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Tayyar M 1
COLONY MORPHOLOGY
A. Colony
1.Clump of organisms growing on the surface of
solid medium.
2.Descendants of a single cell
3.Vary in appearance - characteristic of a genus
B. Characteristics observed :
1.Shape, size
2.Margin
3.Elevation
4.Surface texture
5.Light transmitting property (color)
6.Consistency
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MICROSCOPIC MORPHOLOGY
A. Size
B. Shape
C. Arrangement
D. Cell Structure
1.Basic cell structure
2.Special cell structures
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SIZE
A. Measured in micrometers (µm)
1000 µm = 1mm
B. Ave. Range : Diameter = 0.1 µm
to 2 µm
Length = 0.1 µm to 8
µm
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4.CELL SHAPE, ARRANGEMENT
A. Three groups according to shape. In
addition, variation of arrangement is
characteristic of a genus.
1. Cocci (us)
a. Spherical, oval, egg shaped, bean shaped
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Ignaz Semmelweis
Fungi
Parasite
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Sterilization
Sterilization agent are divided into:
Chemical.
Physical agents.
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Temperature Below 100 C ○
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Temperature around 100 C ○
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Temperature above 100 C ○
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Dry Heat
This is less efficient process as compared to moist
heat, and bacterial spore are most resistant to it.
Spore may require temperature of 140 0C for hours to
get killed.
Example of dry heat is flaming in microbiology
laboratory.
A good example of dry heat is hot oven.
In dry heat it is essential that air circulate between the
objects to be sterilized.
Microbial inactivation by dry heat is due to oxidation
process.
Dry heat used to sterilize glassware, glass syringe,
oils and oily injection as well as metal instruments.
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Time Temperature
Relationship in Heat
Sterilization
Process Temperature Time
Dry Heat 170 ° C 1 hours
160 °C 2 hours
121°C 16 hours
Moist heat 121 ° C 15 minutes
Autoclave 126 ° C 10 minutes
134 ° C 3 minutes
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2- Radiation
two type of Radiation:
A- Ultraviolet radiation
B- Ionizing Radiation
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A-Ultraviolet Radiation: Sterilization
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A-Ultraviolet Radiation:
Sterilization
This technique used for:
sterilizing drinking water
Obtaining pyrogen free water
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B- Ionizing radiation
Includs:
3. X rays
4. Gamma rays
5. Beta rays
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3- Filtration
Filters have been used long time ago
to purify water and sewage.
Application of Filtration:
Sterilization of thermolabile substance
and ophthalmic solution.
Sterility testing of pharmaceutical
products.
Clarification of water supplies
Microbiological evaluation of water
purity
Viable counting procedures
Determination of viral particle size
Air sterilization Tayyar M 28
Types of Filter
Various type of filter available:
Unglazed ceramic filters (chamberland and doulton filters).
Membrane filters
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Sterilization
5- Chemical Agents: chemical
disinfectant are highly toxic for all
types of cells.
They classified into different group
according to their activity or their
site of action.
Table 3.3 and 3.4
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A- Chlorine and Iodine:
Most useful disinfectant
Iodine as skin disinfectant
There function is bactericidal and
they are active against spore forming
bacteria
Different surface acting agent of
iodine such as iodophores used for
sterilization of diary products.
Chlorine, inorganic chloramines,
organic chloramines and
hypochlorite are a good disinfectant.
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B- Hydrogen peroxide (H2O2):
In 3% concentration is harmless, but
very weak disinfectant.
Use for cleansing of the wound.
C- Potassium Permanganate:
Oxidizing agent use to treat urethritis
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D- Formaldehyde:
Act on protein
Use as gas (37% solution in formalin)
Kill bacteria and spore forming bacteria.
Use to sterilize instruments and material
that can be damage by heating. In this
case the instrument are dipped into 2%
glutaraldehyde solution for 15-20 minutes.
This time efficient to kill both vegetative
and spore forming bacteria.
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E- Ethylene Oxide
Alkylating agent
Use in gaseous sterilization
Active against all kind of bacteria and their
spores.
Use to sterilize heat labile substances and
material (heart and lung machine)
Useful for delicate instruments, optics
Use for sterilization of powders and space
craft
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Cystoscope and endoscope can be
sterilized by glutaraldehyde.
Filtration used to sterilize heat
sensitive substances such as
antibiotics, serum and vaccine.
Ultraviolet radiation use to sterilize
small and closed champers.
For large rooms and theatre
fumigation with formalin is the most
effective method.
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Antimicrobials therapy
Origins of antimicrobial therapy in folk
medicine
Cinchona tree used to treat malaria
(quinine)
Moldy bread applied to wounds
(?penicillin)
Foxglove plant for heart disease (digitalis)
Willow bark for pain (aspirin-like
compound)
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Desirable properties of
Antibiotics:
1. Selective toxicity:
Drug must harm microbes without causing
significant damage to the host
2. Bactericidal rather than bacteriostatic
3. Effective against wide range of micro-
organism
4. Should not be allergic
5. Should remain active in various body
fluids
6. Stable and preferably water soluble
7. Desired levels in body should be
achieved rapidly and maintained fro
adequate period. Tayyar M 38
Inhibit cell Wall Formation Damage Cell Membrane
Penicillin Bacitracin Polymyxin
Vancomcin Cycloserine Amphotericin B
Ristocetin Cephalosporin Nystatin
Bacterial cell
Impair DNA Function
Nalidixic Acid Novobiocin
Protein synthesis inhibitor
Grisofulvin Mitomycin
Actinomycin Rifampicin
Nitrofurans 4-Quinolon
Streptomycin Gentamicin
Tetracycline Erythromycin
Chloramphenicol
Metabolites Antagonists
Fucidin
1-Sulfonamides
2-Sulfones
3-P-aminocalisalic acid
4-Isoniazid
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Combination therapy
More than one drug given at the
same time only in the cases where
there is a necessity of achieving high
bactericidal activity.
