1.

) Medical microbiology help physician to deal with

the causative agents ofv infectious diseases, the
ways in which they produce diseases in human
body and essential information for diagnosis and
treatment.
The science of microbiology, virology and
immunology play an important role in finding and
classifications of thousands of microorganisms
and viruses. It also help in development of
identifying the infectious disease caused by
microorganisms and play important role in
diagnosis and treatment of the infectious diseases.

2.) The invention of a microscope was done by

biologist A.Leeuwenhock from
Holland on 1674. His microscope consisted of a
single biconvex lens that magnified object about
x200.
Leeuwenhock discovered and described various
free living and parasitic protozoa, filamentous
fungi and globular bodies from human and animal
feces. He introduced meat and vegetables
infusions in culture medium. He also described 3
morphological forms of animolecules- rod, sphere
and spiral forms.

4.) Robert Koch(1843-1910) a German scientist perfected

bacteriological techniques and introduced methods for
isolation of pure strains of bacteria. He introduced agar as
a setting agent in bacteriological media. He isolated the 1st
bacteria Anthrax bacillus in pure culture on 1876. He also
introduced staining methods. The important discoveries of
Koch was :
1) anthrax bacilli 1876
2) Tubercle bacilli 1882
3) cholera vibrio 1883
He is also known as father of medical microbiology. His
important significance for microbiology and infectious
pathology are:
1)
The organism should be constantly
associated with lesions of the disease.
2)
The organism should be isolated from
the lesion in pure culture.
3)
The isolated organism in pure culture
when injected in susceptible animals should
reproduce the disease.
4)
From lesions produced in experimental
animals the MO must be demonstrated both in
smear and culture.

3.) Luis Pasteur of France (1822-1895) is known as father

of modern microbiology. He was a trained chemist who
was ferment ting wine. While studying fermentation of
grapes in 1857, he noted that alcoholic fermentation of
grapes, fruit and grains was caused by some microbial
agents, which he named as ferments. He derived the
techniques of bacterial cultivation and laid them in 3
principles:
1 Every alternation of either wine or beer depends on
development of microorganisms
2. The MO are brought by the ingredients or apparatus.
3. Whenever beer or wine contains no living
organisms(sterile) it remains unchanged (not
fermentated)
His discoveries and contribution in field of microbiology
are:
1)
Development of methods and techniques
for cultivation of MO
2)
Conclusive evidence of MO in disease
production.
3)
Introduction of sterilization techniques
and development of steam sterilizer, hot-air oven
and autoclave.
4)
Studies on pebrine (silk worm disease),
anthrax, chicken cholera and hydrophobia. He
discovered causative agent of rabies was too
small to be seen by microscope.
5)
Live vaccine-introduced attenuated live
vaccines for prophylactic use.

5.) As for Russian scientist Metchnikoff and Luis Pasture

play important role in development of microbiology. Study
about physiological pathogen began by Luis Pasture. The
immunology pathogenicity began by Metchnikoff. He
specialized on Luis Pastures ideas. He discovered the
opening of cell immunity. He found that inside the
organism has defensive mechanism for living and non
living agents. He found the cell theory in
organism.Machnikoff 1883 discovered phagocyctic
phenomenon.He said that phagocyctic response is the main
difference mechanism against m/o innervation to tissue.He
also found and produced the cellular concept of
immunity.The other is pool eric which found the
phagocytic action.Is the action on foreign
substance(cell)which has been enters to the animal cell
mainly.They are pathogenic m/o can kill them by
phagocytosis.He opened the human organism and then
found Ab.In the 20th century the development of m/o was
very fast.In this century they discovered :
a) bacteriaphages
b)pathogens of chicken embrio
c)genetic theory of cancer
d)vaccinations
Pool eric II was the 1st scientist made the drug for syphilis.
Mint and Machnikoff-relapsing fever.
Lowzimbert virology viral genetic theory of tumor.
Chumachoff different vaccines against poliomyelitis.

6) Ivanovosky (1892) 1st reproduced mosaic disease of

tobacco plant by applying juice of diseased plants after
filtering all bacteria to healthy leaves. He repeated this
many times. Then he found that Mo which cause mosaic
disease can pass through bacteria filter. So that, the MO
must be very small and cannot be seen under normal light
microscope. Then, he found out that the mosaic disease is
caused by MO which is different from bacteria, which is
later designated as viruses.
Development of virology:
Loeffler and Frosch (1898) transmitted foot and mouth
disease of cattle by filter passing virus.
Walter Read (1902) from Cuba proved yellow fever is a
viral origin human disease.
Landsteiner and Popper (1909) demonstrated that
poliomyelitis was due to filterable viruses and transmitted
this disease in monkeys.
Ellerman and Bang (1908) proved virus may cause cancer
in fowl leukemia.
Rous (1911) isolated a virus from fowl sarcoma.
Ruska (1934) introduced electron microscope to study
direct morphological study of viruses.
Goodpasture (1930) developed techniques of viral
cultivation in nonproliferating suspensions of mammalian
cells and in chick embryos.

8)They are many different kinds of microrscopy for studying

bacterial actions as:
a)optic or light microscopy.
b)phase contrast
c)dark field microscopy
d)interference
e)fluorescence
f)electrone microscopy
g)autoradiography
Dark field microscopy
Reflected light is used instead of transmittd light.A dark field coundense
with circular is the main part of microscope.The condenses less system
arranged in a suchaway that no light reaches th eye,unless reflected
fom the object on microscope stage .The basic or object looks luminous
in a dark background use to detect extremely small organism as
spirochaetes.
Phase contrast microscopy
Has a special condenses.The different part of cell and its surrounding
have different refractive power and so when the beam of light passing
thru object due to different refractive power and they have different
intensity of light so some structures are dark n other are light.Used to
study internal structures of unstained living tissues,bacterias and
protozoa.
Fluorescence microscopy
In this ultraviolet light is used.Bacteria stained with fluorescent dye and
give visible as bright object against dark
background.Immunofluorocence combines serology with fluorencence
microscopy by using Ig combined with fluorencence dyes are
Rhedamine and Lissamine.
Elecrone microscopy
In microscope a beam of electrons is focused by a a circular
electromagnetic disc to the lenses of light microscope.When electrons
beams pass thru object the electrons get together and produced an image
on screen.The main thing of a elctrone microscope is formation of
shadow casting due to depositing thin layer of metal on object.The
metal coated object is placed in a pathway of beams.The electrone
beams is directed to form of shadow in uncoated area in other side and a
negative image is made.On fluoresencence viscosing screen a (+)
image is made by (-) image.

7) Subject of medical microbiology

The subject of medical microbiological study is
microorganisms and their characteristics and peculiarities.
Aim:1. Study morphological and physiological criteria of
microorganism
2. Study causative microorganism of clinical disease and
provide treatment.
3. Immunological study and provide vaccination.
4. Study about general characters of microorganism and
their classification.
Parts: 1) General microbiology
Study common character of microorganism
2) Industrial microbiology
Study about microbes which play role in industry for
preparation of object.
3) Agricultural microbiology
Irradiation of some microbes
4) Marine microbiology
5) Veteran microbiology
6) Space microbiology
7) Medical microbiology
Methods using in microbiology:
1) Microscopic method- receive info about shape and
structure of MO
2) Microbiological method- receive info about cultural and
biochemical properties.
3) Immunological / serological- study immune system and
allergic response
4) Biological-for determination of drugs. Use laboratory
animals.

10. FORMS OF BACTERIA.

Bacteria are prokaryotic unicellular MO which divide by
binary fission and do not possess chlorophyll and true
branching. Bacteria belongs to kingdom protista. Their size
varies frm 0.5 to 15 m in length and 0.2 to 2 m in width.
MORPHOLOGY.
There are 3 types :
1) cocci
2) rods 3) spiral shape
1) cocci is divided into :
a) micrococci - irregular arrangement
b) diplococci arranged in pairs
c) streptococci arranged in chains
d) tetracocci in group of 4
e) staphylococci grape like clusters.
f) Sarcina in groups of 8,16,32
2) rods are divided into :
a) non sporing bacteria
b) bacilli diameter of spore is same
c) clostridium diameter of spore is diff.
2) spiral shape can be divided acc. to the no. of curves :
a) vibrio
b) spirillum many curves
ULTRASTRUCTURE.
Constant structures are cellwall, cytoplasmic memb.nucleoid,
cytoplasm,ribosomes, mesosomes.
Inconstant stru. Are spores, flagella. Capsule,
fimbriae.plasmids.episome
CHEMICAL COMPOSITION OF BACTERIAL CELL
- The main part of bacterial cell wall is peptidoglycan.
- They also possess enzymes called autolysins.
- gram +ve cell wall :
a) peptidoglycan layer is thicker and consists of multiple
layers.
b) they also contain teichoic and teichuronic acids.

1)
o
2)

3)

11. EXAMINATION OF COLOURING M/O

SIMPLE STAINING
watery solution of a single basic dye such as methylene blue or
basic fuchsin are used as simple stain
COMPLICATED STAINING
ve staining
bacteria are mixed with dyes(India ink or nigrosin). The
background gets stained leaving the bacterias contrastingly
colourless. Useful in detecting bacterial capsule.
impregnation methods
used for bacterial cell and appendages tht are too thin and
delicate.
It is thickened by impregnation of silver on the surface to
make them visible under light microscope.
differential stains
impart diff. colours to diff. bacteria or their str.
GRAM METHOD
1 staining of heat fixed smear of specimen or bacterial
culture is made with a pararosaniline dye( crystal violet or methyl
violet) for 1 min.
Pour off crystal violet and add dilute solution of iodine, keep
for 1 min.
Wash with water.
Decolourisation with an organic solvent( alcohol) for 10 to 20
sec
Wash with water
Counterstain with a dye of contrasting colour( dilute carbol
fuchsin, safranin or neutral red) for 20 to 30 sec.
Gram +ve bacteria stain violet
Gram ve bacteria stain pink
GRAM +VE & GRAM VE GROUPS OF MO
Gram +ve bacterias :- all cocci except Neisseria and Branhamella
Gram ve bacterias :- all bacilli except Corynebacterim,
Mycobacterium, Bacillus, Clostridium, Actinomyces, Listeria, and
Erysepalothrix

- gram ve cell wall :

a) is a complex str. and thinner.
b) bimolecular layer of peptidoglycan.
c) they have an additional outer membrane which is
much thicker than the peptidoglycan layer.
MAIN DIFF. BTW. PROKARYOTES & EUKARYOTES.
STRUCTURE.
PROKARYOTES
EUKARYOTES
Nucleus
Absent
Present
Nuclear membrane
Absent
Present
Chromosome
One
Many
DNA
Absent
Present
Division
Binary fission
Mitosis
Cytoplasm
Absent
Present
Mitochondria
Absent
Present
Golgi body
Absent
Present
ER
Absent
Present
Lisosomes
Absent
Present
Pinocytosis
Absent
Present
Sterol
Absent
Present
Muramic acid
Present
Absent
Diamino pimelic acid
Present
Absent

PROTOPLAST, SPHEROPLAST
it is the bacteria with defective cell wall
Protoplast : complete removal of cell wall of Gram +ve bacteria
they need isotonic medium to maintain the str.
do not multiply and cannot revert to normal
bacterial morphology.
spheroplast : gram ve bacteria with damaged cell wall.
they are produced by growth with penicillin
they can multiply by binary fission or budding when
placed in suitable environment.
L- FORMS OF BACTERIA
- they are bacteria without cell wall
- they develop spontaneously or by antibiotic treatment.
- They dont have any regular size and shape
- they can grow and multiply on suitable medium.

12. BACTERIAL SPORES

- spores are highly resistant dormant stage of bacteria
formed in unfavourable environmental conditions such as
starvation and dessication.
- 2 types : 1) endospores formed within parent bacterial
cells
2) exospores formed extracellularly frm the ends
of parent cells
- spore forming bacterias are Bacillus and Clostridia
- spores are localized in central, subterminal or terminal.
- Bacillus spores are same size or smaller than bacteria
- Clostridia bigger than the bacteria
- it may be oval or spherical
they are highly resistant to ordinary boiling , heating and
disinfectants
germination may occur in less than 2 hours under optimal
condition and it consists of 3 stages :
a) activation
b) initiation
c) outgrowth
detection :- Gram method spores appear as an unstained
refractile body within the bacterial cell
Ziehl Neelsen method stain with 0.25 0.5 % sulphuric
acid, spores appear red
CAPSULE
it is outer covering of thick jelly-like material tht surrounds
the bacterial cell wall.
It contains 98% water and 2% solids
The solids maybe complex polysaccharide or polypeptide or
hyaluronic acid
It serves as protective covering against antibacterial subs., it
enhances bacterial virulence, and the capsular antigen is
hapten in nature.

Capsulated org. are S.pneumoniae, B. anthracis and C.

perfringens
It is best seen is specimens like pus, blood and sputum
Detection :- Grams method appears as an unstained halo
around stained bacterial body
- -ve staining it appears as a clear halo around the
bacterium
- immunological method stained by serological method,
capsule appears swollen
FLAGELLA
are filamentous, cytoplasmic appendages protruding through
cell wall.
It is unbranched, long, thread-like str. composed of
protein(flagellin)
They are for locomotion
1)
4 types of flagellar distribution :monotrichous single polar flagellum
- eg : cholera vibrios
2) amphitrichous single flagellum attached to each end
- eg : alkaligenes faecalis
3) lophotrichous tufts of flagella at one or both ends
- eg : spirilla
4) peritrichous many flagella around the bacterial body 2)
- eg : typhoid bacilli
- flagella consists of 3 parts ; filament, hook and basal body
- detection : -seen under electron microscope, stained with
phosphotungstic acid, appears as hollow tubes.
3)
INCLUSIONS

13. MORPHOLOGY & ULTRASTR. OF ACTINOMYCETES

- there are 3 types of actinomycetes :
1) Actinomyces
2) Nocardia
3) Streptomyces
1) Actinomyces :
- Gram +ve, non-motile, non-sporing- non-acid fast org.
- strict or facultative anaerobes
- They are part of normal flora in oropharynx and GIT
2) Nocardia :
- strictly aerobic, Gram +ve bacteria, non-sporing, weak acid-fast
org
- they are prevalent in soil
3) Streptomyces :
- Gram +ve, strict aerobes, can form spore
- prevalent in soil and water
PATHOGENIC ACTINOMYCETES
Actinomyces :
A. israelii lives in oropharynx and alimentary tract of man.
Causes human actinomycosis.
A. eriksoni
A. bovis oropharynx of cattle. Lumpy jaw in cattle
A. naeslundii Oropharynx of man and animal. Causes caries and
dental plaque.
A. odontolyticus
A. viscosus human oropharynx. Causes caries and dental plaque.
Nocardia :
N. asteroids causes pulmonary nocardiosis
N. brasiliensis causes subcutaneous nocardiosis
N. madurae
N. otitidis-caviarum - causes subcutaneous nocardiosis
Streptomyces :
S. somaliensis causes mycetoma
ACTINOMYCETES LIKE ANTIBIOTIC PRODUCTORS

1)
2)
3)
-

1)

2)

14. SPIROCHETES.
CLASSIFICATION
has 2 families : 1) spirochaetaceae (Treponema and Borrelia)
2) leptospiraceae (Leptospira)
- general character :- large, motile helical org
MORPHOLOGY
Treponema :
slender cork-screw like filamentous helices.
6 14 m x 0.2 m with 6 -14 evenly spaced coils
Motile
Borrelia :
Gram ve, motile
10-30 m x 0.4-0.7 m with wide and open coils
Leptospira :
spiral org, 5-20 x 0.1 m with many closely set coils and
hooked ends
motile
PHYSIOLOGY
the body consists of a central protoplasmic cylinder bounded
1)
by the cytoplasmic membrane and a cellwall
fine fibrils runs btw the thin peptidoglycan layer of cellwall
and the outer cytoplasmic membrane which are fixed at the
knobs at the 2 poles of the org. These fibrils constitute an axial
filament which constrict and distort the bacterial cell body to
2)
give rise to a spiral conformation.
PATHOGENS FRM DIFF. GENUS
Treponema T. pallidum subsp. pallidum cause venereal
syphilis
- T. pallidum subsp. endemicum - causes endemic
syphilis
3)
- T. pallidum subsp. pertenue causes yaws
- T. carateum causes pinta
Borrelia B. recurrentis causes relapsing fever
- B. vincentii causes vincents angina
- B. burgdorferi causes Lyme disease
3) Leptospira L. interrogans causes leptospirosis

16. MORPHOLOGY & PHYSIOLOGY OF

MYCOPLASMA
- smallest free-living bacteria ( 0.2 m)
- Occur as granules and filaments
- lack cell wall
- require cholesterol or sterols for growth in culture
- fired egg colonies
- facultative anaerobes except M. pneumoniae (grows
aerobically)
- contain both RNA and DNA and can reproduce in cellfree media
- most species utilize glucose
- resistant to penicillin due to lack of cell wall
- reproduce by binary fission or by budding
- Gram ve, motile, non-capsulated
HUMAN PATHOGENIC SPESIES
Mycoplasma pneumoniae
extracellular pathogen of respiratory tract
transmitted by air-borne
treatment is by tetracycline and erythromycin
Mycoplasma hominis
causes puerperal fever, non-gonococcal urethritis, pelvic
inflammatory disease (PID)
transmitted by sexual contact or during birth through
vaginal tract
treatment is by tetracyclines and macrolides
Ureaplasma urealyticum
causes non-specific urethritis or genital infection
transmission and treatment same like 2)

17.Rhicketsia:morphology and physiology

Morphology

-pleomorphic coccobacili appearing as either rods or cocci,nonmotile,non-flagelatted,non-capsulated.

-cell wall-3 layers-peptidoglycansor mucarinic acid and
diaminopimelic acid sometimes lipopolysaccharid also includes 3
layers proteins.
-plasma membrane-trilaminar like gram ve
-has ribosome like particles like intracytoplasmic organells
-is an obligatory intracellular parasite(grows only in cytoplasm of
eukaryotic cell)
Physiology
-they dont grow on artificial media except Riquintaha grows in
blood agar medium
-yolk sac inoculation:they are cultivated in yolc sac of developing
chick embryo
-tissue culture:grow on mouse fibroblast,Hela,HeP2
-Has 3 Ags: 1 specific soluble Ag on surface,2 species capsular
AG stable polysaccharide
Ag divide by binary fission,DNA nad RNA both,sensitive to
many antibiotic,paracitic in
arthropods,found in alimentary canal,destroyed at temperature
56 celcius,susceptible to
tetracycline.
Pathogens
A)Typhus group

1.R.prowazekii
2.R.thypi
3.R.felis
B)Scrub typhus group
Orientia tsutsugamushi
C)Spotted fever group
1.R.rickettsii
2.R.conorii
3.R.akari
4.R.sibrica
5.R.australis

18.Morphology and chemical composition of viruses

-smallest,obligate intracellular,contain only one type of nucleic
acid DNA or RNA genome is enveloped with a sheeth of
proteins called capsid which may hv envelope.capsid + nucleic
acid+nucleocapsid
-capsid is made of large no of polypeptides and are called
capsomers.
-envelope which is a bilayer lipoprotein is largely of host cell
origin
-virus coded glycoprotein substances are exposed on the surface
of envelope and are called peplomers.
-virus symmetry-3 groups---hexonal-cubical-helical and complex
-shape-overall shape of the viruses are spherical,pox-rectangular
or brick shaped,rabies-bullet shaped,tobacco mosaic-rod shaped.
-some r irregular and pleumorphic in shape
Difference of viruses frm other organisms
Methods of viral classification
-toxonomic classification of viruses r based on the following
features:
1. nucleic acid charactheristics:type of nucleic
acid,molecular weight,polarity,no of nuclear acid
strands and segmenta
2. homology of nucleic acid strands
3. morphology shape n size
4. envelope
5. nucleocapsid symmetry
LUKIS CHART pg:492,493

Chlamydia
Morphology
-gram ve,obligate intracellular bacteria
-dont grow in artificial medium
-cell wall contains LPS but does not have a peptidoglycan layer
-possess ribosomes and synthesise their own proteins
-cannot produce their own ATP,rely on cellular ATP for many of
their metabolic actions
-nonmotile,nonsporing,got both DNA,RNA
-They exist in 2 morphological forms:
a)Elementary body( EB)
-is the infectious,extracellular form of organism
-it is a spherical particle,200-300 nm in diameter with an
electron dense nucleoid
-EB is much like a spore and resistant to many harsh
environmental factors.
-it is metabolically inert extracellularly
b)Reticulate body
-replicative form,taken by target cell by phagocytosis
-surface membrane forms a vacuole around particle.
-a metabolically active large reticulate body evolves from small
noninfectious
elementary body 5-6 hours after penetration of target cell.
Physiology
-can cultivate in chicken embryo,tissue culture.HeLa
-resistance: heat lable susceptible to dilute solutions of
formalin and phenol
-Ag:heat stable,complement fixing,heat labile
Phatogens
1.C.psittaci
2.C.trachomatis
3.C.pneumoniae

Tissue cultures and their characteristics

-3 types tissue cultures :
1. organ culture-small bits of organ frm man and animal r
maintained in tissue culture growth medium.organ cultures r
done mainly for highly specialized parasites of certain
organ,e.g.tracheal ring culture for isolation of corona virus
2.
cell culture-for identification and
cultivation of viruses.the growth medium for tissue culture
is basically a balanced salt solution and contains 13 essential
amino acid,glu,salts,buffering system,protein supplement.
3.
explant culturefragments of mixed
tissue grown at explants and embedded in plasma clots.may
also be cultivated in suspension.adenoid tissue explant
cultures were used for isolation of adenovirus.
Cell culture
a)primary cell culture-normal cells obtained frm body,capable
of only limited growthexamples :monkey kidney cell and human
amnion cell culture.
b)diploid cell culture-cells of single type,usually fibroblast
contain same no of chromosomes as the parent cell n diploid.they
used for isolation of fastidious viruses and production of viral
vaccines.the fibroblasts derived frm embryo tissues.
c)heteroploid cultures(continuous tumour cell lines)-cells of
single type capable of infinite growth in vitro.they derived frm
cancer cells.they can be serially cultivated
indefinitely.e.g:HeLa,Hep2,KB cells

