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z
Viruses
Chpt 13 Pg 384 – 414 of Tortora

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 z
LEARNING OUTCOMES

▪ Differentiate a virus from a bacterium.

▪ Explain the difference between enveloped and nonenveloped viruses.
▪ Define viral species.
▪ Describe how bacteriophages and animal viruses are cultured.
▪ Compare and contrast the lytic and lysogenic cycles of bacteriophages.
▪ Define oncogene and transformed cell.
▪ Discuss the relationship between viruses and cancer.
▪ Explain latent viral infections and give an example.
▪ Discuss how a proteins can be infectious.
▪ Differentiate between virus, viroid and prions
 z GENERAL CHARACTERISTICS

▪ The first clue to the existence of viruses was found by Friedrich Loeffler and Paul
Frosch (1898) when he discovered that the cause of foot-and-mouth disease in
livestock was an infectious particle smaller than any bacteria.
▪ Viruses are considered as non-living agents that can infect all life forms (phages
vs. animal viruses)
▪ Viral cultivation differs from bacterial cultivation methods
▪ There are around 1,500 known viruses, however it is estimated that there are
around 400,000 exist
▪ The development and advances in EM allowed for visualization of viruses
 z GENERAL CHARACTERISTICS

▪ Distinctive properties of viruses:

▪ Have a single type of nucleic acids (DNA or RNA)
▪ Have protein coat that surrounds the nucleic acid; which may or may not be
enclosed in an envelope
▪ Multiply in living cells using the machinery of the host cell
▪ Cause synthesis of specialized structures that are ble to transfer viral
nucleic acid to other cells

▪ They have few or no enzymes of their own for metabolism

 z GENERAL CHARACTERISTICS

▪ Host range
▪ Invertebrates, vertebrates, plants, protists, fungi, and bacteria
▪ Most viruses have high host specificity with very few exceptions
▪ Viruses that infect bacteria are called bacteriophages or phages
▪ Phage therapy → using bacteriophage to treat bacterial infection

▪ Viral size
▪ 20 – 1000 nm
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 z VIRAL STRUCTURE

▪ A virion → complete, fully developed infectious viral particle consisting of

nucleic acid surrounded by a protein coat
▪ Viruses are generally classified based on the type of nucleic acid and
differences in the structure of their coat
▪ Nucleic acid → DNA or RNA; single stranded (plus or minus strand) or double
stranded
▪ Capsid
▪ Nucleic acid is protected by protein coat called capsid
▪ Structure of capsid is determined by viral nucleic acid
▪ Accounts for most of the viral mass
▪ Each capsid is made up of protein subunits called capsomeres
▪ In some capsids, the capsomeres is made up of one type of
protein while others are made up of several types
 https://goo.gl/images/nS3zjX

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HIV virus
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 z VIRAL STRUCTURE

▪ Envelope
▪ Not all viruses are enveloped
▪ Consist of lipids, proteins and carbohydrates
▪ Some envelopes are covered with spikes
▪ Carbohydrate-protein (glycoprotein) complex projecting out of the
surface of envelope
▪ Can be used for identification
▪ Influenza virus are able to
clump red blood cells due to
spikes.
▪ Viruses that do not have
envelopes are known as
nonenveloped viruses.
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 z VIRAL STRUCTURE

▪ General morphology
▪ Helical virus
▪ nucleic acid found in the hollow cylindrical capsid
▪ Eg rabies virus and Ebola virus
▪ Polyhedral virus
▪ Capsid in the form of icosahedral (20 triangular faces and 12 corners)
▪ Eg adenovirus and poliovirus
▪ Enveloped virus
▪ Roughly spherical, covered with envelope
▪ Eg influenza virus and human herpes virus
▪ Complex virus
▪ Complicated structure
▪ Eg.  bacteriophage
 Taken from Tortora, Funke
and Chase. Microbiology: An
Introduction. (10 Ed.)
Pearson
 z TAXONOMY

▪ Classification based on type of NA, strategy for replication, and morphology.

▪ Family names end in –viridae
▪ Genus and species names end in -virus.

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 z TAXONOMY

▪ Classification based on type of NA, strategy for replication, and morphology.

