Course of Veterinary Virology

(Vetm 3075)

Samara University,

College of Veterinary Medicine and Animal science

Instructor: Yasin Mohammed

(DVM, MSc)
 Objectives

 At the end of this session you should be able to :

– Define virology
– Understand the evolution of virus
– Understand the mechanism of viral replication
– Differentiate between virus and other microbes
– Describe viral classification methods
– Understand the virus culture system
 1. General virology

History and development of virology

Virology is a science studding;

– Characteristics

– Morphology
– Nomenclature and Classification of viruses
– Diseases caused by them and the measures fighting
against them
 Veterinary virology is one of the specialties of microbiology
concerned with the science of viruses/the study of viruses/.
 Previously: Viruses were considered to be “venoms/toxins/”
that are unique classes of infectious agents which are
extremely small, containing only one type of nucleic acid
(DNA/RNA) and have an absolute dependence on living cells
for replication (unaccepted definition).
 Is a virus equivalent to mere toxin/venom/poison? Why?
 Current/recent operational/accepted/ meaning of
viruses:
Due to filtration experiments, viruses are defined as
“filterable infectious agent/poisonous fluid” which

 are ultramicroscopic infectious entities

 are acellular agents/particles with simple organization

(genome , capsid with or without envelope)

 are obligate intracellular entities/particles

 need living cells for replication

 possess single type of genome (DNA/RNA) enclosed in

protein capsid/ coat and sometimes additional envelope)

 They cannot be observed using a light microscope

 Individuals show no increase in size

 have no metabolic systems, including (reproduction,

biosynthesis, energy generation).

 have no any functional organelle/s/.

 Pass through bacteriological filters

 Viroids

• Viroids are free RNA molecules of low molecular weight

without any protein coat while viruses can have either RNA or
DNA molecules encapsulated in a protein coat.
• Viroids are smaller in size than viruses.
• Viroids infect only plants whereas virus infects all types of
organisms.
 Brief history of veterinary virology

 “The age of virology does not exceed a century”.

 Who contributed to the beginning of field of
virology ?.
 Answer: Many

1. Adolf Mayer (1882): German

 He observed diseased tubacco plants which were
mottled and he referred it “mosaic disease”.
 …

 He made many investigations and he could not

detect any bacteria and fungi excluding their
possibility as in tobacco disease.
 However, he speculated that a" soluble,
enzyme like infectious particle was involved.
• Ivanofsky and Beijerinck (1892-1898) conducted filtration
experiments and suggested that the cause was a previously
unrecognized “infectious agent”.
• The term “virus” was first used by Beijerinck
• Beijerinck also described his finding as “ living contagious
fluid”
• They collected a sample of infected tobacco leaves and
prepared a fluid extract
 …
• Then, they passed the fluid extract through standard filter pore

size of 300nm (0.3µm) that can retain the smallest known

bacteria (prepared a filtrate).

• Bacteria, fungi and other large sized agents remain on the upper

side of the filter paper.

• They injected healthy tobacco plants with the filtrate and

immediately normal tobacco plants were infected by the filtrate.

• They did this experiment repeatedly.

• Finally,

• They concluded that it was a filterable infectious-replicating

agent in the live host (live tobacco plant), thus not a mere

toxin/venom.

• Later on (1935-1939), others further characterized using

electron microscope and crystallography and named as ,

“tubacco mosaic virus”.

Tobacco mosaic virus isolation procedure
• In 1955, Rosalind Franklin proposed the full structure of the
tobacco mosaic virus.
• Many other plant viruses were then identified and
characterized.
• In 1898, J. Loffler and P. Frosch discovered the first vertebrate
virus, foot and mouth disease virus (FMD virus).
• In 1898, Sanarelli discovered Myxoma virus.
• Reed and Carroll discovered the first human virus, yellow
fever virus.
• These early studies provided the definition of viruses as
“filterable agents.”
• What helped the discovery of today’s enormous
types of viruses?
• It is the invention/development of more advanced
laboratory diagnostic techniques
– electron microscope, PCR technique, nucleic acid
sequencing technology, development of invitro cell culture
and other just close to the beginning of 20th century.
 …
• Due to development of these techniques which assist
the study and characterization of viruses, many more
viruses were discovered then just yet.
• Some viruses discovered then include (African swine
fever virus, Fowl plague virus (Avian influenza virus),
Rinderpest virus, Bluetongue virus, Rabies virus, and
others).
 Land marks in development of virology and virus research
Date virologist Discovery

