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6
CHAPTER
CHAPTER OVERVIEW
Viruses:
Are a unique group of tiny infectious particles that are obligate
parasites of cells.
Do not exhibit the characteristics of life, but can regulate the
functions of host cells.
Infect all groups of living things and produce a variety of
diseases.
Are not cells but resemble complex molecules composed of
protein and nucleic acid.
159
160
TABLE 6.1
Properties of Viruses
Obligate intracellular parasites of bacteria, protozoa, fungi, algae,
plants, and animals.
Ultramicroscopic size, ranging from 20 nm up to 450 nm
(diameter).
Not cellular in nature; structure is very compact and economical.
Do not independently fulfill the characteristics of life.
Inactive macromolecules outside the host cell and active only inside
host cells.
Basic structure consists of protein shell (capsid) surrounding
nucleic acid core.
Nucleic acid can be either DNA or RNA but not both.
Nucleic acid can be double-stranded DNA, single-stranded DNA,
single-stranded RNA, or double-stranded RNA.
Molecules on virus surface impart high specificity for attachment to
host cell.
Multiply by taking control of host cells genetic material and
regulating the synthesis and assembly of new viruses.
Lack enzymes for most metabolic processes.
Lack machinery for synthesizing proteins.
INSIGHT 6.1
161
Discovery
Checkpoint
Viruses are noncellular entities whose properties have been
identified through technological advances in microscopy and
tissue culture.
Viruses are infectious particles that invade every known type
of cell. They are not alive, yet they are able to redirect the
metabolism of living cells to reproduce virus particles.
Viral replication inside a cell usually causes death or loss of
function of that cell.
polioviruses could be accommodated by an average human cell. Animal viruses range in size from the small parvoviruses1 (around
20 nm [0.02 m] in diameter) to poxviruses2 that are as large as
small bacteria (up to 450 nm [0.4 m] in length) (figure 6.1). Some
cylindrical viruses are relatively long (800 nm [0.8 m] in length) but
so narrow in diameter (15 nm [0.015 m]) that their visibility is still
limited without the high magnification and resolution of an electron
microscope. Figure 6.1 compares the sizes of several viruses with
procaryotic and eucaryotic cells and molecules.
Viral architecture is most readily observed through special
stains in combination with electron microscopy. Negative staining
uses very thin layers of an opaque salt to outline the shape of the virus
against a dark background and to enhance textural features on the viral surface. Internal details are revealed by positive staining of specific
parts of the virus such as protein or nucleic acid. The shadowcasting
technique attaches a virus preparation to a surface and showers it with
a dense metallic vapor directed from a certain angle. These techniques
are featured in several figures throughout this chapter.
1. DNA viruses that cause respiratory infections in humans.
2. A group of large, complex viruses, including smallpox, that cause raised skin swellings
called pox.
162
Rickettsia
0.3 m
Viruses
1. Poxvirus
2. Herpes simplex
3. Rabies
4. HIV
5. Influenza
6. Adenovirus
7. T2 bacteriophage
8. Poliomyelitis
9. Yellow fever
Streptococcus
1 m
(1)
(2)
Protein Molecule
10. Hemoglobin
molecule
250 nm
150 nm
125 nm
110 nm
100 nm
75 nm
65 nm
30 nm
22 nm
E. coli
2 m long
(10)
(9)
(8)
15 nm
(7)
(3)
(6)
(4)
(5)
YEAST CELL 7 m
FIGURE 6.1
Size comparison of viruses with a eucaryotic cell (yeast) and bacteria. Viruses range from largest (1) to smallest (9). A molecule of protein (10) is
included to indicate proportion of macromolecules.
163
Capsid
Nucleic acid
Fibers
DNA core
(a) Naked Nucleocapsid Virus
Envelope
FIGURE 6.2
A giant among virusesmimivirus. A section through the cell of a host
ameba captures one virus particle inside a vacuole. Note its geometric
shape, dark DNA core, and the fine surface fibers.
Spike
Capsid
Nucleic acid
FIGURE 6.4
Generalized structure of viruses. (a) The simplest virus is a naked
virus (nucleocapsid) consisting of a geometric capsid assembled around
a nucleic acid strand or strands. (b) An enveloped virus is composed of a
nucleocapsid surrounded by a flexible membrane called an envelope. The
envelope usually has special receptor spikes inserted into it.
