Microial Physiology,

Growth and Control

CLASSIFICATION OF MICROORGANISMS

MICROBIAL NUTRITION AND GROWTH (OVERVIEW)

 Growth requirements and classification
 Physical parameters that effect growth and
classification based on growth patterns
 Chemical parameters that effect growth and
classification based on growth patterns
 Population growth -- growth curve
 Population growth – Methods

ENVIRONMENTAL EFFECTS ON BACTERIAL GROWTH TEMPERATURE CLASS OF ORGANISMS

 Temperature Mesophiles ( 20 – 45C)
o Midrange temperature optima
 pH
o Found in warm-blooded animals and in
 Osmotic pressure terrestrial and aquatic environments in
 Oxygen classes temperate and tropical latitudes
 Psychrophiles ( 0-20C)
TEMPERATURE AND MICROBIAL GROWTH o Cold temperature optima
 Cardinal temperatures o Most extreme representatives inhabit
o minimum permanently cold environments
o optimum  Thermophiles ( 50- 80C)
o maximum o Growth temperature optima between 45ºC
and 80ºC
 Temperature is a major environmental factor  Hyperthermophiles
controlling microbial growth. o Optima greater than 80°C
o These organisms inhabit hot environments
including boiling hot springs, as well as
undersea hydrothermal vents that can have
temperatures in excess of 100ºC

pH AND MICROBIAL GROWTH

pH – measure of [H+]
 each organism has a pH range and a pH optimum
 acidophiles – optimum in pH range 1-4
 alkalophiles – optimum in pH range 8.5-11

lactic acid bacteria – 4-7

 Thiobacillus thiooxidans – 2.2-2.8
 fungi – 4-6
 internal pH regulated by BUFFERS and near neutral
adjusted with ion pumps
 Human blood and tissues has pH 7.2+0.2
TEMPERATURE
 Minimum Temperature: Temperature below which
 The acidity or alkalinity of an environment can greatly
growth ceases, or lowest temperature at which
affect microbial growth.
microbes will grow.
 Most organisms grow best between pH 6 and 8, but
 Optimum Temperature: Temperature at which its
some organisms have evolved to grow best at low or
growth rate is the fastest.
high pH. The internal pH of a cell must stay relatively
 Maximum Temperature: Temperature above which
close to neutral even though the external pH is highly
growth ceases, or highest temperature at which
acidic or basic.
microbes will grow
 o Acidophiles : organisms that grow best at low
pH (Helicobacter pylori, Thiobacillus
thiooxidans)
o Alkaliphiles : organisms that grow best at
high pH (Vibrio cholera)
o Most of pathogenic bacteria are neutrophiles
OSMOTIC EFFECTS ON MICROBIAL GROWTH
 Osmotic pressure depends on the surrounding solute
concentration and water availability
 Water availability is generally expressed in physical
terms such as water activity (aw)
 Water activity is the ratio of the vapor pressure of the
air in equilibrium with a substance or solution to the
vapor pressure of pure water ( aw 1.00).
MICROBIAL NUTRITION
 Why is nutrition important?
o The hundreds of chemical compounds
present inside
o a living cell are formed from nutrients.
o Macronutrients : elements required in fairly large
amounts
o Micronutrients : metals and organic compounds
needed in very small amounts
MAIN MACRONUTRIENTS
 Carbon (C, 50% of dry weight) and nitrogen (N, 12%
of dry weight)
 Autotrophs are able to build all of their cellular organic
molecules from carbon dioxide
 Nitrogen mainly incorporated in proteins, nucleic acids
 Most Bacteria can use Ammonia -NH3 and many can
also use NO3-
 Nitrogen fixers can utilize atmospheric nitrogen (N2)

ENVIRONMENTAL FACTORS AND GROWTH

1. Osmotic Effect and water activity
 organisms which thrive in high solute –
osmophiles
 organisms which tolerate high solute –
osmotolerant
 organisms which thrive in high salt – halophiles
 organisms which tolerate high salt – halotolerant
 organisms which thrive in high pressure –
barophiles
 organisms which tolerate high pressure –
barotolerant

HALOPHILES AND RELATED ORGANISMS MICROBIAL GROWTH REQUIREMENTS

 In nature, osmotic effects are of interest mainly in  Source of carbon for basic structures
habitats with high salt environments that have reduced  Source of cellular energy (ATP or related compounds)
water availability to drive metabolic reactions
 Halophiles : have evolved to grow best at reduced  Source of high energy electrons/H, reducing
water potential, and some (extreme halophiles e.g.  power, typically in form of NADH/NADPH
Halobacterium, Dunaliella ) even require high levels of
 salts for growth. CLASSIFICATION OF ORGANISMS BASED ON SOURCES OF C AND
 Halotolerant : can tolerate some reduction in the
water activity of their environment but generally grow
best in the absence of the added solute
 Xerophiles : are able to grow in very dry environments

NITROGEN REQUIREMENTS
 Although many biological components within living
organisms contain N, and N2 is the most abundant
component of air, very few organisms can “fix” or
utilize N2 by converting it to NH3
 N is often growth limiting as organisms must find
source as NH4+ for biosynthesis
 Photosynthetic organisms and many microbes can
reduce NO3- to NH4+
 OTHER MACRONUTRIENTS
 Phosphate (P), sulfur (S), potassium (K), magnesium
(Mg), calcium (Ca), sodium (Na), iron (Fe)
 Iron plays a major role in cellular respiration, being a
key component of cytochromes and iron-sulfur
proteins involved in electron transport.
 Siderophores : Iron-binding agents that cells produce
to obtain iron from various insoluble minerals.

MICRONUTRIENTS
Need very little amount but critical to cell function. Often used as
enzyme cofactors

GROWTH FACTORS
Organic compounds, required in very small amount and then
only by some cells
CLASSIFICATION OF ORGANISMS BASED ON
O2 UTILIZATION
 Utilization of O2 during metabolism yields toxic by-
products including O2-, singlet oxygen (1O2) and/or
H2O2.
 Toxic O2 products can be converted to harmless
substances if the organism has catalase (or
peroxidase) and superoxide dismutase (SOD)
 SOD converts O2- into H2O2 and O2
 Catalase breaks down H2O2 into H2O and O2
 Any organism that can live in or requires O2 has SOD
and catalase (peroxidase)

TOXIC FORMS OF OXYGEN AND DETOXIFYING ENZYMES

 Obligate (strict) aerobes require O2 in order to grow

 Obligate (strict) anaerobes cannot survive in O2
 Facultative anaerobes grow better in O2
 Aerotolerant organisms don’t care about O2
 Microaerophiles require low levels of O2

OXYGEN AND MICROBIAL GROWTH ENVIRONMENTAL FACTORS AND GROWTH

 Aerobes :
o Obligate : require oxygen to grow
o Facultative : can live with or without oxygen
but grow better with oxygen
o Microaerophiles : require reduced level of
oxygen
Oxygen
anaerobes lack superoxide dismutase and/or catalase
anaerobes need high -, something to remove O2
chemical: thioglycollate; pyrogallol + NaOH
H2 generator + catalyst
physical: removal/replacement

 Anaerobes : SPECIAL CULTURE TECHNIQUES

o Aerotolerant anaerobes : can tolerate oxygen Gas Pack Jar is used for Anaerobic Growth
but grow better without oxygen.
o Obligate : do not require oxygen. Obligate
anaerobes are killed by oxygen

Thioglycolate broth : contains a reducing agent and provides

TEST FOR OXYGEN REQUIREMENTS
aerobic and anaerobic conditions
OF MICROORGANISMS
a) Aerobic
b) Anaerobic
c) Facultative
d) Microaerophil
e) Aerotolerant
 CULTURE MEDIA: COMPOSITION
 Culture media supply the nutritional needs of
microorganisms ( C ,N, Phosphorus, trace elements,
etc)
o defined medium : precise amounts of highly
purified chemicals
o complex medium (or undefined) : highly
nutritious substances.
 In clinical microbiology,
o Selective : contains compounds that
selectively inhibit
o Differential: contains indicator
o terms that describe media used for the
isolation of particular species or for
comparative studies of microorganisms. Growth of Staphylococcus aureus on
Mannitol Salt Agar results in a color
TYPES OF MEDIA change in the media from pink to yellow.
Media can be classified on three primary levels
1. Physical State DIFFERENTIAL MEDIA
2. Chemical Composition  Differential shows up as visible changes or variations
3. Functional Type in colony size or color, in media color changes, or in
the formation of gas bubbles and precipitates.
PHYSICAL STATES OF MEDIA  Example: Spirit Blue Agar to detect the digestion of
 Liquid Media fats by lipase enzyme. Positive digestion (hydrolysis)
o Water-based solutions is indicated by the dark blue color that develops in the
o Do not solidify at temperatures above freezing colonies. Blood agar for hemolysis (α,β,and γ
/ tend to be free flowing hemolysis), EMB, MacConkey Agar, …etc.
o Includes broths, milks, and infusions
o Measure turbidity ENRICHMENT MEDIA
o Example: Nutrient Broth, Methylene Blue  Is used to encourage the growth of a particular
Milk, Thioglycollate Broth microorganism in a mixed culture.
 Semisolid  Ex. Manitol Salt Agar for S. aureus
o Exhibits a clot-like consistency at ordinary  Blood agar , chocolate agar, Slenite F broth
room temperature
o Determines motility LABORATORY CULTURE OF MICROORGANISMS
o Used to localize a reaction at a specific site.  Microorganisms can be grown in the laboratory in
o Example: Sulfide Indole Motility (SIM) for culture media containing the nutrients they require.
hydrogen sulfide production and indole  Successful cultivation and maintenance of pure
reaction and motility test. cultures of microorganisms can be done only if aseptic
 Solid (CAN AND CANNOT be converted into a liquid) technique is practiced to prevent contamination by
o Firm surface for discrete colony growth other microorganisms.
o Advantageous for isolating and culturing
o Two Types MICROBIAL GROWTH
1. Liquefiable (Reversible)  Microbes grow via binary fission, resulting in
2. Non-liquefiable exponential increases in numbers
o Examples:Gelatin and Agar (Liquefiable)  The number of cell arising from a single cell is 2n after
Cooked Meat Media, Potato Slices (Non- n generations
liquefiable)  Generation time is the time it takes for a single cell to
grow and divide
CHEMICAL COMPOSITION OF CULTURE MEDIA
1. Synthetic Media
 Chemically defined
 Contain pure organic and inorganic
compounds
 Exact formula (little variation)
2. Complex or Non-synthetic Media
 Contains at least one ingredient that is not
chemically definable (extracts from plants
and animals)
 No exact formula / tend to be general and
grow a wide variety of organisms