For example if patient infected with
endocarditis to prevent emergence
of drug resistance bacteria e.g.
mycobacterium tuberculosis.
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Antifungal Agent
Four antifungal are commonly used belong
to tow different groups:
2. Polyenes: e.g
– Nystatin
– Amphotericin B.
3. Imidazole: e.g.
1. Miconazole
2. Fluconazole (ketoconzole)
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Antiviral Agents
Few antiviral agents are available
these days.
Idoxuridine and acyclovir are
commonly prescribed for herpetic
infections of the oral and perioral
region.
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Drug resistance
Bacteria species developed resistance to
many antibiotics and thus minimize the
activity of these drugs.
Drug resistance is due to several reason
includes:
3. Chromosomal resistance: due to
Spontaneous (random) mutation; when drug
is given to the patients the resistant strain
persist and multiply.
mutation occur at rate of 1in 105 to 1010
5. Extra chromosomal resistance: this
resistance is mediated by plasmid.
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Extra chromosomal
resistance
Plasmid mediated drug resistance also called as
infectious drug resistance.
This kind of resistance transfer form one bacteria to
another by conjugation or transduction.
Transmissible plasmid has two major parts:
R determinant which encode resistance.
RTF (resistance transfer factor) which mobiles the
plasmid to another bacteria.
Because of this phenomena many sensitive bacteria
such as Salmonella typhi, have now become
resistance to commonly used antibiotics.
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Chapter 5
Host-parasite Relationship
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HOST PARASITE
INTERACTIONS
WHAT IS A PARASITE?
MUTUALISM vs. PARASITISM
A parasite may be defined as “
An entity that lives on or
within another organism (The
Host) - Thus living at the
expense of others”
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Host-Parasite Relationships
Parasitic organism
a symbiont that harms or lives at the expense of
its host
Parasitism
Is a life style that has be adopted by many types of
living things.
Parasite
All organism living in or on the bodies of other
organisms and drawing their substance from them
considered to be parasite.
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Symbiotic relation ship
Symbiotic is a general term used to described
any relationship in which two organisms live
together in an intimate association.
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Parasitic organism
Parasites include:
1. Viruses
2. Protozoa
3. Worms
4. Arthropods
5. Fungi
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Parasitic Relationship
The bacteria, fungi, and viruses that are
parasitic are usually described as infecting
their host.
Pathogens: referred to microbe that are
capable of causing infection to a host.
Contaminated host is used to described the
host when it associated with a microorganism.
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Parasitism and
disease
infected host used to described a host when it
invaded by a pathogen or a parasite.
Infection
growth and multiplication of parasite on or
within host
Infectious disease
Part or all of the host body cannot perform
normal functions due to the organism or its
metabolic products
Pathogenicity
ability of parasite to cause disease
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Dynamic of infection
Host-parasite interaction is a
dynamic equilibrium.
When host resistance decrease or
the parasitic virulence increase this
can result in development of the
disease.
There are various sources from which
infection can be acquired by human.
The vehicles can be food, water, air.
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Pathogenic Properties of
Microbes
Bacteria are divided into three group
when it is isolated from human:
Pathogenic bacteria: agents of
disease
Opportunistic pathogenic bacteria:
isolated from patient whose defense
mechanism have been compromised.
Non-pathogens: rarely or never
cause disease.
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Microbial Pathogenicity
attributes
Microbes produce pathogenicity with
help of a wide variety of attributes as
single or group such as:
Factor mediated adhesion
Antiphagocytosis agent
Capsule
M protein
Cytotoxin
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Microbial Pathogenicity
attributes
Survival within phagocytes
Interference with oxidative burst
Prevention of fusion and degranulation
Resistance to lysosomal enzyme
Escape from phagosomes
Siderphore and iron acquisition
Enzyme and toxin
Enzymes
Endotoxin
Exotoxin
Genetic mechanism
R factor Tayyar M 58
Microbial toxin
Toxin are the major toxic factor of microbes.
Exotoxin are the most potent poisons
known.
Exotoxin are composed of two subunit:
Fragment B: the carrier which binds to the
host cell receptors.
Fragment A the fragment that enter the cell
and cause damage to it.
Many organism produce exotoxin which
have activity in the gastrointestinal tract
and called enterotoxin.
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Microbial Toxin
B- Endotoxin: is a component of Gram
negative (G-ve) bacterial cell wall.
Endotoxin composed of lipopolysaccharide
with outer membrane protein.
Most of the toxic effects is due to lipid A
which is part of the lipopolysaccharide.
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Generalized Infection
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Septicaemia
When the bacteria circulate and
multiply in the blood and causing a
fever.
This could be due to:
Endotoxin producing bacteria e.g. E. coli,
Salmonella, Shigella.