(a)
Envelope (not
found in all viruses)
Virus
particle
Nucleic acid molecule(s)
(DNA or RNA)
Central core
Matrix proteins
enzymes (not found in
all viruses)
(b)
FIGURE 6.3
The crystalline nature of viruses. (a) Light microscope magnification
(1,200) of purified poliovirus crystals. (b) Highly magnified (150,000)
electron micrograph of the crystals, showing hundreds of individual viruses.
164
Capsomers
Capsid
Nucleocapsid
(a)
Nucleic acid
(a)
(b)
(b)
Spikes
Nucleic acid
Capsid begins
forming helix.
(c)
FIGURE 6.5
Assembly of helical nucleocapsids. (a) Capsomers assemble into hollow discs. (b) The nucleic acid is inserted into the center of the disc.
(c) Elongation of the nucleocapsid progresses from both ends, as the
nucleic acid is wound within the lengthening helix.
Envelope
Nucleocapsid
(c)
(d)
FIGURE 6.6
Typical variations of viruses with helical nucleocapsids. Naked helical virus (tobacco mosaic virus): (a) a schematic view and (b) a greatly
magnified micrograph. Note the overall cylindrical morphology. Enveloped
helical virus (influenza virus): (c) a schematic view and (d) a colorized
micrograph featuring a positive stain of the avian influenza virus
(300,000). This virus has a well-developed envelope with prominent
spikes termed H5N1 type.
(a) Capsomers
165
Facet
Capsomers
Vertex
Nucleic
acid
100 nm
(b)
Capsomers
Vertex
Fiber
Capsomers
(a)
Envelope Capsid DNA core
(c)
(d)
FIGURE 6.7
The structure and formation of an icosahedral virus (adenovirus is the
model). (a) A facet or face of the capsid is composed of 21 identical
capsomers arranged in a triangular shape. A vertex or point consist of
5 capsomers arranged with a single penton in the center. Other viruses
can vary in the number, types, and arrangement of capsomers. (b) An
assembled virus shows how the facets and vertices come together to
form a shell around the nucleic acid. (c) A three-dimensional model
(640,000) of this virus shows fibers attached to the pentons. (d) A negative
stain of this virus highlights its texture and fibers that have fallen off.
(b)
FIGURE 6.8
Two types of icosahedral viruses, highly magnified. (a) Upper view: A
negative stain of rotaviruses with unusual capsomers that look like spokes
on a wheel; lower view is a 3 dimensional model of this virus. (b) Herpesvirus simplex virus, a type of enveloped icosahedral virus (300,000).
166
A. Complex Viruses
B. Enveloped Viruses
Helical
Icosahedral
(1)
(3)
(5)
(2)
(4)
(6)
(7)
Icosahedral
A. Complex viruses:
(1) poxvirus, a large DNA virus
(2) flexible-tailed bacteriophage
(8)
B. Enveloped viruses:
With a helical nucleocapsid:
(3) mumps virus
(4) rhabdovirus
With an icosahedral nucleocapsid:
(5) herpesvirus
(6) HIV (AIDS)
(9)
C. Naked viruses:
Helical capsid:
(7) plum poxvirus
Icosahedral capsid:
(8) poliovirus
(9) papillomavirus
FIGURE 6.9
Basic types of viral morphology.
167
Capsid head
200 nm
Nucleic acid
Collar
Outer
envelope
Soluble
protein antigens
Sheath
Lateral
body
Tail
fibers
(a)
Tail
pins
Base plate
(c)
FIGURE 6.10
Detailed structure of complex viruses. (a) Section through the vaccinia
virus, a poxvirus, shows its internal components. (b) Photomicrograph and
(c) diagram of a T4 bacteriophage.
(b)
cells, which contain both DNA and RNA, viruses contain either
DNA or RNA but not both. Because viruses must pack into a
tiny space all of the genes necessary to instruct the host cell to
make new viruses, the number of viral genes is quite small compared with that of a cell. It varies from four genes in hepatitis B
virus to hundreds of genes in some herpesviruses. By comparison, the bacterium Escherichia coli has approximately 4,000
genes, and a human cell has approximately 30,000 to 40,000
genes. These additional genes allow cells to carry out the complex metabolic activity necessary for independent life. Viruses
possess only the genes needed to invade host cells and redirect
their activity.