SELECTIVE MEDIA
 Contains one or more agents that inhibit the growth of
a certain microbe and thereby encourages, or selects,
a specific microbe.
 Example: Mannitol Salt Agar [MSA] encourages the
growth of S. aureus. MSA contain 7.5% NaCl which
inhibit the growth of other Gram +ve bacteria
 BINARY FISSION
GROWTH CURVE
 During lag phase, cells are recovering from a period of
no growth and are making macromolecules in
preparation forgrowth
 During log phase cultures are growing maximally
 Stationary phase occurs when nutrients are depleted
and wastes accumulate (Growth rate = death rate)
 During death phase death rate is greater than growth
rate
METHODS USED TO MEASURE MICROBIAL GROWTH
1. Count colonies on plate or filter (counts Live cells)
2. Microscopic counts
3. Flow cytometry (FACS)
4. Turbitity
VIABLE COUNTS
 Each colony on plate or filter arises from single live cell
MICROSCOPIC COUNTS
 Need a microscope, special slides, high power
objective lens
 Typically only counting total microbe numbers, but
differential counts can also be done
TURBITITY
 Cells act like large particles that scatter visible light
 A spectrophotometer sends a beam of visible light
through a culture and measures how much light is
scattered
 Scales read in either absorbance or % transmission
 Measures both live and dead cells
INOCULATION
 Sample is placed on sterile medium providing
microbes with the appropriate nutrients To sustain
growth.
 Selection of the proper medium and sterility of all tools
and media is important.
 Some microbes may require a live organism or living
tissue as the inoculation medium.
INCUBATION
 An incubator can be used to adjust the proper growth
conditions of a sample.
 Need to adjust for optimum temperature and gas
content.
 Incubation produces a culture – the visible growth of
the microbe on or in the media
ISOLATION
 The end result of inoculation and incubation is isolation
 On solid media we may see separate colonies, and in
broth growth may be indicated by turbidity
 Sub-culturing for further isolation may be required.
INSPECTION
 Macroscopically observe cultures to note color,
texture, size of colonies, etc.
 Microscopically observe stained slides of the culture to
assess cell shape, size, and motility.
IDENTIFICATION
 Utilize biochemical tests to differentiate the microbe
from similar species and to determine metabolic
activities specific to the microbe.
 Only counting live cells
 Controlling Microial 
SELECTION OF MICROBIAL CONTROL METHODS
Ideally, agents should be:
o Inexpensive
Growth in the Environment o Fast-acting
o Stable during storage
o Control all microbial growth while being
harmless to humans, animals, and objects

FACTORS AFFECTING THE EFFICACY

TERMINOLOGY OF MICROBIAL CONRTOL OF ANTIMICROBIAL METHODS
 Sterilization  Nature of site to be treated
 Aseptic  Degree of susceptibility of microbes involved
 Disinfection/disinfectants  Environmental conditions that pertain
 Antisepsis/antiseptic
 Degerming SITE TO BE TREATED
 Sanitization  Harsh chemicals and extreme heat cannot be used on
 Pasteurization humans, animals, and fragile objects
 Suffix – stasis/-static  Method and level of microbial control based on site of
 Suffix – cide/-cidal medical procedure

MICROBIAL DEATH RATES RELATIVE SUSCPETIBILTIY OF MICROORGANISMS

ACTION OF ANTIMICROBIAL AGENTS

 Many types of chemical and physical microbial controls
 Modes of action fall into two basic categories
o Alteration of cell walls or cytoplasmic
membranes
o Interference with protein and nucleic acid
structure  Effectiveness of germicides classified as high,
intermediate, or low
ALTERATION OF CELL WALLS AND MEMBRANES o High-level kill all pathogens, including
 Cell wall maintains integrity of cell endospores
o When disrupted, cannot prevent cell from o Intermediate-level kill fungal spores,
bursting dueto osmotic effects protozoan cysts, viruses and pathogenic
 Cytoplasmic membrane contains cytoplasm and bacteria
controls passage of chemicals into and out of cell o Low-level germicides kill vegetative bacteria,
o When damaged, cellular contents leak out fungi, protozoa, and some viruses
 Viral envelope responsible for attachment of virus to
target cell ENVIRONMENTAL CONDITIONS
o Damage to envelope interrupts viral
replication
 Nonenveloped viruses have greater tolerance of harsh
conditions

DAMAGE TO PROTEINS AND NUCLEIC ACIDS

 Protein function depends on 3-D shape
o Extreme heat or certain chemicals denature
proteins
 Chemicals, radiation, and heat can alter or destroy
nucleic acids
o Can produce fatal mutants
o Can halt protein synthesis through action on
RNA
METHODS FOR EVALUATING DISINFECTANTS AUTOCLAVING
AND ANTISEPTICS  Pressure applied to boiling water prevents steam from
 Phenol coefficient escaping
o Evaluating the efficacy of disinfectants and  Boiling temperature increases as pressure increases
antiseptics by determining the ratio of agent’s  Autoclave conditions – 121ºC, 15 psi, 15 minutes
ability to control microbes to that of phenol
o Greater than 1.0 indicates that agent is more
effective than phenol
o Has been replaced by newer methods
 Use-dilution test
o Metal cylinders dipped into broth cultures of
bacteria and dried
o Contaminated cylinder immersed into dilution
of disinfectant for 10 minutes
o Cylinders removed, washed, and placed into
tube of medium for 48 h
o Most effective agent entirely prevents growth
at highest dilution
o New standard procedure being developed
 In-use test
o Swabs taken from objects before and after
application of disinfectant or antiseptic
o Swabs inoculated into growth medium and PASTEURIZATION
incubated  Pasteur’s method
o Medium monitored for growth
 Today, also used for milk, ice cream, yogurt, and fruit
o Accurate determination of proper strength
juices
and application procedure for each specific
situation  Not sterilization; heat-tolerant and heat-loving
microbes survive
PYHSICAL METHODS OF MICROBIAL CONTROL o These do not cause spoilage prior to
consumption
 Exposure to extremes of heat
o These are generally not pathogenic
 Exposure to extremes of cold
 Milk
 Desiccation
o Batch method – 30 minutes at 63ºC
 Filtration
o Flash pasteurization – 72ºC for 15 seconds
 Osmotic pressure o Ultrahigh-temperature pasteurization – 134ºC
 Radiation for 1 second
HEAT- RELATED METHODS
 Effects of high temperatures ULTRAHIGH – TEMPERATURE STERILIZATION
o Denaturation of proteins  140ºC for 1 second, then rapid cooling
o Interference with integrity of cytoplasmic  Treated liquids can be stored at room temperature
membrane
o and cell walls DRY HEAT
o Disruption of structure and function of nucleic  Used for materials that cannot be sterilized with or are
acids
damaged by moist heat
 Thermal death point – lowest temperature that kills all  Denatures proteins and oxidizes metabolic and
cells in broth in 10 minutes structural chemicals
 Thermal death time – time to sterilize volume of liquid
 Requires higher temperatures for longer time
at set temperature thanmoist heat
 Incineration – ultimate means of sterilization
MOIST HEAT
 Used to disinfect, sanitize, and sterilize REFRIGERTION AND FREEZING
 Kills by denaturing proteins and destroying  Decrease microbial metabolism, growth, and
cytoplasmic membranes reproduction
 More effective than dry heat; water better conductor of o Chemical reactions occur slower at low
heat than air temperatures
 Methods of microbial control using moist heat o Liquid water not available
o Boiling  Psychrophilic microbes can multiply in refrigerated
o Autoclaving foods
o Pasteurization  Refrigeration halts growth of most pathogens
o Ultrahigh-Temperature Sterilization  Slow freezing more effective than quick freezing
 Organisms vary in susceptibility to freezing
BOILING
 Kills vegetative cells of bacteria and fungi, protozoan
DESICCATION AND LYOPHILIZATION
trophozoites, and most viruses within 10 minutes at
 Drying inhibits growth due to removal of water; only
sea level
microbiostatic
 Temperature cannot exceed 100ºC at sea level; steam
 Lyophilization used for long term preservation of
carries some heat away
microbial cultures
 Boiling time is critical
o Prevents formation of damaging ice crystals
 Water boils at lower temperatures at higher elevations;
requires longer boiling time
 Endospores, protozoan cysts, and some viruses can
survive boiling
 FILTRATION CHEMICAL METHODS OF MICROBIAL CONTROL
 Affect microbes’ cell walls, cytoplasmic membranes,
proteins, or DNA
 Effect varies with temperature, length of exposure, and
amount of organic matter
 Also varies with pH, concentration, and age of chemical
 Tend to be more effective against enveloped viruses
and vegetative cells of bacteria, fungi, and protozoa
 Major Categories
o Phenols
o Alcohols
o Halogens
o Oxidizing agents
o Surfactants
o Heavy Metals
o Aldehydes
o Gaseous Agents
o Antimicrobics

