VIROLOGY

The smallest infectious agents and the smallest form

of life. Viruses can be viewed only by Electron
microscope.

General charachteristics of viruses:

1- The viral nucleic acid become active only when it
enters to the host cells.
2- Viruses are a live when they multiply in the host.

Differ from other infectious agents:

 They are very small.
 They need host cells for multiplication.
 Contain a single type of nucleic acid, DNA or RNA.
 Contain protein coat.
 Multiply inside living cells using the synthetic
machinary of the cell.
 Can transfer the viral nucleic acid to other cell.
 No ATP-generating system which is very important
for design of antiviral drugs as the drug will be toxic
for the virus and the host cell.
 The virus contains lipid layer so may be the virus
become sensetive to disinfectant or emulsifing agents
e.g. Bile salt and detergents.
HOST RANGE:
2) Animal viruses
3) Plant viruses
4) Bacterial viruses (bacteriophage).

• The specification of the virus to infect the

host cells depend on the requirment for the
viral multiplication found in the host cells.

• The virus interact with specific receptors

site on the surface of the cells-----hydrogen
bonds----by pilli or flagella
(Bacteriophage)--------Receptor sites are
plasma membrane of the host cells (animal
viruses).
CHEMICAL COMPOSITION OF THE VIRUSES:
1) Nucleic acid:
In a centeral core of protein include polymerase enzyme
Viruses are unique contain RNA or DNA, single or double
standed, Surrounded by capsid.
2) Protein Coat (Capsid):
Units that make up the capsid are called capsomers.
Protect the inside nucleic acid core. Complete infective
viral particles called virion which responsible for
infection.
3) Lipoprotein:
The nucleocapsid (nucleic acid + capsid) is covered by an
outer membrane called envelope (animal viruses).
Viruses without envelope called naked viruses.
SIZE:
20-300 nm, size can be determined by:
 Ultrafilter

 High speed centrifuge

 Electronic microscope

Viruses are obligate intracellular parasite because:

1- Defficiency in ATP generating system.
2- Absence of ribosomes which is essential for protein
synthesis.

Viruses envelope may or may not be covered by spikes

which are carbohydrate- protein complexes that project
from the surface of the envelope e.g: Influenza virus.
GENERAL MORPHOLOGY (Shape):
1) Helical virus:
Long rods may be rigid or flexible, their capsid is a
hollow cylinder with helical structure surrounded
nucleic acid e.g. Tobaco mosaic virus.
2) Polyhederal virus:
The capsid is an icosahederal form an equilteral
triangle e.g. Adenovirus, poliovirus.
3) Enveloped virus:
Roughly, spherical, but highly pleomorphic as the
envelope is not rigid. E.g.
Enveloped helical ---------------Influenza
Enveloped polhederal-----------HSV
4) Complex virus:
Bacterial viruses have very complicated structure
e.g. poxvirus, bacteriophage
CLASSIFICATION OF VIRUSES:
Old classification according to hot and organ:
Animal- Bacterial – Plant
Modern Classification:
Morphological classification
Type of nucleic acid
Size of capsid
Number of capsomers
Method of transmission

DNA----envelope: Herpes, poxvirus

Non envelope: adenoviruses
RNA-----Envelope: Retrovirus (HIV)
Non envelope : Polio, Retero virus
isolation, cultivation and identification of virus:
-The requirment of living host cells for multiplication
complicate their detection.
-Cultivation on living cells, plant or animals.
-Bacteriophage growing in bacteria culture.

Gwrwoth of bacteriophage:
-Can be grown on solid bacterial culture or suspension.
-Use solid medium may plaque which use for detection
and counting viruses:
Bacteriophae + host bacteria + melted agar-----poured
on a peteray plate containing a hardened layer of
agar----each virus infect bacterium, multiplies, infect all
bacteria cells----destroyed---plaques on surface agar---
uninfected bacteria---turbid background
PFU---------------Plaque forming unit.
BACTERIOPHAGE CLASSIFICATION:
1) Virulent phage:
-Host cells-----lytic cell
-The phage inject DNA inside host cell leaving the
protein material ouside.
-DNA enter host cell and control the genetic
machine of cell stimulating it to transcript the
DNA of the virus then capsid formed around
DNA and capsid -----rupture of bacteria---
viruses---mature phage.

2) Temperate phage:
-Host cells-----lysogenic as no. 1 but no rupture of
the bacteria due to DNA of virus integrated with
bacterial chromosome.
REQUIREMENTS FOR VIRAL GROWTH
 All viruses are obligate intracellular parasites but they
can survive in certain conditions as non-replicating
particles

 Temperature
Heat - there is great variability in the heat stability of
different viruses but icosahedral viruses tend to be
relatively stable, enveloped viruses are heat labile and
most pathogenic viruses are inactivated at 55-60oC
because their capsid protein is destroyed (an important
exception is the hepatitis virus. Cold - most viruses
can be preserved at sub-freezing temperatures, some
can withstand lyophilisation and can be stored in the
dry state at 4oC or even at room temperature while
enveloped viruses tend to lose infectivity after
prolonged storage at –90oC.
 pH
Most viruses are stable in the pH range 5-9.
Enteroviruses that have to pass through the stomach
can withstand low pHs. All viruses are destroyed by
alkaline conditions.

Radiation
UV produce damaging results on double-stranded DNA
that can cause inactivation of the virus. If conditions
are right the DNA can repair itself.
x-ray, gamma rays and beta particles inactivate
viruses..
 Stabilisation by Salts
magnesium chloride stabilises polioviruses, magnesium
sulphate stabilises influenza viruses and sodium
sulphate stabilises herpes virus. Important in the
preparation of vaccines e.g. non-stabilised polio vaccine
must be stored at <0oC whereas stabilised vaccine
remains potent for weeks at ambient temperature
which is an advantage when immunising in rural areas
 Ether Susceptibility and Lipid Solubility
Enveloped
viruses are inactivated by ether whereas non-enveloped
ones are not (simple efficient test for the presence of
envelopes).
Other organic solvents and sodium deoxycholate also
destroy the envelope.