In chapter 2 you learned that DNA usually exists as a doublestranded molecule and that RNA is single-stranded. Although most
viruses follow this same pattern, a few exhibit distinctive and
exceptional forms. Notable examples are the parvoviruses, which
contain single-stranded DNA, and reoviruses (a cause of respiratory and intestinal tract infections), which contain double-stranded
RNA. In fact, viruses exhibit wide variety in how their RNA or
DNA is configured. DNA viruses can have single-stranded (ss)
or double-stranded (ds) DNA; the dsDNA can be arranged linearly
or in ds circles. RNA viruses can be double-stranded but are more
often single-stranded. You will learn in chapter 9 that all proteins
are made by translating the nucleic acid code on a single strand of
RNA into an amino acid sequence. Single-stranded RNA genomes
168
TABLE 6.2
Order
Family
Genus
Species
dsDNA
Caudovirales
Poxviridae
Orthopoxvirus
Vaccinia virus
neg (ss)RNA
Mononegavirales
Paramyxoviridae
Morbillivirus
Measles virus
pos (ss)RNA
Nidovirales
Togaviridae
Rubivirus
Rubella virus
Adapted from van Regenmortel, M., editor, et al. 2000. Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses. New York:Academic Press.
that are ready for immediate translation into proteins are called
positive-sense* RNA. Other RNA genomes have to be converted
into the proper form to be made into proteins, and these are called
negative-sense RNA. RNA genomes may also be segmented, meaning that the individual genes exist on separate pieces of RNA. The
influenza virus (an orthomyxovirus) is an example of this. An RNA
virus with some unusual features is a retrovirus, one of the few
virus types that can change its nucleic acid from RNA to DNA.
Whatever the virus type, these tiny strands of genetic material
carry the blueprint for viral structure and functions. In a very real
sense, viruses are genetic parasites because they cannot multiply
until their nucleic acid has reached the internal habitat of the host
cell. At the minimum, they must carry genes for synthesizing the viral capsid and genetic material, for regulating the actions of the
host, and for packaging the mature virus.
*sense The sense of the strand relates to its readability into a protein.
*replication (rep-lih-kay-shun) L. replicare, to reply. To make an exact duplicate.
*polymerase (pol-im-ur-ace) An enzyme that synthesizes a large molecule from smaller
subunits.
169
TABLE 6.3
Important Human Virus Families, Genera, Common Names, and Types of Diseases
Family
Genus of Virus
Name of Disease
DNA Viruses
Poxviridae
Orthopoxvirus
Smallpox, cowpox
Herpesviridae
Simplexvirus
Adenoviridae
Varicellovirus
Cytomegalovirus
Mastadenovirus
Papovaviridae
Papillomavirus
Polyomavirus
Hepadnaviridae
Hepadnavirus
Parvoviridae
Erythrovirus
Parvovirus B19
Erythema infectiosum
Picornaviridae
Enterovirus
Hepatovirus
Rhinovirus
Poliovirus
Coxsackievirus
Hepatitis A virus (HAV)
Human rhinovirus
Poliomyelitis
Hand-foot-mouth disease
Short-term hepatitis
Common cold, bronchitis
Calciviridae
Calicivirus
Norwalk virus
Togaviridae
Alphavirus
Eastern equine
encephalitis (EEE)
Western equine
encephalitis (WEE)
Yellow fever
St. Louis encephalitis
Rubella (German measles)
RNA Viruses
Rubivirus
Flaviviridae
Flavivirus
Dengue fever
West Nile fever
Bunyaviridae
Bunyavirus
Hantavirus
Phlebovirus
Nairovirus
Bunyamwera viruses
Sin Nombre virus
Rift Valley fever virus
CrimeanCongo hemorrhagic
fever virus (CCHF)
California encephalitis
Respiratory distress syndrome
Rift Valley fever
CrimeanCongo
hemorrhagic fever
Filoviridae
Filovirus
Ebola fever
Reoviridae
Coltivirus
Rotavirus
Orthomyxoviridae
Influenza virus
Influenza virus,
type A (Asian, Hong Kong,
and swine influenza viruses)
Influenza or flu
Paramyxoviridae
Paramyxovirus
Morbillivirus
Pneumovirus
Parainfluenza
Mumps
Measles (red)
Common cold syndrome
Rhabdoviridae
Lyssavirus
Rabies virus
Rabies (hydrophobia)
Retroviridae
Oncornavirus
Lentivirus
T-cell leukemia
Acquired immunodeficiency
syndrome (AIDS)
Arenaviridae
Arenavirus
Lassa virus
Lassa fever
Coronaviridae
Coronavirus
Bronchitis
Coronavirus enteritis
Severe acute respiratory
syndrome
170
Characteristics used for placement in a particular family include type of capsid, nucleic acid strand number, presence and type
of envelope, overall viral size, and area of the host cell in which the
virus multiplies. Some virus families are named for their microscopic appearance (shape and size). Examples include rhabdoviruses,* which have a bullet-shaped envelope, and togaviruses,*
which have a cloaklike envelope. Anatomical or geographic areas
have also been used in naming. For instance, adenoviruses* were
first discovered in adenoids (one type of tonsil), and hantaviruses
were originally isolated in the Korean Province of Hantaan. Viruses
can also be named for their effects on the host. Lentiviruses* tend
to cause slow, chronic infections. Acronyms made from blending
several characteristics include picornaviruses,* which are tiny
RNA viruses, and reoviruses (or respiratory enteric orphan
viruses), which inhabit the respiratory tract and the intestine and are
not yet associated with any known disease state.