OSMOTIC PRESSURE
 High concentrations of salt or sugar in foods to inhibit
growth
 Cells in a hypertonic solution of salt or sugar lose
water; cell desiccates
 Fungi have greater ability than bacteria to survive
hypertonic environments

RADIATION
 Shorter wavelength equals more energy and greater
penetration
 Radiation described as ionizing or nonionizing
according to effects on cellular chemicals

IONIZING RADIATION
 Wavelengths shorter than 1 nm – electron beams,
gamma rays, and X rays
 Eject electrons from atoms to create ions
 Ions disrupt hydrogen bonding, oxidize double
covalent bonds, and create hydroxide ions; hydroxide
ions denature other molecules (DNA) PHENOL AND PHENOLICS
 Electron beams – effective at killing but do not  Intermediate- to low-level disinfectants
penetrate well  Denature proteins and disrupt cell membranes
o Used to sterilize spices, meats,
 Effective in presence of organic matter and remain
microbiological plastic ware, and medical and
active for prolonged time
dental supplies
 Commonly used in health care settings, labs, and
 Gamma rays – penetrate well but require hours to kill
homes (Lysol, triclosan)
microbes
 Have disagreeable odor and possible side effects
o Used to sterilize meats, spices, and fresh
fruits and vegetables
PHENOL COEFFICIENT
 X-rays require too much time to be practical for growth
control  The phenol coefficient sets the chemical phenol (very
nasty stuff in its pure form) as equal to 1.0 when used
NON-IONIZING RADIATION as a control agent against S. chloraeuis, S. aureus
and
 Wavelengths greater than 1 nm
P. aeruginosa.
 Excites electrons and causes them to make new
 The time required to kill these bacteria by using
covalent bonds
dilutions of the test disinfects, compared to dilution of
o Affects 3-D structure of proteins and nucleic phenol, give a relative strength of the test disinfectant.
acids
 Thus an agent with a phenol coefficient of 3.0 is three
 UV light causes pyrimidine dimers in DNA
times as efficient as phenol as a control agent
 UV light does not penetrate well  Flaw, some chemical agents do not tolerate dilution
 Suitable for disinfecting air, transparent fluids, and well. Example: alcohol at 80% is a superior
surfaces of objects disinfectant, but a low concentration, it has a lower
phenol coefficient than QUATS
DILUTION METHOD
 The same bacteria in the phenol coefficient test that
are dried on a small steel penicylinders are left in
tubes containing dilution of the test disinfectant for 10
min. Microbes on the steel cylinders are cultured to
determine survival
 Demonstration of killing of 59/60 replicates for each
organism on three different lots of product meets FDA
and EPA criteria for hospital disinfectants
 A key feature of the use dilution test is that it can allow
determination of whether the agent killed the bacteria
(bactericidal) or did not kill them but did not let them
grow (bacteriostatic) - this is an important bit of
information
ALCOHOLS
 Intermediate-level disinfectants
 Denature proteins and disrupt cytoplasmic membranes
 Evaporate rapidly – both advantageous and
disadvantageous
 Swabbing of skin with 70% ethanol prior to injection
HALOGENS
 Intermediate-level antimicrobial chemicals
 Believed that they damage enzymes via oxidation or
by denaturing them
 Iodine tablets, iodophores (Betadine®), chlorine
treatment of drinking water, bleach, chloramines in
wound dressings, and bromine disinfection of hot tubs
OXIDIZING AGENTS
 Peroxides, ozone, and peracetic acid kill by oxidation
of microbial enzymes
 High-level disinfectants and antiseptics
 Hydrogen peroxide can disinfect and sterilize surfaces
of objects
o Catalase neutralizes; not useful for treating
open wound
 Ozone treatment of drinking water
 Peracetic acid – effective sporocide used to sterilize
equipment
SURFACTANTS
 “Surface active” chemicals that reduce surface tension
of solvents to make them more effective at dissolving
solutes
 Soaps and detergents
o Soaps have hydrophilic and hydrophobic
ends; good degerming agents but not
antimicrobial
o Detergents are positively charged organic
surfactants
 Quats – colorless, tasteless, harmless to humans, and
antimicrobial; ideal for many medical and industrial
application
o Low-level disinfectants
HEAVY METALS
 Ions are antimicrobial because they alter the 3-D
shape of proteins, inhibiting or eliminating their
function
 Low-level bacteriostatic and fungistatic agents
 1% silver nitrate to prevent blindness caused by N.
gonorrhoeae
 Thimerosal used to preserve vaccines
 Copper controls algal growth in reservoirs, fish tanks,
swimming pools, and water storage tanks; interferes
with chlorophyll
ALDEHYDES
 Compounds containing terminal –CHO groups
 Cross-link with amino, hydroxyl, sulfhydryl, and
carboxyl groups to denature proteins and inactivate
nucleic acids
 Glutaraldehyde both disinfects (short exposure) and
sterilizes (long exposure)
 Formalin used in embalming and disinfection of rooms
and instruments
GASEOUS AGENTS
 Ethylene oxide, propylene oxide, and beta-
propiolactone used in closed chambers to sterilize
items
 Denature proteins and DNA by cross-linking functional
groups
 Used in hospitals and dental offices
 Can be hazardous to people, often highly explosive,
extremely poisonous, and are potentially carcinogenic
ANTIMICROBIALS
 Antibiotics, semisynthetic, and synthetic chemicals
 Typically used for treatment of disease
 Some are used for antimicrobial control outside the
body
DEVELOPMENT OF RESISTANT MICROBES
 Little evidence that extensive use of products
containing antiseptic and disinfecting chemicals adds
to human or animal health
 The use of such products promotes the development
of resistant microbes
POTENCY OF ANTIMICROBIAL SHEMICAL AGENTS
1. Sterilants destroy everything, including endospores
 for sterilizing scalpels, respiratory therapy
equipment, proctoscopes, plastic Petri
dishes, endoscopes
 e.g. gluteraldehye, hydrogen peroxide
2. High level disinfectants do not reliably destroy
endospores
 e.g. iodine, phenol, chlorhexidine, heavy
metals such as silver nitrate
3. Intermediate level disinfectants will kill
Mycobacterium, but do not destroy all viruses or
endospores, even with prolonged exposure
 e.g. alcohols: ethyl alcohol, isopropyl
4. Low level disinfectants will not kill Mycobacterium
 e.g. soaps, detergents
Overview of Human-
Microial Interactions
 Most microorganisms are benign
o Few contribute to health and fewer pose
direct threats to health
 Normal microbial flora
o Microorganisms usually found associated
with human body tissue
 Humans are colonized by microorganisms at birth
 Pathogens
o Microbial parasites
 Pathogenicity
o The ability of a parasite to inflict damage on
the host
 Virulence
o Measure of pathogenicity
 Opportunistic pathogen
o Causes disease only in the absence of
normalhost resistance  As dental plaque accumulates, the microorganisms
 Infection produce high concentrations of acid that results in
o Situation in which a microorganism is decalcification of the tooth enamel (dental caries)
established and growing in a host, whether or  The lactic acid bacteria Streptococcus sobrinus and
not the host is harmed Streptococcus mutans are common agents in dental
 Disease caries
o Damage or injury to the host that impairs host
function  The human gastrointestinal (GI) tract
NORMAL MICROFLORA IN THE
NORMAL MICROFLORA OF THE SKIN o Consists of stomach, small intestine, and
GASTROINTESTINAL
large intestine
 The skin is generally a dry, acid environment that does
not support the growth of most microorganisms o Responsible for digestion of food, absorption
of nutrients, and production of nutrients by
 Moist areas (e.g., sweat glands) are readily colonized
the indigenous microbial flora
by gram-positive bacteria and other normal flora of the
o Contains 1013 to 1014 microbial cells
skin
 Composition is influenced by
o Environmental factors (e.g., weather)
o Host factors (e.g., age, personal hygiene)
o
NORMAL MICROFLORA OF THE ORAL CAVITY
 The oral cavity is a complex, heterogeneous microbial
habitat
 Saliva contains antimicrobial enzymes
o But high concentrations of nutrients near
surfaces in the mouth promote localized
microbial growth
 The tooth consists of a mineral matrix
(enamel)surrounding living tissue (dentin and pulp)

 Functions and Products of Intestinal Flora

o Intestinal microorganisms carry out a variety
of essential metabolic reactions that produce
various compounds
 The type and amount produced is
influenced by the composition of the
intestinal flora and the diet
 Compounds produced include:
 Vitamins
 Gas, organic acids, and
 Extensive growth of oral microorganisms, especially odor
streptococci, results in a thick bacterial layer (dental  Enzymes
plaque)
 As plaque continues to develop, anaerobic bacterial
species begin to grow
NORMAL MICROFLORA OF OTHER BODY REGIONS
 Attenuation
 A restricted group of organisms colonizes the upper o The decrease or loss of virulence
respiratory tract  Toxicity
o Examples: staphylococci, streptococci,
o Organism causes disease by means of a
diphtheroid bacilli, and gram-negative cocci
toxin that inhibits host cell function or kills
 The lower respiratory tract lacks microflora in healthy host cells
individuals  Toxins can travel to sites within host
not inhabited by pathogen
 Invasiveness
o Ability of a pathogen to grow in host tissue at
densities that inhibit host function
 Can cause damage without
producing a toxin
 Many pathogens use a combination of toxins
invasiveness, and other virulence factors to enhance
pathogenicity