 Detergents
Non-ionic detergents solubilise lipid constituents but do
not denature the proteins of the capsid. Anionic
detergents solubilise the lipid constituents and disrupt
the capsids into separated polypeptides

 50% Glycerol
Many viruses remain alive in 50% glycerol for many
years. Vaccinia virus is preserved in 50% glycerol for
many years while bacteria are killed.
 Formaldehyde
Destroys viral infectivity but has minimal adverse affect
on viral antigenicity and is used in the productions of
inactivated viral vaccines

 Photodynamic Inactivation
Heterocyclic dyes like neutral red, proflavine and
toluidine blue intercalate between the bases of
replicating viral nucleic acids and on exposure to light,
they become susceptible to inactivation

 Antibacterial Agents
Antibacterial antibiotics and sulphonamides have no
effect on viruses although some antiviral drugs have
been developed. Oxidising agents are the most effect
disinfectants e.g. hydrogen peroxide, hypochlorite.
Growth of animal viruses in the lab:
1) Living animal:
e.g. mice, rabbit, guinea pig
Styding immunoresponse, a clinical specimen with
virus inoculated the animal ----obseved signgs of
disease or killing animal and examine the tissue.

2) Embryonated eggs:
Conveniant, inexpensive, the viral suspension injected
into the fluid of the egg, viral growth is detected by
death of the embryo, or lession on the membrane of
the egg.
It is the most common method for viral growth and
isolation , and used for preperation of viral vaccine.
3) Cell culture:
The recent method for cultivation and grwoing of virus.
It is homogenous collection of cells.
More convneient than animal or egg.

Prepared by treated the animal tissue with enzyme ----

separate the individual cells.Suspended in solution with
nutrient and grwoth factor required by the cell---grow–
adhere to glass or plastic container----monolayer----by
virus infection---deterioration of cells (cytopathic effect
CPE)---plaques counted.
Cells isolated from human embryo, can live for hundared
generation e.g Hela cells.
Cell culture problems:

- Must be kept from microbial contamination.

- Need antibiotics for preventing contamination.
- Need experience technical worker.
- Identification of viral isolates is not easy so
immunological method mainly used as depend on
antibodies.
Multiplication of animal viruses:
Virus---attachement to the cell---penetration---
uncoating---replication---assembelly---release—
viruses.
 Effect of animal viral infection on host cells:
 CPE: inhibition of DNA or protein synthesis----
Damage of the cell.

Inclusion bodies:
 One type of CPE, a result of accumulation of viruses in
the nucleous or cytoplasm of the host cells or both.
 Other arise at sites of earlier viral synthesis.

 Important: usefual for identification of the particular

virus causing an infection.
 e.g. Rabies----Negri bodies in cytoplasm of nerve cells
and presented in the brain tissue----(Diagnostic test
for rabies, also for measles, vaccinia, small pox,
herpes.
 Polykeryocytes: infection cell accumulation from
myxoviruses.
Viruses and cancer:
Tumor: cell multipication in an uncontrolled way.
Malignant cell multiply in an uncontrolled way.
Most patients die from metastatic ( spread cancer to
other parts of the body) than primary tumor.
Leukomia: blood cnacer, increase white cells in blood.

Virus-cancer relation discover 1908, chicken leukomia

transfer from sick chiken to healthy.
In 1963: virus induced adenocarcinoma (cancer of
glanular tissue) in mice was discovered.

Cancer: The viral particles infect the cells without cancer

Cancer may not developed untill long infection
Cancer do not seem to be contagious.
 Transformation of the normal cell to tumor cell:

Alter the genetic material of the cell----cancerous cell

e.g. chemical, high-energy radiation, viruses
(oncogenic)

Reseptor gene®--------- Oncogen

1,2,3 chromosome
Reprosser protein oncogen blocked

Viral gene (V) Reprosor gene oncogen

The viral protein interfere with the reprossor protein so
the reprosor gene blocked and the oncogen will be
function
Interferons:

-Protect neighboring cells from viral infection.

-It is antimicrobial cytokines which interfer with viral
replication.
Inter. α,β interact with specific receptor or, produced
within 42 hour infection.

Inter. γ with oher receptor, a T-cell product, later

infection.

Protein kinase, 2’,5’ oligoadenylate synthatase----inhibit

the viral RNA translation----inhibit protein synthesis.

Used in chronic hepatitis B and AIDS treatment.

 Mechanism of Action of Interferon:
Interferon itself is not the anti-viral agent. It induces
an anti-viral state by prompting the synthesis of other
proteins that inhibit viral replication. The action is as
follows:
1) The viral nucleic acid stimulates the cell nucleus to
synthesise interferon which can through the cell
membrane into the tissue fluid. Interferon molecules
bind to the cell surface receptors which induce a signal
that is transmitted to the cell nucleus.
2) The binding triggers or the synthesis of several
enzymes responsible for the development of the anti-
viral state.
3) These enzymes block viral reproduction by inhibiting
the translation of viral mRNA while cellular DNA
remains unaffected.
4) Interferon molecules interact with components of the
immune system by modulating humoral (B
lymphocytes) and cellular (T lymphocytes) immunity
increasing antibody secretion and cytotoxicity and
enhancing phagocytic macrophage activity. Interferon
has broad cell-growth regulatory activities