19. Reproduction of viruses(phases of viral host cell

interaction)it has 6 stages,
1.Attachment and adsorbtion
-virusses come in contact withhost cell by random collision.If the
receptors on host cell are specific for the virus only attachment
take splace
2.Penetration
-entering of virus into the host cell by 2 mechanisms:
a)for naked viruses-a process called viropexis which
resembles endocytic as a result the virus is engulfed, a vacuole is
formed
b)for coated viruses-the host cell membrane+te virus
supracapsid forms together and the nucleocapsid is released into
the cell
3.Uncoating/Deproteinisation
- this is the destruction of capsid.It may occur due to action of
lysosymal enzymes in the vacuole or by cellular proteases in the
cytoplasm.The naked nucleic acid is thus formed.
4.Biosynthesis
-synthesis of nucleic acid of virus,capsid protein and also
enzymes required for synthesis and release.Also regulator protein
are synthesized to shutdown the host cells normal metabolism
-site of viral protein synthesis in all cases-host cell cytoplasm
-site of viral nucleic acid synthesis
-in DNA containing viruses-host cell nucleus except pox
virus
-in RNA containing virus-host cell cytoplasm except
orthoxyxovirus,retrovirus and
paramyxoviruses which synthesise paiths in nucleus
-in general the following steps occur:
1.transcription of Mrna frm viral nucleic acid by host cell
RNA polymerase
2.translation of mRNA into early proteins in
cytoplasm.These proteins include
enzymes which initiate synthesis of viral components like
DNA polymerase
20.Phages ( viruses of m/o ):morphology and ultrastructure

-general str maybe characterized by str of Coli-phage of

E.coli.
-bacteriophage is tad-pole shaped with:
Head:hexonal shape n contains core of nucleic acid
covered by capsid.
Tail: cylindrical n is composed of central hollow core
surrounded by protein.
Basal plate at the terminal of tail.
Fimriae/tail fibre or prongs-attached to the basal
plate.fimbriae hv receptors.
LUKIS GAMBAR
Phases of interaction with cell of virulent n moderate
phages
-Virulent phage:virulent/lytic phage undergoes the
virulent/lytic cycle
1.Adsorbtion on the surface of bacterial cell according to
specific affinity of bacterial
receptors to specific phages.attachment occurs by tail.
2.penetration(infection of bacteria by naked phage nuclic
acid-transfection).the tail sheeth contracts so that the blood
plate+tail fibres r held tightly against the bacterial wall.the
halloe core is pushed into the bacterial wall.part of nucleic
acid enters the bacterial wall.
3.synthesis of phage components-immediate transcription
of part of nucleic acid in cytoplasm helps further injection of
nucleic acid.empty head+tail remain outside as ghost or
shell.the viral genome directs protein synthesis which stops
normal metabolism of cell n causes synthesis of viral
structures.
4.maturation+anembly of new virions
5.release-during replication bacterial wall is weakened by
basal plate enzymes,assumes spherical shape

a)DS DNA containing viruses-viral nucleic acid enters nucleus

of host cell.
-Using host cell RNA
polymerase ,transcription of mRNA is achieved .mRNA moves
into cytoplasm-induces synthesis of protein including DNA
polymerase.DNA polymerase enters nucleus causes replication
of viral DNA.
b)SS DNA containing viruses-b4 all other steps,host cell DNA
polymerase is used to make viral DNA double stranded.Mrna is
using newly formed DSDNA as template.
c)SSDNA-occurs in cytoplasm.RNA dependent RNA
polymerase of virus causes transcription of mRNA in
cytoplasm.
d)SS RNA(+ve strand)-the viral RNA codes for all proteins
needed for replication is acts as mRNA itself.This RNA is
infectious.
e)SS RNA(-ve stranded)-they hv viral RNA polymerase.Occurs
in nucleus ( influenza virus)-Transcription of mRNA occurs in
nucleus.nucleic acid r non-infectious.
5. Maturation and multiplication
The viral genoms and protein capsid are assembledin form
daughter virions.This occurs in the nucleus( terpes,adenovirus)
or in the cytoplasm (picorna +pox viruses)
6. Release
-the viruses leaving the host.Can occur by:
a)lysis-destruction of host cell
b)budding-in which enveloped viruses obtain their
supracapsid frm cell or nuclear
membrane.The host cell remains unaffected.
Types of virus-host cell interaction
1-Infectious/productive-form of new viruses-release
2-Integrative-DNA integrate into chromosome to form viruses
3-Abortive-abortion of process at different stages of replication like
anembly

Titer of phage
1.applemon method-pore specimen thru bacteriological
filter.only BP will be filtered.liquid obtained frm this should
be serially diluted into 10 test tubes.m/o are added to the test
tubes.max dilution which causes death of m/o=phage titre
2.Grace method-on Petri plate with agar,m/o r
cultivated.the filtered liquid is diluted and added to Petri
dishes.incubated.sterile zones r found on some Petri
dishes.number of sterile zones r counted and multiplied by
dilution+phage titre
Prophage
Integrated phage nuclic acid into the bacterial
chromosome in the phage cycle of temperate
phages.prophages confer new population to bacteria.for
example,prophage in S.thypi converts into new antigenic
surface str,toxin production by C.diptheria.the prophage
multiplies synchronously with bacterial DNA.the state of
presence of prophage in bacteria without phage productionlymphogeny.the bacteria is called lysogenic bacteria.only
double stranded DNA can be integrated as prophages.some
phages such as P1 do not integrate into chromosome but exist
as autosomal proviruse plasmid.continuous synthesis of
receptor protein by cell is maintained lysogenic state.when RP
level falls-prophage switches to lytic state.this can also occur
due tophysical/chemical factors.

21.bacterial nutrition
1.microbes receive nutritional substances on surface of the cell
2.m/o hv the ability to change the type of nutrition according to
external condition
3.they can use both organic or inorganic substances
4.high speed of metabolic processes.
5.some organic compounds must be degraded into smaller
components b4 being transported into cells.
Types of nutrition
According to source of nutrition,microbes r divided into:
1.autotrophs-autotrophic m/o mk use of inorganic compounds or
natural sunlight.(co2,n2) to synthesize essential substances.
a) phototrophs-derive energy frm sunlight
b) chemotrophs-derive energy frm inorganic
substances(o2,n2,h2) and synthesize organic substances
2.heterotrophs-m/o which require preformed organic substances
for growth
a)saphrophytes-use dead organic matter
b)parasites-use living organic matter
mechanisms of the transport of substances into the cell
1.simple diffusion-small molecules which pass across cell
membrane frm the region of higher concentrationoutside the m/o
into the region of lower concentration inside the m/o.yhis process
doesnt require energy
2.facilitated diffusion-molecule is transported across the cell
membrane with help pf carrier protein.process does not require
energy.occurs according to concentration gradient.
3.active transport-substance is transported against concentration
gradient (concentration of substance is higher inside).this process
requires energy and carrier protein.
4.translocation of radicals-large molecules are broken down by
enzymes into smaller radicals outside the cell and the radicals
pass across cell membrane inside the cell.the molecules r
reformed.this process requires energy.

Nutritional requirements for the growthof the m/o

1.essential elements
a)hydrogen, b)oxygen =in cultural media,found in water
c)nitrogen-needed for formation of proteins,nucleic acids and
other cell constituents.obtained frm ammonia or deamination of
amino acid
d)carbon-used for generation of energy and
biosynthesis.obtained frm carbohydrates by oxidation or
fermentation processing ATP.
e)phosphorus+sulphur-S forms part of coenzymes.P is part of
nucleic acid, ATP, coenzymes .source-free inorganic PO42-.
2.minerals-K+,Ca2+,Mg2+,Na+ required in trace amounts for
enzyme function.
3.organic growth factors-some bacteria require organic
compound which cant be synthesized
by themselves
a)essential growth factors-bacteria cannot grow without such
factors because the home lost ability to synthesize them
b)accessory factors- bacteria can grow without such factors but
presence of accessory factors enhance the growth of m/o.

22.Culture media
- any preparation that contains nutrients essential for
bacterial growth=medium.any medium that has been
successfully inoculated=culture medium
Classification
1.according to origin
-natural media-blood,milk
-artificial media-a)synthetic-prepared frm pure
chemicals
b)semisynthetic
2.according to composition
-simple media-containbasic substances-N,C.for
example:nutrient broth,nutrient
agar,peptone water
-complicated media-for example-enriched medium
containing true enriched matter.when
certain incredients added to a basal medium to study
special characteristics or to provide
special nutrients require for the growth of organism
is called complicated media.for
example:blood serum,egg or meat pieces add to
basal medium.
3.according to physical condition
-solid-stroke,stab cultures
-semisolid
-liquid-blood culture

4.according to assignment
1.
common media-for growth of all m/o
2.
differential media-culture medium contain
certain substances helps to distinguish different properties of
different bacteria.for example=blood agar,McKonkeys agar
3.
selective media-in addition to basal media,they
contain substances like bile salts or deoxycholate citrate
which inhibits or poisons all bacteria except those of
particular type of group of wanted organisms.media contain
substances like bile salts groth of all other m/o except
one.Ex-DEAmedium,McConkeys agar,bile salt agar
4.
special media-for m/o that require special
substances.For example:glucose enriched media
5.
transport media-for transport of specimen
example:stuarts transport medium for urethral
discharge(conococci),bile peptone transport medium for
stool,glycerol saline transport medium for stool(bdysenteriae)
6.
indicator medium-includes indicator (dye) or
reducing substance(ktellurite) color or indicator changes
with bacterial growth
example:wlson+blair medium,sugar medium-hisss medium
when certain indicator(neutral red,bromothymol blue) is
incorporated in culture medium
Main requirements for culture media
1. medium should contain all nutritive
components(O2,N2,C,H,minerals,organic
compounds)
2. moisture is an essential requirement for bacterial
growth
3. sterility
4. optimal pH should be maintained usually 7.2-7.4

23.Growth and division of microbes

-bacteria divide by binary fission.thecell division is initiated
when a bacterial cell reaches a critical mass in its cellular
constituents.bacterial nucleus.2 circular double stranded DNA
molecule sequences in 2 starands.nuclear division precedes
bacterial cell division.replicated DNA distributed to 2 daughter
cells.a transverse septum grows across the cell frm cell
membrane.this is followed by deposition of cell wall material and
finally the 2 daughter cells separate.
Generation time-is interval of tieme between cell divisions or
time require for bacteria to double.example:C.coli-20
minutes.bacterial count.total count-living+dead.viable countcapable of multiplication.
Phases of division
1.
lag phase-immediately after inoculation of culture
medium,bacteria undergoes a period of adaptation.during
necessary enzymes and intermediate metabolites a built up
with increase in its size and meatabolic activity,the bacteria
attains a critical stage for multiplication.the length of lag
phase depends on type of bacterial species,size of
inoculation,quality of culture medium.1-4 hours.
2.
log/exponential phase-the cell division then
proceeds and the members increase.the logarithm of viable
count when plotted against time a straight line. 8 hours.
3.
stationary phase-after sometimes,bacterial growth
ceases almost completely due to exhaustion of nutrients or
accumulation of toxic products.there is a slow rate of cell
death.the total cell count slowly increase but viable count
remains stationary.duration ranges frm few hours to few
days.
4.
decline phase-after aperiod of time in the stationary
phase,the bacterial population starts to die,the number of
total cells remains constant.this phase brought by exhaustion
of nutrients,accumulation of toxins and autolytic
enzymes.duration varies frm few hours to few days.

24.Classification of m/o by the type of

respiration
-according to requirement of oxygen for growth of
m/o,m/o can br divided into
1.
aerobic bacteria-require oxygen for growth
a)obligate aerobes-grow only in presence of O2
-Pseudomonas,Bacillus
b)facultative anaerobes-normally one aerobic but can
also grow in absence of O2,but
less abundantly majority of pathogens are facultative
anaerobes.Vibrio cholera
2.anaerobic bacteria-Cl.tetanus-grow in absence
of O2.obligate anaerobes die in presence of O2
3.Microaerophilic bacteria-grow in presence of
small amounts of O2.Streptococus,Borrelia
Scheme of biological oxidation
1.Aerobes
-obtain energy by process of oxidative phospharilation
where ADP is converted to ATP by the formation of energy
rich phosphate bonds.(34 ATPs are formed)
-ultimate H2 acceptor is O2,H2 donor-organic
substances
-enzymes this process are found in cell wall-mainly
cytochrome oxidase
-organic compound completely oxidized to CO2,H2O
and a little H2O2 is found
C6H12O6 + O2+ADP CO2+H2O+ATP+
(H2O2)
-H2O2 thus formed is broken down by
catalase/peroxidase found in m/o
2.Anaerobes

Cultural conditions
1. all necessary nutrients
2. optimum temperature(36-37)
3. optimum pH(7.2-7.4)
4. moisture
5. sterility of the medium
thermophiles-55-80
mesophiles-25-40
psychrophiles- less than 20

41.BACTERIAL GENE APPARATUS AND THEIR

PECULARITIES IN VIRUSES.GENOTPYE AND
PHENOTYPE OF MICROORGANISMS.
1.genetic material are organized in chromosome which is
presenting cytoplasm.
2.genetic material is haploid but DNA amount found is
constant in microorganism.
3.in MO ,Genetic material is transferred between MO of
same generation
4.presence of special genetic elements plasmid are found
in MO.
5.in viruses either DNA OR RNA are present .
GENOTYPE.
The collection of genes encoding the characteristic of the
bacterial cell which remain unchanged.
PHENOTYPE
The characteristic expressed by the bacterial cell at a
particular time can be attend acc. To environmental
condition .
PERCULARATIES OF VIRAL GENETIC
1.DNA AND RNA
2.cross reactivation :if the viruses enter human cell, cross
changing between the two occurs and a new virus with a
new genetic character new genome of virus.

43.GEENTIC CHANGING IN MO
(RECOMBINATION ).KINDS OF RECOMBINATION AND
THEIR TYPE CHARACTERISTIC .PLASMID :MAIN TYPE
AND THEIR CHARCTERISTIC .

Recombination changing of genes between cells .it s 3

type .
1. Transformation uptake of naked DNA 0.7% of
chromosome
The change depends on usage of free DNA IS induced
into her MO . sequenceof DNA is intergrated only in
complimentery spaces forming MO with new
genome.when MO destroy ,DNA fragmentation can enter
cells tht finds compensatory place and hence MO have
new sequence.
2Transduction
Is transmission og genetic sequence frm one MO to other
with the help of bacteriophage DNA.bacteriopahge enter
blood cells and multiply
Now SP transduction bd intergrated different species of
the doner cell and change sequence of DNA.
Sp - intergarate in sp place and specific sequence or DNA
synthesized.
Absorbtive this do not intergarated to blodd cell DNA
before neutralize inte cytoplasms and change the activity.
Transmission
Nucleotibe between two MO directly wjth the help of
DNA
F(+)- have sex pilli- abilitiy to recombition
F(-)- have no sex pilli
HFR- high frequency recombination f(+) pilli hve
extragenous transport gene- donor cells forms a brigde
between f(+) And f(-) .

a.
b.
c.
a.
b.
c.
d.

42.KINDS OF CHANGING .MUTATION ,THEIR TYPES,

MUTAGENS:KINDS PHISICAL ,CHEMICAL
.BIOLOGICAL.
Kind of changing.
Changes can be 2 type
1.modification
2.mutation
Modification
Is no change in genetic struc. Only suppression or expression of
some genes acc. To environmental condition . they can be :
short time modification
long time modification
Mutation
Heritable variation due to any change in nucleotide sequence of
genes .it can be genotypical or phenotypical
Mutation acc.to mech.
1.Induceable-may be induced by the influence of different
factors like mutagen .
They can be physical ,chemical and biological.
2.Non induceable spontaneaus
Mutation acc. To number of nucleotide
point mutation change in 1 nucleotide ,irreversible
chromosomal change in chromosome .( only in
humans )
gene mutations change in genes .
Acc. To mech of changing
deletion loss of some genes
insersion addition of extranucleotide
inversion change between different nucleotide
transposition /translocation change in position of
nucleotide
Acc . to time .
a. irreversible or direct mutation
b. reversible

44. ROLE OF MUTATION IN EVOLUTION OF

MICROORGANISM, RECOMBINATION AND
SELECTION .
-MO A,B,C antigen first imjected in to rabbits.
-antibodies A, B AND C are produced after days inside
the rabbits it is the non absorbed primary serum.
Those antibody or primary antiserum is added to MO with
antigen A,B,C .

Incubate 1 hours - agglutination

Antibody titre- highest dilution of serum 1/100.
Indirect heamoagglutination test or passive
heamagglutination
Carrier blood is mixed with antigen -ag/RBC and antibody
Therefore it is easy to detect the number of antigen and
antibody.

45. BACTERIAL CHANGEABILITLY AND ITS ROLE

IN DIOGNOSTIC . THERAPHY AND PROPHYLAXIS
OF INFECTIOUS DISIEASE .

Trasformation occur only in few bacterial

species under natural condition for successful
transformation the bacteria must be completed to take
up DNA from environment and incorporate into
genome .cell capable of take DNA from any source .
Transformation normally in 3 stages
In pneumococcus and some gram positive
bacteria and in vading DNA fragment is ciut to 7-10
kb by endonuclease of the membrane and only one
strand enter the cell.
In gram negative heamophilus a membrane
protein binds a sequence about 10 nucleotide and
DNA enters as double strand unside the cell.
Incase of entero bacteria transformation occurs
only after a artificial modification envelope by
conversion to sphero plast or permeabilising the
envelope to DNA by heating the cell presence of cacl
the modified cell surface permit taking up double
straded DNA fragment s.
ADAPTATION
-sometimes phage or plasmid DNA may be taken by
a permeable cell where recombination of invading
DNA with replication of recipient if not necessary
and some become re established in recipient cell and
autonomous replication , the mech. Is same as the
transformation since the introduced viral DNA
genome .this process is called transfection.because it is
result of infection
46.HUMAN MICROBIAL FLORA AND its

-The normal microbial flora is divided into

Resident
Fransicut(facultative flora)
-The normal microflora play important role in body
normal function
-They can be pathogenic when host defensive mech. is
fallen down.
-They colonized and release substance as taxius for the
body maintaing occurs due to the defective immunity in
the host
-The microbial cond. In the interstinal tract is greatly
affected by bad foods ,incorrect drug usage (antibiotic)
because specially the interstinal tract syn.vitamin k by
its normal microflora also several vitamin b types .
-When the normal microflora because changed due to
the failed human defensive mech.they syn. Exotoxius
liberate tissues destroy the own tissues of the organ different disease .
-The normal mircoflora change due to the age .diet ,
hormonal state , health , sanitary cond. , personal hygiene ,
eve na newborn has a contaminated with mother s micro
organisms in the body so theu adapt before they
exposure to the normal environmental changes of the
physical leve l can change the balance between the
microflora pathway
-The microbial flora is controlled by
a)chemical b)mechanical factor c)immune factors
SKIN FLORA ,MUCOUS SURFACE , RESP.TRACT .
ORAL CAVITY .
-many organisms cames and contact with the skin surface
,if they dont have environment to contact they dies .
-gram positive bacteria is most common MO on the skin
*coagulase-negative staphylococcus and s.aueres

Transduction
transmission of piece if host DNA frim cell to
another by bacteriophage .
transduction generally found in cell in infected by
temperate phage intergrate in its chromosome in quiescent
state .
Theraphy
Taking vitamins for infectious diseases before they enter to
the body .
Giving antibiotics tht can interfering with the cell wall syn.
Drug affect in cytoplasmic membrane
Drug inhibit protein syn. And impairment of function of
ribosome aminoglycoside ,tetracycline ,chlormphenicol and
macrolide antibiotics lincomycin inhibit protein synthesis .
In bacterial ribosomes woth out any major effect on
mammalian ribosome
Antimicrobial drug such penicilin ,cephalosporin and
aminoglycoside and other antibiotics .
Macrolides macrolytic lactone ring attach to erythromycin
,sprinomycin , clindamycin and linomycin ,
Penicillin gram positive bacteria -intra muscular and intra
venous
Ampicilin gram positive bacteria intramuscular and oral
Oxacilin gram positive bacteria intra muscular and intra
venous
Amidinopenicilin gram positive bacteria ,intravenous ,intra
muscular and oral ,
Cefotaxime-gram positive and negative bacteria-

intravenous and orally .

RESP.TRACT.
-nose it is the usuall habitant of staphylococci (s.aureus ,
s.epidermiks )sometimes has streptococci (s.puenomoniae
,p.pyogones ) the non pathogenic neisseria is transient in
hear.also corynebacterium and moraxella lacunata .
- throat ,nasopharengeal , oralpharynx ,and tonsils MO
can be find usually in this pathway .coagulase
staphylococci may be found in nasopharynx and tonsils
-euorococci is also present , highly present
c.orynebacterium,actinomycites ,veilovella ,spirochetes
,pnevotella is in tonsils .
-lower resp.tract ,larynx ,trachea , bronchioles ,and lower
airway colinezed MO cytoplasm are aspirated .
-an acute disease of lower airway is by virulent bacteria
in the mouth (s.pneumonia ,s,aureus
ORAL CAVITY
has micrococci ,coliforms ,lactoacccili
struc:viridaus ,nesseria , diphtherioidus and
bacteriodes
Changing Flora due to the Age of Human
- when mens get s old body immunity decrease .when at
tht time MO is very susceptible for body changes and
also for pathogenic MO .
- even the normal body microbial flora can change and
different kind of disease in the body this due to the
changes .
-In its first stage of human like in this during birth of a
baby the body of baby in contact with environmental due
of normal microbial flora so that before the body is
stabilized .often tht development of normal microbial flora
occur in young age has highest concentration of normal
microbial flora .it has normal level

47.MICROBIAL FLORA OF COLON .MAIN .