*tropism (troh-pizm) Gr. trope, a turn. Having a special affinity for an object or
substance.
171
Spikes
2
3
Nucleus
4. Synthesis: Replication and Protein Production.
Under the control of viral genes, the cell
synthesizes the basic components of new viruses:
RNA molecules, capsomers, spikes.
RNA
New
spikes
New
capsomers
New
RNA
FIGURE 6.11
General features in the multiplication cycle of an enveloped animal virus.
other viruses will vary in exact details of the cycle.
Using an RNA virus (rubella virus), the major events are outlined, although
172
Receptor
Host cell
membrane
Receptor
FIGURE 6.12
The mode by which animal viruses adsorb to the host cell membrane. (a) An enveloped coronavirus with prominent spikes. The configuration of
the spike has a complementary fit for cell receptors. The process in which the virus lands on the cell and plugs into receptors is termed docking. (b) An
adenovirus has a naked capsid that adheres to its host cell by nestling surface molecules on its capsid into the receptors on the host cells membrane.
Uncoating step
Host cell membrane
Virus in
vesicle
(a)
Specific
attachment
Free
DNA
Engulfment
Host cell
membrane
Free
RNA
Receptors
Uncoating of
nucleic acid
Receptor-spike
complex
(b)
Irreversible
attachment
Entry of
nucleocapsid
Membrane
fusion
FIGURE 6.13
Two principal means by which animal viruses penetrate. (a) Endocytosis (engulfment) and uncoating of a herpesvirus. (b) Fusion of the cell
membrane with the viral envelope (mumps virus).
173
4. For enveloped viruses, these terms are interchangeable. They mean the release of a virus
from an animal cell by enclosing it in a portion of membrane derived from the cell.
Viral nucleocapsid
Host cell membrane
Viral glycoprotein spikes
Cytoplasm
Capsid
RNA
Budding
virion
(a)
Free infectious
virion with envelope
Viral
matrix
protein
(b)
FIGURE 6.15
Maturation and release of enveloped viruses. (a) As parainfluenza virus is budded off the membrane, it simultaneously picks up an envelope and
spikes. (b) AIDS viruses (HIV) leave their host T cell by budding off its surface.
174
Inclusion bodies
Giant cell
Multiple nuclei
(a)
(b)
FIGURE 6.16
Cytopathic changes in cells and cell cultures infected by viruses. (a) Human epithelial cells (400) infected by herpes simplex virus demonstrate
multinucleate giant cells. (b) Fluorescent-stained human cells infected with cytomegalovirus. Note the inclusion bodies (1000). Note also that both viruses
disrupt the cohesive junctions between cells.
cell. Regardless of how the virus leaves, most active viral infections
are ultimately lethal to the cell because of accumulated damage.
Lethal damages include a permanent shutdown of metabolism and
genetic expression, destruction of cell membrane and organelles,
toxicity of virus components, and release of lysosomes.
A fully formed, extracellular virus particle that is virulent
(able to establish infection in a host) is called a virion (vir-ee-on).
The number of virions released by infected cells is variable, controlled by factors such as the size of the virus and the health of the
host cell. About 3,000 to 4,000 virions are released from a single
cell infected with poxviruses, whereas a poliovirus-infected cell
can release over 100,000 virions. If even a small number of these
virions happens to meet another susceptible cell and infect it, the
potential for rapid viral proliferation is immense.
TABLE 6.4
Smallpox virus
Herpes simplex
Adenovirus
Poliovirus
Reovirus
Influenza virus
Rabies virus
Measles virus
the cell harbors the virus and is not immediately lysed. These socalled persistent infections can last from a few weeks to the remainder of the hosts life. One of the more serious complications
occurs with the measles virus. It may remain hidden in brain cells
for many years, causing progressive damage and loss of function.