ENTRY OF THE PATHOGEN INTO THE HOST-ADHERANCE

 Specific Adherence
 Urogenital Tract
o A pathogen must usually gain access to host
o The bladder is typically sterile in both males tissues and multiply before damage can be
and females done
o Altered conditions (such as change in pH) o Bacteria and viruses that initiate infection
can cause potential pathogens in the urethra often adhere specifically to epithelial cells
(such as Escherichia coli and Proteus through macromolecular interactions on the
mirabilis) to multiply and become pathogenic surfaces of the pathogen and the host cell
 E. coli and P. mirabilis frequently  Bacterial adherence can be facilitated by
cause urinary tract infections in
o Extracellular macromolecules that are not
women
covalently attached to the bacterial cell
 The vagina of the adult female is weakly acidic and surface
contains significant amounts of glycogen  Examples: slime layer, capsule
o Lactobacillus acidophilus, a resident o Fimbriae and pili
organism in the vagina, ferments the  Pathogen Invasion
glycogen, producing lactic acid
o Starts at the site of adherence
o Lactic acid maintains a local acidic
o May spread throughout the host via the
environment
circulatory or lymphatic systems
COLONIZING AND INFECTION
MEASURING VIRULENCE
 The availability of nutrients is most important in
 Pathogens use various strategies to establish virulence
affecting pathogen growth
 Virulence is the relative ability of a pathogen to cause  Pathogens may grow locally at the site ofinvasion or
disease may spread throughout the body
 Measuring Virulence INVASION
o Virulence can be estimated from  Pathogens produce enzymes that
experimental studies of the LD50 (lethal
dose50) o Enhance virulence by breaking down or
altering host tissue to provide access to
 The amount of an agent that kills
nutrients
50% of the animals in a test group
 Example: hyaluronidase
o Highly virulent pathogens show little
 Protect the pathogen by interfering with normal host
difference in the number of cells required to
defense mechanisms
kill 100% of the population as compared to
50% of the population o Example: coagulase
EXOTOXINS
 Exotoxins
o Proteins released from the pathogen cell as it
grows
o Three categories:
 Cytolytic toxins
 AB toxins
 Superantigen toxins
 Cytolytic toxins
o Work by degrading cytoplasmic membrane
integrity, causing cell lysis and death
 Toxins that lyse red blood cells are
called hemolysins
 Staphylococcal α-toxin kills
nucleated cells and lyses
erythrocytes
  Enterotoxins
o Exotoxins whose activity affects the small
intestine
o Generally cause massive secretion of fluid
into the intestinal lumen, resulting in vomiting
and diarrhea
 Example: cholera toxin (Figure
27.24)

 AB toxins
o Consist of two subunits, A and B
o Work by binding to host cell receptor (B
subunit) and transferring damaging agent (A
subunit) across the cell membrane
 Examples: diphtheria toxin, tetanus
toxin,botulinum toxin

 Clostridium tetani and Clostridium botulinum produce

potent AB exotoxins that affect nervous tissue
 Botulinum toxin consists of several related AB toxins
that are the most potent biological toxins known
(Figure 27.22); tetanus toxin is also an AB protein
neurotoxin (Figure 27.23)
ENDOTOXINS
 Endotoxin
 The lipopolysaccharide portion of the cell envelope of
certain gram-negative Bacteria, which is a toxin when
solubilized
 Generally less toxic than exotoxins
 The presence of endotoxin can be detected by the
Limulus amoebocyte lysate (LAL) assay (Figure 27.25)
 Microial Physiology and 
THE ROLE OF ENZYMES
Catalysts – substances that speed up chemical
reaction without being used up in the reaction by
Metaolism 
lowering activation energy.
Enzymes are biological catalysts (e.g. sucrase –
enzyme that breaks down sucrose to fructose and
MICROBIAL METABOLISM glucose)
Microbial metabolism is important in:
 Agriculture
 Food industry
 Bioremediation
 Pharmacology

MECHANISM OF ENZYMATIC ACTION

Lock and Key Mechanism
1. Substrate binds to the active site (substrate-specific)
2. Formation of enzyme-substrate complex.
3. Substrate is transformed into product
4. Product is released and enzyme is unchanged

ENZYME COMPONENTS
 Apoenzyme – protein portion of the enzyme
CATABOLISM VS. ANABOLISM  Cofactor – non-protein component (e.g. Calcium, Zinc,
 Catabolism – breakdown of macromolecules (Energy Magnesium)
is released) o Coenzyme – cofactor made from organic
o Reactions are hydrolytic (use of water to molecule (e.g. Vitamin B1, Riboflavin, Niacin,
break bonds) and exergonic Folic Acid, Vitamin E, Vitamin K)
 Anabolism – build up of macromolecules (Energy is
used up)
o Reactions involve dehydration synthesis
(reactions that release water) and endergonic
 Catabolic reactions provide building blocks for
anabolic reactions and provide energy to drive
anabolic reactions

FACTORS AFECTING ENZYMATIC ACTIVITY

 Temperature – The higher the temperature, the faster
the enzymatic activity but only up to optimal
temperature (35 to 40 0C).
 pH – Extreme pH can cause enzymes to denature.
 Substrate concentration – Higher substrate, higher
chemical reaction (but should have enough active
sites)
 Inhibitors – can fill active site of the enzyme
(competitive inhibitor) or they interact with another part
of the enzyme (allosteric inhibition) changing the
shape of the enzymemaking it non-functional.

CARBOHYDRATE METABOLISM
 The breakdown of carbohydrate molecules to produce
energy: cellular respiration and fermentation.
 Highlights
 o Glycolysis – Production of 4 ATP and 2 ANAEROBIC RESPIRATION
Pyruvate
 Final electron acceptor is not limited to Oxygen.
o Pyruvate transformed to Acetyl CoA
o Psuedomonas and Bacillus use nitrate ion
o Krebs Cycle – Production of 2 ATP, requires
and is reduced to NO2-, N2O, and N2
Oxygen
o Desulfovibrio sp. use SO4 to form H2S
o Fermentation – Production, does not require
Oxygen o Archaeabacteria use CO2 to form methane
 Energy yield is not high unlike aerobic microorganisms
o ETC – production of 32-34 ATPs, requires
Oxygen
Fermentation
 Yields small amount of energy but ensures steady
supply of ATP.
 End products can vary

LIPID AND PROTEIN CATABOLISM

 Microbes also metabolize lipids and proteins.
 Lipase to break down glycerol and fatty acids
(characteristic that is great for bioremediation in areas
where there is oil spills).
 Protease and peptidase to break down protein and
peptides. Amino group, carboxyl and sulfur group
needs to be removed for the final product to enter
Krebs cycle

GLYCOLYSIS
 Also called Embden-Meyerhof Pathway
 Oxidation of glucose to pyruvic acid.
 Preparatory stage : Production of 2 molecule of GP
 Energy-conserving stage: Production of 4 ATPs from
one molecule of glucose, net gain of 2 ATPs

ADDITIONAL PATHWAYS OF GLYCOLYSIS

 Pentose Phosphate Pathway: Breakdown of pentoses
to produce intermediate pentoses for synthesis of (1) ENERGY ANABOLISM
nucleic acid, (2) glucose from photosynthesis, (3) Photosynthesis
amino acids. Used by Escherichia coli, Bacillus  Cyanobacteria use water as Hydrogen donor
subtilis, Enteroccocus faecalis.
 Entner-Doudoroff Pathway: Allows bacteria to
metabolize glucose without the need for glycolysis or
pentose phosphate pathway. Found in Gram-negative  Purple sulfur and green sulfur bacteria use Hydrogen
bacteria (Rhizobium, Pseudomonas, and donor to produce sulfur
Agrobacterium).

AEROBIC RESPIRATION
Krebs Cycle (Tricarboxylic acid cycle)
PHOTOSYNTHESIS
 For all cycle (2 molecules of pyruvate): produces 4 Light-Dependent Reaction: Photophosphorylation
CO2, 6 NADH, 2 FADH2, 2 ATP. Also produces  Occurs within plastids (Chlorophyll)
intermediates for amino acid biosynthesis.
 Excited electron jumps from one electron carrier to the
other within a series of photosystems.
Electron Transport Chain
 Produces NADPH, O2 and ATP needed for the dark
 Chemiosmotic generation of ATP using carrier
reaction
molecules:
o Flavoprotein
o Cytochrome
o Ubiquinone

 Bacteria have diverse ETC but is largely understood in

eukaryotes. Occurs in plasma membrane in
prokaryotes.
 One molecule of glucose produces 34 ATP
Light-Independent Reaction: Calvin-Benson Cycle
 Also called dark reaction.
 Necessary for Carbon dioxide fixation to produce sugar

AMINO ACID AND PROTEIN BIOSYNTHESIS

MICROBIAL METABOLIC DIVERSITY
 Amino acids are used by microbial cells as building
blocks for protein synthesis.
 Amino acids are obtained as intermediates from
metabolic processes.
 Adding amine group to organic compound (amination)
 Joining of amino acids involves dehydration synthesis
and requires ATP

POLYSACCHARIDE SYNTHESIS
 Microbes synthesize sugar and polysaccharides.
 Polysaccharides are used not only as energy stores
but also components of the cell (cell wall).
 Carbon atoms are derived from intermediates
produced during metabolic processes (to produce
glycogen).