1796 Edward Jenner Small pox vaccine developed

1885 Louis Pasteur Rabies vaccine developed

1892 Ivanofski Describe the first “filterable infectious agent- tobacco mosaic virus
(TMV)

1998 Freidrich Loeffler &Paul First demonstration of a filterable animal virus: foot and mouth disease
Frosch virus.

1900 Walter Reed Discovered the cause of yellow fever

1903 P. Remlinger Discovered rabies virus

1904-1908 V. Ellerman et.al First demonstrated a leukemia- causing virus-retrovirus

1915 Frederick T. Discovered bacteriophages

1917 Felix d’Herelle Discovered viruses of bacteria and coins the term bacteriophage

1931 J.furth Used mice as hosts for viruses

 Contd ’

Date virologist Discovery

1938 Max Theiler Developed a live attenuated vaccine against yellow fever

1939 Helmuth Ruska Used an electron microscope to take the first picture of viral particles
1953 W.P. Rowe Discovered adenoviruses
1955 F. L. schaffaer & C. Crystallized poliovirus
EScherdt
1957 Alick Isaacs & jean Discovered interferon ( antiviral agents)
Linderman
1960 C Gajdusek Discovered prions
1963 Baruch Blumberg Discovered hepatitis B virus (HBV)
1967 Theoder Diener Discovered virioids agent of plant disease which have no protein capsid

1970 Howard Temin & David Independently discovered reverse transcriptase in retroviruses
Baltimore
1973 Peter Doherty &m Rolf Demonstrated the basis of antigenic recognition by the cellular immune system
Zinkernagl
1977 WHO many workers Eradicated smallpox
1981 Yorio Hinuma & colleagues Isolated human T-cell leukaemia virus (HTLV) p
1982 Stanley Prusiner Demonstrated infectious proteins (prions), which cause scrapie in sheep

1983 Luc montainer and Robert Announced the discovery of human immunodeficiency virus (HIV), the
Gallo causative agent of AIDS
 Characteristics of viruses

 Complete mature virus particles or virions are simple,

noncellular particles (nucleic acid + Capsid +/- envelope).
 Possess single type of nucleic acid (either DNA/RNA)
 They need living host cell for reproduction
 Lack any functional organelle
 Smaller than other microorganisms (20-400nm)
 Only studied by electron microscope
 Outside the living cells, viruses are inert particles, whereas
they are considered as living in the living host cells
 Lack all metabolism systems (biosynthetic machineries,
energy generation systems, processes for reproduction and
others)
 No cellular structure
 could not be cultivated / can’t be grown/ on artificial
media/nonliving media. Why?
– Have no metabolic systems of their own
– Depend on host cell machinery for energy production, replication
and biosynthesis
 Living medium is required for cultivation of viruses
 Viruses are unaffected by antibiotics
Nonenveloped (naked virus)
Enveloped virion
 Viral structure (Morphology

• Structurally viruses are much simple than

bacteria.
• Structure of a virion
 The nature of viruses

The virions of most viruses are too small to be seen with a light
microscope and can be seen only with an electron microscope.
The units in which virions are normally measured are nanometres
(1 nm = 10−9 m).
Amongst the smallest are parvoviruses, with diameters about 20
nm, while
The microbe-mimicking virus (mimivirus), isolated from an amoeba,
is amongst the largest then herpes and pox virus.
 Viral protein functions

• Facilitate transfer of viral nucleic acid from one host

cell to another cell.
• Protect viral genome against inactivation by host
nucleases
• Enhance virus attachment to host cell surface
• Provide structural symmetry of the viral particle
 Structural components of viruses

Main structural components of viruses include:

1. Genome (total number of genes in a virion)
2.Capsid (composed of capsomeres)
3. Envelope (host derived lipid bilayer membrane)
4. Nucleocapsid (genome +capsid)
5. Surface proteins ( eg, peplomers)
 1. Viral genome

•Viruses contain a single type of nucleic acid, either DNA or RNA, not
both
•Nucleic acids encode genetic information necessary for virus replication
(synthesis of viral components and viral enzymes for replication).
•Viral genomes may have forms like ssDNA, dsDNA, circular DNA,
ssRNA, ds RNA and others
• single or segmented
•circular or linear molecule of nucleic acid functioning as the genetic
material of the virus
The nucleic acid of RNA viruses is usually single

stranded (ss), with the exception of the Reoviruses

and Birnaviruses (dsRNA).