Several viruses remain in a latent state,5 periodically becoming reactivated. Examples of this are herpes simplex viruses (cold sores and
genital herpes) and herpes zoster virus (chicken pox and shingles).
5. Meaning that they exist in an inactive state over long periods.
Both viruses can go into latency in nerve cells and later emerge under the influence of various stimuli to cause recurrent symptoms.
Specific damage that occurs in viral diseases is covered more completely in chapters 24 and 25.
Some animal viruses enter their host cell and permanently alter
its genetic material, leading to cancer. These viruses are termed oncogenic, and their effect on the cell is called transformation. A startling feature of these viruses is that their nucleic acid is consolidated
into the host DNA. Transformed cells have an increased rate of
growth; alterations in chromosomes; changes in the cells surface
molecules; and the capacity to divide for an indefinite period, unlike
normal animal cells. Mammalian viruses capable of initiating tumors
are called oncoviruses. Some of these are DNA viruses such as papillomavirus (genital warts are associated with cervical cancer), herpesviruses (Epstein-Barr virus causes Burkitts lymphoma), and hepatitis B virus. Two viruses related to HIVHTLV I and II6are
involved in human cancers. These findings have spurred a great deal
of speculation on the possible involvement of viruses in cancers
whose cause is still unknown. Additional information on the connection between viruses and cancer is found in chapters 9 and 16.
Checkpoint
Virus size range is from 20 nm to 450 nm (diameter). Viruses
are composed of an outer protein capsid enclosing either
DNA or RNA plus a variety of enzymes. Some viruses also
exhibit an envelope around the capsid.
Viruses go through a multiplication cycle that generally
involves adsorption, penetration (sometimes followed by
uncoating), viral synthesis and assembly, and viral release
by lysis or budding.
These events turn the host cell into a factory solely for making and shedding new viruses. This results in the ultimate
destruction of the cell.
Viruses are capable of lying dormant in their host cells,
possibly becoming active at some later time.
Animal viruses can cause acute infections or can persist in
host tissues as chronic latent infections that can reactivate
periodically throughout the hosts life. Some persistent animal
viruses are oncogenic.
known as the T-even phages such as T2 and T4. They are complex in
structure, with an icosahedral capsid head containing DNA, a central
tube (surrounded by a sheath), collar, base plate, tail pins, and fibers
(see figure 6.10b,c). Momentarily setting aside a strictly scientific
and objective tone, it is tempting to think of these extraordinary
viruses as minute spacecrafts docking on an alien planet, ready to
unload their genetic cargo.
T-even bacteriophages go through similar stages as the animal viruses described earlier (figure 6.17). They adsorb to host
bacteria using specific receptors on the bacterial surface. Although
the entire phage does not enter the host cell, the nucleic acid penetrates the host after being injected through a rigid tube the phage inserts through the bacterial membrane and wall (figure 6.18). This
eliminates the need for uncoating. Entry of the nucleic acid causes
the cessation of host cell DNA replication and protein synthesis.
Soon the host cell machinery is used for viral replication and synthesis of viral proteins. As the host cell produces new phage parts,
the parts spontaneously assemble into bacteriophages.
An average-sized Escherichia coli cell can contain up to 200
new phage units at the end of this period. Eventually, the host cell
becomes so packed with viruses that it lysessplits openthereby
releasing the mature virions (figure 6.19). This process is hastened
by viral enzymes produced late in the infection cycle that digest the
cell envelope, thereby weakening it. Upon release, the virulent
phages can spread to other susceptible bacterial cells and begin a
new cycle of infection.
175
176
Release of viruses
Bacteriophage
Bacterial
DNA
Lysogenic State
Viral
DNA
1
Adsorption
Penetration
Lytic
Cycle
DNA
splits
Spliced
viral
genome
3
Viral
DNA
Duplication of phage
components; replication of
virus genetic material
Maturation
Bacterial
DNA molecule
Capsid
Tail
Assembly of
new virions
DNA
Tail fibers
Sheath
Bacteriophage
FIGURE 6.17
Events in the multiplication cycle of T-even bacteriophages. The lytic
cycle (17) involves full completion of viral infection through lysis and
release of virions. Occasionally the virus enters a reversible state of
lysogeny (left) and is incorporated into the hosts genetic material.