Exotoxin producing bacteria, e.g.
Corynebacterium diphtheriae, Clostridium
tetani
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Pyaemia
Pyaemia
Pyaemia is defined as a condition in
which pyogenic bacteria produce
septicemia with multiple abscesses
in internal organs such as spleen,
heart, liver, and kidneys.
E.g. Lung abscesses which is due to
Klebsiella species, Staphylococcus
aureus.
Multiple abscesses in the body
caused by Mycobacterium
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Host Resistance
There are two major type of immune system
response to infection:
Non specific or natural resistance
Specific resistance or acquired immunity.
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Natural resistance
Components of Natural resistance:
Internal defiance system:
Phagocyte cells
Chemotaxis
Opsonization
Ingestion
Oxygen dependent killing of microbes
Oxygen independent killing of microbes
Destruction of ingested microbes.
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Natural resistance
Components of Natural resistance
Inflammatory response:
Local
Generalized
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Acquired immunity
This is the immunity which a person
a quires during his lifetime.
Based on the pathogen antigen (Ag)
been identified by
1. Antibodies (humoral immunity)
2. Cell mediated which is associated with
macrophages and T lymphocytes
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Classification of Acquired
immunity (Table 5.4).
Immunit Type Outcome of defense
y
Cellular Engulfment phagocytosis
Cytotoxic Destruction of infected
cells
Clumping of organism
Humoral Agglutinatio
n and phagocytosis
Precipitation Clumping of soluble
molecules and
Neutralizatio phagocytosis
Inactivation of toxin
n Blocking of viral
infection
Lyses of parasites
Complement
fixation Phagocytosis of
parasites
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Acquired immunity
Humoral immunity can be
Active immunity: the body synthesis its
own antibodies.
Passive immunity: the body received a
ready made antibody from other
individual.
The acquisition could be:
Natural: immunity for certain disease
provide resistance for subsequent
infection with the same microbes
Artificial: e.g. vaccine.
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Acquired immunity
Passive immunity: could be obtained
by
Natural Passive immunity : Tran-
placental passage of antibodies from
mother to unborn child during the later
part of pregnancy.
Artificial Passive immunity: production
of antibodies in some other individual
(either human or animal) and the
acquisition through parental routes
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Table 5.5 models of
acquisition of active and
passive immunity
Active immunity Passive immunity
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Classification of Acquired
immunity (Table 5.4).
Immunity Type Outcome of defense
Cellular Engulfment phagocytosis
Cytotoxic Destruction of infected cells
Humoral Agglutination Clumping of organism and
phagocytosis
Precipitation Clumping of soluble
molecules and phagocytosis
Neutralization Inactivation of toxin
Blocking of viral infection
Complement Lyses of parasites
fixation Phagocytosis of parasites
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Acquired immunity
Humoral immunity can be
Active immunity: the body synthesis its
own antibodies.
Passive immunity: the body received a
ready made antibody from other
individual.
The acquisition could be:
Natural: immunity for certain disease
provide resistance for subsequent
infection with the same microbes
Artificial: e.g. vaccine.
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Chapter 6
Immunology
The word immune mean free from burden.
Immunity: refer to the ability of an
organism to recognize and defend itself
against infectious disease.
Susceptibility: the opposite of immunity,
it is the vulnerability of the host to be
harmed by an infectious agent.
Specific immunity: is the ability of a host
to mount a defense against particular
infectious agent by physiological response
specific to that infectious agent.
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Immunology
Immunology is the science study
the specific immunity and how the
immune system consists of various
cells.
Immune system consist of various
parts,
Such as :
cells e.g. lymphocyte.
Organs e.g. thymus gland.
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Immunity
The immune system protects an
individual against invasion by foreign
bodies specially microbial agents and
their toxic products.
consequence of immune defenses
includes:
Allergy
inflammatory tissue damage.
Autoimmune diseases
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The duality of immune
System
Immune system of human and all
other vrtebrate divided into
2. Humoral immunity: because it
involves the antibodies that are
dissolved in the blood plasma and
lymph.
These antibodies are produce by
lymphocytes called B-cells.
3. Cellular or cell mediated immunity:
The immunity response depend on
lymphocytes called
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T-cells, which are 82
T-cells do not secrete antibodies into
body fluid's but they do antibody like
molecule that remain attached to cell
surface.
Humoral defense primarily against
bacteria and viruses presents in body
fluids.
Cellular response against
intracellular viruses, fungi, protozoa,
helminthes, transplanted tissue and
cancer cell.
The word antigen come from
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Specific Immune
Mechanism
These mechanism are mediated by B and T
lymphocytes which have receptors for antigenic
determinants on their surface.
The receptros on B cells are Immunoglobulin
(Ig).
The receptors on T cells are Ig like molecules.
Those receptors are only for antigenic
determinants which fits into the receptors like a
key to lock. Tayyar M 84
Lymphocyte that have receptors for
same epitope initiated immune
process which consist of proliferation
of lymphocyte and their functional
differentiation.
The cells in human body may have
receptors for more than 10 million
epitopes.
Immune system dose not initiate
immune response against self-antigen
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Nonspecific Immune
System
These mechanism deal with all
antigen irrespective of their epitopes.
The main component of this system
are physical barriers, phagocytes,
complement and inflammation (Fig
6.1)
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Inflammation Response
Inflammation Response composed off:
2) Blood flow increase.
