S.Y.

2021-2022 / 1st Semester / Midterms

CLINICAL BACTERIOLOGY
Ms. Raquel M. Fernandez, RMT, MSPH
August 25, 2021

CHAPTER 3: MICROBIAL GROWTH AND CONTROL

OUTLINE ▪ Psychotrophs- grows in
I. Requirements for Growth J. Steps in the temperature within 0˚C to 15˚C
A. Physical Pathogenesis of (Alaska)
i. Temperature Infectious Disease ▪ Moderate psychophiles- able to
ii.pH K. Virulence
grow between 20˚C to 30 ˚C
iii.Osmotic pressure L. Virulence Factors
B. Chemical i.Attachment (Canada)
i.Carbon ii.Exoenzymes o Mesophiles
ii.Nitrogen, Sulfur, and iii.Endotoxin ▪ involved in infectious disease
Phophorus iv.Exotoxin process
iv.Trace Elements M. Mechanisms by Which
v.Oxygen Pathogens Escape ▪ live in moderate temperature
C. Organic Growth Factors Immune Responses (between 25˚C to 40˚C)
D. Anaerobic Growth Media III. Host Defense Mechanism ▪ the optimum temperature is between
………and Methods A. Definition of Terms 35˚C to 37˚C (body temperature)
E. Microbial Control Methods B. Host Defense
▪ When culturing bacteria, set the
II. Infection Mechanism
A. Definition of Terms C. Three Lines of incubator at ±35˚C to cater the
B. Classification of Defense Against optimum temperature that the
Infections Infection mesophiles would need.
C. Local vs. Systemic D. Non-specific Host o Thermophiles
Infections Defense Mechanism
▪ live between 50˚C to 60˚C
D. Infection vs. Infectious i.First Line of Defense
Disease ii.Second Line of o Hyperthermophiles
E. Acute, Subacute, and Defense ▪ need 80˚C or higher temperature
Chronic Disease E. Specific Host Defense requirement
F. Latent infection Mechanism ▪ some can tolerate as much as 121˚C
G. Symptoms vs. Signs of F. Two Types of
Disease Immunity (same in autoclave)
H. Pathogenesis of G. Classes of
Infectious Disease Immunoglobulin
I. 4 Periods in the Course H. CMI
of an Infectious Disease I. Lymphocytes pH
• Most bacteria grow best in a narrow pH range neutrality
between 6.5-7.5
• Each bacterial species has its own preferential growth
REQUIREMENTS FOR GROWTH at certain pH
• Lowest pH (acidic) recorded bacteria where it is able to
grow is pH 1 is Chemoautotrophic bacteria
Physical
(cyanobacteria).
• Very few bacteria grow below pH 4
Temperature • Molds and yeasts grow at pH 5 to 6, they can tolerate
• The first requirement needed by the bacteria is the temperature and low pH
correct temperature. • When bacteria are cultured in the laboratory, they often
• Most bacteria grow only within a limited range of produce acids that interfere with their own growth. That
temperatures. is why we need to ensure that the culture medium
• Each bacterial species grows at a particular minimum contains chemical buffers (in growth medium), so when
(lowest temperature which bacterial species will grow), acids are produced by the bacteria it will not lower the
optimum (temperature which bacteria will grow best) pH in the medium, they would die in too much presence
and maximum temperatures (highest temperature on of acidity.
which growth is possible for the bacteria). • Chemical buffers should be included in culture
• Bacteria is very diverse and each of it has its own mediums that we have to provide for them (peptones,
preference for temperature for them to grow. amino acids, and salts)
• On the basis of preferred range of temperature,
bacteria can be classified as:
o Psychrophiles