Other bacteria that are made virulent by their prophages are Vibrio
cholerae, the agent of cholera, and Clostridium botulinum, the
cause of botulism. On page 174 we described a similar relationship
that exists between certain animal viruses and human cells.
The cycles of bacterial and animal viruses illustrate general
features of viral multiplication in a very concrete and memorable
way. See table 6.5 for a summary of their most important
differences.
177
Head
Bacterial
cell wall
Cell
wall
Tube
Viral nucleic acid
Cytoplasm
(b)
(a)
FIGURE 6.18
Penetration of a bacterial cell by a T-even bacteriophage. (a) After adsorption, the phage plate becomes embedded in the cell wall, and the sheath
contracts, pushing the tube through the cell wall and membrane and releasing the nucleic acid into the interior of the cell. (b) Section through Escherichia coli
with attached phages. Note that these phages have injected their nucleic acid through the cell wall and now have empty heads.
FIGURE 6.19
A weakened bacterial cell, crowded with viruses. The cell has ruptured and released numerous virions that can then attack nearby susceptible host cells. Note the empty heads of spent phages lined up around
the ruptured wall.
TABLE 6.5
Animal Virus
Adsorption
Penetration
Occurs in cytoplasm
Cessation of host synthesis
Viral DNA or RNA is replicated and
begins to function
Viral components synthesized
Viral Persistence
Lysogeny
Cell Destruction
Immediate
Immediate or delayed
178
Checkpoint
Bacteriophages vary significantly from animal viruses in
their methods of adsorption, penetration, site of replication,
and method of exit from host cells.
Lysogeny is a condition in which viral DNA is inserted
into the bacterial chromosome and remains inactive for
an extended period. It is replicated right along with the
chromosome every time the bacterium divides.
Some bacteria express virulence traits that are coded for by
the bacteriophage DNA in their chromosomes. This phenomenon is called lysogenic conversion.
(a)
One problem hampering earlier animal virologists was their inability to propagate specific viruses routinely in pure culture and in sufficient quantities for their studies. Virtually all of the pioneering
attempts at cultivation had to be performed in an organism that was
the usual host for the virus. But this method had its limitations.
How could researchers have ever traced the stages of viral multiplication if they had been restricted to the natural host, especially in
the case of human viruses? Fortunately, systems of cultivation with
broader applications were developed, including in vitro* cell (or
tissue) culture methods and in vivo* inoculation of laboratory-bred
animals and embryonic bird tissues. Such use of substitute host systems permits greater control, uniformity, and wide-scale harvesting
of viruses.
The primary purposes of viral cultivation are:
1. to isolate and identify viruses in clinical specimens;
2. to prepare viruses for vaccines; and
3. to do detailed research on viral structure, multiplication
cycles, genetics, and effects on host cells.
(b)
(c)
FIGURE 6.20
Microscopic appearance of normal and infected cell cultures. Microscopic views of (a) A normal, undisturbed layer of vero cells. (b) Plaques,
which consist of open spaces where cells have been disrupted by viral
infection. (c) Petri dish culture of E. coli bacteria shows macroscopic
plaques (clear, round spaces) at points of infection by phages.
179
Inoculation
of amniotic cavity
Inoculation
of embryo
Air sac
Inoculation of
chorioallantoic
membrane
Amnion
Shell
Allantoic
cavity
Inoculation of
yolk sac
Albumin
(a)
A technician inoculates fertilized chicken eggs with viruses in the first
stage of preparing vaccines. This process requires the highest levels of
sterile and aseptic precautions. Influenza vaccine is prepared this way.
(b)
The shell is perforated using sterile techniques, and a virus preparation is
injected into a site selected to grow the viruses. Targets include the allantoic cavity, a fluid-filled sac that functions in embryonic waste removal; the
amniotic cavity, a sac that cushions and protects the embryo itself; the
chorioallantoic membrane, which functions in embryonic gas exchange;
the yolk sac, a membrane that mobilizes yolk for the nourishment of the
embryo; and the embryo itself.
FIGURE 6.21
Cultivating animal viruses in a developing bird embryo.
Cultures of animal cells usually exist in the primary or continuous form. Primary cell cultures are prepared by placing freshly
isolated animal tissue in a growth medium. The cells undergo a series of mitotic divisions to produce a monolayer. Embryonic, fetal,
adult, and even cancerous tissues have served as sources of primary
cultures. A primary culture retains several characteristics of the
original tissue from which it was derived, but this original line generally has a limited existence. Eventually, it will die out or mutate
into a line of cells that can grow continuously. These cell lines can
be continuously subcultured, provided they are routinely transferred to fresh nutrient medium. One very clear advantage of cell
culture is that a specific cell line can be available for viruses with a
very narrow host range. Strictly human viruses can be propagated
in one of several primary or continuous human cell lines, such as
embryonic kidney cells, fibroblasts, bone marrow, and heart cells.
