar parasites after reaching
muscle tissue and organs.
Tachyzoites will form clusters 1. What is the difference between a definitive host
surrounded by a protective and an intermediate host?
wall, developing into a 2. Describe the life cycle of Giardia lamblia that
bradyzoite. yields to disease in humans.
Toxoplasmosis: Disease caused by the proto- 3. What are trophozoites, and what role do they
zoan Toxoplasma gondii. It is play in causing parasitic disease? Include in your
transmitted through contact answer the types and names of parasites which
with infected farm animals, as occur in the form of trophozoites.
well as cats. Most people do 4. Compare and contrast the organisms
not develop symptoms, but the Cryptosporidium parvum and Cyclospora
immunocompromised will suf- cayetanensis in terms of general characteristics,
fer from symptoms that mimic how transmitted, and the diseases they cause.
infectious mononucleosis. 5. Compare the diseases caused by the trematodes
These include rash, headache, discussed in this chapter.
fever, severe muscle pain, and 6. Describe the life-cycle of Taenia, including the
swelling of the lymph nodes. various forms that are involved.
Trematode: A class of parasite also known 7. How is the risk of trichinosis prevented/mini-
as flukes. They have a well- mized?
developed alimentary canal
and suckers on their outer sur-
face for adhesion to tissues.
Critical Thinking Questions
Trichinosis: Disease caused by infection
with the roundworm Tri-
chinella spiralis. Onset of 1. Should we trust the use of microwaving to ensure
symptoms is approximately 2 the safety of fresh meats from the parasite Tri-
weeks, and these include se- chinella? Why or why not? Be complete in your
vere muscle pain, facial swell- answer, providing known evidence for your point
ing, diarrhea, and vomiting, of view.
with death resulting in some 2. What measures could/should be implemented if
cases due to heart failure. we were to significantly reduce the incidence of
Trophozoite: Flagellated form of G. lamblia parasitic diseases in humans due to consumption
after maturation of cysts takes of contaminated raw foods?

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CHAPTER 30

Mycotoxins

Learning Objectives c) Production and occurrence in foods


d) Relative toxicity and mode of action
The reader will be able to: e) Degradation
1. Describe aflatoxins in terms of general struc- 3. Alternaria Toxins.
tures and the conditions under which they are 4. Citrinin.
produced by certain molds. 5. Ochratoxins.
2. Describe the prevalence of aflatoxins in foods, 6. Patulin.
and the mode of action responsible for disease. 7. Penicillic Acid.
3. Describe alternaria toxins, citrinin, ochratoxins, 8. Sterigmatocystin.
patulin, penicillic acid, and sterigmatocystin, in 9. Fumonisins.
terms of the organisms that produce them, their a) Toxin types
structure, the types of foods where found, and b) Growth and production
their mode of action. c) Prevalence in corn and feeds
4. Describe fumonisins, including the conditions d) Physical/chemical properties ofFB J and FB2
under which they are produced, the organisms e) Pathology
that produce them, the foods where they are 10. Sambutoxin.
found, and the disease conditions they cause. 11. Zearalenone.
5. Describe sambutoxin and zearalenone toxins, a) Structure
including the organisms that produce them, the b) Control and production
types of foods where found, and the disease
conditions they cause.
Terms and Definitions

Chapter Outline Aflatoxins: Toxic compounds produced by


some Aspergillus mold spe-
1. General Definitions. cies. They are secondary me-
2. Aflatoxins. tabolites that can cause ab-
a) Structures dominal pain, jaundice, and
b) Requirements for growth and toxin produc- vomiting if ingested. If dose
tion consumed is high enough, it