The small intestine can be contaminated wit different types of
bacteria and parasites .most of these oraganisms are
anearobics as peptostreptococcus ,bifidobacterium
,dorphyromous.drerotella ,zactobscillus .
They play relo in formation of colonization resistance and give
protection of mucous pathway .
Large intestine ,more MO can present than any where in own
body .it is estimated than more 10 bacteria per gram of feces
can be found with anaerobic bacteria in excers by 1000 folds
yeasts .
Non pathogenic parasites can also be seen.the most comman
bacteria is b.lidobacterium, bacteriodes lactobacillus
,eubacterium
Fugobacterium ,veilovella , eubacterium ,pcptococcus and
enterococcus can be seen also pseudomonas ,myoplasms e.coli ,
claudida , present in less amount
DISBACTEROSIS .PROPHYLAXIS.
Occurs using antibiotics or changing of diet or stress or less
vitamin or radiation
It s alteration of normal microbial flora that changing the
balance between MO numbers .in disbacterosis th numbers of
opportunistic pathogenic increase .
This disbacterosis occurs prolong therapy of antibiotic
,hormones ,radiation sickness ,immune suppress ,treatment
infectious of GIT , cancer and immunodeficiency and stress.
STAGES .
1-Decrease of resident without changing a note and MO to
resemble it .
No clinical symptoms
2-Increase count of transient from opportunistic flora .gut
disfunction and local inflammation .
3-Changing microbial flora .

TREATMENT
Administration of vitamin b
Complex to positive normal microbialflora syn. This is
given with antibiotic theraphy .
Vitamin k admistration
Changing the antibiotic
General gnoto biological isolation
Immune theraphy
Admistration of living MO eg.originallly e.coli
,lactobacterium ,genus bascilus .

48.INFECTION AND INFECTIOUS DISEASE .

Main factor necessary for the beginning of infection.
- pathogenic MO
- susceptible host
- number of MO
- environmental and social influences
Differentiation of incetious disease from other human
disease
caused by pathogenic MO
production of immunity
incubation period(absent of clinical symptom
,multiplication of MO adaption )
contagious (contamination )
can be transmited
infection
infection between pathogenic MO and
susceptible host under the special environmental and
social condition .
infectious disease
condition in human characterized by present of
symptom due to present pf MO .
PERIOD OF INFECTIOUS DISEASE
incubation period
prodromal period no clinical symptom ,no
specific symptom are present
manifestation specific clinical are present
decline result could be recovery or death .

49. PATHOGENICITY & VIRULENCE OF

MORGANISM.
Pathogenecity: ability of m.org to cause a disease
Virulence:measurement of degree of
pathogenecity&depends on invasiveness & toxicity of the
organism.
Basic forms of virulence:
adhesion non specific connection by ligands &
R.
subdivided into 4 groups: - pilli (F1)
-cellular factors of adhesion
(F2,F3)
-surface antigen (F4)
-fimbren
special adhesion factors:
bifidobacteria-lipothiac acid
lactobacilli -polysacharide

a)

Influences of physiological conditions:

-conditions of epithelial cells + medium
-presence of inhibitors(antibiotics)
-high concentration of NaCl
Adhesion & colonization in mucous membrane they must
overcome:
-mucous barriers (mechanical barrier)
enzymes for destruction eg: protease
-Chemical barriers
forming of enzymes ; to form neutral/ alkaline medium
around m.org
-normal microfloral barriers

b) Invasions
-causes destruction of host cells/tissue cells when
m.o enters inside of host cells
enzymes that helps invasions eg:DNAase,
protease, histaminase
c) Autophagocytic factors
-causes lysis of m.o :a) cell capsule
b)protein (A protein of
staphylococchi & M protein of
streptococchi)
c) Factors of toxicity:
-exotoxin (antigen) & endotoxin
Methods of measuring virulence
Virulence:measure of ability of m.o to cause infectious
diseases
a)DLM (doses lethalis minimum) min num of m.o which
causes death of 95% of infected animals
b)LD50 (lethalis doses 50)- num of m.o which causes
death of 50% of infected animals
c)DCL -number of m.o causingdeath of 100% of infected
animals
LD50 = A [1 + 50-a ]
b-a

50.MICROBIAL TOXINS, TYPES, UNITS OF TOXIC..

toxin produced by m.o can be exotoxin or endotoxins
property
exotoxins
endotoxins
location
Gm+ & GmGm- structural
compound of
cell wall
extracellular
excreted into medium
chemical nature
polypeptides
lipopolysacharides
stability
unstable(high temp)
stable(sensitive
>60 C)
toxicity
most powerful
< toxicity.can
reduce in large
dose
effect on tissues
on specific tiss eg;
non specific
action
neurotoxin & hemotoxin
fever production
cant cause fever
produces fever,
rapid in high
temp
toxoid conversion
can be converted to
cant be
converted to toxoid.
& usage in medicine toxoid. Can be used in
cant be used in
immunization
immunization
examples
diphtheria, food
salmonella,
endotoxin shock
poisoning, cholera,
tetanus, botulism,
gas gangrene, plague
Units of toxin strength : DLM (minimum lethal dose)
Gene determinants: plasmids, enzymes, lodetoxin HL 1

a)

51.WAYS OF INVASIONS OF M.O IN HUMAN.

contact
-maybe direct or indirect.
-indirect contacts by contaminated objects.eg; towel,
syringe.
-direct by kissing & sex
b) vehicle invasion
spread of disease by by H2O, food, blood or often
biological products.
Eg; waste borne- cholera, typhoid.
Food borne- enteric fever
Blood malaria, syphilis, renal hepatitis
c) vector transmission
-they are arthopods or intervertebral host that transmit
intections
Eg; mosquito, flies
Mechanical- passive transmission of infection by
vector
Eg; house fly carries typhoid
Biological- multiply and grow in incubation period.
Begin the vector transmission
By infection to man. Eg; anaopheles mosquito in
malaria
d) air borne infection
droplets- by respiration eg; TB
dust of bedding (furniture)- Eg; staphylococcus. It
stays in dust
e) transplacental transmission: some diseases as
vortical transmission, from infected
mother to fetus. Eg; rubella
f)iatrogenic transmission: by intermittent and lab stage

Dynamic of development of infections.

Infections maybe localized or generalized.locally infection
can spread by :
-lymph channels(lymphagitis)
-lymph nodes(lymphadenitis)
-blood stream(bacterimia)
m.o can also enter the body through skin; mucosal flora or
exogenous sources.
its period: a) incubation period- absence of clinical
symptom, multiplication of m.o
b)
latent period- does not feel any abnormal
condition
c)
predormal period- common clinical symptoms
d)
clinical form
e)
decline stage.

52. FORMS OF INFECTIOUS PROCESS

a)due to presence of symptoms symptomatic

-asymptomatic
b) due to different forms- abortive
-latent,typical,atypical,chronic,slow,bacterial cleavage.
Persistence of pathogen in organism
Reinfection
-infecting by the same pathogenic m.o after the
illness is recovered
Superinfection
-infection of sick person by the same m.o before
recovery
Secondary degree infection
-when body immunity decrease of the patient by a
primary degree infection disease.
a)new m.o cause and give an infection
b) exogenous and endogenous infections
exogenous infections , the infection occur fm
outside
endogenous, the infection occurs when normal
body microflora is changing its normal(auto infection)
place and this leads to infection development on the
systems.
Local and generalized infections.
Local infections it occurs at the site of invasion when
the patient has a open wound. M.o easily enters the body
at this site. If it lays in superficially ~ local infection
Generalized infection~ this infection spreads through
lymph channel (lymphagitis) blood stream (bacteremia)
54. HOSPITAL INFECTIONS

Disease in patient or medical staff after presence in

hospital independent of the time of the symptom appears
on infectious disease in patients after admission.
Conditions for the beginning:
a)age- new borns & elderly ppl insufficient immunity
b)infected patient can be spread by close contact
c)drug resistance the infection can be resistant against
some drugs
d)susceptible patients- diabetes mellitus,
immunosuppressive patients
e)surgical procedures- eg; medical manipulation systems
by surgical instruments,clothings, injections that enter the
body. Also by nurses , doctors & other medical staff
infection can spread
Hospital strains of opportunistic m.o
-they are stap.epidermidis, stap.aureus, strep.pyogens,
E.coli, klebsiella, pseu.aeroginosa, enterococcus,
enterobacteria, shigella sonnei
Condition for formation
-contacting with other patient & staff
-environmental sources
-by surgical objects~ equipments, blankets, food
-Hospital air~contacting with dust which is infected,
susceptible patients
Surfaces~ blood, body fluid
Characteristics
a)transmission by contacting hand & clothing also by
instruments
b)air borne infections~ droplet of respiratory tract can be
passed to open wounds also dust from floor & bedings
c)oral route

1)
2)
3)
4)

53. ROLE OF M.O, HUMAN ORG, ENVIRONMENT &

SOCIAL CONDITIONS IN THE BEGINNING & DEV OF
INFECTIOUS DISEASE
m.o- m.o that are pathogenic develop infectious disease. They
produce toxins , enzymes, & disturb normal microflora
human organism- has barriers to stop development of infectious
disease like physiological conditions ( presence of inhibitors,
high concentration of NaCl, high pH, immune system) of normal
microflora
environment- may influence the development of resistance & at
the same time increase in the number of m.o
social conditions- unhygienic, sexual contact, eating habits
mixed infection: infection by more than one m.org. They are;
-difficult to diagnose because development of
different disease at same
Time
-difficult prophylaxis & treatment
-high motility (high death rate)
Types of interactions between m.o
m.o maybe from different taxonomic group
-bacteria-bacteria, bacteria-viral, viral- viral, bacteria- fungal,
bacteria- protozoal etc
Main types of interaction are;
independent multiplication-m.o multiply independently
exaltation- one m.o increase frequency of multiplication of the
other
complementation- both m.o depend on each other
indifferentiation- one m.o suppress the multiplication of the other
microbe

55. SPECIALITY OF VIRAL INFECTIONS

Molecular type of interaction
-necessary penetration of virus into susceptible cells
-development of virus inside host cells will damage host cell
-viral infections doent connectwith excretion of virus outside of
the body. Pathogenesis of viral infection depend on the types of
interaction of viral & host factors
a)entry of viruses into susceptible cells
b)viral multiplication & spreading
c)effect of virus on cell function
d)host immuno response
e)viral clearance or establishment of persistant infection
f)viral shedding/spreading
entering of virus: through skin , resp tract, G.I.T & urogenital
tract or conjunctiva or through mucous membrane of respiratory
tract eg: polio virus
some virus introduced directly to the blood eg; HIV, hepatitis B
multiplication & spread: multiply locally & remain confined
there w.out any systemic spreading. Primary multiplication at the
site of entry these virus pass through the lymphatics to local
lymph nodes. After multiplication in local lymph nodes, virus
enter blood & causes primary viraemia. Primary viraemia~central
foci~multiply~enter the blood ~secondary viraemia
(liver,spleen)
viral effect on cells
-cytolytic or cyclocytic growth. Virus replicate in susceptible host
cell releasing new virus which infect new cells, destroy target
cells & tissues & causes diseases
-non cyctocidal productive growth . some viruses are able to
infect cells but do not replicate within the cells.
-viruses nucleic acid & host cell DNA interact causes
transformation of immunity of viral infection

a)

56. IMMUNITY

Infectiveness of viruses
Pathogenic m.o causes infectious diseases eg; plague,
polio virus
Opportunistic m.o may cause infection in a person with
suppressed immunity
(Innate immunity after antibiotic therapy is suppressed)
Saphrophytes, causes disease only in adult with high
suppression of immune system. Eg; patient with cancer ,
influence of radiation ( destruction of T & B
lymphocytes), patients with AIDS, patients with
transplantation
Influences on opportunistic m.o number over the years .
increase in duration of life span . increase in frequency of
invasible manipulation. (injections)
Usage of immunosupressors (hormones , antibiotics,
radiation therapy)
Ecology (chemical , physical influence of life)
Kinds of viral infections
respiratory viruses (resp tract)
eg; influenza A, B, C, para influenza, mumps, measles,
adeno virus, rhino virus, echo virus
b) enteric virus has GIT entering. Eg; polio, coxsackie
virus, rota virus, hepatitis A &B
c) neurotropic virus eg; poilio virus, coxsackie virus~ GIT
rabies virus ~ skin & blood
mumps & measles ~ respiratory
tract

Basic parts of modern immunology

Immunology state of resistance on insusceptibility by
the host to toxic molecules , m.o & foreign bodies cells
Study of immunity immunology (studies the ways &
method of immune status of the body & the way the body
react to against different antigen. When different antigen
comes and react at the surface of the liver , if this is not
destroyed, they lead in formation of a disease. This
condition occurs when the patient is having immune
depression or immune depressing disease. This lead to
immune deficiency and patients get involved in different
immunopathological disease.
Im mmunology the study about cells that regulate the
immune status in body is known as Active natural:follows
clinical or suclinical infections
Artificia: induced by vaccination
Passive natural:tranplacenta passage of Ig G, antibody
protects child For at least first 6 months of life

57. Non-specific Factors of Human Defence:

a) AGE foetus and old person carry high susceptibility of

infectious diseases.
b) HORMONAL INFLUENCE certain hormonal disorders
such as diabetes mellitus, hypothyroidism, and adrenal
dysfunction influence susceptibility to infection.
c) NUTRITION malnutrition predisposes to bacterial
infection such as gram negative bacterial septicaemia,
tuberculosis, herpes viral infection, and measles.
Superficial Surfaces:
Animal body is a close system separated frm the
environment by skin and mucous membrane which are
impermeable to the particulate material of the size of
bacteria.
a) Skin: intact skin provides a mechanical barrier to the
invading microorganisms and also provides bactericidal
secretions. Sebaceous secretions containing unsaturated
fatty acid n free saturated fatty acid have bactericidal and
fungicidal actions. The skin can be freed of transient flora
readily but the resident flora on it cannot be removed even
by washing and application of disinfectant.
b) Nose, Nasopharynx, and Respiratory Tract: inhaled
bacteria arrested in the tortuous nasal passages on the
moist surfaces of the mucous membrane lining. Those tht
pass beyond the larynx are trapped in the bronchial mucosa
and only a few reach the bronchioles and alveoli. Cilia
sweeps the secretion containing the foreign particulate
material towards the oropharynx, where it is swallowed or
cough out. Material tht manage to reach alveoli are
ingested by phagocytic cells present there. Nasal and
respiratory secretions contain muco-polysaccharides which
can neutralise influenza and some other viruses.
c) Mouth, Stomach and Intestinal Tract: saliva possesses

a)
b)
c)

d)

Humoral Factors:
Apart frm specific antibody there are variety of non-specific
antibacterial substances in blood and tissues.
Properdin: it is a euglobulin present in normal serum
and causes lysis of gram negative bacteria with the help of Mg
and complement, it also inactivate some viruses.
Complement: it acts only on microorgm sensitized by
specific antibody, it possesses bactericidal activity.
Lysozyme: bactericidal enzyme of low molecular
weight basic protein. Found in polymorphonuclear leucocytes,
tears and other body fluids. It lyses the mucopeptide of the cell
wall of many gram negative bacteria.
Other Antibacterial substances: betalysin, basic
polypeptides (leukins, frm leucocytes; plakins, frm platelets)
have antibacterial effects. Interferons possess antiviral effect.
Cellular factors:
When agent infect tissue, occurs exudative inflammatory
response, accumulation of phagocytes, outpouring of natural
antibacterial substances and deposition of fibrin. Fibrin entagles
or traps the organism and acts as a barrier to spread of infection.
Role of normal human flora:
a) Beneficial role
-they prevent or suppress the colonization/ invasion of the body
by pathogens by means of bacterial interference, by preventing
them frm gaining a foothold in the body.
-the bacterial flora of intestine are responsible for normal
structure and function of intestinal tract. By their metabolism
they produce vitamins, especially vitamin K and B vitamins.
-antibodies produced in response to commensals cross-react with
pathogens having related or shared antigens.

a)
b)
2)

3)

a)
b)
c)

d)

58. Phagocytic theory of immunity

1) Phagocytosis
Phagocytic cells are classified into:
Microphages: eg polymorphonuclear leucocytes
Macrophages: these are mononuclear phagocytic
cells
Neurophil polymorphonuclear leucocytes: these
are called polymorphs, produced in bone marrow,
circulate in blood for 6 to 7 hrs. Injury to tissue excites
inflammatory response. Polymorphs arrive at scene of
infection.
Macrophages: these are mononuclear phagocytic
cells distributed throughout the body both circulating
in blood and tissues.
Phagocytic action can be divided into four stages:
Chemotaxis: being attracted by chemotactic
substances, phagocytes reach the site of infection.
Attachment (adherence): the infective agent gets
attracted to the membrane of the phagocyte.
Ingestion: phagocyte engulfs the particulate
material into a vacuole
( phagosome) the
membrane of which fuses with a lysosome forming a
phagolysosome. Lysosome contains hydrolytic
enzymes and other bactericidal substances.
Intracellular killing of bacterium: most bacteria
are slaughtered in the phagolysosome by the
hydrolytic enzymes within a few minutes of
phagocytosis.
Indicators for the characteristic of phagocytosis.
Formation of phogolysosome

60.Antigens
1) Main characteristic of antigen

a)
b)
c)
d)
e)

Antigen is a substance usually protein in nature and sometimes

polysaccharide, when introduced in a living animal evokes
specific immune response, either by producing specific antibody
or specially sensitized T cells or both. Properties of antigen:
Foreignness: the immune system possesses
the capacity to distinguish between self and nonself.
Size: larger molecules are highly antigenic
and substances less thn 10 000 daltons molecular weight are
either non-antigenic or weakly antigenic.
Chemical nature: proteins are more effective
in stimulating antibody production than polysaccharide
Susceptibility to tissue enzymes: substances
which can be metabolized and can be converted to soluble forms
by the action of tissue enzymes, can act as good antigens.
Antigenic specificity: antigenic specificity
depends on the specific active sites on the antigenic molecules
(antigenic determinants).
2)Complete and incomplete antigen
Complete antigen or immunogen are used for substances which
possess antigenic properties de-novo, i.e. they are able to
generate an immune response by themselves. These are high
molecular weight (more thn 10,000) proteins, but some are
polysaccharides. The two important properties of a complete
antigen include immunogenicity (capacity to induce formation of
corresponding antibody)
and to react specifically with those antibodies.
Hapten or incomplete antigen
Is a low molecular weight (less than 10,000), usually non-protein
substance, unable to induce an immune response by itself but can
become immunogenic only when covalently linked to proteins
(called carrier proteins) in vitro or in vivo. However haptens can
react specifically with its corresponding antibody.
Carrier molecule of haptens: it may be serum protein such as

59. Complement:
1) Chemical structure
Complement system consists of approximately 20 serum proteins
which include components of complement ( C1 to C9), properdin
system and regulatory proteins. Some of these proteins are
enzymes, some are control molecules while others are structural
proteins without any enzymatic activity. Complement in general
is heat-labile and inactivated at 56C for 30 minutes. The C
proteins constitute about 5% of the serum proteins and are
structurally unrelated to immunoglobulins. They are
glycoproteins.
2)Routes of activation.
a) Classical pathway: begins when an antigen and antibody
combine to form an immune complex. The complement
components are grouped under three functional units: C1 the
recognition unit: C4, C2 and C3, the activation unit: and C5 to
C9, the membrane attack unit.
b)Alternate or by-pass pathway of C: generally activated by
nonimmunological means such as inulin, zymosan, bacterial
endotoxins, yeast walls, cobra venom. The mechanism can also
be triggered immunologically by IgA, IgE and IgG.
3)Role in the antiinfectious defence
Lysis of foreign cell by classical pathway. Immune adherence and
opsonization. Fixation of C36 on surface of cell for easy
phagocytosis. Chemotaxis, C5a, C5b, 6, 7 fragment 4 B9
increase permeability capillary C5b, 6,7, C8,9
4)Sources of preparations
C1 intestinal epithelium
C2 to C4 macrophages
C5 to C8 spleen
C3 C6 C9 liver
5)Practical using
Complement activity is tested with sheep RBC tht are coated
with sheep RBC serum of rabbits, incubated with dilution of
serum sample, the reciprocal activity of serum tht lysis 50% of
RBC is called haemolytic activity of complement.
a)
01.mplex: relatively large molecules and combine with
specific antibodies forming visible precipitate.
b)
02.ple: low molecular weight simple chemical
substances. When they combine with specific antibody, no
precipitate is produced.
3)Specificity of antigens
Antigenic specificity: depends on the specific active sites on the
antigenic molecules(antigenic determinants).
Species specificity: tissues of all individuals in a particular
species possess, species specific antigen.
Isospecificity: alloantigens or isoantigens are found to be present
in some but not all members of a species which are able to
produce alloantibodies or (isoantibodies) in individuals who are
free frm the antigens.
Organ specificity;organ specific antigens are confined to a
particular organ or tissue.
Autospecificity: the autologous or self antigens are ordinarily not
immunogenic but under certain circumstances lens proteins,
thyroglobulin and others may act as autoantigens.
4)Group antigens
a) foreign antigen: antigens are cell walls, pili, enzymes, toxins
dust, pollens
b) autoantigens: antigens are thyroglobulin, DNA, corneal tissue
c) isoantigens: blood group antigens (ABO, Rh)
d) heteroantigens: heterophile antigens, Cross-reaching microbial
antigens
5)Species antigens
Tissues of all individuals in a particular species possess, species
specific antigen. Thus human blood proteins can be differentiated
frm animal protein by specific antigen-antibody reactions.
However, antibody produced against human serum proteins,
show some degree of cross reaction with proteins frm related
species.