LIPID SYNTHESIS
 Cells synthesize fats by joining fatty acids and glycerol.
 Fats are necessary in cellular structure (plasma
membrane).
 Fatty acids are built up when fragments of Acetyl CoA
are added to each other.
 Fats are linked via dehydration synthesis reactions
that require energy.
 Micro ial growth
REQUIREMENTS FOR GROWTH: PHYSICAL
 Temperature – microorganisms have varying
temperature ranges
o Psychrophiles
o Psychotrophs
o Mesophiles
o Thermophiles
o Hyperthermophiles
 May induce diseases (e.g. catheters in hospitals have
CONTROL OF BIOFILMS
 Use of antimicrobials when surface cleaning.
 Use of lactoferrin (binds with iron making it unavailable
 pH – microorganisms grow best in a narrow pH range
(pH 6.5 and 7.5)
o Very few can grow at pH below 4 except for
acidophiles that can survive at pH 1.
o Bacteria grown in the lab produce acids that
can interfere their own growth – use of
peptones and phosphate salts acts as
buffers.
 Osmotic pressure – Spontaneous net movement of
solvent molecules through selectively permeable
membrane from a region of high water potential to a
region of low water potential.
o Hypotonic solution – solute concentration in
environment is higher than in cell.
o Hypertonic solution – solute concentration
in cell is higher than in environment.
o Isotonic solution – solute concentration in
and outside cell is equal.
Anaerobic Microorganisms
 Oxygen needs to be removed
REQUIREMENTS FOR GROWTH: CHEMICAL
 Carbon – the structural backbone of living matter.
Chemoheterotrophs get most of the carbon from
organic materials.
 Nitrogen, Sulfur, Phosphorus – Necessary for amino
acid and protein synthesis.
 Trace elements – inorganic elements are used as
cofactors (e.g. Iron, Molybdenum, Zinc, Copper).
 Oxygen – have varying effects on growth of bacteria.
 Obligate aerobes – require Oxygen to survive. Comes
with catalase that converts the Hydrogen peroxide to
water and Oxygen.
o Facultative anaerobes – can use Oxygen
when present but can grow in its absence (E.
coli)
o Obligate anaerobes – strictly cannot survive
in the presence of Oxygen. Does not possess
catalase.
o Aerotolerant anaerobes – fermentative
bacteria that cannot use Oxygen for growth
but tolerate it due to SOD (Lactobacili)
o Microaerophiles – grow where Oxygen is
lower in concentration than in air
BIOFILMS
 Microorganisms exist in communities called biofilms.
 Appear as thin slimy layer that adheres to a surface.
 Bacterial colony release signaling chemical called
inducer as a means to communicate to other cells and
induce them to aggregate near the source.
biofilm and can cause nosocomial infections)
to bacteria).
CULTURE MEDIA
 Sterile nutrient material used in the laboratory to grow
microorganisms.
 Inoculum – Microbes introduced into the culture
medium to grow.
 Culture – microbes that grow and multiply on a culture
media.
 Microorganisms require certain condition to grow in
culture media.
 Agar is the most common ingredient of solid Culture
Media but broth can also be used.
 Agar is heated up to liquefy and poured into sterile
petri plates.
INGREDIENTS OF CULTURE MEDIA
 Should be chemically defined – contain exact chemical
composition specific for a type of bacteria that needs
to be grown.
o Chemoheterotroph – culture media should
contain organic compounds as source of
carbon and energy.
 Fastidious organisms – microorganisms that require
complicated (e.g. Leuconostoc mesenteroides).
GROWTH REQUIREMENTS
o The use of Palladium crystals to remove free
Oxygen by combining it to Hydrogen to form
water.
o Presence of lid with O-ring.
o The use of expensive Carbon dioxide
incubator.
o Other safeguards are in place depending on
the type of microorganisms you are working
with (e.g. Ebola – BSL-4 laboratory)

CULTURE MEDIA
Liquid vs. solid – Broth and agar. Used depending on
your goal.
 Rich vs. minimal – Rich media supplies a wide range
of ingredients while minimal media contain basic
metabolites for microbial growth.
 Chemically defined vs. complex – Chemically defined
has a known composition and quantity of nutrients
(therefore reproducible) (e.g. glucose salt broth) while
complex come with water soluble extracts of plants or
tissues (e.g. brain heart infusion, tryptic soy agar).
 Selective, Differential, and Enriched – Selective
medium inhibits growth of many microorganisms
except for a few. Differential medium contains
ingredients that can distinguish one microbe to the
other. Enriched medium provide extra nutrients to help
fastidious organisms grow
OBTAINING PURE CULTURE
 Colony – distinct group of microbes that grow on solid
agar (with distinct size, margin, texture, color).
o Arise from a single spore or vegetative cell
from a group of similar microorganism
 The use of streak plate method to generate colonies
on the plate
GROWTH OF BACTERIAL CULTURE
 Microorganisms undergo binary fission
 Microbial growth is logarithmic
 Growth is divided into 4 phases: lag, log, stationary,
and death
MEASURING COLONY NUMBER
 Pour Plate Method – inoculum is poured first into the
petri plate before agar is poured.
PHYSICAL METHODS OF MICROBIAL CONTROL: OTHERS
o Problem: Molten agar might be too hot.
 Spread Plate Method – inoculum is streaked on
solidified agar
Phenols and Phenol Derivatives
Control of Microial
growth
ACTIONS OF MICROBIAL CONTROL AGENTS
 Alteration of membrane permeability – plasma
membrane regulates the passage of molecules in and
out the cell.
 Damage to proteins and nucleic acids – Proteins and
nucleic acids are building blocks to life
PHYSICAL METHODS OF MICROBIAL CONTROL: HEAT
 Heat – application of heat (moist heat or dry heat) to
kill microbial life.
o Moist heat sterilization – Autoclaving for 15
PSI (1210C) for 15 to 20 minutes to kill off
endospore.
o Dry heat sterilization – direct flaming of wire
loop (until it glows red) or the use of hot air in
the oven to sterilize glassware at 1700C for 2
to 4 hours.
 Others (food): Pasteurization (700C) or Ultra-High-
Temperature Treatments (140C for 4 seconds)
 Filtration – passage of liquid or gas through a pored
PHYSICAL METHODS OF MICROBIAL
material to screen out microbes.
CONTROL: FILTRATION
o Use of High-efficiency particulate air (HEPA)
filters.
o Size of the pores vary (0.22um to 0.45um)
 May not filter all microbes (e.g. Mycoplasma, virus,
viroids, and proteins
 High Pressure – High pressure changes the molecular
structure of protein and carbohydrates.
 Radiation – Use of high energy electron beams
(ionizing radiation) to denature DNA.
 Desiccation – Removal of water (drying) kills off
microorganisms (e.g. Freeze drying)
CHEMICAL METHODS OF MICROBIAL
 First used by Joseph Lister to control surgical infection
CONTROL: PHENOLS
in operating room.
 Examples cresols, bisphenols, triclosan
 Essential oils also contain phenols and terpenes
 Phenols kill microbes by injuring the plasma membrane
 (attacks lipids) or inhibiting lipid synthesis.
 Can effectively kill Mycobacterium
 Good surface disinfectants
 CHEMICAL METHODS OF MICROBIAL  Endospores are resistant to some biocides
CONTROL: BIGUANIDES  Cysts and oocytes of protozoa and helminths are
Biguanides resistant to chemical disinfection.
 Effective against gram-positive and gram-negative  Viral envelop also influence the resistance of virus
bacteria. against biocides (non-enveloped are more resistant)
 Have broad spectrum activity that affects bacterial  For prions, no effective sterilization method has yet
membrane. been employed. Everything potentially contaminated
 Examples of biguanides include chlorhexidine and needs to be destroyed via incineration
alexidine.
 Used as surface disinfectants for preoperative skin
preparation in patients

Halogens
 CHEMICAL
ExamplesMETHODS OF MICROBIAL
include Chlorine and Iodine.
CONTROL: HALOGENS
o Iodine – impairs protein synthesis and alters
cell membranes by forming complexes
withfatty acids and amino acids. Great for
surface disinfection
o Chlorine – Forms hypochlorous acid (an
effective oxidizing agent) when added to
water.
 Can both be used to disinfect drinking water
EVALUATING A DISINFECTANT
 Disk diffusion assay – observance of zone of inhibition

CHEMICAL METHODS OF MICROBIAL CONTROL: ROH

Alcohol
 Can kill bacteria and fungi but not endospores and
non- enveloped virus.
 Can denature protein and dissolve lipid.
 Good as surface disinfectant but not for open wounds.
 Ethanol and Isopropanol (70%) are effective in killing
off most microorganisms

 Heavy metals have oligodynamic (antimicrobial) action

CHEMICAL METHODS OF MICROBIAL
against microbes.
CONTROL: HEAVY METALS
o Copper
o Silver
o Zinc
 Used in treating water, aquarium (can kill fish
parasites and algae), killing biofilms

Soaps
 CHEMICAL METHODS OF MICROBIAL
Applying soap mechanically removes bacteria.
CONTROL: SOAPS
 Soap breaks lipid in the plasma membrane.