3) Phagocytes activated.
4) Capillary permeability increased.
5) Complement activated.
6) Clotting reaction.
7) Regional temperature increased.
8) Specific defense activated.
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Nonspecific Immune
System
The phagocytes engulf and destroy
the infectious agent.
The complement is system of enzyme
that attack foreign substances and
lymphokines are liberated by
lymphocytes and act as functional
communication system among the
immune cells.
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T cells on stimulation, are differentiated into:
A- Regulator cells.
B- Effector cells which differentiated into
1. Helper cells
2. Suppressor cells
The effector T cell produce cell mediated
immune (CMI) response, that involve:
cytotoxic T cell which specifically destroy body
cells infected with the antigen.
Delayed hypersensitivity T cells which further
activate phagocyte cells to kill microbes.
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The activated T cells use a variety of
soluble factors (Lymphokines) to
activate macrophages for killing the
ingested cells.
B cells produce specific antibody
against the presenting epitopes.
The B cells response known as
humoral immune response or
antibody mediated immunity.
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The antibodies are secreted into
circulation where they produce a
powerful and specific effector
mechanism.
These may inactivate the antigen by
neutralizing its infective (as with
virus) or toxins and by removing the
antigen from the system.
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The antibody binds to the antigen and
the resulting complex is much more
attractive to phagocytosed
(opsonization) than the free antigen.
Also antibody attract the antibody
dependent cytotoxic cell (ADCC) to
attack and destroy the infected cells
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Both B and T cells retain antigenic memory for
the antigen after they get in contact with it.
The second attack by the same antigen results in
a faster and more vigorous immune response.
The antibody, complement and polymorphs
provide protection against most extracellullar
oragnism
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The antibody, complement and polymorphs
provide protection against most extracellular
organism.
T cells, soluble lymphokines and macrophages
deal with intracellular infection.
immunopathological tissue damage occur due
to inappropirate hypersenstivityn reaction to
foreign particle.
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Anatomy and Biology of
Immune System
The tissue in which the immunological
response system called lymphoid.
Lymphoid tissue divided based on
their function into
1- Primary lymphoid organ.
2- Secondary Lymphoid organ.
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Primary lymphoid organs are
A- Thymus.
B- Bone marrow.
In which proliferation and differentiation of
lymphocytes takes place independent of
antigenic stimulation.
Which mean that stem cell that give
lymphocytes are presents in these tissue.
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The mature lymphocytes are seed into the
peripheral system where they exposed to
antigens.
Organs that receive and maintain functioal
lymphocytes are called secondary lymphoid
organs.
The maturation of T cells takes place in thymus
whereas pre-B cell originate and mature in bone
marrow.
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Secondary Lymphoid
organs
Secondary Lymphoid organs include
2. Lymph nodes.
3. Spleen.
4. Gut associated lymphoid tissue
(GALT).
5. Appendix.
6. Tonsils.
7. Adenoids.
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Biology of immune
Response
Immune response are divided into
2. Primary immune response.
3. Secondary immune response.
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• Primary immune response
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• Primary immune response
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The nature and extent of immune
reposne to an antigen is determined
by:
2. Dose of antigen.
3. Nature of antigen.
4. Route of administration.
5. Adjuvant.
6. Nutritional status of the host.
7. Presence of immuno-deficiency
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Secondary Immune
Response
After second injection of the same antigen
Antibody in the circulation will drop
sharply as they form complexes with the
antigen.
After 2-3 days a marked increase in the
antibody occur for several days, this
increase is higher than what is seen in
primary response.
Figure 7.2 page 31.
This response called memory anamnestic
or booster response.Tayyar M 109
Secondary immune
response
Secondary Response differ from the
primary response in the following:
2. Shorter latent period.
3. More rapid production of antibody.
4. More abundant production of antibody
5. Antibody response maintained a higher
levels of longer periods
6. Antibody produce has more capacity to
bind the antigen.
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Immunosuppression
This define as reduction in the activity of
macrophages, B and T cells.
Which could be measured as lowered
capacity to phagocytose or to produce Ig or
lymphokines.
This cause the presence of not effective
immune response.
Immunosuppression could be due to
chemical (corticosteroids) and physical
(Radiation) agents, biological agent as well
such as antibiotics (Actinomycin D,
chloramphenicol and mitomycin C.
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Antigen and Antibody
The word antigen come from Antibody and
generator.
Antigen is a substance that catalyzes
lymphocytes into specific adaptive
response.
Lymphokines are called interleukins.
Antigen may alter the behaviour of
phagocytosis, either directly during
phagocytosis or indirectly in response to
lymphokines (interleukins)
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Antigen Characteristics
The important Characteristics of
antigens are:
2. Macromolecular size >10.000
Dalton molecules are more
antigenic than antigen with less
molecular weight.
3. Complex molecule.
4. Solubility in body fluids.
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Classification
Antigen can be classified into various
group:
foreign (outside the host).
Drugs
Environmental antigen
Autoantigen
Thyroglobulin
Corneal components.
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Antigen can be classified into various
group:
Isoantigen: is a type of antigen which is
present only in subsets of a species
Histocompatibility tissue antigen.
Blood group antigen.
Heteroantigen: (definitions not
necessary) antigens occurring on the
surface of tissue cells of many different
species and showing extensive
interspecies cross relationship.