One way to detect the growth of a virus in culture is to observe
degeneration and lysis of infected cells in the monolayer of cells. The
areas where virus-infected cells have been destroyed show up as clear,
well-defined patches in the cell sheet called plaques (figure 6.20b).
Plaques are essentially the macroscopic manifestation of cytopathic
effects (CPEs), discussed earlier. This same technique is used to detect
and count bacteriophages, because they also produce plaques* when
grown in soft agar cultures of their host cells (figure 6.20c). A plaque
*plaque (plak) Fr. placke, patch or spot.
develops when the viruses released by an infected host cell radiate out
to adjacent host cells. As new cells become infected, they die and release more viruses, and so on. As this process continues, the infection
spreads gradually and symmetrically from the original point of infection, causing the macroscopic appearance of round, clear spaces that
correspond to areas of dead cells.
180
Discovery
INSIGHT 6.2
Artificial Viruses Created!
Newspapers are filled with stories of the debate over the ethics of creating life through cloning techniques. Dolly the cloned sheep and the cattle, swine, and goats that have followed in her footsteps have raised
ethical questions about scientists playing God, when they harvest genetic material from an animal and create an identical organism from it,
as is the case with cloning.
Meanwhile, in a much less publicized event, scientists at the State
University of New York at Stony Brook succeeded in artificially creating
a virus that is virtually identical to natural poliovirus. They used DNA
nucleotides they bought off the shelf and put them together according
to the published poliovirus sequence. They then added an enzyme that
would transcribe the DNA sequence into the RNA genome used by
poliovirus. They ended up with a virus that was nearly identical to poliovirus (see photograph). Its capsid, infectivity, and replication in host
cells are similar to the natural virus.
The creation of the virus was greeted with controversy, particularly
because poliovirus is potentially devastating to human health. The scientists, who were working on a biowarfare defense project funded by the
Department of Defense, argued that they were demonstrating what could
be accomplished if information and chemicals fell into the wrong hands.
In 2003, another lab in Rockville, Maryland, manufactured a working bacteriophage, a harmless virus called phi X. Their hope is to create microorganisms from which they can harness energyfor use as a
renewable energy source. But the prospect of harmful misuse of the new
death of the embryo, defects in embryonic development, and localized areas of damage in the membranes, resulting in discrete,
opaque spots called pocks (a variant of pox). If a virus does not
produce overt changes in the developing embryonic tissue, virologists have other methods of detection. Embryonic fluids and tissues can be prepared for direct examination with an electron microscope. Certain viruses can also be detected by their ability to
agglutinate red blood cells (form big clumps) or by their reaction
with an antibody of known specificity that will affix to its corresponding virus, if it is present.
technology has prompted scientific experts to team with national security and bioethics experts to discuss the pros and cons of the new technology, and ways to ensure its acceptable uses.
What basic materials, molecules, and other components would be
required to create viruses in a test tube?
Answer available at www.mhhe.com/talaro6
Checkpoint
Animal viruses must be studied in some type of host cell
environment such as cell cultures, bird embryos, or
laboratory animals.
Cell and tissue cultures are cultures of host cells grown
in special sterile chambers containing correct types and
proportions of growth factors using aseptic techniques to
exclude unwanted microorganisms.
Virus growth in cell culture is detected by the appearance of
plaques.
Eggs are used to cultivate viruses for vaccines. Inoculation
of animals is an alternate method for viruses that do not
readily grow in cultures or embryos.
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Medical
INSIGHT 6.3
Uncommon Facts About the Common Cold
The common cold touches the lives of humans more than any other viral
infection, afflicting at least half the population every year and accounting for millions of hours of absenteeism from work and school. The reason for its widespread distribution is not that it is more virulent or transmissible than other infections, but that symptoms of colds are linked to
hundreds of different viruses and viral strains. Among the known
causative viruses, in order of importance, are rhinoviruses (which cause
about half of all colds), paramyxoviruses, enteroviruses, coronaviruses,
reoviruses, and adenoviruses. A given cold can be caused by a single
virus type or it can result from a mixed infection.