1)

2)

6)Type antigens
Organ specific antigens are confined to a particular organ or
tissue. Certain organs like brain, kidney, thyroglobulin and
lens protein of one species share specificity with tht of
another species. Brain specific antigen are shared by man
and sheep.
7)Autoantigens
The autologous or self antigens are ordinarily not
immunogenic but under certain circumstances lens protein,
thyroglobulin and others may act as autoantigens. These
antigens normally remain sequestered in the body and do not
come in contact with the general blood circulation or tissue
fluids for which these are not recognised as self antigens.
Antigens which are absent during embryonic life but appear
later eg sperm also considered as nonself antigens. These
self antigens act as autoantigens producing autoimmune
disease in the following ways:
whenever these antigens are released into the tissues
following injury of the lens, damage to the thyroid or testis,
the lens protein, thyroglobulin or sperm act as autoantigens
and antibodies are produced against them.
The antigenic specificity of self antigen may be
modified either through a drug or bacterial product and thus
may become immunogenic.

a)
b)
c)

61.Antigenic structure of bacterial cell

1) indication
Bacterial antigen can be identified as 3 types:
somatic antigen (O)
flagella antigen (H)
capsule antigen (K)
2)position of antigen
Somatic antigen present in cell wall
Flagella antigen present in flagella
Capsule antigen present in capsule
hapten in nature and specific for bacteria
3)characteristic
Somatic antigen (O)
Chemically lipopolysaccharide, heat stable, cant be destroyed by
C2H5OH
Flagella antigen (H)
Consist of protein, heat lable, may be destroyed by formalin
Capsule antigen (K)
A, B and L classes, heat stable and heat lable antigen,
mucopolysaccharide, may be destroyed by acetone.
4)preparation
5)practical using
Mainly antigen is used in production of Ig (Ab) to certain
constitution.
6)group and species antigens of microbes.
7)Antigenic structure of viruses
a) surface antigen
b) nucleocapsid antigen
c) hidden antigenic

a)
b)
c)
d)
a)
b)

62.Immune system of a men

1)immunocompetent organs, cells
there are many immunocompetent organs
lymph nodes (peripheral lymph nodes)
thymus
spleen
bone marrow
immune organs classified into:
primary central (thymus, bone marrow)
secondary peripheral (lymph nodes, Peyers patches)
The time of differentiation of immune cells in central lymphoid
organs is genetically predominant and doesnt depend on foreign
substance. Biological subs of orgm can influence on differentiation
of immune cells. In norm foreign subs cannot enter to central
organs, as antigen but they can penetrate to secondary immune
organs. They have immune cells.
Cells: a) T-lymphocytes Ts, Th (Th1, Th2, Th3), Tk
b) B-lymphocytes
c) Large granules lymphocytes (NK, K-cells)
d) Macrophages
lymphocytes has 20-30% in all blood cells. Lymphocytic synthesis
in bone marrow. The 1st cell is stem cell. Then lymphoid stem cell
2) their function
a) are able to regulate immune response by
Th- immune
regulation
Tssuppress humoral immune response
Tkthey kill target cells but cannot kill
free living cells
b)they can produce different biological active substances:
Th Th1 secretes IL2/ IFg / TNF
Th2 secretes IL4 / IL5 / IL6 / IL7
Th3 secretes IL1 / IL2
Ts- produce biological active substance
c)large granulocytes produce Ab-dependent cytotoxicity (cellular)
only if they can interact with Ab or target cells. If they cannot they
produce Ab- independent cellular cytotoxicity.
*they can recognize Ag without little restriction.

3)T-lymphocytes and their function

migrate in thymus. 1st in cortex. It is called corticothymocytes
the main marker is corticothymocyte CD4 and CD8. then cells
begin to differentiate. Then they express TcR for Ag.
Then they go to medulla. Thymocytes interact with epithelial
cells, they have information about all Ag. Can recognize cell and
non cell Ag.
When they go to medulla they form CD4 and CD8
CD4 produce Th
CD8 produce Ts
CD4 and CD8 are MHC1 and MHC2
CD4 recognize MHC class 2 Ag
CD8 recognize MHC class 1 Ag
4)Cell and humoral immunity
a)Humoral immunity
on exposure to an antigen for the 1st time, Ab is produced in a
characteristic pattern depending on the nature, dose and route of
administration of antigen (parental or oral). When 1st introduced,
the Ag selects appropriate B cells involving cellular interactions.
b)Cell mediated immunity
the term cell-mediated immunity refers to a form of acquired
specific immune response mediated by sensitized T-cells. This
form of immunity can be transferred frm immunised donor to
nave recipient with intact lymphocytes, but not with antisera.
The specific cell mediated responces are exhibited by two types
of lymphocytes, CD4 T-cells and CD8 T-cells.

63.Mchanism of immune answer.

1)interaction between T & B-lymphocytes & macrophages.
-Ag is taken up by ADC and are presented to T cells
-T cells starts to proliferate
-AMI Th cells recognise the ag on the macrophage n produces
cytokines in the presence of IL1
-they produce 4, 5, 6 and 10 which activate B cells, they undergo
clonal proliferation and differentiation and produce
immunoglobulins and burgery cells
-Ig are produced by the plasma cell tht are produced frm B cells.
They also form B memory cell
-in the thymus independent activation of B cells and triggers B
cell blast formation to a lesser degree of thymus dependent
-T cytotoxic cells meet with ag complex with MHC1 cell
-they release cytotoxic subs and causes the death of the ag
-at the same time a small quantity of T memory cells are also
formed
2)Their role in cell & humoral immunity
-macrophage have a very important role to play with both CMI
and AMI
1)they are ag presenting cells both for CMI and AMI
2)they secrete
a) interleukins- IL1 and IL6 causes proliferation of T cells
b) TNF Tumour cidal activity
3)cooperation btw macrophages T cytotoxic, macrophages
infected the ag can be recognized by T ag and destroy
4)plays a role in delayed hypersensitivity
5)extracellular effect on mast cells, if its too big they attach along
with monocytes, neutrophils or eosinophils.

64. Antibody
1)immunoglobulins, their main characteristic
Ab is a specialized serum protein. They confined due to response
to an Ag. (specific Ag has its specific Ab). Then it reacts with Ag
(Ab+Ag=Ab+Ag complex)
There are
a) humoral Ab
b) body secreting Ab
c) body fixed Ab
Ig, they are protein in nature. It has Ab-globulin, plasma proteins
of myeloma, cryglobulinaemia, macroglobulinaemia and
naturally occurring subunits of Ig.
2)structure
An Ab molecule is made up of 2 identical heavy and 2 identical
light polypeptide chains held together by disulphate (S-S) bonds.
The longer chains are called heavy (H) and the shorter ones
light (L) chains. Each L chain is attached to H chain by a
disulphide bond. The 2 symmetrical H chains are held together by
1 to 5 S-S (disulphide) bonds, depending on the type of Ig.
3)basic classes of immunoglobulin, their physical, chemical &
biological properties.
a) immunoglobulin G
main serum Ig making up to 80% of total amount (12 mg/ml
blood). Consist of 2H chains linked by disulphide bond. Has 2
identical ag binding sites called divalent. Appears 2 weeks after
infection n persist for longer period of time. Predominant ab in
2ndary immune response. Participates in most of immunological
reactions such as precipitation, complement fixation and
neutralisation of toxin and viruses. Passes thru placenta and
provides natural passive immunity to newborn. Not formed in the
fetus. Half life is 23 days.

b)immunoglobulin A
principal Ig tht appears in the sero-mucous secretions such as
milk, saliva, tears, nasal fluids, sweat, colostrums and in
secretions of respiratory, intestinal and genital tracts. Protects
exposed mucous membrane. Made of 2 H2L2 units and 1
molecule each of peptide (J chain) and secretory component
(protein).
c)immunoglobulin M
is a pentamer consisting of 5 H2L2 units and 1 molecule of J
chain, which joints the Fc fragments of the basic subunits. Called
macroglobulin. Cannot pass thru placenta. If presence of IgM ab
in serum of foetus or newborn indicates intrauterine infection.
d)immunoglobulin D
is a 7S monomer, present on the membranes of a proportion of
unstimulated B lymphocytes of bl and serve as recognition
receptors for antigens. The corresponding ag interacts with the ag
receptors on B cells and lead to specific stimulation, either
activation and cloning to form ab or suppression. Two subclasses
of IgD, IgD1 and IgD2 are known.
e)immunoglobulin E
found in serum of patients with certain types of allergies. These
ab are also referred to as regains. Is a 8S molecule with short life
of 2-3 days. Inactivated by heat, distributed extravascularly.
Does not pass placental barrier or fix complement. Got unusual
affinity for the surface of tissue cells, particularly mast cells and
basophils of the same species. Attaches itself to mast cells by Fc
fragment.
4)specificity of antibodies
The antigenic determinant (epitope) makes contact with an area
on the hypervariable region of the antibody (called paratope). The
molecules are held together in lock and key arrangement by
spatial complementarity and not by covalent bonding.

65.Production of antibodies
1)cells and their interactions
Antibodies are produced rapidly in the system against the
antigenic determinants of infecting orgm. Ag combines with
specific ab in the observable manner and the reaction btw ag and
ab is specific. This specificity is the basis of many serological
reactions in vitro and in vivo.
Antigen-antibody interactions:
The union of antigen and antibody occurs in 2 stages:
1. Primary interaction.
No visible effect and the reaction is rapid, it is reversible. The
forces of interaction for better fitting of the molecules include,
electrostatic force (attraction btw oppositely charged ionic grps),
hydrogen bonding (formation of reversible hydrogen bridges),
presence of hydrophobic grps and Van der Waals bonds
(complementary electron cloud formation on the combining sites
of antigen-antibody). Examples of tests are Coombs test and
fluorescent ab tests
2. Secondary stage
Precipitation, agglutination or alternatively in the activation of
nonantibody component such as serum complement or histamine
frm mast cells. Complement fixation, toxin-antibody reaction,
neutralisation and immobilisation test.
2)practical usings
They are used in some important tests such as:
a) precipitation
b) agglutination
c) immune lysis reaction
d) ELISA
e) Complement fixation
f) Immunofluorescent test

a)
b)
c)

a)
b)
c)

66.Complete & incomplete antibodies

Complete Ab have two active eg, IgM
Incomplete Ab has 1 active . due to blocking or
inactivation of other .. eg, Rh, Ab/Ig G
1)Detection
Direct Coombs test
it detects the presence of incomplete Ab absorbed on
RBC surface
Indication for test autoimmune haemolytic anemia,
erythroblastosis foetalis
Red cells washed in saline 3 times by centrifugation to
remove proteins except globulin. When washed RBCs
mixed with AHG in presence of bovine albumin, clumping
of RBCs occur
Indiret Coombs test
it detects incomplete ab present in serum
detection of antiRh ab in serum of Rh negative
pregnant women of Rh positive husband
patients serum mixed with saline washed Rh positive
group O ( or tht of same grp of patient) red cells and
incubated at 37C for 30 minutes. Red cells washed in saline
three times. Washed cells mixed with Coombs serum.
Agglutination occurs in positive cases.
2)autoantibody, their role in pathology.
an example of auto ab is Rh factors. It can cause pathology,
during blood transfusion or transplacentally.
Eg, if Rh ve pregnant women gives birth to Rh +ve
baby (father is Rh +ve). Rh incompetability late phase. Then
the 2nd baby can be in danger.
3)theory of Ab formation.
a)clonal selection theory
-clonate of B cells are present to produce Ab against every
Ag
b)instruction (templastic theory)
-ag act as template

IgA contains glycine rich polypeptide called the secretory

component which is produced by mucosal or glomerular
cells.the secretory component protect IgA frm denaturation
by bacterial proteases.
Phases of their formation.
IgA is largely synthesized locally by plasma cels n only a
little amount may b derived frm serum.IgA is dimerised
with the epithelial cells to acystein rich polypeptide called
J-chain and Fc fragment.the dimeric IgA combines with
another protein,secretory component synthesized by local
epithelial cells to then the complex is transported acroos
the cytoplasm and secreted into body fluids.
Functions.
-covers the microbs and inhibit their adherence to mucosal
cells.
-aggregated IgA can bind to polymorphs and activate
alternate compliment pathway.
-Promote phagocytosis n promoting intracellular killing of
microbs.

67. Localized immunity:

Local immunity-is the immune response at the site of primary
entry of the microorganisms.
Main mechanism.
In poliomyelitis and influenza, systemic immunization with killed
viruses elicit humoral antibody response which neutralise the
viruses when they enter the blood stream .Humoral antibodies do
not prevent multiplication of viruses at the site of entry [eg.gut
mucosa,resp.tract]as the antibody titre in local secretion is not
high enough to kill the viruses.
Natural infection or administration of live oral polio vaccine and
intranasal influenza vaccine provide local immunity of gut
mucosa n nasal mucosa.IgA plays important role in local
immunity.
IgA provides the principal component in external secretions such
as mucous of respiratory,intestinal,urinary n genital
tracts,tears,saliva,and milk.
IgA in external secretions is synthesized by plasma cells residind
in the mucosal tissue or secretory organ.Its production is
stimulated more effectively by local than by systemic infection or
ag administration.
This mucosal or secretory immune system can neutralise viruses
and can inhibit attachment of bacteria to epithelial cells.
B:Specialty of the structure of secretory Ig;phases of their
formation n functions.
Secretory IgA is made of 2 H2L2units n one molecule each of
peptide[j chain] n secretory component[protein].
IgA is demerised within the epithelial cells of the glands,intestine
and resp.tractwith cysteine rich polypeptidecalled Jchain.
The dimeric IgA combines with another protein ,secretory
component synthesized by local epithelial cells and then the
complex is transported across the plasm n secreted into body
fluids.

68.A.Transplantational immunity:kind of grafts,human HLA

complex.
Transpalntational immunity is the immune response provided by the
transplantation[HLA] antigens.They are present in all mammalian
nucleated cells and is the reason for rejection of exogenous grafts.
Kind of grafts.
1.Autograft[autogenic graft]-tissue of one site engrafted to another site
with the same individual.
2.Isograft=syngraft[syngenic graft]-graft placed on another individual of
the same genetic constitution.
Eg.monozygous twins.
3.Allograft[homograft or allogenic graft]-grft transfr btw.two genetically
different members of same species.
4.Xenograft[xenogenic graft]-graft btw.members of different species.it is
formerly called heterograft.
Human HLAcomplex.
They are alloantigens present on the surface of leukogenic and are called
Human Leukocyte Antigen.The cluster of genes encoding for them is
called HLAcomplex.
HLAcomplex of genes is located in the short arm of chromosome
6,consist of 3 loci grouped into 3 classes.
Class 1-HLA-A,HLA-B,HLA-C:major class responsible for tissue
rejection.
Class11-HLA-DP,HLA-OQ,HLA-DR:regulates immune response.
Class111-C2,C4,FactorB:Complement system.
-HLA antigens are widely distributed with tissues except RBC which the
twin onlyABH antigens.
-HLAsystems is very useful in tissue typing and maturing prior to
transplantation.
-It has application in patternity determination.

-Diseases showing strong association with HLA include Reiters

syndrome,Juvenile Rheumatoid Arthritis,Coeliac disease and
Graves disease.
HLA is also called MHC.[major histocompatibility complex].
-MHC provides strength and suppression of immune system.
-MHC controls production of time complement component.
-MHC stimulates antibodies and blasts transformation of cells.
-MHC eradicates grafts.
-MHC provides reaction of graft with host.
B .Priciples of donor and recipient selection.
1.By cross matching can check the match donor and recipient
components.
2.Take organ frm donors only after death.
3.In this case must have a limited time for organ transplantation.It
depends frm graft.
C. Overcoming of transplantational
barrier[immunosupresors].
-Tissue compatibility agents.
-Using of immunosuppresors.
a.Corticosteroids-with high dosage cause depletion of
lymphocyte.
b.Antimetabolites-Folic acid antagonists,anloues of purine
cystosin and uracil interfere the synthesis of DNA n RNA or both
and inhibit cell division and differentiation.
c.Cyclospmine-inhibits selectively T-cell activity.
d.Antilymphocytic serum-primarily effective against T-cells and
specifically on cell mediated immunity.they may also inhibit
thymus dependent A antigens.It reacts on circulating lymphocytes
and mainly used for the prevention of graft.

C .Interferons.
Conditions of production:They are produced upon a potent insull
like injection with virus,bacterial endotoxin
It starts being produced in an hour and is max in 6-12hrs.
The potent inducers are-Togavirus,vesicular stomatitis virus.
Type-alpha;produced by B cells,macrophages [virus activated]
Beta;fibroblast [virus activated]
Gamma;T cells [antigen activated]
D.Mechanism of antiviral action.
-It bind to receptors on injected cells and upregulates some
cellular genes and downregulates others to inhibit replication of
viruses.
-It can block sites of virus attachment.
-stabilise viral capsid with endosomal membrane.
-inhibits protein synthesis by [2,5-A Synthetase]
-protein kinase inhibits ribosomes assembly.
E.Inductors of interferon.
Viruses ,double stranded RNA ,bacterial endotoxins ,tilorone
,some synthetics-Bpoly I:C.
F.Practical using.
1.Viral infections-keratitin,genital,respiratory viruses.
2.adjunctive therapy-in breast carcinoma,osteosarcoma.

69.A .Antiviral immunity:non-specific factors of defense.

Non-specific factors includes macrophage action that leads to
phagocytosis of the infected cells and release of interferon by the
injected cell itself.
B .Role of phagocytosis and antibodies.
It includes the humoral[AMI]and the cellular mechanism[CMI].
AMI.
-Caused by B cells.
-B cells can recognise the epitope of a free virus and this leads to
the neutralization of the virus.
-IgM and IgG neutralise virions on the mucosal surface.
-Abs can cause aggregation of virus particles and the complex is
readily phagocytosed.
-Activation of complement can cause opsonization of viruses.
-Complement also can inactivate some viruses.
-Classical and Alternative pathways can be activated by some
viruses.
CMI.
-Participating cells:CD4,CD8,NK cells,macrophages{nonspecific}
1.CD8 or target cells can recognise specific Ag on the target cell
and release cytotoxic substances killing the infected cells.
2.ADCC-Antibody Dependent Cell Mediated cytotoxicity.
These cells recognise Ic portion of virus bound Igs that all in
turn bound to infected
cells.Toxic substances are released into the target cell,killing it.
These cell are macrophages,monocytes,lymphocytes,NKcells.
3.Sensitised Th and Tc cells produce lymphokines that eliminate
viruses [eg.IF gamma]

70.Mechanism of connection btw antibody and antigen and

reactions of immunity.
1.Primary interaction.
-There is no visible effect and the reaction is rapid.
-The antigen antibody reaction is reversible.
-the forces of interaction includes,electrostatic force,hydrogen
bonding and Van der Waals bonds.
Eg.Coombs test and Fluorescent antibody test.
2.secondary interaction.
-visible demonstrable effect such as precipitation,agglutination
test.
Eg.Precipitation,Agglutination,CFT,Toxin antitoxin
reaction,Neutralisation and Immobilisation tests.
B.Type of antibody.
1.polyclonal-heterogenous antibody that are produced in
response to a single antigen by several different antibodies.
2.monoclonal-antibody produce by single antibody forming cells
or clone directed against a single antigen or antigenic
determinant.
C.Monoclonal antibody:scheme of preparation,practical
using.

Practical using.
1.Diagnostic use identification of bacteria ,viral and other
antigen.
2.Test for vaccines-identification and participation of microbial
products.Both for vaccine and industrial use.
3.pure antibody.
4.store for future use.
5.Therapeutic uses.

71.Reaction of neutralizing of toxin:mech and ingredients.

-when an antitoxin combines with a toxin the biological effects of the
toxin are neutralized.
-toxin antotoxin neutralisation can be done in vitro and in vivo.
1.Neutralisation in vivo.
a.Toxigenecity test in vivo.
Homologous antibodies prevent the biological effect toxin.toxigenecity
test of c.diphteriae consists of intradermal inoculation of bacterial toxin in
guinea pig previously protected by ADS.No biological effect of toxin is
seen in the controlled animal but unprotected animal dies.
b.Shick test.
When 0.2ml of diphtheria toxin[1/150MLD] is injected intradermally.
There is no reaction at the site of injection of the person contains
circulating antitoxin[0.01 unit or more per ml of blood]
Eg.immune to diphtheria.
11.Neutralisation test in vitro.
a.Agar gel precipitation test.
It is employed to defect production of c.diphteriae.
b.Nagler reaction.
cl.welchi toxin is neutralise by antitoxin when the organism is grown in
serum or egg yolk medium containing antitoxin.
c.streptolysin o neutralisation.
d.virus neutralisation.
Mostly done with typing viral isolates.
Can be done in cell cultures,egg embryos and animals.
B.Usage for the measuring of the level of antitoxic immunity[name of
reaction,description of the method].
Shick test.
-when 0.2ml of diphtheria toxin [1/150MLD]is injected intradermally.
-there is no reaction at the site of injection of the person contains
circulating antitoxin[0.01 unit or more per ml of blood].
-immune to diphtheria.
C.Usage for diagnostic aim.
-used to detect toxicity of c.diphteria[alex test n shick test]
-Neglers test for cl.perfringens.

C. Preparation of antibody and antigens.

Has 7 test type;
1. Ring test-antigen solution layered over a column of whole
serum [Ab] in a narrow Test tube.
2 .Slide flocculation-a drop each of Ag solution and serum
placed on a slide ,mixed By shaking.
3. Tube flocculation-serial dilutions of antigen made in test
tube to which fixed amount serum added.
4. Immunodiffusion-antibody and antigen solution placed in
wells opposite each other And allowed to diffuse Ag and Ab
for few days.
5. Single diffusion-antibody incorporated in agar gel on a slide
or petridish and Antigen are placed in well cut on the agar
gel.Ag diffuse radially And forms a ring shaped band.
6. Double diffusion-Ag and Antisera are placed in different
wells cut out in agar gel And allowed to diffuse to each other
[Ouchterlony technique], Lines of precipitate formed may be
identical when two adjacent Antigens are identical.
7. Immune electrophoresis-for determination of the presence or
absence of serum proteins and detection of unusual patterns
such as human myeloma protein.
C. Practical usage.
1. Identification of bacteria
2. Identification of bacterial component with infected tissue.
3. Detection of unknown antibody.
4. Identification of human blood or seminal fluid.
5. Standardisation of toxins and antitoxins.
6. Testing for food alteration.

72.Precipitins and reaction of precipitation.

Precipitin : are antibodies which bring about the formation
of a minute deposit [precipitate]upon interaction with a
specific antigen.
Reaction of precipitation : is a specific interaction of the
antigen [precipinogen] and antibody [precipitin] with the
presence of an electrolyte with the formation of a deposit or
precipitate. Mainly IgG is used.
B.Mechanism and ingredients.
Multivalent antigens combine with bivalent antibodies in
optimal proportion in zone of equivalence.The free antigen
binding sites in antibody and antigen determinant groups
remain available after initial combination of antigen and
antibody molecules,which enables the complexes to link up
to form a large lattice in the zone of equivalence.The lattice
consists of alternating antigen and antibody
molecules.When the size of lattices exceed critical
volumes ,they settle out of solution spontaneously.In the
zone of antibody excess ,all the functional determinants of
the antigen molecules are taken up with antibody,so that
very little linking occurs between Ag-Ab complexes.In the
zone of antigen excess ,less precipitate forms because of the
inability of the Ag-Ab complexes to link up to other
complexes which results in failure to form a large aggregate
or lattice.