Heterophile antigen
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Antigen determinant sites (Epitope):
2. Small part of an antigen molecule
is inducer of B and T cell response.
3. Also it is the part of the antigen
with which the antibody reacts.
The part of the antibody that comes
in contact with epitope is known as
paratope.
One antigen molecule may have a
large number of epitope.
Number of epitope per antigen
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Haptens: are small molecule to be
antigen by themselves.
Those haptens are capable of
reacting with antibody by injection a
hapten-carrier complex.
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Monoclonal antibody
Antibodies or immunoglobulins were first
identified in 1890.
It start with the observation by Behring
and Kitasato, that rabbits immune to
tetanus have substance in their blood
capable of destroying tetanus toxin.
They called this molecule antitoxin.
The first treatment application was in 1891
when both scientist treat children with
diphtheria with antitoxin isolated from
diphtheria toxin-immunized rabbit.
Recently this immunoglobulin
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has been 118
Antibody
Immunolglobulin (Ig) molecules with
symetrical structer, theose Ig have
“Y” shaped in solution.
glycoprotein's consist of four
polypeptide chains:
1. Two identical so called heavy chain
with molecular mass of approximately
50-55 kDa.
2. Two light identical
Tayyar M chains (25kDa) 119
The structure of
antibodies
http://www.path.cam.ac.uk/~mrc7/igs/mikeimages.html
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Antibody Biology
Antibody molecule are divided into
several parts:
Antigen binding fragment F(ab).
Compose of two parts:
Constant part: common to each
immunoglobulin class
Variable domain: unique for every
immunoglobulin.
Constant region (Fc-fragment
crystallizable)
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Basic str. of Ab
Prototype structure of lgG,
: revealed by chemical & enzymatic methods
& alkylation
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FIGURE 4-3
Classes (Isotypes) of
antibody
In human there are five classes of Ig based on
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Characteristic of
Immunoglobulin E
1. Has little concentration in serum.
2. Synthesized by very few plasma cell.
3. These antibody remain bound to
basophiles when injected into skin.
4. Contact with antigen release vaso-active
amine.
5. They protect the external mucosal
surface by local recruitment of plasma
factor and effectors cells by activating an
acute inflammatory response
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Characteristic of
Immunoglobulin D
The first antibody to be produce in
small amount.
Its production switch on the
synthesis of IgM.
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Antimicrobial Action
Antimicrobial action of
Immunoglobulin includes: (table
8.3)
2. Opsonization for phagocytosis
3. Complement activation, enhancing
phagocytosis and inducing lyses
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General structures of the five major classes
of secreted antibody
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The most abundant type of antibodies
found in blood, IgG are 150 kDa
glycoprotein's consist of four polypeptide
chains:
2. Two identical so called heavy chain with
molecular mass of approximately 50-55 kDa.
3. Two light identical chains (25kDa)
– The hinge of the Y-shaped Ig is rich in
proline (amino acid) and highly
vulnerable to attack by proteolytic
enzymes. Tayyar M 132
Discovery of
Viruses
Before 1892 bacteria were the smallest infectious agents.
In 1892 Dmitri Iwanowski (Russian scientist) reported that
filtration did not sterilized tobacco plant extracts because the
liquid that come through the filter was still able to cause mosaic
disease.
He was not able to see any bacteria under his light
microscope.
He also was not able to cultivate from it using standard
bacteriological media.
Six years later Beijerinck reported the same observation.
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Are the virus alive?
Virus are non living organism. As they are:
2. Not able to carry on metabolism.
3. Viruses lack most cellular structures including
cytoplasm, ribosome's, a nucleus or nucleoid.
But viruses are
5. Able to evolve and adapt to a changing
environments.
6. They contain some macromolecules similar to
those in the cellular organisms.
Virus are packaged bits of genetic information that
tell the host cells to make more viruses.
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Virology
Virus is a Latin word mean poison.
Virus as identified by Luria and
Darnell in 1978: viruses are entities
whose genomes are element of
nucleic acid that replicate inside
living cells, using cellular synthetic
machinery and causing the synthesis
of specialized elements that can the
viral genome to other cells.
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General properties of
viruses
Consist of either DNA or RNA but neither
both.
Obligate intracellular parasites.
Fail to grow on artificial media
Smallest infectious agents
Nucleic acid is enclosed in a protein shell.
Entire infectious unit is also called virion.
Not inactivated by antibiotics.
Divided by replication
Mostly limited hostTayyar
range.
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Virus classification based
on their host
Human and animal viruses
Insect viruses
Bacterial viruses (bacteriophage)
Plant viruses
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Viral Modern
classification
Modern classification based on their
genetic material (nucleic acid)
content (figure 28-1):
1. Double strand DNA (dsDNA)
2. Single strand DNA (ssDNA)
3. Complex DNA
4. ssRNA negative sense.
5. ssRNA positive sense
6. dsRNA
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Morphology of viruses
The viruses are composed of
nucleic acid and protein.
The genome contain all
necessary information for viruses
multiplication.
The genome surrounded by
protein coat called capsid.
Both genome and the capsid are
called nucleocapsid.
Viral envelope is a coat
surrounding the nucleocapsid
made of lipid.
Complete viral particle called
virion figure 28.2. Tayyar M 140
Capsid are aggregate of arranged
repeating subunits call capsomers.