Charisse Patricia Paronda – BSMT 3A

 CHAPTER 3: MICROBIAL GROWTH AND CONTROL

Osmotic Pressure
• Organisms require water for growth
• The growth of the cell is inhibited as the plasma membrane
pulls away from the cell (addition of salts)
• Hypertonic or hypotonic environment
o Hypertonic environment
- Adding salt
- Will cause the cellular water (the water
from the bacterial cell inside) to pass out
through the plasma membrane whether it
is high salt concentration, from rigid low
concentration to higher region of
concentration (high solute
concentration).
- If the bacterial cellular water passes out
to the plasma membrane the bacterial Nitrogen, Sulfur, and Phosphorus
cell will shrink and will die • Nitrogen makes up the 14% of the dry weight of the bacterial
o Hypotonic environment cell. Sulfur and Phosphorous are about 4%
- Exposed to distilled water, the water • These chemicals are needed by the bacteria for synthesis
tends to enter the cell (from a region of of protein
lower concentration to a higher • For protein synthesis (Nitrogen and Sulfur)
concentration). It will enter the bacterial • For DNA and RNA synthesis
cell and the bacterial cell will swell, when
• K, Mg, and Ca are also needed as co-factors for bacterial
it cannot hold the water coming inside, it enzymes
will burst.
- Bursting means lysing of the cell, the Trace Elements
bacterial cell will die.
• Some microbes have relatively weak cell wall maybe by the
• (Fe) Iron
increase of osmotic pressure, it can also inhibit bacterial
• (Cu) Copper
growth. It is very important to provide the correct amount of
• (Mo) Molybdenum
water, hypotonicity, and hypertonicity of the environment
• (Zn) Zinc
should also be carefully considered.
• They are naturally present in tap water
• Obligate or extreme halophiles (30% salt)
• Halophiles - salt loving bacteria that can tolerate 30% salt Oxygen
• Obligate halophiles - strictly needed for a hypertonic • Metabolic systems require oxygen for aerobic respiration
environment for them to be able to live (metabolic pathways, final electron acceptor is oxygen)
• Facultative halophiles - 2-5% salt concentration is what • Microbes that use molecular oxygen produce more energy
they need in order to grow from nutrient than microbes that do not use oxygen (more
• Bacteria have diverse requirement ATP, more energy is produced when you use molecular
oxygen than those do not use oxygen)
• Oxygen is important for living organisms. Oxygen is also
Chemical
poisonous, a toxic gas. We normally produce the toxic
• Sources of C, N, S, P, trace elements, O2, and organic forms of oxygen during respiration in small amounts.
growth factors. Therefore, we also produce toxic gas as well, this is the way
how oxygen is toxic. It will instigate the chemical reactions
Carbon inside the body that can produce free radical, super active
• All organic compounds are made up of Carbon (even molecules that can destroy our cells. But our bodies are
humans) designed to capture and be able to use nutrients like oxygen
• C is the structural backbone of living matter to our advantage.
• Half of the dry weight of typical bacterial cell is carbon • Obligate aerobes – strict aerobes, oxygen is a requirement
• Chemoheterotrophs, chemoautotrophs, photoheterotrophs, • Facultative anaerobes – part time anaerobes, basically
and photoautotrophs aerobes their energy will decrease in the absence of
• Chemotrophs (chemical energy source), Phototrophs oxygen, they can use oxygen to their advantage
(light energy source) • Obligate anaerobes - strict anaerobes, they can live
• Largely, bacteria are classified under chemoheterotrophs. without the presence of oxygen
• Aerotolerant anaerobes - anaerobes that can tolerate
oxygen, they cannot use oxygen to their advantage

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 CHAPTER 3: MICROBIAL GROWTH AND CONTROL
• Microaerophiles - type of bacteria that can only tolerate o Most aerobic respiration produces traces of
small amount of oxygen, around 5-10% of oxygen. Oxygen hydroxyl radicals but they are transient (go
in the air is around 21%. away).
• Capnophiles (capnophilic)- needs carbon dioxide when o In phagolysosome, once we ingest
they metabolize and live pathogens, they are killed by exposure to
these toxic forms of oxygen. These are
contained in the lysosomes.

Organic Growth Factors

• Vitamins which functions as co-enzymes
• Other organic growth factors: amino acids, purines,
pyrimidines
• May be synthesized by bacteria themselves or should be
obtained from the environment

Anaerobic Growth Media and Methods

• Reduced media is used such as sodium thioglycollate that
chemically combine with dissolved oxygen and deplete the
oxygen in the medium

Anaerobic System

• Toxic Forms of Oxygen- can destroy cell membrane.

• Singlet oxygen (1O2-)
o is normal O2 boosted into a higher energy
“Anaerobic Jar System”
state and is extremely active
• Culture the bacteria, put those culture plate or broth
• Superoxide free radicals (O2-)
inside and then put a gas pack, pack of chemicals
o formed in small amounts during the normal
(sodium bicarbonate & sodium borohydride)
respiration of organisms. SOD neutralizes
• Cut off the edge of packet, put water. Once it is filled
this.
with water, hydrogen and carbon dioxide will be
o Oxygen combined with hydrogen the SOD is
produce by this pack of chemicals. The palladium
the one that will neutralize the superoxide free
catalyst will allow the oxygen in the jar combined with
radicals on order to convert them into simpler
the hydrogen produced by this packet of chemicals.
chemicals, it will be converted by the SOD into
Oxygen and hydrogen, water is formed. When they
hydrogen peroxide and oxygen. But still,
react, you will see droplets of water inside the jar. It
peroxide is still a toxic form
means the reaction is taking place and the oxygen is
o This can start several chemical reactions; this
being removed. Carbon dioxide is the only element
is the form we normally respire
remains inside
O2 + O2 + 2H+ → H2O2 + O2
• Peroxide anion (O22-)
o Toxic from which is neutralize by catalase
converts it to water and oxygen;
2H2O2 → 2H2O + O2
o Peroxidase also convert it to water but no
oxygen, thus (in case catalase is absent)
H2O2 + 2H+ → 2H2O
• Hydroxyl radical (OH-)
o Intermediate form of oxygen and probably the
most reactive. It is formed in the cellular
“Candle Jar Extinction”
cytoplasm by ionizing radiation.