73.Agglutinins and reaction of agglutination : mechanism of reaction

and ingredients.
Agglutinins are an antibody capable of clumping the corresponding
microbs by producing visible conglomerates.
Agglutination is a reaction between antigen antibody in which an
antibody combines with a particular antigen in the presence of electrolytes
at optimal pH and temperature resulting in visible clumping of the particle.
Since the patients body produces Abs continuosly throughout the disease
process a serum sample would contain the Abs. Addition of the specific Ag
to this Ab leads to a reaction between them and seen clumping by the naked
eyes.
Mechanism.
1. Slide agglutination test.
This is an widely used method to detect known antigen.A suspension of
antigen is made in saline on a slide and a small drop of antiserum is added
and the slide is rocked for a minute. Clumps are formed in a few minutes.
2.Tube agglutination test.
For determination of antibody titre.Serum is diluted in saline solution in a
series of test tubes. Equal volume of standard antigen suspension [0.5ml
cell suspension] is added to all tubes and then incubated .highest dilution of
serum at which agglutination occurs is recorded as antibody titre.
B. Preparation of polyvalent and monovalent serums .
C. Reaction of passive agglutination.
a precipitation reaction can be converted into agglutination reaction by
certain carrier particles ,such as latex particles ,bentonite ,fixed ,
staphylococcal cells and RBC by coating with soluble antigen .The polysac
antigen get absorbed to the cells by simple mixing while protein antigen
require tanned red cells .This is added to the serum with the Ab and
agglutination is seen .Instead of antibody ,when the antigen is adsorbed
onto the carrier particles for estimation of antigens ,it is called reversed
passive agglutination.
D. Practical use.
1. Identification of unknown culture.
2. Blood grouping and cross matching.
3. Serological diagnosis of enteric fever [Widal test],typhus[Weil-Felix
reaction],Brucellosis and infectious mononucleosis [Paul- Bunnel test]
4. Streptococcus MG agglutination test or diagnosis of primary atypical
pneumonia.

74.Reaction of immunofluorescent [direct and indirect

methods ]
Direct.
blood film or smear from clinical material is fixed in
ethanol for 5 minute and dried.
Then fluorescent tagged antibodies are brought into
contact with antigens fixed on the slide allowing them
to react.
The excess of antibody is washed of and the
preparation is examined under ultraviolet light
microscope.
Site of union of conjugated antibody with its antigen
appears as pale green fluorescent areas on the slide.
This is a sensitive method to diagnose rabies by
detecting rabies virus antigen in brain smears.
Indirect.
useful in detection of antibodies in serum or other body
fluids.
The unlabeled unknown antibody is applied to the
known antigen on a slide.
After incubation , the slide is washed with buffer to
remove unfixed excess serum , leaving behind only
antibody globulin which coats surface of the antigen.
The smear is treated with a fluorescent tagged antibody
to human globulin.
The slide is washed to remove excess fluorescent
conjugate and examined under u .v light.
In positive case, the Ag-Ab complex will fluorescent.

75:Reaction of immune lysis:

1)Reaction of hemolysis:Based on this reaction test, sheep RBC and rabbits
antibodies complement.Rabbits antibody are determined
by injecting sheeps RBC into rabbits and take rabbits
serum after few days.The result is hemolysis and releasing
Hb.Lysisof erytrocytestake part due to the action of
complement.Complement acting on Ag-Ab complexes of
hemolytic system produce membrane injury of RBC.
2)Reaction of complement system:The presence of complement system is necessary for
complete reaction of certain lysis Ab.Ag-Ab complex fixes
with complement.The coupling of Ag-Ab complex does
not have any visible effect.So an indicator system is
used,sheeps erytrocytes are used,coated with anti-sheep
RBC-Abis used.Complement lysis Ab coated cells,Ag
must be quantitated.Test serum must be inactivcated by
breaking at 56c for 1\2 hrs.noncomplementary effect by
some non specific inhibitors.Complement is observed from
a guinea pigs serum.Freshly drawn guinea pigs serum is
used.
Ag-Ab are mixed and are measured the amount of
complement and are intubated at37c for 1hrs.If Ag-Ab
match,this complement willremain free.An indication
system containing sensitized sheeps RBC is added to next
mixture and intubated at 37c for 30mins.

B . Elisa test .
- This test s for measuring Ag-Ab reaction.
- The test is done on solid phase .
- The test can be performed as direct or indirect assay depending
on whether the
enzyme is conjugated to the primary or secondary antibody.
- The enzyme [horse radish peroxidase ,alkaline phophatase ]
give rise to a colour
change in addition of specific substrate only when the Ag
and Ab react
specifically.
-

double antibody techniques for detection of antigen

[sandwich ELISA ].
Indirect method for detection of antibody.
C. Radio immunoassay .
- add labelled antibody with isotope to antigen.
- complex formed if specific after some time.
- radioactivity measured by gamma spectrometer to find out
amount of antigen.
D. Practical use.

The result of this is:-Absence of hemolysis (+ve result)it indicates

that complement was used or fixed up during intial reaction,present
hemolysis(-ve result).it indicates that complement remain and was
not fixed during lysis of RBC.
T
Test
A m N
System
u
serum
g l a
b
C
e
l
n
o
.
Sheep
RBC :
Rabbit
hemoly
sis
0.5 :
0.5
0.5
1

0
.
5

1:0
0
.
5

0
.
5

0
.
5

0
.
5

0
.
5

0
.
5

0.5
2

1:0

h
e
m
o
l
y
s
i
s

1:0

76Application of reactions of immune answer for

diagnostic of viral infections
1)ELISA(using HIV virus as an eg)
ELISA is the most widely used direct solid phase test in
which commercial test kits are available.Ag is prepared
from HIV cultivated is continuous T-lysis cell line or by
re combination technique.Ag is prepared from as abovge.
Ag is coated on surface of microtitre walls to which test
serum is added.HIV antibody serum is washed away.Then
anti human goat Ag linked to a suitable enzymes (alkaline
phosphatase)is added and intubated for 1hrs at 37c.A
suitable solution is added to split the substrate and left at
room temperature.Till the positive control develops a
colour,it is read visually or ELISA readers.
2)WESTERN BLOT
In this test,HIV is broken down (separated) into protein
fragments in a polyacrylamide gel.The separated protein
.The separated protein are blotted electrophoretically frm
the gel into a strip of nitrocellulose sheet .This sheet are
incubated with test sera .Ab to HIV protein,present in test
serum combine with all or any fragment of HIV. The strips
are washed and treated with enzymes :conjugated
antihuman gamma globulin and allowed to react.Then a
suitable substrate is added that produces colour bands.The
positive of colour bands on strip indicates the fragments of
Ag to which serum Ab has combined.

78.The valuation of im .status:Immune statud s is a complex of parameter reflecting state of

im.sys of whioch person at which movement of time.
Main valutions are-for diagnostic purpose
-for admin of treatments
2)Tests for the valuation of factors non specific defence:A)Lab investigation:
-determination of Tlym,Blym,Ig,hemolytic activity of coplements
-func. Of activity of jy-specific Ab against infection
-funtional activity of phagocytic cells
-conc of cytokines-ELISA
-determine components of compliments
B)Objective method:
-state of lymph nodes,tonsils,rashes,heart rate.
3)Humoral and cell mediated immunity:Humoral-on exposure to an Ag for first time Ab is produced in a
characteristics pattern depending on the nature,dose and route of
admin.whn 1st introduced,the antigen selects appropriate B cells
involving cellular interactions already described.
Cell-refers to a form of acquired specific immune response
mediated by sensitized Tcells.this form of immunity can be
tranfered frm immunized donor to nave recipient with intact
lymphocytes but not with antisera.the recognition of antigen on
target cell by a t-cell is triggering effect.the specific cell
mediated responses are exhibited by two types of lymphocytes
,cd4 Tcells and cd8 Tcells.
4)Hypersensitivity:Or allergy refers to a condition in which immune response results
in excess rx leadind to tissue damage,disease or death.There are
two main types immediate and delayed.Classification:-Type 1-4.
type1-3 are antibody mediated (immediate type) and ab
mediated,type4 are cell mediated(delayed type) and T cell
mdiated.

77.Pathology of immune answer.

1)Classification:
a))Hypersensitivity or allergy-hyper response of immune
system.condition of human body changes leading to tissue
damage,disease or death.
b))IM-deficiecy- pathology of absence or slow funt. Of immune cells.
2)IM deficiency conditions :
Classification of deficient cells:-T-dependent,B-dependent,complement
dependent,combined B and T cells ,myeloid cell.
Acc to origin:A)primary IM-def-organic origin,absence of thymus,bone marrow or
under development
-absence of gamma globulin.
B)secondary IM-def-develop during life by external infl.
Defect of nutrition-def in Ab production and T-cells.
Primary drugs-IM suppress antibiotics,
Infection-m.o suppress im.resp or destroy im sys cells.
Tumour-causes degeration of im cells.
2)Primary immuno deficiency(PID)
Its conjenital and genetically determined.it occurs as a result of defect in
almost any stage of determination. in the whole IM sys.In pathology PID
the may be in stems cels ,Tcells,compliment sys or phagocytosis.
3)Secondary immunodeficiency
Secondary immune results as a secondary consequences of
malnutrition,certain acquired diseases or as consequences of certain
theraputics measures which depress immune sys.
1)depression of humoral immune response :Results whn B cells are depleted ,excess of production of
abnormal immunoglobulins,normal immunoglobulins level is
decreasesd.immunosuppresants are responsible.
2)depression of cell mediated immunity:CMI depresses in AIDS hogkins dis
Principles of their treatment :-transplantation of organs
-admin of ig preparation
-antibacterial treatment

79.ROLE OF MEDICAL MICROBIOLOGY IN

PROPHYLAXIS INFECTIOUS DISEASES.
1) Role- immunisatioin against infectious disease. Immunization
aim is to produce artificially a degree of resistance sufficient to
prevent a clinical attachment of the natural infection to the
recipient. Immunization involves active or passive immunization.
2) drugs for prophylaxis
- antigenic preparations
vaccines ~ induce production of specific antibody.
a) live attuenuated org. eg; measles, mumps, rubella, BCG
b) killed (inactivated) vaccine. Eg; pertesis, typhoid,
cholera, rabies
c) toxoids. Eg; diphtheria & tetanus
injection of exogenous antibodiespresent in human or animal
serum to give immediate protection to an anticipated infection.
This type of immunity is very short lasting.
Human sera ~ pooled Ig
~specific Ig
Sera are biological medicines which contain ready prepaired
antibodies. It stimulates acquired passive immunity.this is the
specific treatment.
Diagnostics: antiserum~ prepared with animal antigens
Main directions of applied immunity
-immunogenetics
-immunomorphology
-immunopathology
-immunohematology
-immune of ontology
-anti cancer immunology
-vaccinology
-immunology of grafts

80. PROPHYLAXIS BY VACCINES

The vaccines are prepared with certain concentration of antigen
& are applied to creation in an organism of artificial active
immunity.
2)kinds of vaccines
-alive
-killed(inactivated)
-chemical
-toxoid
-associated vaccines
3) their properties
-alive vaccines
Attenuation (reduce of its strongness) of vaccines is done by
aging of culture, cultivate at high tempreture (anthrax) passage
through diff animals, drying, by continuous cultivation in
pressure of antagonistic substance (BCG) and by repeated
subcultivation in artificial media. Eg; tuberculosis (BCG),
plague, anthrax, brucellosis, yellow fever, parotidis
-killed vaccines
m.o killed by heat , formalin, phenol, alcohol, uv &
photodynamic inactivation . eg; cholera, rabies, wood cutter
encephalitis, lepro spirosis
- toxoid
Bacterial exotoxin inactivated by heat & formalin. They loses
toxigenicity & have immunogenicity. Toxin mix with 0.3%
formalin & filtrate it. Incubate at 37C until toxicity has
disappeared.antigenicity of toxoid is increased by absorption on
to a mineral carrier [Al(OH)3 /AlPo4]. Alternatively toxoid is
mixed with a suspension of other bacteria containing
polysaccharide, endotoxin such as pertusis component of
DPT(diphtheria, pertusis , tetanus).toxoids absorbed in to a
mineral carrier remains longer in a depot after injection & serves
better antigen . eg; tetanus toxoids, diptheris toxoids , staph
native, staph absort

- chemical vaccine
different antigen components that is reduced vaccines interact to
live and killer cells that contain microbial bodies.
-associated vaccines
Contain antigen indifferent nature absorbed (DPT) vaccines ,
inactivated pertusis vaccines , diphtheria, & tetanus toxoid
absorbed on Al(Oh)3 also drive combined eropean thypus
vaccine E (ACDVE) contain dry alive culture of ricketsia
prozekii of virulence strain E mixed with soluble ag of virulence
ricketsia, cholera vaccine contain cholerogenic toxoid & o
antigen of cholera vibrio
-Recombined vaccines
Against hepatitis B intergration of subunit of hepatitis B
virus into saprophyte cells which is multiple on artificial media
giving antigen against hepatitis B.
4)Main direction of important improvent of vaccines
a)artificial
bio organic vaccines complexes providing strong immune
response on antigen against genetic disposition of P.R genes
b)gene engineered vaccines
genes providing antigen arte intergrated into replicative cells
(virus,m.o,plasmid) &inject the human by virus , m.o or plasmid
due to presence of gene producing antigen the immune response
develops
c)some vaccines are produced by cloning the surface antigen
gene in yeast cells
d)cetain vaccines has to be conjugated to protein to make it
immunogenic
e)subunit vaccines

vaccines of 3rd generation

adjulant are combined vaccines. They sometimes enchance
antibodies production . eg; pertusis component of DPT vaccines
acts as an adjulant for toxoids of diphtheria and tetanus. They
contain antigen against some infectious disease
eg; measles , mumps , rubella vaccines(DPT)
vaccine therapy
-vaccines can be udsed for therapy . it is called immunotherapy.
For patient with chronic infection . autovaccines are used. They
are prepared form strains from patient.
Gene vaccines
Its used for treatment of chronic gonorrea. Tit stimulates immune
response against gonococcus.

81.Alive vaccine
1)Method of preparation:Attenuation of live vaccine is done by ageing of
culture,cultivate high temperature,drying,continued
cultivation by presence of antagonist substance(BCG) and
by repeated in subculture in artificial media.
2)Connservation of alive vaccine:Tuberculosis,BCG,plague,small
pox,antrax,rubella,influenza,yellow fever, and parotids.
Specificity in using:In alive vaccine suspension of living organism with reduce
virulence,mimic natural infection with Ab,production
without symptoms.A single dose of live vaccine induce
long lasting immunity and it reinforced by subsequent
booster dosage.some vaccines are oral.sometimes related
organism with shared Ag are used in preparation of live
vaccine.Bovin tubercle bacillus employed and produce
BCG vaccine.Alive vaccine should not be administered in
pregnant womens,AIDS patients,leukemia, n etc.
3)Dead vaccine:M.O are killed by heat,formalin,alcohol,
Conservation:The inactivated vaccines process are common to the
original pathogen which do not replicate.The inactivated
vaccines need large dosage,usually 3 doses.generally the
second dose six weeks after first dose,and the third dose
after six months of second dose.used for patient who are
tyhoid,cholera,rabies,hepatitisB,.

82.Anatoxin,their preparation:In this case antiserum,is applied to antitoxic sera is raised

in horses by active immunization.In pratically.animal sera
has foreign proteins which recipient forms Ab resulting
in:-rapid elimination
-hypersensiytivity
e.g-tetanus,dipheteria,snake bite,
2)Native and adsorbed anatoxins:Native,e.g:-dipteria,gas gangrene,botulism.Antiserum is
applied to antitoxic Ab prepared in animal.The antitoxic
sera are mixed in horses by active immunization.in that
antitoxic Ab filtrate and prepared as vaccines
Adsorbed,in this case antitoxin Ab in mineral solution,e.g:ALPO4

No 90;meningococci
Morphology;
N.menigitidis
Gram ve cocci,arranged in pairs,coffee bean
shape,non motile,nonsporing noncapsulated
Physiology;
Gro,wth occurs in medium enriched in blood
agar,serum.on solid media colonies are
small,translucent,round,convex,bulish gray
Strict aerobes does not grow in anerobic medium,
Catalase and oxidase and indole
Glucose and maltose are formed,produces acid but
without gas
Antigenic structure
13 serogroups are identified on the basis of capsule
polysaccarides.Group A<B<C<D are usually
responsible for meningitis.
Factors of virulence;
Meningococci surrounded by large capsule,that
encloses them with great ability to defense
mechanismin circulation and may grow in high
numbersin blood.meningococci contains endotoxins
in membrane is produced and released into blood .
Forms pili
Is an extracellular parasite.
Diseases.
Seen mostly in nasopharynx or asymptomatic
carriers.

89;Streptococcus pneumoniae
Morphology;ovovid,Gram +ve,cocci occurs in
pairs,capsulated,nonmotile,nonsporing
Physiology;
grow only in enriched media(blood agar)
-small circular,raised and smooth colonies
facultative anerobes,lack catalase and oxidase
ferment in few sugars,by formic acid only bile soluble
destroyed by 52 in 15 minutes and antiseptic sensitive
antigenic structure;-specific capsular polysaccharide.used for
serological classes.there are 90 types of classes
Role in human pathology;
Gives pneumonia in man due to pneumococcal infection
The capsule of mo prevents phagocytosis by neutrophils and
macrophages
Pneumococci synthesizes neuramidase,leucoccydins,hemolysins
It is a human pathogen.the source of infection is resp.tract by
patients or carriers.transimitted by each other by inhalation or
contaminated droplets
The mo colonize in nasopharnyx to ororpharnys to the lung
sinuses to the meninges
Can also give pericarditis,peritonitis
Mainly by air borne mechanism,but for food poisoning is due to
food
Laboratory diagnosis;
a)bacterioscopical and bacteriological methods-take
sputum,pus,cerebrospinal fluid from patient and slecet media for
inoculatin(bood agar and gentamycin),isolate and identify pure
culture
use disc diffusion method to detect antibiotic sensitivity
b)agglutination test,radioimmuneassay
c)test of capsule swelling-specimen are treated by specific serum
which gives swelling of the capsule and rise to presence of
pathogen in the group.
Treatment;penicillin,cephalosporins,erythromycin,
Prophlaxis;vaccines

Labdiagnosis;specimens include CSF,blood and biopsy

samples,sputum
Using microscopical method it is stained by Grams
technique.
Bacterioscopical method include inoculation of clinical
specimen on blood agar or chocolate agar
platesidentification of pure culture is made by morphology
staining techniques,biochemical reactions,serologicalk
staining using polyvalent or monovalent serum
Antibiotic sensitivity test may also be done
If mo is in small number immunoflouresecnce test may be
done
Radioactive Ag biding test
Treatment and prophylaxis;
Treatment;penicilinG
-Chloramphenicol or broad spectrum
caphalosporins used
prophylaxis;polyvalent vaccines that has capsule
polysaccarides of group A and C
immnune stimulatants to excite immunity of
older childrens.

84.methods of prophylaxis and therapy for of toxemic

infections.drugs,directions
prophylaxis;giving antitoxins in combination with
chemotherapeutics.
active immunisation of toxoid induces antiboby
production
infections can transmit from food,animals,faeces,poor sanitary
conditions that is used for treatment of patients,and carriers must
control thier infections
boiling of water,sanitary conditions and personal
hygiene,,avoidance of contact with infected animals or humans abd
excrete or control on anal,oral or sexual contact..
therapy;using antibiotics,penicilins like Benzylpenicilin,oral
penicilin,ampicillin,amoxycillin,carbenicilin, which effects on both
Gram +ve and Gram -ve organisms.
cephalosporins;cephalosporium acremononium and
chemically similar to penicilin,are bactericidial drugs act same
way as penicilins
Gram +ve and Gram -ve bacteria.
aminoglycosides;contains amino sugars.examples are of
Kanamycin,Neomycin,Paramycin are closely related drugs.
tetrcyclins are isolated from Streptomyces
aurafacieus was introduced in 1948.
other antibioitics;Macrolides-macrocyclic lactonewing to
which sugars are attached to erytromycin and spiromycin
lindamycin,Linomycin,Vacomycin,Quinolones,Bacitracin,Chloram
phenicol
Antitubeculous drugs such as
Isonoazid,Parazinamid
Metradiazole effective against anerobic
bacterias and protozoas.