The capsomers are made of protein.
Capsid and viral envelope has two
function:
Protect the genome from external harmful
factors or agents
Introduce the viral genome into the host cells.
Viral enveloped present in most viruses
families except.
The lipid of the viral enveloped is derived
from the host cytoplasmic membrane.
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Size of viruses:
Viruses vary in diameter from 300 to
20nm.
Box-viruses are the lagest viruses.
The picorna and parvo-viruses are
the smallest.
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Viral proteins
Viral protein different on their size as well as
number of protein species.
Viral envelope may posses glycoprotein's which are
presents as
1. Spikes or projection (rabies virus)
2. Matrix protein, present in the inner surface of the
envelope and provide extra-strength.
– In addition there are two types of viral proteins-
heamaglutinins and enzymes.
– Protein found in the coat are called structural
protein.
– Heamaglutinins and enzymes are called non-
structural protein. Tayyar M 143
Preservation of viral
infectivity
Viruses are very fragile and more sensitive
to environmental changes than other
organism.
1- Temperature: increase temperature
destroy viruses rapidly:
General roles:
viruses survive for seconds at 60ºC
At 37ºC for minutes, hours at 20ºC, days at 4ºC, and years
at minus 70ºC or below.
2- pH viruses prefer physiological pH and any alteration
mat cause deterioration of viral infectivity.
3- Lipid solvent: ether or chlorophorm, detergent and
any lipid solvent come in contact with viruses,
especially enveloped viruses
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Viral Replication
The virus is intracellular parasite, make use of
host metabolic process for its own benefit.
Viral replication consist off:
Attachment or adsorption
Penetration
Uncoating
Transcription
Translation
Genome replication
assembly
Release
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Attachment or Adsorption
For a virus to infect a host cell, that cell must
have receptors for the virus on its surface and
also be capable of supporting viral replication.
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Penetration
Enveloped viruses enter the host cell in one of four
methods:
• Translocation: when the whole non-enveloped virus enter
the host cells by moving across cell membrane.
• Virus enters by endocytosis (formation of an invagination
of the plasma membrane that engulfs the virus).
• Viral envelope fuse with the host cell cytoplasmic
membrane and the nucleocapsid is released into the
cytoplasm.
• Direct fusion of the viral envelope with the surface
membrane of the cell.
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Uncoating
Uncoating is the release of the viral genome
from the remainder of the virus.
The viral capsid is removed by the action
of host components and enzymes.
Viral genome replication starts at this stage
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Transcription
Positive sense (polarity) single
stranded RNA bind to ribosome's to
synthesize protein.
negative sense ssRNA use RNA
dependent RNA polymerase to
produce positive since RNA (mRNA).
While DNA virus use DNA dependent
DNA polymerase to perform this
function
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Translation
Viral mRNA uses the ribosome's to
synthesis protein in the same ways
the host mRNA did.
Two type of protein are synthesized
upon time of appearance
Early protein
Late protein
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Genome Replication
The viral genome directs the host cell's
metabolic machinery (ribosome's, tRNA,
nutrients, energy, enzymes, etc.) to synthesize
viral enzymes and viral parts.
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Assembly
Virion assembly may take place in
the cytoplasm or cell nucleus of the
host.
The process of acquiring envelope is
through the process of budding
through the cellular membrane.
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Release
There are three machinery of
mature virions release from the
host cell and start their life cycle
again.
2. Cell lysis
3. Cell degeneration
4. budding
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Duration of viral replication between 5-15
hours in DNA viruses and 3-10 hours in
RNA viruses.
Viral infections is not always productive
Sometimes the virus protein are not
synthesized or their assembly is defective,
such process called an abortive viral
infection.
Some viruses fail to replicate in a
responsive host cell, these are known as
defective viruses.
Some replicate in the presence of helper 154
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Naked Virus Replication
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Cultivation of viruses
Since viruses are obligate
intracellular parasites, a living cell
system needed to grow them such
as.
1. Laboratory animals
2. Embryonated eggs
3. Cell lines (tissue culture)
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Recognition of Viral Growth
in Cells
Viral growth in cells can be detected
by one of the following:
Cytopathic effect (count dead cells)
Haemagglutinin and haemadsorption
Immunofluorescence
Interference
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Adenovirus
Adenoviruses has a dsDNA genome, non
enveloped virus.
Adenovirus infection occur world wide.
By age of five almost all children have
been infected with at least one serotype.
Few serotypes are pathogenic to man.
Adenovirus infections of human include:
Acute respiratory disease
Sore throat
Pneumonia
Meningoencephalitis
Swimming pool conjunctives.
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Herpes viruses
The family Herpesviridae consist of
more than 80 viruses.
All Herpes viruses are enveloped
virions with dsDNA genomes.
Herpes simplex virus:
Exist in two serotype HSV-1 and HSV-2.
HSV-1 infect region above the waist.
HSV-2 infect genital and anal regions.
Latency for HSV-1 in trigeminal ganglia
HSV-2 latency in sacral ganglia.
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Herpes simplex virus
Disease caused by herpes simplex
virus
2. Stomatits.
3. Herpes labialis
4. Eczema herpeticum.
5. Traumatic herpes
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Varicella Zoster Virus
Varicella Zoster Virus
Etiological (responsible) agent for two disease
of man, Varicella and herpes zoster.