▪ Order names ends with ~ales
▪ Family names end in –viridae
▪ Genus and species names end in -virus.
▪ Eg:

▪ Order: Herpevirales
▪ Family : Herpesviridae
▪ Genus: Herpesvirus
▪ Species: Human herpes virus HHV-1, HHV-2, HHV-3
https://goo.gl/images/3awWpc
 z TAXONOMY

▪ International Committee on Taxonomy of Viruses (ICTV)

▪ 7 orders, 96 families, 22 subfamilies, 420 genera, 2618 species have been
identified
▪ The orders are Caudovirales, Herpevirales, Ligamenvirales, Mononegavirales,
Nidovirales, Picornavirales and Tymovirales
Taken from Tortora, Funke and Chase. Microbiology: An Introduction. (12 Ed.) Pearson
Taken from Tortora, Funke and Chase. Microbiology: An Introduction. (12 Ed.) Pearson
Taken from Tortora, Funke and Chase. Microbiology: An Introduction. (12 Ed.) Pearson
Taken from Tortora, Funke and Chase. Microbiology: An Introduction. (12 Ed.) Pearson
 ISOLATION, CULTIVATION &
z
IDENTIFICATION

▪ Because viruses require living host cells to replicate, thus cultivating of

viruses has to be done in vivo

▪ There are generally several ways viruses are often cultivated:

▪ Using bacteria for bacteriophages
▪ Living animals
▪ Embryonated eggs
▪ Cell cultures

▪ Easiest virus to be cultivated are the bacteriophages

 ISOLATION, CULTIVATION &
z
IDENTIFICATION

▪ Growing bacteriophages
▪ Can either be grown in bacterial suspension (liquid media) or bacterial
cultures on agar
▪ Most common uses bacterial cultures on agar
▪ Phages are mixed with bacteria and added to molten agar. The mixture will
then be poured in Petri dishes and incubated
▪ Phages will infect bacterial cells, multiply and release new virions thus
causing bacterial cell lysis
▪ This lysis can be seen as clear zones → plaques
▪ Each plaque originates froma single viral particle and expressed as
plaque forming unit (PFU)
 ISOLATION, CULTIVATION &
z
IDENTIFICATION
Plaque Assay

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 Plaque Assay

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 ISOLATION, CULTIVATION &
z
IDENTIFICATION
▪ Growing animal viruses (in living animals)
▪ Eg. Mice, rabbits, guinea pigs
▪ Animals are inoculated with the targeted virus and observed for clinical
sign and symptoms
▪ Then the animals would be killed so that infected tissues can be
harvested and examined
▪ Advantages:
▪ Large quantities of viruses can be cultivated
▪ Immunological aspect of the infection can also be studied
▪ Disadvantages:
▪ Not all human viruses can grow in animal cells
▪ Animal rights issues
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 ISOLATION, CULTIVATION &
z
IDENTIFICATION

▪ Growing animal viruses (in embryonated eggs)

▪ A hole is drilled in the shell of embryonated eggs and the viral
suspension is injected into the egg’s fluid (or where it is most appropriate
for the virus).
▪ Viral growth is signalled by death of embryo, or formation of typical
pocks or lesions on the egg membrane
▪ Disadvantages:
▪ If used for viral vaccine development → may cause allergic reaction
to vaccine due to egg proteins that may be present in the vaccine
https://www.youtube.com/watch?v=Eq9JIq9HvMg

How we grow flu inside an egg

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Moving embryo inside egg

 ISOLATION, CULTIVATION &
z
IDENTIFICATION

▪ Growing animal viruses (in cell

cultures)
▪ Tissue culture → cells grown in
culture media in the laboratory
▪ Replaced embryonated eggs as
preferred viral cultivation
technique
▪ When viruses infect these cells,
they cause cells deterioration
called cytopathic effect (CPE)
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Taken from Tortora, Funke and Chase. Microbiology: An Introduction. (12 Ed.) Pearson
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 ISOLATION, CULTIVATION &
z
IDENTIFICATION

▪ Viral identification
▪ Not an easy task
▪ Viruses require electron microscope for visualization
▪ Serological method such as western blotting are the most commonly
used method
▪ Detection of viruses based on reaction towards antibodies
▪ Observing cytopathic effect can also help in identification
▪ Molecular method
▪ RFLP restriction fragment length polymorphism
▪ PCR – polymerase chain reaction
 z VIRAL MULTIPLICATION

▪ Viruses do not contain enzymes for energy production or protein synthesis.

▪ They hijack the host cells and use them as a factory to produce their
progeny; using the host cell metabolic machinery to produce viral enzymes
and components.

▪ Multiplication of bacteriophage
▪ Lytic cycle eg. T-even bacteriophage → lysis of host cells
▪ Lysogenic cycle eg. - bacteriophage
 z VIRAL MULTIPLICATION

Lytic Cycle

1. Attachment → Sites on the phage’s tail fibers attach to complementary

receptors on the bacterial cells
2. Penetration → Phage lysozyme opens a portion of the bacterial cell wall
while the tail sheath contracts to push the core through the cell wall. The
phage DNA then enters the bacterial cells while the capsid remains
externally
3. Biosynthesis → transcription of phage DNA → mRNA → proteins
necessary for phage multiplication. DNA is replicated and viral component
are produced
4. Maturation → phage DNA, capsids and other viral components
assembles to form complete virions
5. Release → phage lysozyme breaks the bacterial cell wall, releasing the
fully assembled virions.
Lytic Cycle of a T-Even Bacteriophage