Anaphylactic reaction;type if Ig E mediated hypersensitivity

reaction,which develops quickly after introducyion of large shocking dose
of antigen following one or more small sensitising doses.rare event in
man and is observed in hypersensitive individuals by insect
stings,injection of foreign serum or penicilin
factors influencing anaphylaxis;1)sensitisation-may occur in routes as
injection,ingestion,inhalation or contact.minute doses of antigen can
sensitise susceptible animals.
2)waiting period-interval of 2-3 weeks
between sensitising and shocking dose is required.
3)shocking or eliciting dose-shocking dose
is effective when adminisreed ivly .the ag-ab complex stimulates mast
cells for prompt release of
mediators that causes clinical
manifestations of anaphylaxis.
mechanism;IgE formed in response to antigenic stimulation,mast cells are
present in large numbers in submucosal layers of respiratory and
gastrointestinal tract .IgE are bound to mast cells of tissues and basophils
of blood.on subsequent exposure to large dose of same antigen,it
combines with fab fractions of Ig E
Chemical mediators;signals for release are got once bound to portion of
antigen and mediators are relased;
primary mediators histamine and serotonin
secondary mediators;SRS,prostaglandins and platlet
activating factor.
type ii:cytotoxic reaction.
mediated byantibody directed toward antigen persent on the surface cell
or other componenets present on the tissue cell resuklting in
damage.antibody is directed against epitope that can be a microbial
products passively adsorbed onto a cell.Both Ig G and M are prodused in
type ii reaction,it may be a complement mediated lysis.demonstartion of
this types are demonstared in direct antiglobulin and agglutination test
iimmune disease;ag-ab medaitora takes place in the blood.the complex
get adsorbe into the tissues especially the vascular endothelium and
causes the relase of Hagemans factor which induces kinin-kalikren
system coagulation.,chemotaxis is mediated is mediated by mast
cells,basophils.can cause serum sickness

85;Allergy and its types.Main pecularities

Hypersinsitivity or allergy refers to a condition in immune response
results in excessive or exxagerated reactions leading to tissue
damage,disease or even death.
Allergy is the altered reactivity of an animal to repeated contacts with a
foreign antigen.
hypersensitivity reactions are of two main types immediate and delayed
based on time required by host to respond to the shocking dose of the
antigen.Immediate form is mediated by humoral antibody and manifesta
itself in few minutes to hours which reaches a peak after 48-72 hours and
mediated by t cells.
also widely accepted are 4 types of 1 to iv.first 3 are antibody
mediated(immediate)and type iv is cell mediated(delayed type).2
additonal types are recenly introducedtyoe v is antibody mediated and
type viis both antibody and cell mediated
HYPERSENSITIVITY
]
---------------------------------]------------------------------IMMEDIATE TYPE
DELAYED TYPE
(antibody mediated)
(t cell mediated)
]
]
]--------------]-------------------------]
type 1
type2
type 3
(anaphylaxis) (cytotoxic)
(immune complex)
main pecularities;
it appears in minutes and resides in hours
induced by antigens or haptens in any route
is and antibody mediated reaction
passive transfusion is possible with serum
desensitization is easy but short half life

86.hypersensitivity of delayed type;main specialities and

mechanism..
main specialities;supress slowly
last long
induced by Ag or hapten intradermally
cell mediated reaction
cannot be transformed to serum
sensitisation difficult
mecahnism;provoked my intracellular microbial injections or
haptens and involves macrophages and lymphocytes mainly has 2
stages1)sensitization-the rpimary exposure to the antigen
2)stage of allergic manifestation-the sensitization
of t cells affects on leucocytes,macrophages and tissue cells.there
are 2 types of delayed hypersensitivity-infectious type and contact
dermatitis type
role of antimicrobial therapy;

infectional allergy;'when tuberculin is injected intradermally to a

sensitised person,an ertyhema and swelling appears afetr 12 hours
and becomes maximum in 24-28 hours.this site ins infiltrated by
mononuclear cells (lymphocytes and macrophages) subsidises
CDA cells release P lymphokines and accumulation of
macrophages occur.failing to kill intracellular pathogens.Some
macrophages are transformed into epitheloids express MHC class 1
and 11 molecules.Cells recognizes the MHc Ag in macrophages
and release TNF B(kills macrophages) and IL2.this substances kill
other tissue cells too.The dead macrophage release Ag and can no
longer be recognized due to lack of MHC1 and cause chronic
granuloma.

87.staphylococci:morphology,structure,classification(species

morphology;Gram +ve grapes arranged classes,non sporing,nonmotile,non-flagellated,single cells or part cells,noncapsulated(but some has slight layers),grown in simple
media,also growth in nutritional media and nutritional broth and
agar.form turbidity and surface of agar they give colonies.Staph
aurues give 95% of infection,form yellow pigment,they can
synthesize enzyme coagulase,.they give clotting plasma,they can
grow in antibiotic,neurobiotic,the most selective media for
isolation has 5-10% Nacl ,majority of microorganism grow in
concentrated of Nacl 0.5%,facultative anerobe can ferment sugars
as mannite produce acid but not gas.
Classification;staph.aureus
satphy.epidermidis
staph.saprophyticus
Factors of virulence;
a)plasma coagulase factor-gives clotting of plasma,free coagulase
can be seen by incubation,bound coagulase connected with cell
membrane with microorganism
Staph gives production of enzyme lectihinase or phopholipase
11.it gives fermentation of egg yolk .due to this around
microorganism can see zone of fermentation of thi substance
b)the nuclease distract the nuclear Dna and RNA
c)the mo synthesizes strong toxins hemolysins and leucocydins
also synthesizes enterotoxin of many types.heat stable
also synthesize epidomylitic toxinsand gives diseases of 'scalded
skin syndrome'in children
also gives toxic shock syndrome

88.streptococci;morphology,physiology,.
morphology;Gram +ve,arranged in chains,round or
spherical shape
-non motile,nonsporing,non flagelatted,noncapsulated(only Strep.pneumonia capsulated)
physiology;facultattive anerobes and obligate anerobes.
most important are strep.pyogensgives pus
inflammation reaction
grows in solid media can be cultivated in liquid
media by sugar broth.gives small transluscent colonies in
sugar broth.
classification on hemolysis;
a)alpha hemolysins-incpmplete
hemolysis,gives rise to green pigmented ring
b)beta hemolysins-gives complete
hemolysis.surrounded colony has clean transparent
ring(Strepto.pyogens)
c)gamma hemolyisn-no
hemolyisis.in this there are no pathogenic microorganism
antigenic structure;
classified according to C antigen in the cell
wall.it has serological types as
a)A-H b)K-V
group of A strep.has majortiy of
pathogens.they are divided into m antigen which has 16
serological types

staphy.diseases
a)food poisoining
b)scalded skin snydrome
c)toxic shock syndrome
d)also affects the skin,respiratory tract,cns,intestine and
cardiovascular diseases.
skin infection-abcess,sepsis in wounds
bone-osteomyelitis
respiratory tract-tonsilitis,pneumonia in lungs
cvs-endocarditis
CNS-meningitis
intestinal-enterocolitis
mo travels orally,skin infections and spreads all over the body
lab diagnosis;
specimens can be pus,sputum,blood or other bodily fluids
-incubate specimen using 2 medias as blood media and selective
media of growth
-if blood is used,incubat in sugar broth,inoculate on sugar media
18-24 hours incubation
-examin production of coagulase by incubation of 6 hours and
then inoculate microorganism
-biological and biocemical properties can be determined
bymannitol fermentation,phage typing and antibiotics sensitivity
test using antibiotics
treatment and prophlaxis;
treatment;-depending on antibiotics sensitivity test which needs
time(gentamycin,Lindomycin)
-fast treatment of Fluloxacilin drugs
prophlaxis-non spcific;toxoids
hospital infections;staph has high resistance to antibiotics so can
develop hospital infections quicklydue to fast
mutliplication.vaccination is possible.

factors of virulence;
a)hemolysins-O hemolysins
-S hemolysin
b)ertytogenic toxins-by Strep.aureus
c)streptokinase and fibrinolysins
d)M proteins for adhesion of mo
e)neuromidase
strep.disease
gives 2 groups of infection-non suppurative and
suppurative infection
Strep.pyogens gives respiratort tract diseases like
tonsilitis,scarlet fever skin infection;wound
infection,genital infection
also non suppurative disease-rheumatic fever and kidney
disease of acute glomerulitis
autoimmnu disease
lab.diagnosis
examination of blood in suitable media(sugar broth)
smears-Gran +ve spherical or oval shaed
culture-blood agar medium 37 degrees overnight with
co2..serological test of hemolytic streptococci done for
classification
antigen detection test-ELISA and agglutination test
treatment;
a)antibiotics like
penicilins,streptomycins,cephalosporins,gentomycins
prophylaxis
vaccines and non spcific ones like penicilins.

No 91;gonococci;morphology and physiology.

Morphology;Gram ve cocci(gonococci)
Arranged in pairs(diplococci)
The sides are flagellated together ,coffee bean shape,non
motile,non sporing,capsulated in most species

Condition of cultivation
Requires enriched medium like lysed blood or chocolate
agar for growth.inoculated medium incubated in moist
anerobic condition at temperature 35-36c.enriched agar
medium with lysed blood and antibiotics.
Factors of virulence.
1)capsule-loosely asscociated with the cell wall and easily
stripped off.synthesis is dependent upon environmental
and nutritional factors.capsule inhibits phagocytosis
2)pili-produce altered appearance of colonies in culture
with sharply defined borders.helps in attachement of
organisms to host cells
3)proteinsconatins of porin,protein ii and protein 111
which is associated with the protein 1in cell surface in
formation of pores.
4)lipopolysaccharides-for endotoxic effects
5)other proteins-Ig A1 proteases
classification of microorganism and diseases
g:neisseria
s:N.gonorrhea

N0 92;Escherichia:morphology and
physiology.Antigenic
Morphology and physiology;
Gram ve,non sporing,non capsulated,measuring 1-3
micrometer.Most strains are motile some immotile
Are aerobis and facultative anerobic and grow on ordinary
culture media at 37c in 8 to 24 hours.on Mc-conkey
agar,they are rose pink due to lactose fermentation.
Break down most sugars(glucose,mannitolsucrose,malsotse)
Has slightl hogher resistance to heat.killed at 55c for one
hour.
Sensitive to antibiotics like
amphenicol,cephalosporins,tetracycline
Antigenic structure
Based on the O H and K antigens detected by agglutinations
reactions
-Somatic antigen(O antigen)
are hea stable..acidic components are hexuronic acid
-Surface antigen(K antigen)
looked as capsular structures.are destroyed by heating at
121c for one hour.it was previously desribed into 3 classes
L,A, and B on the basis of i)effect on heat on agglitinability
ii)antigenicity
iii)antibody bindingpower of bacterial strains carrying them
-Flagellar antigen(H antigen)
thermoliabile and they are 50 antigens so far described.
-Fimbrial antigen(F antigen)
thermoliabile protein and heating this organisms at 100c
detaches its fimbriae.these antigens may cause confusion is
serum agglutination test.it has no significance in antigenic
classification of E.coli

diseases of gonorrhea is a clinical condition in which

semen flowed in male organ without erection.it is sexually
transmitted and has an incubation period of 1-2
days.primary infection begins in the columnar epithelium
of urethra
-acute urethritis is characterized by vaginal discharge.it
may also involve rectum or oropharynx
other complications that may arise may also be opthalimia
neonatorum-conjunctivitis of the new born,gonococcal
vulvovaginitis-occurs in perpubertal girls
lab diagnosis;
the discharge in acute stage contains large number of mo
and diagnosis is readily made
it is difficukt to detect mo from chorinic cases or from
patients with arthrirtis
speceimens colleceted are urethral and cervical
discharge.the meatus is first cleaned by a gauze soaked in
saline and sample is collected and is subjected to cultural
study and smear examination
in chorninc cases the main syndrome is dysuria
microscopical methods can also be performed and
biochemical test where N.gonorrhea is oxidase positive
and ferments sugars woth production of acid only.
Treatment;divided according to resistance;
a)low level resistance- an high level resistant both of this
are penicillin resistant.in cases of this 2
resistant,cefuroxime,cefotaxime,erythromycin,tetracycline,
ciproflaxin can be used.
Prophylaxis;safe sexual contacts.

Physiological role:
Acts as microbial flora in intestineHemolytic E.coli in faeces in
pathological condition.
Diseases:
Urinary tract infection:
E.coli is predominant coliform bacteria responsible for 60-80%
infection of urinary tract.bacteria normally present in colon of
both infected and non infected person.
Diarrhoea and dysentery:
Produce diarrhea with different pathogenic mechanism
-Enteropathogenic E.coli-cause enteritis in infants
-Enterotoxigenic E.coli-produce heat labile enterotoxin or heat
stable enterotoxin or both.
-Enteroinvasive E.coli-produce illness in patients of all ages
-Vero cytotoxin producing E.coli-cause hemorrhagic colitis
pyogenic infection
-may cause infection,peritonitis,biliary tract infection,septicemia
and meningitis
Labaroty diagnosis:
-specimens-depending on tupe of lesions,cultured and examined
-microscpoy-Gram staining
-fresh specmen should be inoculated directly on Mc-Conkey agar
plate and blood agar plate an incubated overnight at 37c
-Biochecmical rwactionMost strains produce acid and gas from
large number of carbohydrates
-Agglutination testTreatment:REHYDARATION THERAPY
Prophylaxis-tak jumpa sorry.

102:Clostridium botulinum:
Characteristics:
Morphology:gram+ve,bacilli,non-capsulated,motile by
flagella,give oral,subterminal,bulging spores,optimum
t=35c,cultivate in sugar blood agar,serologic type A-G,spores
highly resistance,t=100c till 20mins
Types of toxins:
they synthesize exotoxins,have lable toxins,destroy several hours
at t=100c.exotoxins act on CNS,very strong biological
toxins,death of man only need 0.001mg of toxins.have several
types:A,B,C,E,but spores are not destroy(synthesize
exotoxins),they give botulism or food intoxication .Toxins can be
destroy by pressure cooking&boiling for 20min.Neurotoxins is
present in blood,peripheral nerve
Onset of disease:
Toxins produced intracellularly&appear in medium only in death
of cells.This is botulism not by wound injection,by food
intoxication occurs due to presence of toxins.There are in canned
food.Exotoxins enter man thru food coz has anaerobic media in
canned food.Spores vegetation forms.Multiplication only in
surrounding area.eg:if 2ppl eat the same food but 1
affected&another not bcoz they only develope in certain part of
food.Spores contaminating meats,vegetables&fish can be
injected.Temperature in canning not enough to kill
microorganism.Canned food have anaerobic condition,release
exotoxins,food born botulism,rapid paralysis.Clinical
manifestation:synthesis neurotoxins,no vomiting,diarrhea,pain
but cause hallucination,constipation,difficult in breathing,double
vision

103:Non spore forming anaerobes bacteria:

Classification,morphology,physiology:
Anaerobes,capsulated,non sporing,culture media with blood
Classification:gram ve:bacteriodes,fragilis,prerofella melanin
genicus,fusobacterium neurophorum,F.nucleatus cocci,vellonella
Gram +ve:pepticoccus niger,peptostreptococcus akna
Gram +ve arrange in pair
Gram +ve bacilli:lactobacillus,rod shaped,not
motile,biochemically active,pleomorpic,not
flagellated.Microaerophilic CO2,ph=6,grow on blood
media,nutrient agar,after 48h of incubation
Gram-ve bacilli:Bacteroidaceae family,non motile,small oval
pleomorpic rods,obligate anaerobes,robetson cooked medium,vit
k
Basic pecularities of clinical
Use lactobacillus,coumousals in intestine sym vit b12,vit k,vit
b7.In vagina convert glycogen to lactic acid(maintain high ph in
vagina)injection of internal organ,depression
Disease:oppotumestic pathogens.Cns:brain
abscess.Mouth:gingivitis,dental abscess.Resp:lung
abscess,bronchioeatasis.Abdominal:appendicitis,peritonitis.Femal
e genitals abscess.Skin:cellulites,gangrene
Lab diagnsis:
Take pus,should eb done fast,since in 20min m/o will
die.immediately transfer and inoculate in anaerobis media.Direct
smear exam,culture BA+vit K+neomycin anaerobic condition
Treatment:Fucidin,TC.Metravidozole.Direct abscess.Simply
open cavity,evacuate pus,clean.Treat with antiseptics

Lab diagnosis:
Take vomitlus,stool,food or blood,check concentration of
toxins&its type.Take different antitoxins against
A,B,C,D,E,F,G serology type&do the test,use passiave
agglutination test&precipitation in gel.Clinical
diagnosis,serological,biological method are aim at
revealing of toxins in clinical
specimen(blood,food),serology include passive
agglutination test&precipitation test in gel.Clinical
specimen is inculate into laboratory animals(mice)with
antitoxins in order to determine the toxins neutralization
test in vivo bacterioscopical&bacteriological methods are
useful if Clos.Botulinum is present in the clinical
experiment
Treatment&prophylaxis:
Treatment:removal of unabsorbed toxins fr
stomach&intestinal tract,neutralize toxins by giving
polyvalent serum to type A,B,E
Prophylaxis:vaccine containing toxoid fr A,B,E
types,active immunization is not used

104:Pathogens of diphtheria:
Pecularities of morphology&culture qualities.
Morphology:gram+ve rods,non sporing,not motile,not
capsulated,pleomorpic,inclusions present at the end of
rods,metacentric granules
Anaerobes,for growth serum containing medium,for isolation
Loeffleris serum media
Toxin production:
Genetically determine the toxigene can be transfer fr TBC to
another by lysogenic phages.Exotoxins:they are specific for
epithelial of upper resp. tract 2 sub-units.Lethal factor:provide
toxicity.Binding factor:provide transfer into cells
Diphtheria in man:
Source:sick person by air borne.local effect:BC multiply at site of
entery
,produce toxins,toxins cause necrosis,concentrated upper resp
tract bleeding.General effect:diffused in blood,toxaemia,necrosis
of parracervical cells of liver,kidneys,adrenals&polyneuritis
Immunity:
Alum precepitated toxoid:active:ADT- alum precipitated
toxoid.DADT-durity precipitated toxoid.TAF-toxoid precipitated
floccules.Children>10years immunized by
toxoid.Passive:succeptiple person given 500-1000units of
antitoxins after skin test.Combined immunity:active
immunization in endemic area stated at 3months old by DPT
vaccine.Baster dose given at school at 6years old&adult given
vassice by IM
Treat:DC,EM.Prop:DPT
Methods of its discovering
Specimen fr sick person,m/o fr place of localize,eg
wound,nerves,mucus
Bacteria-carriage:
Carrier-fr nasal cavity

Specific prophylaxis&therapy:
Slide nelson staining or MB method or alberts stain (show
typical chines letter pattern),inoculate into blood fellulite medium
isolate pure culture,identify biological varieties.Biochemical
properties:check production of cystinase&abscess of
urease.Identify production of precipitation test.After incubation a
fixed into paper 100k like lines of precipitation
Epidemiology:
Natural source-sick person,carrier,m/o secreted by
coughing,sneezing with saliva,root of transmission:air-borne,by
contact with infected saliva
105:Mycobacteria
morphology:gram+ve,rods,acid fast
physiology:anaerobes,all except M.Leprae,grow on ordinary
media,artificial culture rate of grow depend on optimal
temperature.
Pathogens of leprosy:Mycobacterium leprae
Characteristic:gram+ve,rods,straigut(slightly curved are polar
odies&intracellular element way present.Not sporing,not
capsulated,not flagellated.Growth on lowerstein-jeuson
medium.Bacilli multiply on food pads&granuloma develop at site
especially they cultivate in animals,mice,rats,where lymph
nodes,spleen,liver.Resistance remain viable in numed
environment,moist,soil,exposured to direct sun light
Lab.diagnosis:specimen:mucous,exudation fr ulcer,CSF,skin.
Slide stain by Zeild-nelson,check red dots,test on mice,lepra-non
pathogenc,TB-pathogenic.Leprosy:chronic granulomatous
disease,human involve skin,peripheral nerves,nasal
mucosa.Disease:classify into A
types:Lepromatosis,tubeculoid,diumorphous,indeterminent
Treatment:isolate in special camp,leprosy:face
changing.Antilepramatous preparation&immuno
stimulators.Additional
drugs:Rifrasulfan,Solosulfan,Deosafan,Lampran,Ethron
amide.Properties:long term chemoprophylaxis
BCG.Vassine:induce leprosy positivity

Treatment:Europian type:drugs of 1st time treatment,has

bacteriostatic action,drug os 2nd time,eg
paracinamids.American type:bacteriosidal
drugs(pitasinamine)bacterial drugs that kill
tuberculosis(isoniasine,streptomycine)bacteriostatic
drugs(proteonamide)treat in1-6months
Prophylaxis:disease of poor people,people with poor
hygiene&nutrition.Non-specific;use
mask,gloves.Specific:vaccination by BCG

106:Mycobacterium tuberculosis
morphology:stain by violet colour.gram+ve,rods shape,bacteria,hard to
stain by gram method,use acid fast staining,they r thin,thicker or chain
like organized rods.
Physiology:aerobes,optimal temperature is 37c.The growth is form by
presence of CO2 in medium.2 medium use ofr tubuculosis,Loveustein
gensen medium(contain egg yolk,potatoes as nutrients),Tinn2
medium&cotton medium.1st variable growth 8-10days,slowly growing.In
lipid medium grow like pilicals.In solid medium grow like rough
colonies.On semisolid medium grow on the surf of medium(bovis grow
under the medium can diffused them)
Classification:mycobacterium tuberculosis,m agricanum,m bovis,m
arium:they r main group that gives tuberculosis
Pathogenic m/o:m necrotic,m ultura
Lab diagnosis:(A)take sputum fr patient&biopsy.Time of tuberculosis is
8-10days,so 1st must use differentiation test.Conclusion is given by
microscopical examination,in this must find acid fast bacilli in
sputum.But this is difficult because m/o is inside mucosa,so used
enrichment test b4 this.Take mucous,tube+alkaline solution(NaOH),the
mucous get layers&separate,m/o leaves mucous due to
neutralization.Seperate the medium fr m/o by 2methods.1-add medium
benzol oxylate&shake,m/o comes to medium,take the layer.2ceutrifigutation.Ientify by microscope.Make pure
culture.Sputum+acid,cultivate not more than 8days(acid is put because
they r acid fast&resist to acid.So all other m/o r dead&only tuberculosis
is live.After8days take colony,inoculate in gusons medium,examine
biochemical properties.In3-4 weeks diagnosis is given.(B)Sckolchicov
rush methods:based on mycobacteria to syn coat factor.Take sputum,to
slid,after drying,slid+acid(to kill other m/o),put slide to liquid
blood,incubate3-4days 37c,myco multiply in blood noritible colonies.
(C)biological test:inject to rabbits,identify presence of tubuculosis
bacilli(D)Florencent microscopy:stain slid by Ab,can see florencent by
microscope in the slide.(E)allergy test:due to hypersensitivity delayed
type.Inject specimen and after 24hour,see redness.Check diameter of
redness,less then 20mm is normal.Greater is pathology,do x-ray&check
clinical feature.

107:Pathogenic fungi:
Class acc to kinds of multiplication.Perfect fungi:have
sexual and asexual-miscellanieoces fungi,which may
produce mycerium.Sacromycetes.Imperfect:no sexual
spores,eg:dermaloucycetes,caudida fungi
Morphology&physiology:mycelium with septa may
substare cell surface or inside tissue or media or air
mycelium(above media)ph>7 aerobic majority,mesophiles
37c.colorless colonies.Later stage pigmented.Cultivate on
liquid/solid media.Common media:seuberoud
media,mucor&aspargillias-place of accumulation of
exopores.Mycosis acc to synthesis:1systemic,deep:coccidiasis,cryptococcosis,blaslomycosis,pa
racoccidisis.2-SC:sporotricuosis,motaramycosis.3superficial:dermatomycosis,epidermamycosis.endogenous
infection under certain condition
Lab diagnosis:skin
scraping,nails,hairs,pus,stool,sputum,urine,CSF,blood.Prep
are drop with alcohol&glycerol wout staining.check under
microscope,examine bochem properties.Test Ab
titre,IF,passive hemagglutination.Reaction of
hypersensitivity delayed type(skin allergy test),biological
test.
Treatment&prophylaxis:no special properties,treatment
Ab act on eukaryotic cells by big side effect medicasion
under superfivision

108:Candida fungi:
morphology:2forms:1-spherical or oroid budding cells. 2elongated filamentous cells,join end to end
physiology:they ar a part of normal flora of human body
Difference fr yeast:ability to form pseudomycelium by a
chain of elongated budding cells joined end to end.
Condition that assist the beginning of candidiasis:main
disease that posses development of candidiasis as
DM,immunodeficiency,malignant,long term broad
specimen antibiotics,premature babies,stress also chang the
diet.This condition gives changes in body states from its
normal condition so it gives decrease immunity&easily
fungal infection development than in normal.
Role of fungi in human pathology:?