Varicella follows exogenous contact with the
causative virus.
Zoster reflects endogenous activation of the
Varicella Zoster virus infection that has
survived in latent form following an attack of
Varicella.
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Varicella is contagious disease of seasonally
epidemic propensities the follows primary exposure
of Varicella Zoster a susceptible person, most often
child.
The disease known as chickenpox.
Herpes zoster is an endemic sporadic disease, most
frequent in elderly people, characterized by
Unilateral
Painful
Vesicular or eruption localized.
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Cytomegalovirus
Cytomegalovirus
Most of CMV infection is
asymptomatic.
In rare cases the infection is fatal.
Some times CMV cause permanent
brain damage.
CMV might transfer through placenta
causing cytoplasmic inclusion
disease (CMID) of newborn
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Epstein-Barr virus (EBV)
Epstein-Barr virus cause glandular
fever.
Characterized by:
Fever
Lymphoadenopathy
Sore throat
Malaise
Fatigue
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Acquired Immunodeficiency
virus (AIDS)
RNA enveloped viruses.
HIV can be transmitted by any of the
following routes:
1. Sexual intercourse
2. Blood transfusion
3. Transplacenta
4. IV drug
5. Breast milk
6. Accidental needle-stick injury
Chronic infection with HIV has diverse
symptoms, due to the destruction of
immune system.
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Rubella
Rubella
Known as German measles.
Clinical features of rubella virus:
1. Rash
2. Lymphoadenopathy with fever.
3. congenital malformations in children if
the mother contracts this infection
during the first trimester of pregnancy.
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Hepatitis Virus
Hepatitis virus refer to primary infection of the liver
by one of the four etiological agents:
1. Hepatitis type A (HAV)
2. HBV
3. Non-A, non-B hepatitis (NANB)
4. HDV
The newer type are known as:
1. HEV
2. HCV
3. HGV
4. HFV
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HAV
ssRNA non-enveloped virus.
Clinical feature:
1. Fever
2. Headache
3. Fatigue
4. Vague pains
5. Anorexia
6. Nausea
7. Vomiting
8. Dark urine
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HBV
Circular ssDNA, enveloped virus.
Clinical feature similar to those in HAV.
HAV does not persist in the host.
HBV may give rise to persistence infection
of liver (prolonged carrier state to chronic
infection).
HBV may lead to hepato-cellular
carcinoma.
HCV also could cause hepato-cellular
carcinoma.
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Medical Mycology
Medical mycology is the study of
human diseases caused by fungi and
mycosis.
Around 100 or more species of fungi
are recognized as human pathogen.
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Fungi Differences from
Bacteria
1. Have larger size, more complex
morphological development.
2. Fungi can be unicellular or multicellular.
3. Having ridged cell wall compose of
chitin, mannan and other polysaccharide.
4. Presence of true nuclei with nuclear
membrane and pair chromosome.
5. Presence of sterols in the cytoplasmic
membrane.
6. Reproduction can be asexual or sexual or
by both these modes.
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Medical Mycology
Medical mycology is the study of
human diseases caused by fungi and
mycosis.
Around 100 or more species of fungi
are recognized as human pathogen.
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Fungi Differences from
Bacteria
1. Have larger size, more complex
morphological development.
2. Fungi can be unicellular or multicellular.
3. Having ridged cell wall compose of
chitin, mannan and other polysaccharide.
4. Presence of true nuclei with nuclear
membrane and pair chromosome.
5. Presence of sterols in the cytoplasmic
membrane.
6. Reproduction can be asexual or sexual or
by both these modes.
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Dermatophytes
Dermatophytes are group of closely
related filamentous fungi that infect
only superficial keratinised tissue, i.e.
skin, hair and nails.
They caused dermatophytoses, or
Ringworm or Tinea.
Tinea (moth) used to describe the
dermatophytes infection that give
the hair of the head a moth-eaten
appearance
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Epidermophyton is the most
important genera of dermatophytes.
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Candidiasis
Candidiasis is the disease which affect the
skin, the mucosa, the skin appendages
and the internal organs as well as cause
septicemias due to yeast fungi of the
genus Candida.
7 out of 81 Candida species are
pathogenic.
The most common is Candida albicans.
Candida reproduce by budding.
They are able to ferment a number of
sugars. Tayyar M 176
Predisposing Factors for
Candidiasis
1. Impaired immune 1. Pregnancy
defences.
2. Spontaneous 2. Menopause
hormonal changes 3. Premature birth
Corticosteroids
3.
4. Obesity
4. Immunosuppression
5. Long term antibiotic 5. Child birth
therapy 6. Diabetes
6. Oral contraceptive mellitus
7. Preexisting lesion of
skin or mucosa or 7. Adrenal
internal organ insufficiency
8.
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Clinical features
Candidiasis
1. Candidiasis of Thrush
visera Cheilosis
2. Bronchopulmonar Intertrigo
y Candidiasis Onychia
3. Septicemia Paronychia
4. Intestinal Erythema
Candidiasis mycoticum
5. Granuloma infantile.
6. Vaginitis
7. Balanitis
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Fungi Diagnosis
Microscopic examination
Culture
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Aspergillosis
Aspergillosis name given to a
number of different diseases states:
Allergic aspergillosis
Aspergilloma
Invasive aspergillosis
Aspergillosis is secondary infection in
tuberculosis, carcinoma etc.