The genome may be linear (most RNA viruses) or /

circular

 Viral RNAs can be

1. Monopartite (unsegmented/single) or

2. Multipartite (segmented):
 …

Retroviruses, Paramyxoviruses, Rhabdoviruses,

Picornaviruses, and others have monopartite

genomes.

Orthomyxoviruses, Bunyaviruses, and Arenaviruses

have multipartite genomes

• Viruses with ssRNA are divided into two groups:

• If the RNA of the genome has the same polarity as

the viral mRNA and can thus function directly as
messenger RNA.
– it is called a plus-strand (or positive-strand) or “sense”
strand RNA viruses.

• Examples of positive polarity RNA viruses:

Picornaviruses, Caliciviruses, Coronaviruses, etc,.
•
 If the genome RNA has the polarity opposite to that
of the mRNA, and therefore can not be translated
into proteins until it has first been transcribed into a
complementary strand,
 it is called a minus-strand (or negative-strand) or
“antisense” RNA strand and
 …

 The viruses are antisense or minus-strand viruses.

 Examples of negative polarity RNA viruses:

Paramyxoviruses, Rhabdoviruses, Orthomyxoviruses,
etc,.
 2. Capsid

• Capsid is a protein coat enclosing and stabilizing viral nucleic

acids
• It is a protein coat of N.A., act as a protective container.
• The arrangement of capsomers determines the virus symmetry.

Capsid composition and organization

 Capsid is composed of many smaller protein units called
capsomeres
 Many capsomeres organize/assemble/ to form capsid protein
/coat
 Cont’d…

Function of capsid:

Protect the viral genome( nucleic acid) from physical destruction

and enzymatic inactivation by host nuclease

Assist transfer of viral genome to cross host plasma membrane

Provides binding sites

Facilitates the assembly and packing of viral genetic information

Provides the structural symmetry to the virus particle

 Cont’d…

 It participates in attachment of virions to susceptible

cells (for naked viruses).

 It determines the antigenicity of the virion.

 Basic morphology of viruses

• The capsid of a virion can take on a variety of

geometric shapes characteristic for many viral
families.

• Capsid is constructed from morphological units

(capsomeres)

• Each capsomere is made from clusters of structural

units (protomers)
 Cont’d…

 Based on the arrangement of morphologic units

(capsomeres), the viral architecture can be grouped into

three shapes/symmetries

A) Icosahedral

B) Helical and

C) Complex symmetry
 Cont…

Icosahedral symmetry

Viruses have 20 equilateral triangular faces

12 corners or apices and

30 edges. Eg Birnaviridae, reoviridae

 Cont…
• The vertices of the icosahedron are pentameric
capsomeres (12 pentamers (pentoses)
– Example. The bacteriophage

• Therefore, there are 60 protomers in the simplest

capsid.
• In more complex capsids, the faces and edges are
constructed from hexoses (hexamers).
• Icosahedral viruses may be naked or enveloped.
• Icosahedral naked virion means a virus with only two
structural components (nucleic acid and capsid)
• These groups of viruses lack additional lipid membrane
over the capsid.
• Structural components of naked icosahedral virions
• (Genome + capsid (icosa capsid))
• Most DNA viruses are naked icosahedral viruses.
• Example
– Adenoviruses, Polyomaviruses, Parvoviruses, Papillomaviruses and
others

• Icosahedral virions can be enveloped (possess an

additional lipid membrane of the capsid):
– Examples of enveloped viruses include Herpesviruse,
Hepadnavirusese ……
 B. Helical symmetry

• This is an other type of cylinder shaped capsid

symmetry in which the protein structural units are
arranged as a helix (spiral path), hence the term
helical.
• In helically symmetric nucleocapsids, the genomic
RNA forms a spiral within the nucleocapsid.
Helical capsid symmetry
Influenza virus
Helical enveloped RNA Viruses
• Very few RNA nucleocapsids are naked (Tobacco
mosaic virus)
Enveloped viruses
Retroviruses (HIV)
 C. Complex symmetry
• Some viral particles do not exhibit either simple
icosahedral or helical symmetry.
• Rather, they are more complicated in structure ,
hence complex type of symmetry,
• In some cases, these can be mixtures of
arrangements containing both helical and
icosahedral symmetries.
• Examples of viruses with complex type of symmetry.
• Poxviruses, Bacteriophages
Bacteriophage
Complex symmetry (poxvirus)
 3. Envelope