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 CHAPTER 3: MICROBIAL GROWTH AND CONTROL

INFECTION

Definition of terms

• Pathogenesis is the step of mechanisms involved in the

development of the disease
o Path- refers to disease
• Pathogenicity – the ability to cause disease
• Pathogen – is a microorganism capable of causing
“Anaerobic Chamber” disease (may be bacteria, virus, fungi, etc.)
• It has a containment for the mixture of oxygen provided • Pathology is the study of the structural and functional
in cylinders of gases (nitrogen, oxygen, etc.). It is tightly manifestation of the disease
closed and sealed. • Infection
• For greater mobility for culturing anaerobes, it is the o Injurious contamination of body or parts of the
best system for anaerobic environment setting. body by bacteria, viruses, fungi, protozoa, and
rickettsia or by the toxin that they may
produce.
Capnophiles
o May be local or generalized and spread
throughout the body
o Once the infectious agent enters the host, it
begins to proliferate and reacts with the
defense mechanisms of the body producing
infection symptoms and signs (pain, swelling,
redness, functional disorder, rise in
temperature, pulse rate, and increase in
WBC)

Classification of Infection
• Primary infection
o Initial infection with organism in host
o First time that the host will encounter a
microbe, virus, or any microbe is the primary
infection
Microbial Control Methods
• Reinfection
o Subsequent infection by same organism in a
host (after recovery)
o After recovery, the same organism will attack
the host
• Superinfection
o Infection by same organism in a host before
recovery
o Infection by the same organism before the
host could hardly recover
• Secondary infection
o When in a host whose resistance is lowered
by preexisting infectious disease, a new
organism may set up infection
o The host could have a bacterial infection in the
beginning, because of the
immunocompromised situation of the patient,
another infection set up by maybe a virus is
now on its way
• Focal infection
o It is the condition where due to infection at
localized sites like appendix and tonsil,
general effects are produced.
o Only in a certain area of the body

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• Cross infection
Chronic Disease
o When a patient suffering from a disease and
new infection it set up from another host or • Has a slow onset and last a long time.
external source • Examples: Tuberculosis, leprosy, syphilis
o One patient with the disease, the infection is • It will take a long time for a patient to recover from these
transmitted to another host set of illness.
• Nosocomial infection Subacute
o Cross infection occurring in hospital
• Diseases that come on more suddenly than chronic
o Hospital acquire infections
diseases but less suddenly than acute diseases.
▪ Examples: facilities, contaminated
• Somewhere in between acute and chronic illness
surfaces, contaminated materials
• Example: Bacterial endocarditis
• Subclinical infection
o It is one where clinical effect are not apparent
o The infection is in the patients but there are Latent Infections
no signs and symptoms that would tell you
that the patient is infected with the certain • A disease that may go from being symptomatic to
microbe asymptomatic and then back to being symptomatic.
• The causative agent or pathogen remains inactive for a
Local vs. Systemic Infection period of time and then becomes active again when it
changes its mind and produce symptoms again
• Example: Syphilis – for three weeks it will present signs and
Localized
symptoms, and after 2 to 6 months will have the primary
• An infection that remains localized at one site chancre, and then develop rashes, lesions, fever within 2 to
• Remains confined to a certain site 6 months’ period. Later on, after five years or few years, no
• Examples: Pimples, boils abscess symptoms will be present. Only to find out that after other
Systemic/Generalized years again, symptoms will reappear and the body is
already destructed (spinal cord and organs are really
• An infection that has spread throughout the body
destroyed)
• Example: Tuberculosis – it may spread to different
organs of the body
Symptoms vs. Signs of Disease
Primary vs. Secondary Infection
Symptoms
• Primary infections- the first disease
• some evidence of a disease that is experienced or
• Secondary Infections- the second disease
perceived by the patient only
• This is only known by the patients
Infection vs. Infectious Disease • Examples: Aches, pains nausea, itchiness
o Symptomatic disease (clinical disease) – a
• Infectious disease is a disease cause by a microbe disease in which the patient is experiencing
and the microbes that cause infectious disease are symptoms
collectively referred to as pathogens. o Asymptomatic disease (subclinical disease) – a
• Infection, according to many microbiologists, means disease that the patient is unaware of because
colonization by a pathogen. he/she is not experiencing any symptoms
• A person can be infected with a certain pathogen but
not necessarily have the infectious disease. Signs
• Infection means there are pathogens colonizing a • some type of objective evidence of a disease
certain area, but it does not actually involve in the • Examples: Fever, pulse rate, blood pressure, test result,
disease yet. A person may be infected but do not have respiratory rate, bradycardia, tachycardia
the illness yet. Meaning a person is infected, maybe not
presenting any signs and symptoms. Pathogenesis of Infectious Disease

Acute, Subacute, and Chronic Disease Questions:

• Why would infection would not necessarily result to
infectious disease?
Acute Disease
• Why would a certain infection would not lead to infectious
• Has rapid onset usually by a relatively rapid recovery
disease?
• Examples: Measles, mumps, influenza
• What are the possible reasons why a person maybe infected
• These are acute diseases you would recover from 1 to
with a certain pathogen but not necessarily have the
2 weeks
infectious disease?