Taken from Tortora, Funke and Chase. Microbiology: An Introduction. (12 Ed.) Pearson
Lytic Cycle of a T-Even Bacteriophage

Taken from Tortora, Funke and Chase. Microbiology: An Introduction. (12 Ed.) Pearson
 z VIRAL MULTIPLICATION

Lysogenic Cycle

1. During lysogenic cycle, phage DNA integrates into the bacterial

chromosome → prophage
2. Prophage is replicated every time the cell divides
3. In the presence of certain determining factors, prophage may exit the
chromosomal DNA of it’s host and initiates the lytic cycle
4. Lysogenic phage can transfer bacterial genes from one cell to another by
transduction (generalized or specialized)
▪ Three important outcome for lysogenic cycle
▪ Lysogenic cells are immune to reinfection by the same virus
▪ Phage conversion – changes of the host cell characteristics
▪ Specialized transduction
 z VIRAL MULTIPLICATION

Phage conversion

▪ Host cell may exhibit new

properties brought upon the phage
▪ Eg. Corynebacterium diphtheriae
causes the disease diphtheria by
the toxin that they produce. The
synthesis of this toxin is only
possible when the bacterial cell
carries a lysogenic phage.

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Taken from Tortora, Funke and Chase. Microbiology: An Introduction. (12 Ed.) Pearson
 z VIRAL MULTIPLICATION

▪ Three important outcome for lysogenic cycle

▪ Lysogenic cells are immune to reinfection by the same virus
▪ Phage conversion
▪ Specialized transduction
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 z VIRAL MULTIPLICATION

▪ Multiplication of animal viruses

▪ Varies depending on the type of viruses
▪ Generally go through five steps:
▪ Attachment
▪ Entry and uncoating
▪ Biosynthesis
▪ Maturation
▪ Release
z
Replication of DNA-containing viruses
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 z
DNA viruses

▪ Adenoviridae
▪ Adenoviruses – acute respiratory disease (common cold)
▪ Poxviridae – all diseases caused by poxvirus (smallpox, cowpox)
▪ Herpesviridae –
▪ Human herpes virus (HHV) 1 and 2 from the genus Simplexvirus →
coldsores
▪ HHV3 from genus Varicellovirus → chickenpox
▪ HHV4 from genus Lymphocryptovirus → mononucleosis
▪ HHV5 from genus Cytomegalovirus → CMV inclusion disease
▪ HHV6 and 7 from genus Roseolovirus → roseola
▪ HHV8 from genus Rhadinovirus → Kaposi’s sarcoma in AIDS
patients
 z
DNA viruses

▪ Papovaviridae – Papillomavirus →
warts. Some can transform normal
cells causing cancer
▪ Hepadnaviridae –
▪ Hepadnaviruses causes
hepatitis
▪ Differ from other DNA viruses
because the synthesize DNA by
copying RNA using viral
enzyme reverse transcriptase
z
Replication of RNA-containing viruses
 z
DNA viruses

▪ Picornaviridae
▪ Picornaviruses such as enteroviruses and poliovirus
▪ Smallest viruses
▪ Contain sense strand (+ strand) – can act directly as mRNA
▪ + strand → - strand → more + strand
▪ Togaviridae
▪ Enveloped viruses
▪ Includes arthropod-borne arboviruses from the genus Alphavirus
▪ Contain a single + strand RNA → - strand → more + strand
 z
DNA viruses

▪ Rhabdoviridae
▪ Rhabdoviruses such as rabies virus from genus Lyssavirus
▪ Contain a single - strand RNA → + strand → more – strand → more + strand
▪ Reoviridae
▪ Affect the respiratory and digestive system oh humans
▪ Contain dsRNA which will be digested upon entry → mRNA → capsid proteins
and – strand → + and – strand RNA pair up →dsRNA
Taken from Tortora, Funke and Chase.
Microbiology: An Introduction. (12 Ed.)
Pearson
 z RNA viruses that use DNA

▪ Includes retroviruses and oncogenic RNA viruses

Taken from Tortora, Funke and
Chase. Microbiology: An
Introduction. (12 Ed.) Pearson
 z VIRUSES AND CANCER

▪ The earliest relationship between cancer and viruses was discovered I the early
1900s via the transfer of chicken leukemia and chicken sarcoma by cell-free
filtrates.
▪ Transformation of normal cells into tumor cells
▪ Oncogenes – genes which in certain condition can transform a normal cell
into a tumor cell
▪ Viruses that can activate these oncogenes are called oncogenic viruses or
oncoviruses
▪ Approximately 10% of cancer cases are known to be virus-induced
▪ Tumor cells undergo transformation and many of them produces virus-
specific antigen on their cell surface →tumor specific transplantation antigen
(TSTA) or an antigen in their nucleus called the T-antigen
▪ Transformed cells tend to be irregularly shaped and exhibit certain
chromosomal abnormalities (unusual number of chromosome, fragmented
chromosome)
 z VIRUSES AND CANCER