• Many animal viruses have host derived lipid bilayer outer

membrane enclosing the nucleocapsid
• These type of virus groups are referred to as enveloped
viruses
(Orthomyxoviruses, Paramyxoviruses…….)
• Those viruses have no outer lipid membrane are called
naked viruses
(eg, Adenovirus, Picornavirus,… )
• This membrane is derived from host cell during virus budding process

• Depending on the virus type and site of replication

The membrane may be derived from

– Plasma membrane
– Nuclear membrane
– endoplasmic membrane/Golgi apparatus

• depending upon the location in the cell where of the virus replicates
(composition of each membrane differs).
• Enveloped virus groups include Poxvirus, Herpesvirus, Paramyxovirus,
orthomyxovirus, etc.,.
 Cont’d…

• It is a lipid or lipoprotein enclosed the capsid

• lipid component derived directly from host cell but protein

component is virus-coded.

• In addition to capsid and internal proteins, the two types of proteins

associated with the envelope are:

i. Spike -virus specific protein- frequently glycoproteins present in

the from of spike – like projections on the surface, which attach to
the host cell receptors during the entry of the virus into the cell.
II. Lipoprotein- derived from the host cell or nuclear membrane in
a process called budding.
Some viruses contain Matrix protein which is found as an
interior layer inside the envelope.
Unlike enveloped viruses, viruses with out envelope are
named as naked viruses.
 Enveloped viruses are sensitive to ether due to their lipid content.
Treatment by ether will result in loss of infectivity.
• The infectivity of most enveloped viruses depends on the integrity of
envelope.
• Viral envelope contains glycoprotein which is virus encoded.
– It is responsible for the interaction with the cellular receptors and represents an

important viral antigen.

• The presence of an envelope confers instability on the virus as they

are more sensitive to heat, drying, detergents and lipid solvents as

alcohol and ether.

 4. Peplomers (surface protein)

• These are glycoprotein projections having enzymatic

/adsorption/ and or haemagglutinating activity
Exercises
1.What are the main components of
1. Naked viruses
2. Enveloped viruses
2. Define a virion?
3. How do you differentiate viruses from other
microbes?
 ….cond

•Generally, viruses are infectious agent with both living

and non living characteristics

•Viruses are usually termed as “at the edge of life” because they lack any
functional organelles except nucleic acid

•They possess attributes of both aliveness and inanimate things.

1. Living characteristics of viruses

•They reproduce at fantastic rate, but only in living host

cells

•They can mutate

 Cont’d…

2.Non –living characteristics of viruses

–Do not contain cellular organelles

–They carry out no metabolism on their own and must replicate using the

host cell’s metabolic machinery. In other words, viruses don’t grow and

divide, instead new viral components are synthesized and assembled

with in the infected host cell

–The vast majority viruses possess DNA or RNA but never both

•  
 Forms of Life
Prokaryotes (unicellular) and Eukaryotes (multicellular) organisms

☺Are viruses classified under the class of prokaryotic or eukaryotic organisms ?

• Eukaryotic cells have a true nucleus, bound by a double membrane.

• Prokaryotic cells have no nucleus.
• Eukaryotic DNA is linear; prokaryotic DNA is circular (has no ends).

• The cytoplasm of eukaryotic cells is filled with a large, complex collection of

organelles, many of them enclosed in their own membranes; the prokaryotic cell
contains no membrane-bound organelles which are independent of the plasma
membrane. This is a very significant difference, and the source of the vast majority of
the greater complexity of the eukaryotic cell.
Unicellular micro organisms can be arranged in order of decreasing size and
complicity:

 Protozoa, Fungi, Bacteria, Rickettsia, Mycoplasma & Chlamydia.