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Etiological Agents
Common specie causing aspergillosis
are:
Aspergillus fumigatus
A. niger
A. flavus
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Parasitology
Medical Parasitology deals with the parasites
which infect man and disease produce by them.
Parasite: an organism which is dependent on
another organism for its survival. It obtain
nutrient and shelter from the host.
Host: an organism which harbours the parasite
and is usually larger than the parasite.
Ectoparasite: parasite which lives on the outer
surface of the host.
Endoparasite: parasite which lives within the body
of host.
Commensal: a parasite which does not damage
the host while deriving its advantage. A
commensal is capable of independent life
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Symbiosis: A parasitic relationship which
results in great advantage to each other
as compared to disadvantage.
Obligate parasite: A parasite which is
completely dependent upon the host.
Pathogen: a parasite which is able to
produce disease.
Intermediate host: a host in which the
intermediate stage of the parasite
develops.
Definitive host: a host is which sexual
reproduction takes place or the adult form
of the parasite resides.
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Paratenic host: a host which acts as a
transporting agent for the parasite and in
which parasites dose not undergo any
development.
Infection: refer to the presence of parasite
in or on the tissue of the host.
Infestation: presence of arthropods on the
skin of the host.
Incubation period: the time interval
between the entrance of the parasite into
host and the beginning of disease.
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Zoonosis: disease which are
transmissible between man and
animals.
Virulence: refer to the degree of
pathogenicity.
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Mode of infection in
parasite disease
Infection can be acquired by various
routes.
1. Through animals :
A. pig : e.g. porktapeworm
B. Cat : e.g. toxoplasmosis
2. Autoinfection: threadworm
3. Water-borne: e.g. Amoebiasis, giardisis.
4. Vector borne: e.g. mosquito (malaria)
5. Contaminated food: Ascaris, Amoebiasis
6. Vertical transmission: malaria, toxoplasmosis
7. Penetration through skin: Hookworm,
Schistosomiasis Tayyar M 186
Ascariasis (roundworm
infection)
Etiological agent: nematode called Ascaris
lumbricoides.
Habitat: the adult worm live in the lumen of the
small intestine mainly jejunum.
Morphology: the mature worm is cylindrical with
tapering ends, resembling an ordinary
earthworm.
Characteristics of eggs:both fertilized and non-
fertilized egg could be found in the faeces of
patient suffering from ascaris.
A mature female can liberate more than 200,000
eggs daily.
The characteristic of this eggs help in their
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Life cycle:
Infection occur from ingestion of the empryonated eggs.
The infective larva hatch out in the small intestine and penetrate
the intestinal wall to enter the portal circulation.
From the liver these are carried to the heart and via the
pulmonary artery to the lungs.
In the lungs these break out of the capillaries into alveoli and
undergo another moulting to become the fourth stage larva.
These larvae move up to bronchi and are either swallowed with
saliva or crawl over epiglottis to enter the digestive system.
In the intestine these again moult to become sexually mature
worms.
The lifespan of an adult worm is about one year after which it is
spontaneously expelled.`
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Pathogenicity
A) Hinder normal development in children
B) Fever, abdominal pain, urticaria.
D) Pneumonitis
F) Appendicitis
H) Malnutrition.
Treatment : Mebendazole is the drug of choice.
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Malaria
Aetiological Agents: four specie of
Plasmodium could cause Malaria in
human:
P. vivax
P. falciparum
P. malariae.
P. ovale
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Life cycle
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Entamoeba Histolytica
This microorganism cause
amoebiasis.
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Normal Microbial Flora of
the Human Body
The normal microbial flora refers to the
population of microorganism that inhibit the
skin and mucous membranes of healthy
normal persons.
Normal flora could be of two type:
Residential flora: fixed type of
microorganism regularly found in a given
area at a given age.
Transient flora: this consist of nonpathgenic
or potentially pathogenic microorganism
Function of resident flora:
Maintain prober health by perform various
function.
Benifictal function of resident flora:
Help in absorption of nutirent
Synthesise vitamin espically vitamin K
Prevent colonization by pathogens having
related to shared antigen.
Bacteriocins produced, inhibit growth of
• Harmful function of residental Flora
Oppurtinistic pathogens
Produce disease if introduced in large number
in forigne location with predisposing factor.
Penicillinaze produced can antagonize therapy.
Excessive production of endotoxin may cause
shock.
May transfer organism from one part of the
world to the another.
Ubiquitous presence creates problems in
establishing accurate diagnosis.
For microbiological diagnosis purpose
the specimen must be obtained from
the suspect lesion and not from an
adjacent non-involved area.
The site of sampling should be
properly prepared.
Human body divided into
Site normally sterile. Sites with normal flora:
Bone Marrow Skin
Tissues
Bacteriological examination of air is required
following conditions such as:
Surgical theaters
Premises where food articles are prepared and packed
Premises where pharmaceutical preparation are made
Hospital words when nosocomial infection have
occurred.
Hospital associated
infections
Hospital is the place which help
people in getting cured of illness.
Some times hospital become potential
transmittance of disease.
Infection which acquired in the
hospital called hospital infections or
nosocomial infections
Source of hospital infection:
Exogenous route :
From other patient
From a member of the hospital staff.
Out patient
Hospital staff