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 CHAPTER 3: MICROBIAL GROWTH AND CONTROL
factors that destroy the pathogen) that destroy the
newly arrived pathogen
Why infection does not always occur?
o Importance of presence of microbiota in the
• The microbe may land at an anatomical site where
body
it is unable to multiply
o Pathogen might have landed on the skin but
there is the normal microbiota that made the 4 Periods in the Course of an Infectious Disease
bacteria not succumb to the infection because
it counterattacks those invading microbes that Incubation Period
might land on the skin. • The time that elapses between the arrival of the
o Some other reasons are the factors in the skin pathogen and the onset of symptoms
that would not promote the multiplication of • Incubation period will depend on the nature of the
the bacteria (presence of oils, chemicals) that
microbe that arrived in the body/system. (short or long)
would not allow the microbe to survive those
• This the time when you are not yet experiencing
chemicals on those conditions.
symptoms
• This time the pathogen had just arrived
• Many pathogens must attach to specific receptor
• Factors influencing the incubation period:
site before they are able to multiply and cause
o Over-all health and nutritional status of the
damage
host- the healthier you are, the lesser chance
o This is the reason why viruses that are in
or the longer it will be for the incubation
animals cannot transfer to humans because
period. The pathogens will have a difficult time
they don’t have specific receptor sites on
creating or trying to find the chance to invade
human cells.
the body
o Same with the animals to animals, the
infection cannot transfer because of the
o Virulence of the pathogen- mechanism or
unavailability of receptor sites.
factors in the pathogen will make it able to
cause disease.
• Antibacterial factors that destroy or inhibit the
growth of the microbes maybe present at the site ▪ Examples: Fungi (low virulence),
where the pathogens are Corona virus (highly virulent)
o Possibility of antibiotics, antimicrobials and
the likes are present that would not promote o Numbers of the pathogens that enters the
survival of the pathogen in that area or site. body- for some disease, it will take a several
numbers of the pathogen before they could
• The individuals’ nutritional and overall health status initiate a disease.
often influences the outcome of the pathogen/host ▪ Ex. Tuberculosis – only one live
encounter tuberculosis bacilli can cause
o If the resistance is high and immune to a tuberculosis in an
certain pathogen, there are chances on not immunocompromised patient
getting the disease. If you are overall healthy, ▪ For some disease, it will take a 106 of
(getting enough sleep, exercising) have good bacteria to be able to cause diarrhea
immunity, the body will not be succumb in the for example.
pathogen. ▪ The number of pathogens will
depend as it enters the body
• The person maybe immune to that particular
pathogen and is the result of prior infection or
Prodromal Period
vaccination
• the time during which the patient starts to feel
o Possibility of being immunized or vaccinated
something is wrong but do not experience yet the
with that particular pathogen.
actual symptoms of the disease
• Signs and symptoms are present but not specific
• Phagocytic white blood cells may engulf and
(nonspecific signs and symptoms)
destroy before it has an opportunity to multiply,
• Ex. Fever, feeling heavy
invade and cause disease
• The period wherein you know something is wrong in
o Presence of phagocytes that had a chance to
your body but don’t know the cause.
engulf the pathogen before it could create
havoc to the host

• The indigenous microbiota at the site may produce

antibacterial factors (bacteriocins – antibacterial

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 CHAPTER 3: MICROBIAL GROWTH AND CONTROL
• INVASION/SPREAD of the pathogen
Period of Illness o The spread of the pathogen will create the signs
and symptoms and at this time the host defenses
• The time when the person experiences the symptoms will be evaded by the pathogen
of the disease. Communicable diseases are mostly
easily transmitted during this stage • EVASION of host defenses
• Specific signs and symptoms are present that actually o It will create damage to the host, leaving the host
identify having illness defenseless

• DAMAGE to host
Convalescent Period
o The host defenses have been evaded by the
• The time when the person recovers. Person may pathogen so it will now create damage to the host
recover from the illness but there may be permanent
damage of tissues of the affected area Virulence
• This 4th period may be either convalescent period or • It is a measure of the degree of pathogenicity; different
death/disability (depending on the outcome) species or different strains of microbe vary in durability to
• This is the time when the person recovers cause disease.
(convalescent) • Virulence factors are the phenotypic characteristics of
• The person may recover from the illness but there may microorganisms that enable it to cause disease
be permanent damage of the affected areas (disability) • Virulent Strains – are capable of causing disease
or death • Avirulent strains – not capable of causing disease

Virulence Factors

Attachment

• To cause the disease, the microorganism must be able to

anchor themselves to cells after gaining entrance into the
body. This is where attachment factors are very important
• Receptors and Integrins
o Molecules on the surface of the host cell that a
particular pathogen is able to recognize and attach
to which are often glycoprotein molecules
TAKE NOTE: As the period of infectious disease continue, the o Surface of the host cell is the particular site where
number of the microorganisms initially in the incubation period the pathogen will be able to attach.
are below. It will peak at a certain point (including the period of o Receptors is where the pathogen will attach
illness), and then decline (the number of microorganisms). As • Adhesins and Ligand
the period progresses at the point of period of decline, the o Used to describe the molecule on the surface of
number of microorganisms will also decline, up to the period of the pathogen that is able to recognize and bind to
recovery or convalescents, wherein the number of a particular receptor
microorganisms will be at the lowest. o Therefore, the receptors and integrins are in the
host. While the adhesins and ligands are on the
pathogens.
Steps in the Pathogenesis of Infectious Disease
o Ex. Streptococcus pyogenes – they have adhesins
• ENTRY of the pathogen into the body
called protein F, it is on their surface that will
o We have several portals of entry in the body.
adhere to the protein on the host cell called
Examples: Skin and Mucous membrane
fibronectin (lock and key). Adhesion on the
pathogen and then surface/receptor protein on the
• ATTACHMENT of the pathogen to some tissues
host cell.
o Once it has gain entrance to the body it will attach
o Ex. HIV – they have the GP120 (glycoprotein 120)
to some tissues
it is an adhesin, it must attach to the CD4+ cells or
T helper cells of the host cell.
• MULTIPLICATION of the pathogen
• Bacterial Fimbriae (Pili) / Common pili
o After attaching, it will lodge into the tissues and
o Long thin, hair-like, flexible projections composed
cells of the body and multiply. The multiplication of
of primarily of an array of proteins called pilin (by
the pathogen will lead to the spread and invasion
nature it is protein) which enable the bacteria to
of the different body parts or tissues of the body
attach to the surfaces and cause infection