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can result in liver necrosis. kidneys, causing necrosis of
Consumption of low doses for the tubules in rats, dogs, and
extended periods of time may swine. May cause a renal dis-
lead to liver cancer. order in humans called Balkan
Alternariol: Mycotoxin produced by Alter- Endemic Nephropathy.
naria species, not proven to be Patulin: Mycotoxin produced by Peni-
harmful to humans. cillium expansum, carcino-
Ames Test: Assay used to determine the genic to mice. It is acid-stable
mutagenicity of chemical com- and heat-stable. It is suspected
pounds. It is based on the ge- to be carcinogenic to humans,
netic change of a Salmonella but it has not yet been proven.
Typhimurium culture, from Primary
one that requires certain nutri- Metabolites: Compounds produced by mi-
ents to grow, to one that is able croorganisms that are essential
to grow without the nutrient for growth, and exerted during
being supplied. active growth.
Citrinin: Mycotoxin produced by Peni- Sambutoxin: Mycotoxin produced by Fusar-
cillium citrinum and P. citreo- ium sambucinum and F. oxy-
viridae. It is heat-stable and sporum. It causes hemorrhag-
causes tubular damage in kid- ing in the stomach and intes-
neys of swine as ochratoxins, tines of rats, and it is toxic to
yet it is somewhat less toxic chick embryos.
than ochratoxin. Secondary
Cyc1opiazonic Metabolites: Compounds produced at or
Acid: Compound produced by some near the end of exponential
A. flavus strains, thought to growth phase. They have no
contribute to the toxicity of apparent usefulness to the or-
aflatoxin. ganism in terms of growth or
Fumonisins: Mycotoxins produced by metabolic capacity.
Fusarium species. There are at Trichothecene: Mycotoxin produced by
least seven fumonisins, with Fusarium sporotrichioides,
FBI being associated with also known as T2 toxin. It is
esophageal cancer in humans. responsible for alimentary
Hyperestrogenism: Symptoms mimic those caused toxic aleukia, a condition that
by having very high levels of causes a burning sensation in
estrogen. Females get enlarged the mouth, followed by acute
and edematous genitals result- gastroenteritis. This is followed
ing in abortions, and males by destruction ofthe bone mar-
undergo a feminizing effect, row, development ofleukemia,
having enlarged mammary anemia, yielding to total atro-
glands and atrophied testes. It phy of the bone marrow and
is caused by consumption of lung hemorrhaging, ultimately
zearalenone mycotoxin. resulting in death.
Ochratoxin: Mycotoxin produced by As- Zearalenone: Mycotoxin produced by
pergillus ochraceous and some Fusarium graminearum, also
Penicillium species. Type A is known as F-2 toxin. It causes
most toxic and is heat-stable to estrogenic syndrome in swine.
169C. It affects the liver and It is heat-stable.

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Basic Knowledge Questions 6. What is sambutoxin?
7. What is hyperestrogenism and what organism is
1. What is the difference between primary and sec- responsible for causing it?
ondary metabolites?
2. List the various types of aflatoxins, and compare
them in terms of toxicity and other characteris-
tics. Critical Thinking Question
3. Draw the chemical structures of citrinin, ochra-
toxin, patulin, and penicillic acid. 1. There are several mycotoxins that are known to
4. List and describe the various fumonisins, and list affect only animals. Design an experiment that
the organisms that produce them. would prove/disprove the role of fumonisins in
5. Describe the disease syndromes caused by con- causing esophageal cancer in humans.
sumption of trichothecenes.

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CHAPTER 31

Viruses and Some Other Proven and


Suspected Foodborne Biohazards

Learning Objectives Chapter Outline

The reader will be able to: 1. Viruses.


1. Define viruses and describe the methods used to a) Detection methods
detect them in foods. b) Incidence in foods and the environment
2. Discuss the prevalence ofviruses in nature and in c) Destruction in foods
foods, and how they can be eliminated from d) Hepatitis A virus
foods. e) Norwalk and related viruses
3. Describe the main characteristics of Hepatitis A f) Rotaviruses
virus, of Norwalk and related viruses, and of 2. Bacteria and Prions.
rotaviruses. a) Histamine-associated (Scombroid) poisoning
4. Discuss the role of bacteria in Scombroid food b) Aeromonas
poisoning. c) Plesiomonas
5. Discuss the major characteristics and the ill- d) Bacteroides fragilis
nesses caused by Aeromonas, Plesiomonas, e) Erysipelothrix rhusiopathiae
Bacteroidesjragilis, Erysipelothrix rhusiopathiae, f) Klebsiella pneumoniae
Klebsiella pneumoniae, and Sreptococcus iniae. g) Streptococcus iniae
6. Define the term "prion" and describe how these 3. Prion Diseases.
structures may be responsible for "mad cow dis- a) Mad cow disease
ease" in cattle, and related illnesses in humans. b) Cruetzfeld-Jakob Disease
7. Describe the toxigenic phytoplanktons that cause 4. Toxigenic Phytoplanktons.
paralytic shellfish poisoning, ciguatera poison- a) Paralytic shellfish poisoning
ing, and poisoning caused by domoic acid. b) Ciguatera poisoning
8. Describe the main characteristics of Pfisteria c) Domoic acid
and its suspected role in causing foodbome ill- d) Pfisteria piscicida
ness.