 Contr asti ng proper ti es of Unic el lular MOs & Vir uses

Property Bacteria Rickettsia Mycoplasma Chlamydia Viruses

> 300 nm in diameter + + + + −

Grow on non living media + − + − −

Binary fission + + + + −

Contain both DNA & RNA + + + + −

Nucleic acid infectious − − − − +

Ribosomes + + + + −

Metabolism + + + + −

Sensitivity to antibiotics + + + + −
Cont.
 Reasons for studying viruses
1. Some viruses cause disease
Mild infection ………….lethal & devastating infection

2. Some viruses are useful

 Phage typing of bacteria: Some groups of bacteria, such as some Salmonella
species, are classified into strains on the basis of the spectrum of phages to
which they are susceptible.
 Sources of enzymes: A number of enzymes used in molecular biology are
virus enzymes. Examples include reverse transcriptases from retroviruses
and RNA polymerases from phages
 Anti-bacterial agents: phages were used to treat some bacterial infections
Viral nucleic acid
• Viral DNA or RNA can be double stranded (ds) or single stranded (ss), circular or linear,
segmented or non-segmented.

• Most viral genomes are quite fragile once they are removed from their capsid.

• Genome of all DNA viruses consists of a single molecule, which is ds except Parvovirus &
Circoviruses, circular (Papovaviruses & Circoviruses) or linear (poxviruses).

• RNA viruses may be ss or ds, segmented (Orthomyxoviruses), linear (Picornaviruses).

• The NA of viruses of certain families of both DNA and RNA viruses is infectious.
Virus genome
 Viruses have 2 phases in their life cycle:

 Outside the host cell (in env’t)- virion is metabolically inert/passive.

- It is the transmission phase of virus
- Extracellular form and susceptible
 
 Inside host cell (intracellular): it is the reproductive form
- Metabolically active (virus contain active viral genes)
-viral proteins that assemble to form new virions.

• Many viruses can reproduce themselves when only when the NA genome enters the cell,
i.e their nucleic acid is infectious.

 Larger sized viruses like herpes virus & pox virus are more independent of cellular
functions than the smaller viruses.

= larger viruses are more susceptible to antiviral chemotherapy because more virus-specific
processes are available as targets for drug action.
 
 Viral reaction to Physical & Chemical agents
1) Temperature (heat and cold)

 Most viruses are inactivated if incubated for 30 min. at 56°C.

 There are heat resistant virus as serum hepatitis.

 Preservation at 4°C for only several days, while preservation for

month, or years must be at -70°c or lower in liquid nitrogen.
 Some viruses are sensitive to repeated freezing and thawing.

 Enveloped viruses are much more heat labile, rapidly dropping in titer
at 37oc.
 Viral infectivity is generally destroyed by heating at 50 to 60 oc for 30
minutes.
2) Salts

• Viruses are preferentially stabilized by certain salts (MgCl2, Na2SO4, MgSO4)

3) Radiation

• Viruses are inactivated by UV light, x-ray and high-energy particles.

• Visible light is destructive to many viruses .

• UV light usually destroy them much more rapidly.

• Ionizing radiation (x-rays or -rays) causes break, in the nucleic acids and therefore inactivate it.

4) pH

• Viruses remain viable at PH values 6.5-7.5, but high acidity or alkalinity destroys many viruses except
enteroviruses which can resist acidic environment.

• All viruses are destroyed by alkaline conditions.

5) Ether and chloroform
• the infectivity of enveloped viruses is destroyed readily by these lipid solvents
• Enveloped viruses are inactivated by lipid solvents while non enveloped viruses are
resistant
• Viruses inactivated by ether: Herpes, Orthomyxo, paramyxo, Rhabdo, Retroviruses
• Viruses resistant to ether: parvovirus, Papovavirus, Adenovirus, Picornavirus,

6) Detergents
• Detergents can solubilize viral envelop & disrupt capsids
• detergents are used commonly by virologists to solubilize viral envelopes and to
liberate proteins for use as vaccines

 
 
7) Antibiotics & Other Antibacterial agents

• Antibiotics have no effect on viruses i.e. viruses are resistant to antibiotics because
they have no metabolic activity and due to their structure

8) Antiviral drugs

• Not common and only effective prophylactically or in the early stages of disease
Literally viruses can be grouped as:
Typical viruses: these are particles composed of an
internal core
Atypical viruses: There are virus-like agents
Types of atypical viruses:
1- Defective viruses
2- Pseudoviruses
3- Viroids
4- Prions