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 CHAPTER 3: MICROBIAL GROWTH AND CONTROL
o Bacteria like Neisseria gonorrhea (gram negative o Ex. Streptococcus can produce streptokinase
diplococci) can anchor to the inner walls of the that will be able to dissolve blood clots
urethra causing urethritis. But that Neisseria • Hyaluronidase
gonorrhea that are not fimbriated can be flushed o Called spreading factor because it enables
out when the host urinate. The fimbriated strains the pathogen to spread throughout the
are therefore more virulent than the non-fimbriated connective tissue by breaking down
strains. They are more likely to cause disease is hyaluronic acid
the virulent factor for them. o Hyaluronic acid – this is a polysaccharide
• Obligated Intracellular Pathogens cement that holds the tissues together
o Rickettsia, Chlamydia, Ehrlichia must live o Once the bacteria have this enzyme the tissue
within the host cells in order to survive and will fall apart. When the tissue is destroyed,
multiply the pathogen can spread throughout the
o When the pathogen is inside cell, antibodies connective tissue.
will not work for them because they are inside o Hyaluronidase is secreted by
the host cell Staphylococcus, Streptococcus, and
• Facultative Intracellular Pathogens Clostridium spp.
o Pathogens that are able to survive • Collagenase
intracellularly and extracellularly. These o Breaks down collagen which is the supportive
microorganisms are able to survive within protein found in tendons, cartilage and bones
phagocytes. enabling the pathogen to invade tissues
o Once the bacteria are ingested by WBCs o If the bacteria can excrete collagenase, it can
(macrophages or neutrophils), the invade tissues. This is one virulence factor of
phagosome will fuse with the lysosomes Clostridium perfringens
(contains superoxide hydrolytic enzyme • Hemolysins
peroxide) which will cause destruction of o Enzymes that cause destruction of RBCs.
bacteria. o Provides iron to the pathogen
o Once ingested, facultative in the cellular o The pathogen lyses the RBCs so that it can
pathogens could opt to leave the cell and go capture the iron
outside the host cell. o The Hemolysins could be alpha-beta or
• Capsule gamma hemolysis
o They can evade phagocytosis because they • Lecithinase
are sticky or slimy o Enzyme produced by Clostridium perfringens
• Flagella which breaks down phospholipids collectively
o They enable to invade aqueous area of the referred to as lecithin. This enzyme is
body that non-flagellated bacteria will not be destructive to cell membrane of RBC and
able to do. It is hard to be targeted by host other tissues
defense mechanism because moves around. o This is how Clostridium perfringens cause gas
gangrene.
Exoenzyme
Endotoxin
• enzymes that are liberated by the bacteria outside.
• Necrotizing Enzymes • Integral part of the cell walls of gram-negative
o Bacteria produce proteases and lipases which bacteria producing septic shock, chills, fever,
cause destruction of tissues prostration, and septicaemia during gram negative
o Ex. Proteases and Lipases – streptococcus infection. The cell wall contains lipopolysaccharide
pyogenes this strain will cause destruction of (LPS) called LIPID A, which is actually the endotoxin.
soft tissues called flesh eating necrotizing
fasciitis Characteristics of Endotoxins
• Coagulase
• Proteins polysaccharide lipid complex heat stable
o Enable S. aureus (positive for coagulase) to
• Forms part of cell wall (don’t diffuse into medium)
clot plasma and thereby form a sticky coat of
• Obtained only by cell lysis
fibrin around themselves for protection
• They have no enzymatic action
against phagocytes, antibodies, etc. so that
• Effect is non-specific action
they may not be engulfed or not be
• No specific tissue affinity
recognized by the phagocytes
• Active only in large doses 5 to 25 mg
• Kinases or Fibrinolysins
• Weakly antigenic
o Substance that will dissolve the fibrin clot that
the host will attempt to form (escape from • Neutralization by antibody infective
clots) • Cannot be toxoided
• Produce in gram negative bacteria