Antibiotics – Certain types of antibiotics are used in food

CHEMICAL METHODS OF MICROBIAL
preservation.
CONTROL: OTHERS
 Nisin (a bacteriocin) is added to food to inhibit growth
of gram-positive endospore bacteria. Nisin produced
by Lactococcus lactis.
Aldehydes – Glutaraldehyde used in disinfecting surgical tools
and surfaces.
Peroxygens – Destroy cell walls via oxidation (steal electrons
disrupting metabolic activities). Also good in killing anaerobic
bacteria (do not have catalase)
 Example is Hydrogen peroxide

MICROBIAL CHARACTERISTICS MATTERS

Microbial characteristics matter
 Most gram-negative bacteria are susceptible to
antimicrobial agents compared with gram-negative
bacteria due to the porin on the cell wall (selective
integrins).
 Mycobacterium have a very waxy and lipid-rich cell
wall that makes it difficult to break.
 Human Microe o Microbial antagonism: competition of
microorganism for food sources and territory.
o Pneumocystis jirovecii is antagonistic against
Interaction – Indigenous Microflora other bacterial species

INDIGENOUS MICROFLORA: MOUTH

Bacteria found:
 Streptococcus
 Lactobacillus
THE HUMAN MICROBIOME  Actinomyces
 The human microbiome is the aggregate of all  Bacteroides
indigenous microorganisms living on or within human  Veillonella
tissues and biofluids.  Neisseria
 Microbial population is established before birth  Haemophilus
(Enterobacter and Priopionibacterium in placenta)  Fusobacterium
 Treponema
INDIGENOUS MICROFLORA: SKIN  Staphylococcus
Bacteria found:  Corynebacterium
 Propionibacterium  Candida (fungus)
 Staphylococcus
 Corynebacterium
 Micrococcus
 Acinetobacter
 Brevibacterium
 Candida (fungus),
 Malassezia (fungus)

 Secretions like oil and sweat can have antimicrobial

properties (making it hard for microbes to establish
residency)
 Other barriers: keratinized layer, low pH and low
moisture content in many areas

INDIGENOUS MICROFLORA: EYES  Mouth is abundant in moisture and is warm enough to

Bacteria found: support growth of microorganisms.
 Staphylococcus epidermidis  Constant presence of food also makes it ideal to
 S. aureus support diverse microbial populations.
 Propionibacterium  Biting, chewing, tongue movement, and salivary flow
 Neisseria can dislodge microbes.
 Corynebacterium  Saliva contains antimicrobial substances (lactoferrin,
 streptococci Hydrogen peroxide, lysozymes)
 Micrococcus
INDIGENOUS MICROFLORA: DIGESTIVE SYSTEM
 Conjunctiva is a mucous membrane and contains Bacteria Found:
similar microflora found on the skin.  Escherichia coli
 Tears and blinking eliminate some microbes and  Bacteroides
inhibit others from colonizing  Fusobacterium
 Lactobacillus
INDIGENOUS MICROFLORA: RESPIRATORY TRACT  Enterococcus
Bacteria found:  Bifidobacterium
 Nose: Aerobic diphtheroids in the nose  Enterobacter
 Throat: S. epidermidis, S. aureus, diphtheroids,  Citrobacter
Streptococcus pneumoniae, Haemophilus, and  Proteus
Neisseria
 Klebsiella
 Lower respiratory tract: Neisseria, Candida albicans,
 Candida (fungus)
diptheroids, streptococci, staphylococci, Pneumocystis

 Nasal secretion can kill/inhibit microbes.

 Mucus and ciliary action in the upper respiratory tract
can mechanically remove microbes.
 Microbial antagonism occurs among normal microbiota
in the respiratory tract.
  Large intestines contains the largest numbers of ANTAGONISTIC SUBSTANCES PRODUCED BY INDIGENOUS MICRO
resident due to the availability of moisture and food
resources.
 Mucus and periodic shedding of the lining prevent
many microbes from attaching to the lining of the
gastrointestinal tract.
 Mucosa produces several antimicrobial chemicals (bile)
 Indigenous microflora produces bacteriocins that can
inhibit the growth of alien microorganisms

INDIGENOUS MICROFLORA: URINARY AND

REPRODUCTIVE SYSTEMS
Bacteria found:
 Bladder – Proteobacteria, Actinobacteria, EFFECT OF ANTIBIOTIC THERAPY TO MICROBIOME
FirmicuteStaphylococcus, Micrococcus, Enterococcus,
Lactobacillus, Bacteroides, aerobic diphtheroids,
Pseudomonas, Klebsiella, and Proteus
 Urethra - Lactobacilli, Streptococcus, Clostridium,
Candida albicans (fungus)
 Vagina – Trichomonas vaginalis (protozoan), Candida
albicans, Lactobacillus, Bacteroides, Pseudomonas,
Bifidobacterium, Enterobacter, Gardnerella, Prevotella,
Atopobium

Nutritional Determinants
ENVIRONMENTAL DETERMINANTS OF
 Different parts of the body produce different sources of
INDIGENOUS MICROFLORA
energy (carbon sources) for microbes.
o Skin – lipids
o Respiratory mucosa – proteins and mucins
o Cecum and ascending colon – carbohydrates
o Descending colon – proteins
 The human body provides microbiota with complex
compounds that the microbes need to hydrolyze the
molecules to smaller units to be used up in
metabolism. Many microorganisms work in
cooperation with several species to digest complex
carbon sources.
 Some bacterial communities utilize toxic substances to
allow other bacteria to thrive (e.g. Malassezia spp
metabolizes lauric acid that is toxic to
 Lower urethra in both sexes has resident populations. Propionibacterium acnes).
 Mucus and periodic shedding of the lining prevents
microbes from attaching to the lining. Physico-Chemical Determinants
 pH and urea in urine is antimicrobial  Homeostatic mechanisms ensure fairly constant
 Vagina has acid-tolerant population. environment at sites colonized by microbes.
 Cilia and mucus expel microbes from cervix of the  Variables that affect indigenous microflora include:
uterus into the vagina where they are killed by acidity. temperature, pH, water activity, atmospheric
composition, salinity, light.
 Host and indigenous microbiota display symbiotic
RELATIONSHIP BETWEEN HOST AND
relationship: Commensalism, Mutualism, Parasitism A. Temperature: microbes that can colonize the body are
INDIGENOUS MICROFLORA
 Normal microbiota prevent overgrowth of harmful mesophiles (25 to 400C)
microorganisms through competitive exclusion or B. pH: pH in the body varies on different tissues (acidic:
microbial antagonism (production of bacteriocins) stomach, cecum, skin, duodenum) (alkaline:
 Indigenous microflora population that are known subgingival region, tear film, ileum) (neutrophiles: the
pathogens (opportunistic bacteria) are kept in check rest of the body)
by variables like pH. C. Atmospheric composition: Vast majority of indigenous
 Indigenous microflora can also wreck havoc to the microflora are obligate aerobes and facultative
body if they find their way in other tissues. anaerobes (skin and oral cavity)
 Alteration of their environment cause opportunistic D. Water activity: Availability of water for microbial activity
bacteria to increase in number but there are some microorganisms (e.g
Staphylococcus) that can survive and colonize dry
regions of the skin.
E. Salinity: High salt concentration is detrimental to
microbes (osmotic pressure). Skin has high salt
concentration.
F. Sunlight: Sunlight contains damaging UV radiation.
Skin and eyes are exposed to UV light but little
evidence exerts light with inhibitory effect on skin
microflora.
Mechanical Determinants
 Certain regions are subjected to mechanical forces.
 Flow of saliva in the oral cavity removes bacteria from
the mouth.
 Intestinal secretions create hydrodynamic shear forces
to prevent bacteria from attaching to mucosal
surfaces.
 Peristalsis and other gut movements remove
unattached microbes.
 Periodic flushing of urine removes microbes on
mucosal surfaces
 Ciliary action of the upper respiratory tract removes
microbes
Biological Determinants
 Innate immune system can control indigenous
microflora: monocytes, macrophages, NK cells,
dendritic cells
 Release of cytokines
 Production of antimicrobial peptides (e.g. lysozyme,
lactoperoxidase, collectins)
 Studies have shown that the immune system of the
host is tolerant to indigenous microflora
Age
HOST CHARACTERISTICS AFFECTING
 Microbiota communities differ in young and
INDIGENOUS MICROFLORA
individuals.
 Infants have immature immune system, limited
(milk-based). Fecal microbiota is dominated
Bifidobacterium
 Adults are dominated by Bacteroides sp.
colonization of the urinary tract is also observed
Host Genotype
 Colonization of bacteria is associated with host
genotype (genes, genetic information)
 Helicobacter pylori (vacA type s1a strains) are found
only in individuals with Asian descent.
Gender
 Males have higher risk of being colonized by
Helicobacter pylori.
 Males have higher density of microbes due to
increased production of sebum andsweat
ROLE OF INDIGENOUS MICROFLORA TO HUMAN HEALTH
Host Development
 Indigenous microflora can affect the development of
host tissues.
 Absence of indigenous microflora can affect anatomy
and physiology of host.
 Indigenous microflora changes gene expression in
host to improve absorption andprocessing of
biomolecules and micronutrients.
 Intestinal microflora stimulates the growth and
differentiation of crypt cells.
Improve Host Nutrition
 Food items that are not easily digested by enzymes in
the stomach (cellulose, hemi-cellulose, inulin, guar,
and karaya) but is digested in the colon.
 Intestinal microflora is capable of amino acid
fermentation produces acetic, propionic, and butyric
acid that is taken up by colon epithelial cells for
synthesis of biomolecules.
 Intestinal microflora produces vitamins that is used up
by the host.
Detoxification
 Microbial metabolism can detoxify potentially harmful
dietary constituents
 Gram-positive bacteria (Lactobacillus, Clostridium, and
Bifidobacterium can detoxify Heterocyclic aromatic
amines.
 Other harmful dietary constituents metabolized by
bacteria
o Phytoestrogens from soybeans
o Methylmercury from fish
Disease Pathogenicity:
Principles of Epidemiology
EPIDEMIOLOGY
The study of the factors influencing the frequency and
distribution of diseases, when and where the disease occur and
how they are transmitted in a population.
Use of epidemiological studies
 For policy development, implementation, and
old
evaluation of treatment and management of diseases.
o What are the health problems in the
diet community?
by o Where are the occurring?
o Which population are at risk?
and o Are there diseases that have increasing
incidence?
HALLMARKS OF EPIDIMOLOGICAL STUDIES
 John Snow (mid-1800s) – investigated deaths in
London between 1848 to 1849 and associated it with
cholera outbreaks.
 Ignaz Semmelweis (1846-1848) – observed maternal
death at Vienna General Hospital. Determined the
association between medical students not washing
their hands before handling women in childbirth
(reduced the mortality rate to only 2%).
 Florence Nightingale (1858) – recorded statistics on
epidemic typhus in military and civilian populations.
Observed that unsanitary conditions and poor food are
killing soldiers.
 Robert Koch (1884) – developed the germ theory of
disease.
EPIDEMIOLOGY OF DISEASES
A review on Koch’s Postulate:
1. The same pathogen must be present in every case of
the disease.
2. The pathogen must be isolated from the diseased host
and grown in pure culture.
3. The pathogen from the pure culture must cause the
disease when it’s inoculated into a healthy, susceptible
Laboratory animal.
4. The pathogen must be isolated from the inoculated
animal and must be shown to be the original organism.