▪ DNA oncoviruses
▪ Adenoviridae, Herpesviridae, Poxyviridae, Papovaviridae and
Hepadnaviridae
▪ Papillomaviruses → uterine cancer (cervical cancer)
▪ All cervical and anal cancers are caused by human papillomavirus
(HPV) – a vaccine is recommended for 11-12 yrs old girls and boys
▪ Epstein-Barr (EB) virus → Burkitt’s lymphoma
▪ Hepatitis B (HPB) → liver cancer
 z VIRUSES AND CANCER

▪ RNA oncoviruses (Retroviridae)

▪ The virus ability to produce tumors is related to the presence of
reverse transcriptase
▪ The DNA synthesized from the viral RNA becomes incorporated as a
provirus into the host’s cell’s DNA
▪ A provirus can remain latent, can produce viruses or can transform
the host cell
 z VIRUSES AND CANCER

▪ Viruses to treat cancer

▪ Viruses that can infect cancer cells thus causing lysis → oncolytic
viruses
▪ Includes adedovirus, vaccinia virus and Simplexvirus
▪ Viral therapy may provide treatment for some cancers

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 LATENT VS PRESISTANT VIRAL
z
INFECTION
▪ Latent viral infection
▪ Virus remain in host cells (provirus) for long period of time without producing
infection

▪ Eg. Cold sores and shingles

shingles

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 LATENT VS PRESISTANT VIRAL
z
INFECTION
▪ Persistant viral infection / chronic
Taken from Tortora, Funke and
viral infection Chase. Microbiology: An
Introduction. (12 Ed.) Pearson

▪ Viral disease processes that

occur over a long period of
time and generally are fatal
▪ Caused by conventional
viruses
Taken from Tortora, Funke and Chase. Microbiology: An Introduction. (12 Ed.) Pearson
 z PRIONS

▪ Infectious proteins first discovered in 1980s

▪ Small proteinaceous infectious particles (very resistant to inactivation)
▪ Inherited and transmissible by ingestion, transplant, and surgical
instruments
▪ Causes spongiform encephalopathies eg, sheep scrapie, Creutzfeldt-
Jakob disease (CJD), mad cow disease
▪ PrPC: Normal cellular prion protein, on cell surface. Involved in cell death.
▪ PrPSc: Scrapie protein; accumulates in brain cells, forming plaques.
▪ It is a result of altered protein due to mutation of the normal gene PrPC or
contact with the altered protein PrPSc
 z
PLANT VIRUSES AND VIROIDS

▪ Plant viruses similar to animal viruses in their

▪ Morphology
▪ Nucleic acid
▪ Some plant viruses can even multiply in insects
▪ Plant viruses can infect plants of economic importance such as beans (bean mosaic
virus), corn and sugarcane (wound tumor virus), and potatoes ( potato yellow dwarf
virus)
▪ Pathological effect on plant
▪ Color change
▪ Deformed growth
▪ Wilting
▪ Stunted growth
▪ Some viruses produce no symptoms as the plant serve as only reservoirs of infections
 z
PLANT VIRUSES AND VIROIDS

▪ Plant cells generally protect themselves from diseases by inpenetrable cell

thus viruses can only gain entry via wounds or plant parasites (nematodes,
fungi and insects)
▪ Once infected, disease can be spread by pollen
▪ Plant viruses are cultured in
▪ Protoplast (plant cells with cell wall removed)
▪ Insect cell culture
▪ Some plant disease are caused by viroids →short pieces of naked RNA
▪ 300-400 nucleotides
▪ No protein coat
▪ Eg of disease caused by viroid → potato spindle tuber disease
 Fruit distortion on eggplant fruit
Grapevine fanleaf virus
caused by Tomato bushy stunt virus

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Tomato plant leaf with Tobacco Mosaic Virus

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 z
LEARNING OUTCOMES

▪ Differentiate a virus from a bacterium.

▪ Explain the difference between enveloped and nonenveloped viruses.
▪ Define viral species.
▪ Describe how bacteriophages and animal viruses are cultured.
▪ Compare and contrast the lytic and lysogenic cycles of bacteriophages.
▪ Define oncogene and transformed cell.
▪ Discuss the relationship between viruses and cancer.
▪ Explain latent viral infections and give an example.
▪ Discuss how a proteins can be infectious.
▪ Differentiate between virus, viroid and prions