Congenital syphilis:treponema can pass thru placenta

barrier.Transmission occur in 10th week
Lab diagnosis:specimen:exudates fr chancle,blood,lymp.T.pallid
can be shown on dark field microscope slides made by silver
impregnation,floresence microscopy,serological
methods.VDRL(venual disease research lab):on a special slide
with depression of 14mm 0.05ml decomplimented serum is taken
a drop of cardiolipin is added. It is rotated at 180 levolution 4
minutes.microscope low magnification,see
clumps,positive,crystals-negative.RPR(rapid plasma regain
test)same as VDRL but unheated serum can be used. Carbon
added to make reaction visible.Compliment fixation:Inactivated
serum+serum fr guinie pig,37c/1hour.Reagin containing serum
fixes compliment.Add sheep RBCs,positive,no
hemolysin.Negativehemolysin.Immunofloracence,ELISA,passive
hemagglutination,Buccle&Lizhenburg test.Reaction of
immobilization:diluted serum mix with compliment actively
motile Nichols strain Ab cause immobilization in positive
reaction.
Immunity:no specific only prophylaxis.Treatment of eases,sex
hygiene,use condom and antibiotics
Treatment:penicillin group:no resistant for hourly 1
month,prolonged forms of penicillin once daily.Doxycidine
ointment with salts of heavy metals as they are very effective
against m/o
Pecularities of syphilis:no immunity after infection.Non-sterile
immunity also called charra immunity.

109:Pathogenic spirochete:
Class:family:spirochetales is divided into 2family.1spirochetaceae:Treponema,Borrelia,SPirochaeta,Cistispira.2Leptospiraceae-Leptospira
Treponema pallidum:gram-ve,spiral shape
rods,endoflagillated,activity motile,non capsulated,non
sporing.Axial filament located outside cell wall 8-12 constant
arrives inconstant L or V.
Cultivation:cant grow in artificial media.it can be in rabbit
testis.The nichots strain has been maintain by serial passages in
rabbit testicales.Since1912,can be cultivated in Robertson
cooked.
Factor of virulence:protein of external
membrane&lipopolysaccharides.Antigenic structure
specific&non specific Ag.Ag-group specific Ag&species specific
Ag-polysaccharides.non-specific:lipoid hapten Ag
Pathogenesis of syphilis:can be acquired or congenital
Acquired syphilis:has 3stages.Incubation2-4weeks or even
5years.1-Papille appears on genital area.Ulceration forming.This
ulcer like other ulcer is painless. It isflat,dull,red,exudates a
serous fluid.Lesion can also occurs in lips,moth,nippe,fingers.The
fluid exudates is highly infectious,ulcers heals in3-6weeks
forming a thin scar.2-after 2-6month generalized syphilis
occurs,rash appears on skin&mucous.Papillae&condyloma are
formed specially around anus.Various inflammating&painless
lymphadenopathy developes.Symptoms disappear in 8-12 weeks
in untreated cases.This follows of latent phase.3-after 10-15years
tertiary lesion appear.Gamma appear on skin,mucous
membrane,bone,tongue,liver,testis,aorta,heart.Syphilitic
mesoaortitis is a serious complication leading to severe
anemia&hemorrhage.General paralysis occur due to eurosyphilis

110:Spirochete of relapsing fever:

Borrelia:3-8 inconstant curves,gram-ve,anaerobes,B.Recurrentis
epidemic relapsing fever are transmitted by body louse,pediculus
humans&humansoccurs in unhygiene&poor social economics
condition.Natural reservoir is man&vertors louse.Incubation 310days.m/o invade into blood during fever subsides bcame Ab
reaction with the most in blood&are killed.Again m/o with new
Ag structure enter circ&multiply causing fever.Endemic typhus is
cause by B.Caueasia transmitted via ticks.Manifestation not as
serious no epidemic type
Diagnosis:blood,smear.Giemsa or leishman stain.m/o are seen in
the cell of blood
Romnavosky method of staining:biological inoculate 1-2ml of
patient blood in mouse in 2-4 days slide of mouse blood show
m/o
Epidemic:source fr man,louse vector,causative agent is
B.Recurrent,no of relapses 3-10,manifestation intense
Endemic:source is animal,ticks vector,causative agent is
B.duttoni,B.persica,B.caucasia,no of relapses 5-6,manifestation
milder.
Role of Minch:he establish infectivity of relapse fever&typus
fever.Discover that disease was transmitted by blood,sucking
insects.He did tests on himself.
Role of Mechanicoff:he was founder of study of immunity of
organism to infectiour disease.He establish intracellular digestion
brought about by mesodermas cell.He named such cells in human
body as phagocytosis.Both systems are placed seperatly in a
thermostat to 2hours,they are mixed&placed for 60-80min. if it is
positive within a day the erythrocytes settle to bottom&the
supernatant liquid is tansparent&colorless.In neative reaction the
compliment will not combine with the complex of 1st sys while
the liquid becomes pink&any precipitation of erytrocte.This test
is use with diagnosis of glanders,syphilis,typhus fever,Q
fever&other rickettsioses,viral disease

Q 111.
pathogens of lyme disease : morphology
Bacteria
-b.Burgdorferi
Morphology
GRAM negative , motile , size 4-5 x0.2-0.5
mm
Pathogenesis contains toxic lipopolysaccharide inflammatory
prop. Incubation 3-30 days in skin, due to tick bite then move to
lymph nodes, reach blood then week then months and then cause
cardio n neurological plb . n increase of IgM n immune complex
the dev atritis n immune complexes deposition in synovial tissue
Epidemiology : - smalls mammals are reservoirs of the
bacteria .the tick feed on Infected mammals n then lava cause
dev of tick parasite in mammals, - its transmitted by tick (exodus
ricinus/dammini) to men
Clinical stages : Early stages migrain , skin red ,fever chills
,malaise, muscle pain
Late stages - 2 years later neurological n cardiac sympthoms
Meningitis , encephalitis ,heart block
-2nd stage of late stage is arthritis for months to years
Lab diagnosis : 1.staining- wright or gimsa staining on blood .
if + will be sliver starain. 2.culture and isolation on blood
3.serology- immunoflorescence test or ELISA
4..PCR
Treatment
: early stages - doxycycline , amoxicillin ,
erythromyocin
Late stage parentral admin triaxone

Q112
Genus :Leptospiraceae
Species : L.icterohaemorrhagiae
L..interrogans- pathogen in man n animals
L.biflexa in pools ditches fresah water
Morphology
- spiral shape 5-20x.01 mm
- narrow diameter closely set coils so its stained poorly in
fluorescent n sliver impregnation techniques
-motile rotating n flexing in axis
Pathogenesis
Transmission: men 2 men by contaminated water with urea or
faeses
Entery site : skin , mucus then in cubate for 1-2 weeks and go in
to blood n then organs
Exp :liver cause hyperbilirubinaemia n jaundice
Vascular involvement leading to coagulation
Renal failure due to tubular necrosis
Clinical disease
Subclinical l.intergos in packing house workes n veterinarians
Anicteric leptospirosis after 1-2 week or incubation period like
influenza
1 month headache , nausea ,
vomiting
Icteric leptospirosis - most severe hepatic in volment jaundice
Fever ,myalgias, diffuse rashes headache

Lab diagnosis
Specimens: blood 1st week, urine 2nd n 3rd week , serum of
antibody detection n CSF
Detection : 1.Dark ground microscopy : blood n CSF shows DG
illumination
2.PCR detection of leptospires in urine
Serological: using titers and the result is 1 week the titer is high
and towards the the 5th
Week it will decline
TEST : microscopic test , ELISA test IgM n IgG, C.T.F indirect
Hem agglutination
Confirmation test : using antigen :make suspension with live of
filled formalin killed
A series of dilution of the patients serum
is tested against the antigen
MAT (microscope agglutination test )
Treatment
Penicillin is drug of choice : doxycyline
,chlortetracycline,piperacillin,cefotaxime
Propylaxis : avoid contact with contaminated water . natural host
are dogs pigs mouse

Q113. MYCOPLAMA
Character
-Two types free living n self replicating .size 1-5 nm x
50nm
-Have ridgid cell walls and are pleomorphic .resistant to
penicilline
- it penetrates cell membrane but some are normal
microflora of the body
- are gram - , non sporing , non capsulated , non flagellated
Classification
-Family is devided to two type Mycoplasma (glucose or
atrginine ) and ureaplasma (hyrolyses urea)
-mycoplasma can be pathogenic .got 80 types
Species that cause human diesese:
M.pneumoniae,M.hominis,M.genitalium,M.urealyticas
Perculaties
Morphology grow on fluid media , spherical shape .
gram - ,
Cluture-in infusion of peptone broth with 2% agar
Pathogenic species
M.pneumoniae-transmitted by infected person
M.hominis- by arthritis ,postpartum sepsis ,pelvic infection
M.genitalium- acute chronic men gonococcal uterharitis
M.urealyticus- genital in fection . vaginaosis

Myco.pneumoniae
Adhere to the respirtory epithelium n interact with cilia on
epithelium cell surface
May develop atypical pneumonia .may also infect the
myocardium , cns ,skin ,joints
Clinical syndromes 15-25days sympthoms of
pneumonia,pharyngitis tracheobronchitis ,otitis media.
Immunity IgA in limited role in the external area , IgG in
blood
Epidemiology-secretion from infected person
1.transmission .1week to 2 weeks can
see on set of sympthom
2 susceptibility . crowded areas favours
transmoission in children n teens
3.distribution .due to climate
4.incidence. based on poplation n
military recruits
Diagnosis
1.compliment fixation test .depending on the titer
2.assay for specific test IgM (immunoflorescent test , IFA,)
3.DNA probes n PCR for development of P1 adhesin
Therapy
Antiboiotics tetracyclines,erythomyocin
Prophylaxic- non spesific

Clinical syndromes
Happen in spring n summer multiply in tick gut n comes
in feaces.3-5 days n contact with .
When louse feed on the human so it inro duced to skin
and then incubate there fot 2 weeks
Cause headache , fever , muscular rash, late then
myocardial n CNS involvement
Lab diagnosis
1.microscope
2.cultural isolation from guineapig,yolk sack or chicken
embrio
3.immunofluorescent test with polyvalent anti serum
4.PCR
5.Serological test complement fixation test with specific
antigen
- indirect fluorescence test IgM n IgG
- direct fluorescent
Treatment n prophylaxis
General measure celaning up jungle exe
Vaccination ,chemoprophylaxis
Drug : doxycycline n chloramphenicol

Q114.Rickettsia
-gram -, rod shape , binary fussion, nonmotile, non flagellated
-non capsulated,non sporing
-cell wall contain muramic acid , sensitive to antibiotics
Factors of virulence is call walls
Classification
Famili:rickettsiae
Genus: R.rickettsii n rochtunaea n coxiella
2 morphological form
Regetative-nucleoid, cytoplasmic ,cell membrane n wall
Cystic from thick coat protect against environment factors
Kinds of typus
1.epidermic typus- R.prowazekii, R.typhi
2.kind of fever- R.rickettsi,R.sibrica,R.akari
3.scrub typus- O..tustsugamushi
Louse brone typhus
By R.prowazekii
Dev-by like q fever . vector is tick mite suck on blood of infected
men
Pathogenesis
1. bite or crushed on the surface of skin go thru endothelial
of skin .in microvessles and then incubate 7days . in this time
there will be increase of hyper
sensitivity and have lymphadenitis near bitten area.later some get
killed by macrophages
2.7-10days later go thru blood n replicate on endothelial cells and
cause vasculitis

Q115. Q fever
By Coxiella burnettii
Morphology
Gram - , ploemorphic coccabacilli , non sporing nonflagelated
,non motile , non sporing
Physiology
Related to virus are small and obligated intracellular parasite
Contain enzymes of bacteria DNA n RNA
Sensitive to antibiotics
Source
Wild animals like from slaughters house like cattle , sheep
Poultry by ixodid ticks
Pathogenesis
Alveolar macrophages becomes infected n lead to rickettsia
species contain plasmids
Cause fever n influenza like syndrome there no rash in the
sympthom
Transmission
Present in large quantities of milk products
So drinking milk inhaling aerosol
Enters thru skin , mucose, lungs
They stay in environment in dry state for long periods 1 month
Can stand 60 degrees
Laboratory
Serological CTF indirect immunofloracent assay
Treatment prophylaxis
Oxyciline n tetracycline, coctrimoxazole , rifampicin
Treatment general hygine , insecticides
Pasturization of milk
Vaccine

Q116.Chlamydia
Morphology
Small, non motile, gram non sporing
Exist in two forms elementary body and reticular body
Physiology
Aerobic or faculatively aerobic
Intracellular parasites
Look like virus have both DNA n RNA
Multiply by binary fussion
Have metabolic active enzyme
Elementary forms -spherical electron dense nucloide
Recticular forms matahbolicly active n particales
Have surface membrane .cells make vacuole around he particles
The particles enlarge and reproduce by binary division
and the cell breaks and liberate from host n cycle from 24-48 h
intermediate forms condensation of reticular body which look
like bulls eye
Pathogenic species
C.pasittaci -cause psittacosis n ornithosis
C.trachomatis- A,B ,C endemic tracoma
L1,L2,L3,--Llymphogranuloma venum
D-K nongonoccal , uretritis mucopulurent cervicitis
C.pneumonia- acute respriratory syndrome
Lab diagnosis
Specimen blood , lung tissue , sputum . a single
Cell culture
Serology Cf testing n microimmunofluroscence test
Mouse impregnated into yolk sac for 6-8days old egg or tissue .
mice will die in 10days
Smears made from elementary bodies 1:32 titer
No specific available
Treatment
Tetracycline and erthromycin

Q118.Rabies virus
Virus two kinds :
Characteristic
- Bullet shaped when seen by electron microscope size 76175nm
- Contain a single strand RNA chromosome nucleoprotein
Large non structural protein
- got outer lipoprotein envelope contain protruding
haemaglutinating peplomer spikes
10nm long
Epidemiology
Wild animals like dogs , jackels horse cattle .
Virus transmits by sliva so plp get it when they are bitten
Pathogenesis
Replication n spreading -1st invades in the muscle and then
lead to nerve fibers
And grow in the grey matter if the brain
Incubation period 1-3 months
Then there will be degeneration in the cortex , midbrain ,basal
ganglia pons medulla
Clinical features
Headache ,malssiae low grade fever and anxiety in 1-10days
,infected person have Hydrophobia fear of water so they are
thirsty ,and then cause muscular spasm
Patient then develop coma , disorientation ,delirium .
Death from neurological and pulmonary sympthoms

Q117.Tarchoma and unveneral uretitis

Pathogens :Chalamydia .tharchomatis serotype A, B,
Ba ,C ,universal (serotype D-K)
Morphology
Small , nonmotile , gram - , obligative intracellular
parasites , exist 2 forms
Physiology
Aerobic or faculty aerobic . are small and obligate
intarcellular parasite .they wwere previously called viruses
,have both DNA n RNA
Does binary fussion .have metabolic enzyme
Clinical picture
Endemic tharcoma most printable from of blind ness
.sexually transmitted infection or urogenital
trach,lymphogranuloma venulum ,epidermitis ,reiters
syndrome, proctitis,
Salpingitis
Ways of transmission
By close contact .mother to infant , sexual, Eye, fingers
touch

Laboratory diagnosis
Diagnosis in men sample of sliva , test by
immunoflourescent test
Antibodies to virus develop slowly presence of anti bodies
in serum in an unvaccinated rabies infection .
PROPHYLAXIS
1.prexposure .Handle animal which has potential of
infection .two intra derminal vaccine and HDCV ( human
diplod cell strain vaccine ) given in 4 weeks of interval .lit
is follod by yearly booster
2.postexposure .cleanest first with water and soap n with
ammonium detergent
Then given rabies antiserum The wound is left
unsaturated .rabies can be prevented if treatment is
iniciated in a day of biting .
Bitting animals should be kept strict for 10day
Hyperimmune- human rebis antiserum is the safest
antiserum . half of the antiserum locally

a)
b)

Q119. Orthomyxoviruses
Structure
Size n shape : 80-110nm
Genome nucleic acid of eight have negative sense single strand
RNA polymerase
With the transcription with PB1,PB2
Envelope
Contain two glycoprotein , hemagglutinin(HA) , and
neuramidase (NA) spikes that project to the outside , and lined
by the matrix , membrane , haemagglutinin
Formation HA and NA spikes coded by viral genome
n are synthesized in early period of replication cycle
Immunogenecity both HA n NA molecules
Classification
Diff in the nucleoprotein (NP) and the matrix protein(M)
Types A,B,C.influenza A virus has sub type H1-H15
Epidemiology
Type A and B tend to cause epidemics that recur every 1-3
years .
Type C virus usually does not cause epidemicis
Influenza shows remarkable ability to undergo antigenic
variation due to frequent changes in the antigenicity of HA n
NA .
1.Antigenic Shift and Drift are responsible for the remarkable
epidermics of influenza virus antigenic refers to the minor
antigens changes ineither the haemaglutinin or neuromidase
or both .
2.Antigenic shift refers to major antigenic changes in HA(H2
to H3)
Transmission is by person to person via respirtory secretion
It remain contageneous 24h b4 the clinical symptoms till 48h
after on set of symptom
Subtype 9NA(N1-N9).

25.Methods for the cultivation of obligate anaerobes:

principles apparatuses
a)displacement of oxygen gases:
such as hydrogen,nitrogen,helium or carbonate.insert gas to
sealed or leaded with inoculated media but this method rarely
produces complete anarebiosis
b)absorption of oxygen by chemicals
-pyrogallic acid:
In large tube NaOH and pyrogallic acid added.the tube placed
inside air tight jar loaded with inoculated media.
-chromium(Cr) and sulphuric(H2SO4)acid.(Rosentha medium)
This method also used for producing anarobiosis.
-Gas pak
It is a disposable pocket-when water is added hydrogen and
carbon dioxide and liberated hydrogen combine with oxygen in
presence of catalyst.
Method-after inoculated media is placed inside air tight jar/gas
pak with water is added.
c)Malntosn-Fildes jar
palladnised asbestos lept inside the jar as catalyst-combination of
hydrogen + oxygen-water reduced methyline blue is use as
indicator
[anaerobic-colourless Oxygen present-blue colour]
Biological method
By incubating aerobic and anaerobic organism together two
different plates are placed on over other and sealed.it is slow and
ineffective
d)by incorporating reducing agent in media two most widely
used anaerobic liquid
-thioglycollate broth
Contain nutrient broth and 0.1% thioglycollate
-Robertsons cooked meat media.
It persist the grow of even strict anaerobics

Clinical disease
Incubation period varies from 1-4 days
Fever chills ,headache , dry cough ,myalagia .fever last for
another 3 days .an uncomplicated case usually resolves
within 7 days .
Lab diagnosis
Isolation of virus use baboon kidney cells see
haemagglutination results following addition of
erythrocytes to influenza virus containing media
1.immunofloroscent test .to see viral antigens
2.ELISA n RIa technique can also be used
3.SEROLOGICAL with acute antibody titer since normal
individuals already have influenza virus antibodies .
Treatment n prophylaxis
Amantadine is rimantadine are effect in prophylaxis and
treatment of influenza a illness during the 1st 24-48hours.
1.compozitions of vaccines .type An B viruses are grown
in allantoic cavity of chick embryos and inactive by
formalin. This vaccine includes 3 type of strand Type A
(H2N3)n(H1N1) n Type B.

26.Significance of identification of morphological and..

1.Microscopical morphology
-smears prepared from bacterial colony or liquid culture is
examined by staining method.
Gram stain:Gram +
Gram
Acid fast method: acid fast
Non acid fast
Motility preparation.in normal saline:motile
Non motile
(B)colonial morphology
-Colour :pigment and haemolysis cause change in colour
-Shape: circular,irregular and radiate
-Surface: smooth,wary,rough,granular
-Size: in millimeter
-Elevation: flat,elevated,loss convex,convex,umbonate,
ecitire,undulate,crerated,lobate,ciliate
-Opacity-translucent,transparent,opaque
(C) growth in liquid media
In peptone water,nutrient broth,Hisss serum bacterial growth
appears
-turbid
-deposit formation
-surface pellicle formation aerobes grow on surface of media.

27. Microbial enzymes.classification by the biological

role.
The bacteria has ability to produce enzyme during its
growth.mainly bacteria produces products during these life
time as: 1)bacteria toxins 2)enzymes 3)pigments
1)classification by biological role.substrate specificity
They are classified into 5 groups.(microbial enzymes)
a)tissue degrading enzymes:collagenase,clostridium
perfringens produce this enzyme
-this remains bonds between collagen and it spreads bacilli
in tissue.
b)cagulase,staphyloccocus aureus produce
c)thyalusonidase,staphylococcus aureus,streptococcus
pyogenous and clostridium produce this enzyme.the
hyaluridinic acid id hydrolysed and spread its action.
d)streptokinase,by streptococcus pyogenus dissolves
coagulated plasma and spread streptoccoci in tissue
e)haemolysis,streptococci pyogenus produce this
enzyme.it dissolves RBC
f)leukocidines,streptococci pyogenus produce this
enzyme.it kills tissue cells on RBC.
Bacteria produce enzymes which is not toxic but keeps but
keeps them in stand to the host enviroment.this gives the
ability to live and to be alive in the host organism.thi also
gives ability to produce disease in human.