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Terms and Definitions: blurred vision, and temporary
blindness.
ASP: Amnesic shellfish poisoning, Diatom: Single-celled alga with silicon-
caused by domoic acid pro- based walls.
duced by the diatom Dinoflagellate: Member of the algae family, it
Pseudonitschia pungens. is aquatic in nature, photosyn-
BSE: Bovine spongiform encephal- thetic, and some produce toxin.
opathy, also known as "mad Consumption of particular di-
cow disease," refers to an ill- noflagellates by certain fish
ness in cattle belonging to the result in various seafood toxin
family of diseases called syndromes.
"tranmissible spongiform en- Domoic Acid: Toxin produced by the diatom
cephalopathies." It is caused by Pseudonitschia pungens. It
a prion, and results in causes a disease from con-
uncoordination and brain dam- sumption of mussels infected
age. with the dinoflagellate. It ex-
CJD: Creutzfeldt-Jakob disease, re- cites the brain cells until they
fers to an illness belonging to die from overstimulation, re-
the family of diseases called sulting in amnesia.
"transmissible spongiform en- Mad Cow Disease: Common term used for the
cephalopathies." It is caused by degenerative disease called
a prion infecting some indi- bovine spongiform encephal-
viduals. It is thought that a opathy. The disease is thought
harmless configuration of the to be caused by prions infect-
prion is naturally found in ing the cattle.
people. Certain unknown con- Paresthesia: Tingling, especially around the
ditions are thought to cause a mouth.
conformational change in the Prion: Small proteins that, upon as-
prion to a form that causes suming a specific conforma-
holes in brain tissue, ultimately tional shape, can cause brain
resulting in death of the in- damage in animals and hu-
fected individual. It is also sus- mans. They aggregate into
pected that a new variant of the fibrils, causing nerve cell de-
prion, termed nvCm, is found generation.
in cattle and that it can cause RT-PCR: Reverse-transcription poly-
cm by infection from animals merase chain... reaction, refers to
to humans. a method of identification of
Ciguatera: Food poisoning due to con- foodbome viruses when only
sumption of tropical herbivo- a few plaque forming units are
rous fish which themselves available (viral particles).
have ingested the dinoflagel- Foodborne viruses contain
late Gambierdiscus toxicus. RNA instead of DNA as their
Onset of disease is within 5 genetic material. In order to
hours, resulting in diarrhea, identify them, the samples con-
nausea, vomiting, and abdomi- taining viral RNA must be re-
nal pain. Several hours later, acted with the enzyme reverse
neurological symptoms de- transcriptase in order to syn-
velop, such as hot/cold inver- thesize DNA from the RNA
sion, metallic taste, dizziness, molecules. At this point, the

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polymerase chain reaction as- and burning sensation in the
say can be applied in order to mouth are some of the symp-
augment the number of copies toms due to the presence of
of viral DNA. With an in- histamine produced from the
creased number of DNA mol- break down of histidine.
ecules, gel electrophoresis can SRSVs: Small round structural viruses,
then be applied to detect the of which the Norwalk virus is
viral DNA and thus the particu- a member. These viruses be-
lar organism in the sample. long to the family Calicivi-
Saxitoxin: Toxin produced by the di- ridae.
noflagellate Gonyaulax caten-
ella. It causes paralytic shell-
fish poisoning due to con- Basic Knowledge Questions
sumption of mussels, clams,
and other shellfish that have 1. What is the incidence of viruses in foods in the
ingested the dinoflagellate. U.S.?
May cause death due to respi- 2. List and describe the three groups of Norwalk
ratory paralysis within a few and Norwalk-like viruses.
hours. 3. What is histamine food poisoning caused by?
Scombroid 4. What organisms does Aeromonas resemble and
Poisoning: Disease caused by consump- in what way?
tion of scombroid fish such as 5. What are prions and how do they cause disease?
mahi-mahi, sardines, and an- 6. Describe some of the disease syndromes caused
chovies. These contain high by toxins produced by dinoflagellates.
levels of free histidine in their 7. What is Pfiesteria piscicida?
muscles. This compound
serves as a substrate for the
enzyme histidine decarboxy-
Critical Thinking Question
lase produced by various spoil-
age bacteria. Rash, edema,
nausea, vomiting, diarrhea, 1. Describe the evidence that supports or contra-
abdominal cramps, headache, dicts the theory that prions harbored in cattle are
palpitations, tingling, flushing, responsible for cm in humans.

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