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• Highly potent, e.g. 3kg botulinum can kill all the inhabitants
Exotoxins
of the world
• They are also poisonous substances but they are • They are generally formed by Gr+ bacteria and also by some
producing within the bacterial cells and it is released Gr- organisms like Shigella, V. cholorae, and E. coli
from them. They are usually named from the target - Usually produced primarily by gram positive
organs that they affect. • Exotoxin is specifically neutralized antitoxin
• Neurotoxin • Can be separated from culture by filtration
o most potent exotoxin produced by • Action is enzymatic and it has specific tissue affinity
Clostridium tetani (causative agent of tetanus) • Cannot cause pyrexia (high fever) in a host
and Clostridium botulinum • Can be toxoided
o Toxin that will affect the nerves
o Tetanospasmin - neurotoxin of C. tetani; it Mechanisms by which Pathogens Escape Immune
will create the condition of spastic paralysis Response
o Botulinum toxin – blocks nerve impulses
producing a different kind of paralysis called • Antigenic Variation
flaccid paralysis o Pathogens are able to periodically change their
• Enterotoxins surface antigens
o Cause diarrhea and sometimes vomiting o Ex. Influenza viruses – by the time the host has
o Toxin that is produced in the intestine produced antibodies, they will come up with new
o Toxin B produced by C. difficile damages the antigens and had already shed the old ones. Even
surface of the colon leading to if the antibodies are present already, it will not be
pseudomembranous colitis effective anymore because they had changed their
• Toxic shock syndrome toxin (TSST) antigens already. Once they change their antigens,
o Produced by strains of S. aureus and S. there are new strains of influenza.
pyogenes which primarily affect the integrity o Ex. Trypanosomes – these are the blood
of capillary walls. flagellates. They can keep up their antigenic
• Exfoliative Toxin variations for 20 years. They never presented the
o Also called epidermolytic toxin, causing sloughing same appearance twice.
of the epidermal layers of the skin leading to SSSS
(staphylococcal scalded skin syndrome) cause by • Camouflage and Mimicry
Staphylococcus aureus or by some Streptococcus o They can mimic or coat and hide themselves.
pyogenes o Ex. Schistosomes - they are able to hide their
o It is usually seen in newborn babies (skin reddish foreign nature by coating themselves with host
and sloughing). The epidermal layer is removed in proteins (camouflage)
the process. o Molecular mimicry – pathogen surface would
• Leucocidin resemble the host antigens. It will not be
o Toxin that will destroy leukocytes, produced by recognized by the immune capabilities
Staphylococcus and Streptococcus
• Diphtheria toxin • Destruction of Antibodies
o Toxin produced by Corynebacterium which o Some bacteria can produce molecules or enzymes
inhibits protein synthesis killing mucosal epithelial that could destroy our antibodies produced.
cells and PMN’s (polymorphonucleus) o Ex. Heamophilus influenzae and Neisseria
o This is the one that causes diphtheria. If a gonorrhea – can produce the enzyme called IgA
Corynebacterium is not able to secrete exotoxin protease, that will counter act and destroy the
therefore it is non-pathogenic. antibody IgA that we can produce.
o This is why after identifying Corynebacterium, the
next step to do after it is isolated and identified is HOST DEFENSE MECHANISMS
to do toxigenicity test (if the Corynebacterium is
not positive in diphtheria toxin then it is not at all Definition of Terms
pathogenic) • Resistance – the ability to ward off disease
• Endotoxin is a lipid a component of the gram-negative outer o Nonspecific Resistance- Defenses that protect
membrane, and exotoxin that can be produce by the against all pathogens like virus, bacteria and fungi
bacteria and release it outside. o Specific Resistance- Protection against specific
pathogens
Characteristics of Endotoxins • Susceptibility – vulnerability or lack of resistance. If you are
• Heat labile protein susceptible, you are prone to contract disease and different
• Diffuse readily into the surrounding medium infections brought by different microbes or pathogen.

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Host Defense Mechanism Non-specific Host Defense Mechanism

Non-specific Host Defense Mechanism First Line of Defense

• Serve to protect the body from a variety of foreign

substances or pathogens (first and second line of
defense)
• Would cater to all types of pathogens. It will not
recognize if it is a virus, microbe, or bacteria.
• Will serve for all substances (toxins or pathogens)

Specific Host Defense Mechanism

• Are directed against a particular foreign substance or

pathogen that has entered the body (third line of
defense)
• If the first line of defense did not work, the second line
of defense will be activated. And when the second
defense did not work, the third line defense will be • Physical or Mechanical Barriers: Intact Skin and Intact
activated. Mucous membranes
o Mucous Membranes: Line gastrointestinal,
Three Lines of Defense Against Infection genitourinary, and respiratory tracts.
o Two layers: Outer Epithelial and inner
connective layer.
o Epithelial layer secretes mucus which maintains
moist surfaces
▪ Although they inhibit microbial entry, they
offer less protection than skin.
o Several microorganisms are capable on
penetrating mucus membrane such as Treponema
pallidum (spirochete), papilloma virus,
Enteroinvasive e. coli, and E. histolytica.
First Line of Defense • Chemical Factors such as digestive enzymes, acidity of
stomach (pH 1.5) and alkalinity, acidity of vagina, lacrimal
• Are non-specific natural barriers which restrict entry of apparatus.
pathogens. It is naturally found in human. o Sebum: Oily substance produced by sebaceous
glands that forms a protective layer over skin.
Second Line of Defense
Contains unsaturated fatty acids which inhibit
growth of certain pathogenic bacteria and fungi.
• Macrophages and neutrophil (phagocytic leukocytes) o PH: low, skin pH usually between 3 and 5. Caused
by lactic acid and fatty acids.
• Proteins that are anti-microbial against different
o Perspiration: produced by sweat glands. Contain
pathogen
Lysozyme and acids
• Innate that provide rapid response against invading
o Lysozyme: Enzyme that breaks down gram-
pathogens once it breaches the first line of defense
positive cell walls. Found in nasal secretions, saliva
(entering the skin or mucus membrane), the second
and tears.
line of defense will be activated.
o Gastric Juice: Parietal cells secrete mixture of
Third Line of Defense hydrochloric acid enzymes, and mucus. Ph
between 1.2 to 3 kills many microbes and destroys
most toxins. Many enteric bacteria are protected
• It has antigen specific immune responses. The invaders by food particles.
that will pass in two levels of non-specific defenses will ▪ Helicobacter pylori neutralize stomach
be attack specifically. acid and can grow in the stomach,
• B and T lymphocytes causing gastritis and ulcers.
• Y shape antibodies
• It develops in the process.