28. Methods of sugarlitic

I.Methods of detection of prolytic and sugarlytic enzymes
Can differentiated E.coli from S.paratyphi A.when use H2S
E.coli can destroy protein by H2S and s.paratyphi A cannot
destroy protein by H2S.
II.Culture media
Consist of water or source of hydrogen and
oxygen,electrolytic/Nacl nad others , peptone or complete
digested proyein of animal or vegetables, yeast/protein degrading
products,CHO, inorganic salts and growth factor.Blood /
neutralize media uses sheep.Agar derived from seaweed.
Types: agar 1-2% ,increase solidition or due to mole
a)aerobic
b)anaerobic
Due to nature
a)natural-eggs,meat,blood
b)artificial-derivatives of nature
c)synthetic-prepared by chemical
Due to nutritional factors
a)simple-meat,peptone
b)special-add more nutrition.glucose and indicator and
heat by peptone agar.
Special media
There are shown in petridishes oxygen tubes as methods of
inoculation.
a)streak culture
b)conpet culture
c)stroke culture
d)stab culture
e)powr cor plate method

Advantages:-special colony forms

-quantitative bacteria: count and relative
preposition of diffrent bacteria can
make
-by taking colonial morphology can identify
bacteria
Liquid media
Distribute in tube with cotton wool and screw capped
bottles of flask.

29.Differentiation media. Enumerate main kinds

2 main kinds of media:culture media and synthetic media.
Culture media
water,electrolyte
mid peptone:complex mixture of digest proteins by enzyme
action.commercial preparation of peptone.(bactopeptone)contain peptone,protease,polypeptides amino acids and
inorganic salts.
Meat extracts/yeast extracts
Blood
Enriched media-5%-10% defibrinated horse or sheep blood
used.sometimes serum is also required.
Gas
Derived from sea weed contain mainly carbohydrate,small
amount of protein like material sometimes traces of long chain
fatty acids and a variety of impurified including inorganic salts.
Synthetic media
Prepare from pure chemicals.added separately for preparation of
product of unknown composition.they may classified into
various ways.based on physical state.
They may be liquid and solid on the basis of presence of
molecular oxygen and reducing substance in the
media,aerobic,anaerobic ciulture media.
Simple media.
Nutrient broth.contains peptone 1% meat extract 1% NaCL 0.5%
and distilled water and PH of 7.4-4.5.2%-3% agar is added to
nutrient broth it called nutrient agar.simplest medium and
routinely used i8n laboratory for diagnostic purpose.reduced
concentration of agar 0.2-0.4% enables motile bacteria to form
spreading or swarming growth.

Advantage:-when bacteria prevent in less numbers

inaculation.they grow only in liquid.
-use for biochemical test
-large inoculation can be tested -useful
enrichment like selenite f
Disadvantage:-culture tubes and bottles inaculated by
touching by a change loop

31.Influence of environmental factors on microorganism

1)water
Moisture ia required for bacterial growth.About 80% of bacterial
cell consist of water..
2)oxygen-divided into sub groups according to their requirement
for molecular oxygen.
a) aerobes-require oxygen for growth
Obligate aerobes-grow only in the pressure of oxygen
.eg:pseudomonas,
Bacillus.
Facultative anerobes-ordinary aerobes but can also
grow without oxygen
Eg;vibrio cholera. Most pathogenic bacteria are
facultative anaerobes.
Microaerophiles-can grow with oxygen and carbon
dioxide.eg:clostridium,
Bacteriodes.
3)carbon dioxide
Small amount of of carbon dioxide is required by all bacteria
usually its available endogenously as a product of cellular
metabolism or by atmospheric carbon dioxide.
4)temperature
The optimal temperature range for growth varies with different
bacterial species.they are grouped as follow:
a)psychrophile-below 20 degrees eg.soil+water saprophyles
b)mesophile-between 25 to 40 degrees eg:majority of pathogenic
bacteria
c)thermophile-between 55 to 80 degrees eg:bacillus
stearothermophilus.

32 STERILIZATION, DISINFECTANT, ASEPTIC.

STERILIZATION : is process by means of which an article, surface or
medium is made free from all living m/o including spores.
DISINFECTANT : is a process of destruction of vegetative forms of
pathogenic organisms which are capable of producing infection but not
necessarily resistant spores.
ASEPTIC : technique that is employed in preventing infections from
gaining access to an uninfected tissues.
ANTISEPTIC : Substances which either kill m/o or inhibit their growth.
BY HEAT
MOIST HEAT kills by coagulating and denaturing their enzymes and
structural proteins a process that requires participation of water.
Sterilization by saturated steam under pressure ( 121 C ) takes 15
minutes.
DRY HEAT : kills the m/o by destruction of oxidation of intracellular
components. Achieved by exposure of the organism to a dry heat at 160C
for two hours.
PROCEDURE OF STERILISATION BY DRY HEAT
1- RED HEAT metallic objects such as inoculating wires, tips of
forceps and needles are held in flame of a Bunsen burner for instant
sterilization
2- FLAMING glass slides, scalpels, needles, cotton wool plugs are
passed over flame. These are not allowed to get redheat.
3- INCINERATION is employed for destruction of infective
materials. Soiled dressing, bedding, pathological materials like sputum
and stool and carcasses are reduced to ashes by burning.
4- HOT AIR OVEN for glassware. Temperature either 160C for 60
minutes, 170C for 40 minutes or 180C for 20 minutes.
Uses: glasswares like syringes, petri dishes, test tubes, flasks,
pipettes.
surgical instruments like forceps, scalpels, scissors.
oily fluids which are impermeable to water such as oils and fats
chemicals such as powders which would clump or form into cake
in presence of
moisture
PRECAUTIONS :
1- substances to be sterilized should be absolutely dry.
11- rubber goods, fabrics or any inflammable or volatile substance should
not be put inside oven
111- instrument should not be overloaded and must have space for
circulation
1V- oven should be let to cool for 2 hours before opening.

5)hydrogen ion concentration

The optimal PH necessary for growth of most pathogenic
bacteria is similar to physiological ph 7.2-7.4
Some bacteria such as lactobacilli,thiobacillus thioxidans
(acidophilic) grow at acid ph 3.0
Cholera vibrios and alkaligenous faecalis (alkalophilis)
grow at alkalius ph 10.5
6)light-bacteria prefer to grow in
darkness.photochromogenic mycobacteria produce
pigment when incubated in presence of light.
7)osmotic pressure-bacteria can withstand a wide range of
external osmotic pressure due to the mechanical strength
or cell wall..0.5% NaCl is added in most culture media to
create a suitable environment for bacterial growth.
8)symbiosis for antagonism
Symbiosis-groth of 1 organism fascilitates the growth of
other organism.
Eg:groth of staphylococcus helping growth of influenza
Antagonism-growth of 1 organism I inhibits the growth of
other organism.eg:pseudomonas aeuruginosa hampering
the growth of gonococci of anthrax bacilli.

Procedure of sterilization by moist heat

AT TEMPERATURE BELOW 100C
1 VACCINE BATH: serum or body fluids are sterilized at 56C
for an hour and bacterial vaccines are sterilized by a special water
bath at 60C for 1 hour.
2 PASTEURISATION OF MILK : maintain a temperature of
63C for 30 minutes or 72C for 20 seconds.
3 FRACTIONAL STERILISATION : serum or egg medium is
tubed and placed in special racks and slow solidification of serum
or eggs is carried at 80C to 85C for one hour and for sterilization
of the medium the process is repeated for three consecutive days.
AT TEMPERATURE OF 100C
1 boiling: spores withstand boiling. Boiling for 10- 30
minutes kills most of vegetative forms
2 steam at atmospheric pressure at 100C for 90 minutes :
90 minutes sterilization time include loading and heating time
from 0- 100C
3 TYNDALLISATION : culture medium is steamed for 30
minutes each on 3 successive days. Vegetative cells are destroyed
at first exposure, in the intervals between the heatings the
remaining spores germinate in the favorable nutrient media which
are killed at subsequent heating
AT TEMPERATURE ABOVE 100C
Autoclave:- it is a modified pressure cooker or boiler which may
be horizontal or vertical. Made of stainless steal with a
supporting frame. Steam circulates within the jacket and is
supplied under high pressure to the closed inner chamber where
goods are kept for sterilization. It is done at temperature of 121C
and chamber pressure of 15lb per square inch for 15-20 minutes.
Also can be done at 126C for 10 minutes or at 133C for 3
minutes.

FILTRATION
The fluid to be filtered is sucked through the filter into a
receiving flask by negative pressure with the help of an exhaust
pump. Useful for making bacteria free preparations of substances
which get damaged by heat process.
Types of filters:- a) bacteriological filters - earthen ware candles,
asbestos disc filters, sintered glass filters
b) membrane filters & c) sand filter
Uses:- hydrated fluid, pharmaceutical preparations ( antibiotic
solutions ) and blood products.
RADIATION
IONISATION RADIATION - include gamma ray, X- rays and
accelerated electrons.
Uses: rubber or plastic disposable goods, disposable syringes,
surgical catgut, bone and tissue grafts, adhesive dressings
NON-IONISING RADIATION - a) infra red radiation b)
ultraviolet radiation
GAS VAPOUR STERILISATION
1- ETHYLENE OXIDE - plastics, polythene tube, artery and
bone grafts, endoscopes, vaccines and culture media.
2- FORMALDEHYDE GAS - used for decontamination of
surgical safety cabinets
3- HYDROGEN PEROXIDE VAPOURS - instruments.

Enumerate main subjects of each group

a)Acids, alkalis
- strong acids, alkalis kill bacteria but weak
organic acids inhibit growth.This is due to toxicity of anion.
b)Halogens- Cl2,I2 is commonly used as skin disinfectant. They
destroy bacteria & also have effect on spore producing bacteria.
c)Chloroform
- either liquid or strong vapour kills
vegetative forms.
d)Salts- Salts of heavy metals such as mercury chloride, silver
nitrate have toxic effect on bacteria,due to bacterial proteins.
e)Antiseptic of phenol- used mainly to prevent bacterial spread
in hospitals
cresols:used as solutions of cresols in soap,toxic to skin,used to
sterilize glass ware in labs.
chlorohexidine:
Hexachlorophane:has bacteriostatic action,used as soap
product.
f)Formaldehyde - effective against all types of bacteria.Used
in solution/gas
g)Glutaraldehyde
- new disinfectant, kills
spores,bacilli(tuberculosis)
h)soaps,dyes,alcohols have bactericidal action,remove
bacteria from skin
i) Aerosols & other gaseous Cl2,SO2 ,Propylene Glycol is
used.

33: INFLUENCE OF DISINFECTANTS ON THE

MICROOGANISMS
Influence of disinfectants on the microorganisms
1) Chemical groups of disinfectantsThe process of destruction of vegetative forms of pathogenic
organisms which are capable of producing infection but not
necessarily resistant spores.
Majority of these chemical substances have no effect on spores.
Few of them are able to destroy spores.
Uses: destruction method is applied above, sterilization is not
effected as bed pans and cooking items do not to be sterilized.
They only need to be disinfected by solutions.
The disinfectants are not used directly on the skin.
Gases:
Formaldehyde: This is used to disinfect
a)woolen blankets
b)footwear of persons(fungal infections)
c)operative theatre & hospital
ideal condition for this gas is low temperature, dry conditions
Chemicals:
a)wide spectrum of action
b)rapid action
c)effective even in pressure,
Organic subst.,pus,blood
d)no hypersensitivity contactdermatitis
Chemical Disinfectants
1)acids & alkalis
2)halogens
3)Chloroform
4)salts
5)antiseptics of phenol
6)formaldehyde
7)glutaraldehyde
8)various dyes & alcohols
9)aerosols & other gaseous

34: MICROBIAL FLORA OF H2O

Microbial flora of H2O:Water can be divided into
i. table water
ii.environmental water (lakes,sea,rivers)
the water can be contaminated by
pathogenic microbes
such as shigella,cholera,vibrions etc
Analysis
a)collection of sample
b)methods of analysis
1)presumptive coliform
a)multiple tube method
b)membrane filter method
2)differential method
Coliform can be used as sanitary important
Coli index/litre amount of bacteria coliform in 1 litre of water
Coli titre smallest amount of water containing 1 coliform
Microbial flora of air
Characterised by number of microbial cells in 1 m 3 of air
Measuring
1) On an open Petri dish,wait 1 hour for
fermentation on the
medium(not very effective)
2)By passing some amount of air thru Muller pump. After 24hours
at temp. 37 C, count the no. of colonies on agar medium.
Microbial flora of soil
The best place for microorganisms is 20cm below the
surface,because nutrition(organic waste) is waste, oxygenation is
present & absence of sunlight(UV)
Soil borne diseases are :Gas gangrene- Clostridium perfringens
Botulism - Clostridium botulinum
Mo that may cause disease may be alive for 2 months.

Sanitation microorganisms normal microflora present in

respiratory tract of humans.
1. These microbes cannot multiply through air.
2. Time of life must be same as that of pathogenic mo.
3. Their isolation and examination be easy.
4. They must be normal flora.
These are:Streptococci
} in
Staphylococci
} larynx
Absence of mo in air of operating theater is impossible.
Microbial no. before operation cannot be more than 500 & after
operation(1500)3 times.
LIVING TIME OF PATHOGEN IN ENVIRONMENT
In H2O
Salmonella Typhi - 2day 3months
Shigella
- 5 -9 days
V.Cholera El Tor - many months???
Tularemia pathogenfor days 3months
- 7 -150 days
In Soil
Salmonella Typhi
- 2 -3 weeks(max 12 months)
Shigella
- 1.5 5 weeks(max 9 months)
V.Cholera
- 1-2 weeks(max 4 months)
V.Cholera El Tor
- 4 weeks (max 6 months)
M.tuberculosis
- 13 weeks
(max 7 months)
Brucella
- 0.5 3 weeks(max 2 months)
E.Coli/
Clostridium Perfringens

36.INTERACTION BTW MO IN ASSOCIATION

SYMBIOSIS
I. Symbiosis relationship btw 2 or more mo in which
participants are of material aid & bent to one another
a)Commensalism 2species live together. 1 can get benefits
from this & the other either gets benefit or harmed
b)Parasitism 1 organism(parasite) gets benefits from the
relationship, while the others(host) is harmed.
c)mutualism 2 species living together. Both benefit from
the relationship.
II. Metabiosis - when one mo produce requirements that are
necessary for the other.
III. Synergism when one population cause positive
influence for others
IV. Antagonism one mo suppress life of

35.BACTERIOLOGICAL EXAMINATION OF H2O

1.Collection of samples :- in sterilized bottles, from tap
water,water from reservoir(sea,river,lakes).
Tubes in incubation for 24 hours & check growth of
coliform bacteria. Isolate the culture & innoculate 24hours
in MacConkeys agar with indicator & lactose(only E.coli
can ferment lactose).pH will change the colour of
indicator.
Then oxidation test is employed.
Microbial filters are used spores cannot pass through
such filter papers
All microorganisms in water will be on surface if
microfilters present.If we put filter in MacConkeys agar
for 24 hours & colonies are observed.
Then again oxidation test is employed.
Presence of E.coli is checked

NO.37 CHEMOTHERAPY
Chemotherapy
- treatment if infectious diseases by chemothereapeutic drugs.
Chemotherapy 1stly observed & used by Alexander Fleming to
observe staphylococci growth & it ( - ) the surrounded the
surrounding air-borne fungi.so he understood that one mo can
( - ) the growth of the other.
- But by Florin & chain produce 1st antibiotic as penicillin by
penicillium notatum
Then all scientists around the world was checking this
penicillin.They came back to Russia &did experiment & said that
both penicillium is effective.In Russia.they used
Cristasiu(penicillium crustosum)
Group of chemo.preparations
a) historically according to their origin.
natural
artificial
i. synthetic
e.g. penicillin
ii. Semisynthetic(the modification of natural antibiotic structure)
b) due to chemical synthesis.
1.-lactam(the -lactam ring as basic structure)
e.g. penicillin benzylpenicillin,aminopenicillin,carboxipicillin
2. tetracyclines
3. aminoglycosides i. 1st generation streptomycin
ii. 2nd generation gentamycine
iii. 3rd generation
4. macroslides (erythromycines,actinomycines)
5. Rifampicin (tuberculosis treatment)

c) due to spectrum of action

i.antibacterial ii.antiviral iii.antifungal iv.anticancer
d) class due to origin
- plants
- microorganism
-actinomycetes
- fungi
- animal tissues
e.g. plants (onion,garlic,mint)
fungi penicillin,cephalosporins
mo cholicine,polimycine
animal tissue ( IF ) leukocyte
lysozymes
chicken eggs
erithsia erythrocytes
actino all mycines
e.g. erythromycines
lincomycine
vancomycine

39.COMPLICATIONS AFTER USE OF ANTIBIOTICS

Many complications can occur after administration of antibiotics to
the patient.This is due to wrong administration of antibiotics.
1)Because all humans has a normal microbial flora in our body.They act
in different places of the body to give different functions to
prevent,protect against pathogenetic mo entering.If not humans
could get different kinds of pathogenic mo from
enteringeffectdeath in short time
So the normal microbial flora of body protect us from these
pathogenic mo.
Especially in stomach,intestine,there arre specialized microbial
flora(normal) that prevent multiplication of pathogenic mo because
GIT pathway is a very easy pathway for entrance of pathogenic
mo,e.g. thru bad food,drinks,contaminated H2O.Entering of
pathogenic microflora prevented,multiplication stopped and excreted
via faeces
This mechanism can be changed;effected due to antibiotic action of
GIT.So all the time with antibiotics,normal GIT flora must be
introducedto the body.This is done to prevent dysbiosis which causes
changing of the normal microflora from normal to abnormal.This
leads to different diseases as which spread by pathogenic mo.
2)Prolonged admin. of antibiotics leads to stomach or intestinal ulcers.
3)Allergy reaction before admin. antibiotics must do antibiotic
test\
i. disc diffusion
Ii .tube diffusion
if not different complications may arise e.g. allergyanaphylactic
shock in severe cases
4)Toxicity also in high dosage of antibiotics could have toxic
action
Prophylaxis : detect sensitivity before antibiotics
Treatments :
a) toxicity give a/biotics substitute(as detoxification,stop usage
a/biotics)
b) allergy must admin. antihistamine to stop anaphylactic shock
c) If disbalance gives correct a/biotics,give probiotics
d)Tolerance same treatment as allergic reaction

38.ANTIBIOTICS
class. of it due to chem. nature
- same as question 37
mechanism of action
a)block cell membrane:
Polymyxin B & polymyxin E bind selectively with surface
membrane of Gram (-) bacteria,are rich in phosphatidyl
lethanolamine act as cationic defrigens.( - ) of cell membrane
function leads to escape of macromolecules & ions from the
cells,resulting cell damage & death(act on permeability)
e.g.polymyxin B/ EI
b)drug ( - ) of protein synthesis & impairment of function: of
ribosomes,
e.g. Aminoglycosides
Tetracyclines
Chloramphnicol
c)drugs inhibiting synthesis of nucleic acids
e.g. Rifampin
( - ) oriculation of DNA mole:
e.g. quinolones,actinomyctes
d)drugs with antimetabolic action:
e.g. Sulphonamides,Sulfones,pAS,INH
act as antimetabolic drugs during bacterial metabolism:folic acid
is derived by pABA.The sulphonamide having a structural
similarity to pABA
e)IF suppress protein synthesis of virus
f)anticancer & antitumor drugs suppress the oxy metabolic
effects

40.DRUG RESISTANCE OF MICROORGANISMS

Drug resistance is of 2 types, primary & acquired
1)Primary resistance
:some bacteria posses an inate
property of resistance ro certain drug,e.g. resistance of E.coli to
penicillin
2)Acquired resistance:Acquired resistance results from either
from mutation or gene transfer.Spontaneous mutation is less
frequent but transfer of gene either through chromosome or
plasmid plays an important role in the spread of drug resistance
from bacterium to bacterium,both within and outside of species.
Mechanism of transmission
1)Permeability:MO change their cell wall permeability to drug by
possible alteration in chem. Nature of outer membrane e.g.
tetracycline resistance by Ps.aeruginosa.
2)Altered structural target:Aminoglycosides attach with 30S
subunit erythromycin with 50S subunit of ribosome.In
chromosomal rsistance,bacteria develop an altered receptor.
3)Production of enzymes:Staphylococcci resistant to penicillin
produce a beta-lactamase that destroys the drugs.
4)Altered metabolic pathway:By altering metabolic pathway of
drug,bacteria bypasses reaction inhibited by drug
Role of plasmids
Inactivation of antibiotics occurs by plasmid coded enzymes e.g.
beta lactamase destroying beta lactam ring responsible for the
antibacterial action of penicillins.
Method of transfer of plasmid & genetic material
1)Transduction: Plasmid DNA enclosed in bacteriophage is
transferred from one bacterium to another of same species.
2)Conjugation: R-factors carrying genes for drug resistance are
transferred by conjugation from donor to recipient.This is
commonest way of spread of multi drug rsistance among different
genera of Gram-negative bacteria.
3)Transpostion: Transfer of a short segment occurs between
plasmid to plasmid or between plasmid to
chromosome(transposons)which may encode a/biotic resistance.

4)Transformation: Transfer of naked DNA carrying genes for

drug resistance occur in bacterium by process of transformation.
Methods of examination of antibiotic sensitivity
1)Disc diffusion method
Stokes method: outer side of plate is inoculated with standard
organism and the test organism in the middle of the plate and he
zone of inhibition around the disc is compared.
Result: zone of inhibition above 12-15mm in diameter considered
significant.
Primary sensitivity testing: Urine or fluid specimen containing
bacteria is directly inoculated on a solid medium & medicated
discs are placed on the surface of the plate & incubated.
Concentration of medicated discs: Routinely used
concentration per disc is as follows:penicillin-10 units,ampicillin10 mcg,amoxycillin-10mcg,tetracyclines-30mcg & etc.
2)Tube dilution method
Laborious procedure but useful in assessment of therapeutic dose.
Method: Serial tube dilutions of the drug in broth are inoculated
with the bacterium under test and incubated.
Observation: Lowest concentration of drug inhibiting bacterial
growth represents minimum inhibitory concentration(MIC)
(bacteriostatic effect).Minimum bactericidal concentration(MBC)
is then determined by subculture from the tubes on MIC test into
solid media.Bacterial growth on solid media indicates presence of
surviving bacteria.The lowest concentration of drug that kills all
bacteria is found out from appropriate tube which will not show
any growth on subculture.
Ways of overcoming resistance
-carry out specific drug sensitivity tests to attain specific
therapeutic dose & not overdose of antibiotics which in the long
term may lead to drug resistance in MOs.