LIMITATION OF KOCH’S POSTULATE
 Many microorganisms are recalcitrant due to their
unique culture requirements (e.g. Treponema
pallidum, Mycobacterium leprae) while some can only
multiply in living tissues (e.g. viral pathogens).
 Many pathogens do not have defined signs and
symptoms (e.g. Mycobacterium tuberculosis is
implicated in the disease of the lungs, bones, skin, and
other internal organs).
 Ethical consideration also imposes exception to the
Koch’s postulate (e.g. HIV does not have any other
known host other than humans and it is not ethical to
inoculate humans with it).
DEVELOPMENT OF DISEASE
 Incubation period – interval between initial infection
and first appearance of signs or symptoms. The time
of incubation depends on the growth rate, microbial
load, and host resistance. Pathogens also become
infective once theystart replicating inside the host.
 Prodromal period – a short period following
incubation period. Characterized by early and mild
symptoms of disease. Symptoms may be generic with
other diseases.
 Period of illness – The disease is most severe and
signs and symptoms become evident.
 Period of decline – Signs and symptoms subside.
Body’s immune system overcome the pathogen.
Patient is still vulnerable to secondary infection.
 Period of convalescence – Individual regains
strength and body recovers,

Alternatives to Koch’s Postulate

 Alternative method of detecting microbes are in place
while still following the principles of Koch’s postulate
(e.g. inoculation of tissues into animal models)
SPREAD OF INFECTION: RESERVOIR
 Some pathogens present a clear-cut distinguishable
 Human Reservoirs – Humans harbor pathogens and
signs and symptoms
transmit it in/directly to others. Human reservoir can be
symptomatic or asymptomatic (carriers) (e.g. Adults
CLASSIFICATION OF INFECTIOUS DISEASES
carry Bordetella pertussis can transmit it to non-
Definition of terms
vaccinated infant, “Typhoid Mary”).
 Symptom – changes in body function (pain and
 Animal Reservoirs – Refers to wild and domestic
malaise). Subjective changes not apparent to
animals that harbor pathogens that can cause
observer.
diseases to humans (zoonoses).
 Signs – objective changes that can be observed and
 Non-living Reservoir – Anything that harbors
measured.
pathogens (e.g. soil and water) (e.g. Clostridium
 Syndrome – a specific group of symptoms or signs
botulinum and ringworm come from soil).
that accompanies by a particular disease. Diagnosis
requires laboratory tests.
TRANSMISSION OF DISEASE
 Communicable disease – contagious disease that can Contact transmission – disease is spread by in/direct contact
be transmitted to individuals via an infectious agent or droplet transmission
directly or indirectly (e.g. chicken pox, small pox,
 Direct contact transmission – person-to-person
tuberculosis)
(animal-to-human) transmission via physical contact
 Non-communicable disease – disease that does not (e.g. Hepatitis A, measles, and most STI, rabies,
spread from one host to the other (e.g. Clostridium anthrax).
tetani)
 Congenital transmission – disease from mother to
fetus (newborn). Pathogen is able to cross the
OCCURRENCE OF DISEASE
placenta or through direct contact via vaginal
 Incidence of disease – number of people in a secretions (e.g. Herpes, Rubella, Enterovirus).
population who developed the disease for a particular
 Indirect contract transmission – (fomite) the agent of
time period. Also an indicator of the spread of disease
disease is transmitted via non-living object.
 Prevalence – number of people in a population who
develop a disease at a specified time regardless on Droplet transmission – pathogens are carried by droplets
when it first appeared. (e.g. coughing, sneezing)
o HIV AIDS – incidence in the US in 2014 was
37,600 prevalence in the same year is 1.1 Vehicle transmission – transmission of disease via air, water,
million
or food.
 Frequency of occurrence – how often a particular
disease occurs (
 Vectors – refers to arthropods that carry diseases via:
o sporadic – occurs occasionally
 Mechanical transmission – passive transport of
o endemic – constantly present
pathogens from insects (e.g. Houseflies carry
o epidemic – occurrence of the disease within a
short period of time shigellosis)
o pandemic – widespread occurrence of  Biological transmission – active process of transmitting
disease). pathogen (e.g. biting)
 Microial Pathogenicity
and Modes of Transmission
I. MODES OF TRANSMISSION
 PORTALS OF ENTRY
 FACTORS AFFECTING PATHOGENICITY
PATHOGENICITY AND VIRULENCE OF MICROORGANISMS
DEFINITION OF TERMS
 Pathogenicity – ability of microorganisms to overcome
host defenses and cause disease.
 Virulence – degree of pathogenicity.
 Portals of entry – points of the body which pathogens
use to gain entrance to the human body.
PORTALS OF ENTRY
Mucous Membrane
 Membrane that lines the body cavities (e.g.
respiratory, digestive, urogenital tracts, and
conjunctiva).
o Respiratory tract – most frequently portal of
entry. Microbes are inhaled from the nose or
mouth via aerosol or dust particles.(e.g.
common cold, pneumonia, tuberculosis,
influenza, and measles)
o Gastrointestinal tract – can gain access via
contaminated food and water. Only a small
population survive as most are destroyed by
the HCl in the stomach and bile enzyme in
the intestines (e.g. typhoid fever, dysentery,
giardiasis, cholera, shigellosis).
o Genitourinary tract – entry portal of pathogen
responsible for sexually transmitted infections
(e.g. STI, genital warts, chlamydia)
Skin
 Serves as the first line of defense against pathogens
(especially unbroken skin).
 Pathogens can still enter the skin via hair follicles and
sweat gland ducts.
 Some pathogens have enzymes that can break
through skin (e.g. hookworm larvae)
 Can enter via exposed mucous membrane (e.g.
conjunctiva)
 Common for organisms to take on the parenteral route
(e.g. presence of punctures, injections, surgery, bites,
cuts and any skin break-offs)
FACTORS INFLUENCING PATHOGENICITY
Preferred portal of entry
 Pathogens have preferred portals of entry (e.g.
Salmonella typhi prefer the oral route, Streptococcus
pneumoniae can be pathogenic when inhaled but not
when swallowed.)
 Some pathogens (e.g. Yersinia pestis and Bacillus
anthracis can initiate disease from more than one
portal of entry)
Number of invading microbes
 Large microbial load can overcome the host’s
defenses.
 The virulence of a microbe is often expressed as the
ID50 (estimated number of organisms required to
produce infection in 50% of normal adults exposed by
a given route).
 Bacillus anthracis – ID50 varies on portal of entry:
o Skin - 10 to 50 endospores
o Inhalation – 10K to 20K endospores
o Gastrointestinal route – 250K to 1M
endospores
 Vibrio cholerae
o 1 x 108 cells via ingestion
o If stomach acid is neutralized with
o bicarbonate, the ID50 decreases.
Adherence
 Pathogens have means of attaching on host tissues
 Pathogens have ligands (adhesins) that bind to
surface receptors on the cells.
o Adhesins are found on the glycocalyx, pili,
and fimbrae
o Adhesins are made up of glycoproteins and
lipoproteins
o Surface receptors are made from sugar
(mannose)
o Some pathogens adhere only to specific
kinds of cells (e.g. Staphylococcus aureus
only binds to the laminin and fibronectin on
skin cells)
 Biofilms also allow microbial communities to aggregate
together on a surface
o release of chemoattractants to attract more
microbes to the biofilm
o Secrete glycocalyx for other bacteria to
attach to the surface of the biofilm
 Biofilms in the body: dental plaque
 Evolutionary advantage for the microorganisms to
resist disinfectants and antibiotics (significant in
nosocomial infections as they reside on catheters and
stents)
II. HOW PATHOGENS PENETRATE HOST DEFENSE
 COMPONENTS OF CELL WALL
 HOW BACTERIA EVADE HOST DEFENSE
Capsule
MICROBIAL PROPERTIES
glycocalyx (aFOR PENETRATING
 Made from polypeptide slime) HOST
DEFENSE
 Bacterial cell wall increases the virulence of pathogens.
 May also resist host by impairing phagocytosis.
 May also be responsible to the virulence of some
pathogens (e.g. Streptococcus pneumoniae is virulent
when capsule is present).
 Bacteria that produce capsules that are virulent:
Klebsiella pneumoniae, Haemophilus influenzae,
Bacillus anthracis, and Yersinia pestis.
Cell Wall Components
 Cell walls contain substances that contribute to
virulence
o Streptococcus pyrogenes – has heat-
resistant and acid-resistant proteins (M
proteins)
o Neisseria gonorrhea – use Opa protein to
effectively attach to host cells
o Mycobacterium tuberculosis – contains
mycolic acid to resist digestion by phagocytes
and allow pathogens to multiply inside
phagocytes
o Gram-positive bacteria – peptidoglycan
inactivates lysozymes
Enzymes
 Coagulase – enzymes that coagulate fibrinogen (turn
to fibrin) in the blood. Fibrin clot protects pathogens
from phagocytosis.
 Bacterial kinase – breaks down fibrin and digests clots
to isolate the infection (e.g. Fibrinolysin/Streptokinase)
  Hyaluronidase – hydrolyzes hyaluronic acid
(connective tissues). Produced by Clostridia that
causes gangrene. Involved in tissue blackening of
infected wounds
 Collagenase – produced by Clostridium and breaks
down protein and collagen.
 IgA protease – enzymes that destroy IgA antibodies
(e.g N. meningitidis, N. gonorrhoeae)
 Invasin – rearrange cytoskeleton arrangement on
plasma membrane causing membrane ruffling.
Antigenic Variation
 Ability of the pathogen to alter surface antigens before
the immune system mounts a massive attack on
pathogens.
o N. gonorrhoeae – has copies of Opa
encoding gene that result to bacterial cells
having varying antigens over time.
o Influenza virus can alter its surface proteins
Hemagglutinin-neuraminidase (to allow to
stick to potential cells)
TOXIN NOMENCLATURE
 Toxins are named after the type of host cell that they
attack (e.g. hepatotoxin – attack liver cells, neurotoxin
– attacks glial cells)
 Toxins are also named after the type of disease they
produce (e.g. diphtheria toxin – causes diphtheria)
 Toxins are also named for specific bacterium that
produces them (e.g. botulinum toxin – produced by
Clostridium botulinum)
DIRECT DAMAGE: EXOTOXIN
 Exotoxin – produced mostly by gram-positive bacteria
but can also be present in gram-negative bacteria.
Toxin produced in bacteria and released after lysis.
 Comes with enzymatic action – small amounts of
exotoxins are harmful (e.g. only 1 mg of botulinum
exotoxin can kill 1 million guinea pigs)
 Exotoxins are soluble in water and can diffuse into the
blood to be transported all throughout the body.
 Promotes the production of antitoxins in the body.