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• Microbial Antagonism: By indigenous microflora; and over • Wandering macrophages: Originate from monocytes that
all nutritional status and state of health leave blood and enter infected tissue and develop into
o More appropriate term is microbiota phagocytic cells.
o It is bacteria that inherently present in our body • Fixed macrophages (histocytes): Located in liver,
that provides protection against invading nervous system, lungs, lymph nodes, bone marrow and
pathogens (no need to activate). several other tissues.
o Part of our health immune system

Second Line of Defense

• Transferrin and lactoferrin: Are tie up iron (iron binding

protein), thereby preventing pathogens access to this
essential mineral.
o Once iron is no more available, it will not be
accessible to the bacteria (since bacteria needs
iron)
o Prevent pathogen access
• Fever: That augments host defense by stimulating
leukocytes to deploy and destroy invaders, reducing
available free plasma iron, inducing of 1L - 1, which causes
proliferation, maturation, and activation of lymphocytes in
the immunologic response.
o Elevated body temperature also slows down the
rate of growth of certain pathogens and can even
kill some especially fastidious pathogens.
o Temperature is very important factors for growth
(either kill or let them grow)
• Interferons: Are small, antiviral proteins that prevent viral
multiplications in virus infected cells and serve to limit viral
infections.
• Inflammation: Localizes an infection and prevent the
spread of microbial invaders, neutralizes toxins and aid in
• When injury happens, the cells nearby will send off chemical
the repair of damaged tissues. signals in form of chemotaxis to call out phagocytes to the
• Phagocytosis: Cell eating. Microbes are being engulfed. site of injury
• Complement system: Involves approximately 30 different • Dilation and increased permeability so that phagocytes can
blood proteins that interact in a step-wise manner known as squeeze in in the site of injury
the complement cascade.

CONSEQUENCE OF ACTIVATION OF THE COMPLEMENT

SYSTEM:
• Initiation and amplification of inflammation.
• Approximately 30 activated directly by pathogen.
• Indirectly by antigen antibody reaction.
• It generates active components in order to fight of the
invading pathogens.
o Attraction and activation of leukocytes.
▪ It sends signals to call out the leukocytes
to be deployed in site of invasion.
o Lysis of bacteria and other foreign cells.
▪ Attacking the pathogens with protein
materials PROCESS OF PHAGOCYTOSIS
o Increased phagocytosis by phagocytic cells. 1. Chemotaxis and adherence of microbe to phagocyte
▪ Labeling the pathogen by opsonization or (the adherence of the microbe to the phagocyte will
coating them by antibodies depends on the virulence factor)
PHAGOCYTOSIS 2. Ingestion of microbe by phagocyte
• Derived from the Greek words “eat and cell”. 3. Formation of phagosome
• Carried out by white blood cells: macrophages, neutrophils, 4. Fusion of the phagosome with a lysosome to form a
and occasionally eosinophils. phagolysosome
• Neutrophils: predominate early in infection 5. Digestion of ingested microbe by enzyme

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 CHAPTER 3: MICROBIAL GROWTH AND CONTROL
6. Formation of residual body containing indigestible o Interferon alpha: produced by B-
material lymphocytes, monocytes, and macrophages
7. Discharge of waste materials. o Interferon beta: fibroblasts and other virus
infected cells.
INFLAMMATION
o Interferon gamma: activated by T-
• triggered by tissue damage due to infection, heat, lymphocytes and NK cells.
wound, etc.
• Four Cardinal Major Symptoms of Inflammation Specific Host Defense Mechanism
1. Redness- Rubor
• Immunology is the scientific study of the immune system
2. Pain- Dolor and immune responses.
3. Heat- Calor • The immune system is the third line of defense against
4. Swelling- Tumor pathogens; it is a specific host defense mechanism.
but may also observe: • Two types of acquired immunity
5. Loss of function 1. Active Acquired Immunity
PURPOSES OF INFLAMMATION a. Natural active acquired immunity: that is
acquired in response to the entry of a live
1. Destroy and remove pathogens
pathogen into the body (i.e., in response to an
2. If destruction is not possible, to limit effects by actual infection). It has long duration.
confining the pathogen and its products. b. Artificial active acquired immunity: that is
3. Repair and replace tissue damaged by pathogen and acquired in response to vaccines. Its duration
its products. for many years; but must be reinforced by
boosters.
2. Passive Acquired Immunity:
a. Natural passive acquired immunity: that is
acquired by a fetus when it receives maternal
antibodies in utero or by an infant when it
receives maternal contained in colostrum. Its
duration from 6 months- 1 year.
b. Artificial passive acquired immunity: that is
acquired when a person receives antibodies
contained in anti-sera or gamma globulin. Its
duration from 2-3 weeks.