TYPES OF EXOTOXINS
 A-B Toxins: Consist of the enzyme component (A part)
and binding component (B part) (e.g. Diphtheria toxin)
o Genotoxin – a type of A-B toxin produced by
gram-negative bacteria that disrupts cell
division
 Membrane-disrupting toxins – causes lysis to host
cells (e.g. luekocidins and hemolysin produced by
staphylococci and streptococci)
 Superantigens – altered proteins that provoke intense
immune response. Bacterial proteins that combine
with protein on macrophages to stimulate T cells to
release cytokines (for regulation and immune
response) (e.g. staphylococcal toxins that cause food
III. HOW PATHOGENS DAMAGE CELLS poisoning and toxic shock syndrome)
 MECHANISM ON HOW MICROBES DAMAGE HOST
CELLS

MICROBIAL PROPERTIES THAT DAMAGE HOST CELLS

Ways on how pathogens damage the host cells
 Using host’s nutrients
 Causing direct damage in immediate vicinity of invasion
 Producing toxins that damage tissues far from site of
infection
 Producing hypersensitivity reactions

SIDEROPHORES
 Pathogens secrete siderophores.
o Proteins that take iron way from iron-
transport proteins from host (e.g. hemoglobin,
lactoferrin, ferritin, and transferrin)
 Free iron enters the bacterium to be used as cofactor
or in the oxido-reduction mechanism.
 Some pathogens, when low in iron, release toxins to
host cells to release iron from the host. DIRECT DAMAGE: ENDOTOXIN
 Toxins that are part of the bacterial cell (and not a
DIRECT DAMAGE: TOXINS metabolic product)
 Many pathogens grow in host cells and are released  Made from lipids
when cells rupture (e.g. virus, protozoans)  Released during bacterial multiplication
 E. coli, Shigella, Salmonella, and Neisseria  Common in gram-negative bacteria (e.g. lipid A is an
gonorrhoeae can induce host epithelial cells to engulf endotoxin)
them by a process that resembles phagocytosis
 Most use toxins to damage cells
o Toxigenicity – capacity of microorganisms to
produce toxins (toxin is transported via blood
or lymph)
o Toxemia – presence of toxins in the blood
detected through laboratory tests
o Intoxication – symptoms caused by the
presence of toxin and not by microbial growth
(e.g. Clostridium perfringens)
 ENDOTOXIN ACTION
 Endotoxin exert their effects by stimulating
macrophages to release cytokines (high
concentration of cytokines can result to endotoxic
shock)
 May activate blood clotting proteins to
obstruct capillaries.
 Endotoxin can also be released via phagocytosis of
pathogen by macrophages (e.g. release of tumor
necrosis factor). TNF bind to capillaries increasing the
permeability of tissues causing leakage (drop in blood
pressure)
o Haemophilus influenza type B in CSF causes
weakening in the blood-brain barrier.
ENDOTOXIN
 Endotoxins do not promote the formation of antitoxins.
 Released by Salmonella typhi and Neisseria
meningitidis.
 Presence of endotoxin can be detected with Limulus
amebocyte lysate (LAL) assay.
o Use of the hemolymph (blood) of an
Atlantic horseshoe crab
IV. PATHOGENICITY OF VIRUS
 MECHANISMS ON HOW VIRUS INFECTS HOST
CELLS
VIRAL LIFE CYCLE
VIRAL LIFE CYCLE
 DS DNA integrate into cell’s genome as a provirus and
remains latent until the host physiology is stressed
(e.g. herpes virus)
 SS RNA virus (Class IV) serve directly as mRNA.
 Retroviruses uses reverse transcriptase to turn RNA to
ds DNA. Viral DNA is integrated into chromosome
where it is transcribed by host cell into viral RNA
BALTIMORE CLASSIFICATION SYSTEM
 Baltimore classification is a system used to classify
viruses based on their manner of messenger RNA
(mRNA) synthesis. By organizing viruses based on
their manner of mRNA production, it is possible to
study viruses that behave similarly as a distinct group.
Seven Baltimore groups are described that take into
consideration whether the viral genome is made of
deoxyribonucleic acid (DNA) or ribonucleic acid
(RNA), whether the genome is single- or double-
stranded, and whether the sense of a single-stranded
RNA genome is positive or negative.
VIRAL MECHANISM FOR INVADING HOST CELLS
 Cytopathic effect of viruses – visible effects of viral
infection and used to diagnose viral infection.
o Cytopathic effects can vary by virus.
 Cytocidal effect – cytopathic effects that result in death
of host cells.
 Noncytocidal effect – cytopathic effects that result to
cell damage but not death.
TYPES OF CYTOPHATIC EFFECTS
 Inhibits macromolecule synthesis and cellular
processes (e.g. Simplex virus irreversibly halts
mitosis)
 Host cells are hijacked by virus to release enzymes
from lysosomes.
 Virus release inclusion bodies (proteins/nucleic acid
granules) or replication factories necessary for viral
replication inside the cell. (e.g. Negri bodies released
by rabies virus)
 Virus causes cells to aggregate into a multinucleated
cell called syncytium (e.g. common cold, measles,
mumps)
 Virus may result to the change of the host cells’
function (e.g. measles virus reduce the production of
cytokine when attached to its receptors)
 Viral infections induce antigenic changes on the
surface of infected cells by incorporating their genome
in host cell eliciting auto-immune response from the
host.
 Induce chromosomal changes (e.g. oncogenes can be
activated by virus, HPV) or may transform host cells to
malignant type (does not stop from dividing).
 o Fomites
o Infected blood
o Vectors
 Necessary for the pathogen to spread through the
population and is important in the epidemiology of
diseases.

V. HOW PATHOGENS DAMAGE CELLS

 HOW EUKORYOTIC PARASITES INVADE CELLS

FUNGI
 Produce metabolic products that are toxic to humans
o ergot produced by Claviceps purpurea that
causes hallucinations (e.g. ergotism)
o Aflatoxin from Aspergillus flavus often
associated on peanuts (carcinogenic)
 May provoke allergic response to host (e.g.
Trichothecenes that causes Athlete’s foot)
 May secrete enzymes (e.g. Candida albicans and
Stachybotrys produce protease.
 May produce mycotoxins (neurotoxins) phalloidin and
amanitin (e.g. Amanita phalloides)

PROTOZOANS
 May produce metabolic wastes that are toxic to
humans.
 Can invade and rupture host cells (e.g. Plasmodium)
 Can hijack host cells and change its physiology (e.g.
Toxoplasma enters macrophages and prevents
digestion.
 Can attach to host cells and digest cells and tissue
fluids.
 Can produce many antigens to fool the immune
system (e.g. Trypanosoma)
 Few species produce neurotoxins (e.g. dinoflagellate
Alexandrium produce saxitoxin)

HELMINTHS
 Evokes cellular damage by using host cells to produce
large parasitic masses (e.g. Wuchereria bancrofti)
 Helminth infection often associated with:
o Low lipid/cholesterol levels (Schistosoma
mansoni)
o Increased creatine phosphokinase, lactate
dehydrogenase, aldolase, and
aminotransferase (e.g. Pinworms or worms
attached to muscle tissues)

PORTAL OF EXIT
 Portals of exit include: secretions, discharges, or
shedding of tissues.
o Expelled via coughing and sneezing (e.g.
pathogens that cause tuberculosis, whooping
cough, mumps, chickenpox, flu,
meningococcal meningitis)
o Expelled via feces (e.g. pathogens that cause
dysentery, cholera, typhoid fever,
salmonellosis)
o Expelled via saliva (e.g. rabies, mumps, and
most pathogens infecting respiratory tract
o Urogenital tract can be a portal of exit (e.g.
STI)
o Skin shedding can spread ringworm, Simplex
virus and warts.
o Drainage from wounds