Two Types of Immunity

Humoral Immunity: Antibodies and antigens:

COMPLEMENT ACTIVATION • an antibody is a protein produced in response to

• Consequences of Complement Activation: foreign substance (antigen) that will react specifically
1. Cytolysis: Due to the formation of a membrane with that substance. The reaction between the antibody
attack complex (MAC) which produces lesions in and the antigen will lead to destroy the antigen or the
microbial membranes. pathogen that carry the antigen or inhibit it.
o The bacterial cell will lies because of the • Antibodies are produced by WBC (plasma cells) and
lesions that be created by the MAC. may present in the blood and the body fluids or
2. Inflammation: Complement components (C3a) attached to surfaces of cells. They are also called
trigger the release of histamine, which increases immunoglobins (Ig)
vascular permeability. • Antibodies are glycoproteins produced by B
o It will attract macrophages and neutrophils. lymphocytes, that once it is transformed to plasma
3. Opsonization: Complement components (C3b) cells, it become antibody secreting. It will bind to
bind to microbial surface and promote specific antigen in the antigenic determinant.
phagocytosis. • Antibodies are very particular and specific, and it reacts
INTERFERONS like lock and key. It is also called immunoglobulin.
• Antiviral proteins that interfere with viral multiplication. • Antigens are substances that stimulate the animal body
o They serve to limit the viral infection, very to produce antibodies that will specifically react with
small protein around 30,000 heat stable and the antigens.
resistant to low pH • Antigen is always complex macromolecules (mostly
• Have no effect on infected cells. glycoproteins)
• Host specific, but not virus specific • Antigen is antibody generating molecules and digenic
o Unable to save the virus infected cell but once or immunogenic
release, they attached to membranes of that • The best antigens are the foreign proteins because of
surrounding cell and prevent viral replication the complexity of the confirmation proteins
o They are effective against variety of viruses, • Bacteria are full of antigens. It is a mosaic of antigen
and they are specific only in the species of determinants (epitopes-antigenic site)
animals • Antigens can be haptens- small molecules that may act
• 3 types of Interferons as antigens only if they are coupled with a large carrier
molecule such as proteins.

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the peripheral blood. Several types are T-helper,
Cytotoxic T-cells, etc.
Classes of Immunoglobulin
• IgG: This is the greatest percentage of antibody molecules
in the blood, these globulins combine to small antigen and
combine and neutralize toxins (antitoxins). Long lived and
crosses the placenta.
• IgM: is a pentamer, has 10 antigen binding sites, first
antibodies formed in the primary response to antigens.
Does not cross the placenta.
• IgA: Known as secretory antibodies because they are found
in breastmilk, respiratory and intestinal mucin, saliva, tears,
and vaginal secretions protect these parts of the body from
infectious agents. It exists in three for monomer, dimere,
trimere. 10-20% of the total serum immunoglobulin is
compose of IgA
• IgD: It is found on the surfaces of the B-lymphocytes where
it acts as a specific antigen receptor.
• IgE: Its activities involved in both the resistance to parasites
infections and hypersensitivity. It is found on the surfaces of
basophils and muscles.
All antibodies are made up of proteins, globular glycoproteins. It
can be found in the blood, saliva, colostrum, lymph. The amount
in type of antibody produces during antigenic stimulation
depends on the nature of the antigen, the site of the stimulus the
amount of antigen, and the number of times of exposure.

CMI (Cell Mediated Immunity)

• Does not involved production of antibodies

• Controls intracellular pathogens
• Cells that participate are macrophages, T-helper cells,
cytotoxic T-cells, NK cells, Killer cells, and granulocyte.
• Major player in CMI are B cells and T cells.

Typical CMI Response

• Step 1: A macrophage engulfs and partially digests a

pathogen. Fragments (antigenic determinants) of the
pathogen are then displayed on the surface of the
macrophage.
• Step 2: A T-helper cell binds to one of the antigenic
determinants being displayed on the macrophage
surface. the T-helper cell produces lymphokines which
reach an effector cell of the immune system.
• Step 3: The effector cell binds to a target cell.
• Step 4: Vesicular contents of the effector cell are
discharged.
• Step 5: Toxins produced by the effector cells enter the
target cell causing disruption of DNA and organelles.
The target cell dies.

Lymphocytes

• B-lymphocytes: 10-15% of lymphocytes in Peripheral

blood. B cells migrate to lymphoid tissues where they
produce antibodies that circulate through lymph and
blood. They live about 1-2 weeks.
• T-lymphocytes: T-cells are phagocytic cells, it engulfs
the antigen or the pathogen that carries that antigen, it
destroys the infected body cells, and it rejects the
foreign tissue. About 70%-80% of the lymphocytes in

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