The name implies a condition caused by cold weather or drafts. But
studies in which human volunteers with wet heads or feet chilled or exposed to moist, frigid air have failed to show that cold weather alone causes
colds. Most colds occur in the late autumn, winter, and early springall
periods of colder weatherbut this seasonal connection has more to do
with being confined in closed spaces with carriers than with temperature.
The most significant single factor in the spread of colds is contamination of hands with viruses in mucous secretions. The viruses usually
invade through the mucous membranes of the nose and eyes. The most
common symptom is a nasal discharge, and the least common is fever,
except in infants and children.
Finding a cure for the common cold has been a long-standing goal
of medical science. This quest is not motivated by the clinical nature
(Insight 6.3). If one also takes into account prominent viral infections found only in certain regions of the world, such as Dengue
fever, Rift Valley fever, and yellow fever, the total could easily
exceed several billion cases each year. Although most viral infections do not result in death, some, such as rabies, AIDS, and Ebola,
have very high mortality rates, and others can lead to long-term
debility (polio, neonatal rubella). Current research is focused
on the possible connection of viruses to chronic afflictions of
unknown cause, such as type I diabetes, multiple sclerosis, and
various cancers.
Dont forget that despite the reputation viruses have for being
highly detrimental, in some cases, they may actually show a beneficial side (see Insight 6.1).
The diseases associated with prions are known as transmissible spongiform encephalopathies (TSEs). This description
recognizes that the diseases are spread from host to host by direct
contact, contaminated food, or other means. It also refers to the effects of the agent on nervous tissue, which develops a spongelike
appearance due to loss of nerve and glial cells (see figure 25.29b).
Another pathological effect observed in these diseases is the
buildup of tiny protein fibrils in the brain tissue (figure 6.22a). Several forms of prion diseases are known in mammals, including
scrapie in sheep, bovine spongiform encephalopathy (mad cow disease) in cattle, and wasting disease in elk, deer, and mink. These
diseases have a long latent period (usually several years) before the
first symptoms of brain degeneration appear. The animals lose
coordination, have difficulty moving, and eventually progress to
collapse and death (figure 6.22b).
Humans are host to similar chronic diseases, including
Creutzfeldt-Jakob syndrome (CJS), kuru, fatal familial insomnia,
and others. In all of these conditions, the brain progressively deteriorates and the patient loses motor coordination, along with
sensory and cognitive abilities. There is no treatment and most
cases so far have been fatal. Recently a variant of CJS that appeared in Europe was traced to people eating meat from infected
cows. Several hundred people developed the disease and nearly
100 died. This was the first indicator that prions of animals could
cause infections in humans. It sparked a crisis in the beef industry
and strict controls on imported beef. Although no cases of human
disease have occurred in the United States, infected cattle have
been reported.
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Brain
cell
Prion
Fibrils
(a)
(b)
FIGURE 6.22
Some effects of prion-based diseases. (a) Cow in the early phase of bovine spongiform encephalopathy. Symptoms are a staggering gait, weakness,
and weight loss. (b) An isolated brain cell with prion fibrils on its surface.
in the viral life cycle you learned about earlier in this chapter.
Azidothymide (AZT), a drug used to treat AIDS, targets the nucleic acid synthesis stage. A newer class of HIV drugs, the protease inhibitors, disrupts the final assembly phase of the viral life
cycle. Another compound that shows some potential for treating
and preventing viral infections is a naturally occurring human cell
product called interferon (see chapters 12 and 14). Vaccines that
stimulate immunity are an extremely valuable tool but are available for only a limited number of viral diseases (see chapter 15).
We have completed our survey of procaryotes, eucaryotes,
and viruses and have described characteristics of different representatives of these three groups. Chapters 7 and 8 explore how microorganisms maintain themselves, beginning with microbial nutrition (chapter 7) followed by metabolism (chapter 8).
Checkpoint
Viruses are easily responsible for several billion infections
each year. It is conceivable that many chronic diseases of
unknown cause will eventually be connected to viral agents.
Other noncellular agents of disease are the prions, which are
not viruses at all, but protein fibers; viroids, extremely small
lengths of protein-coated nucleic acid; and satellite viruses,
which require larger viruses to cause disease.
Viral infections are difficult to treat because the drugs that
attack the viral replication cycle also cause serious side
effects in the host.
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184
Multiple-Choice Questions
1. A virus is a tiny infectious
a. cell
b. living thing
c. particle
d. nucleic acid
c. fungi
d. all organisms
8. The nucleic acid of animal viruses enters the host cell through
a. injection
c. endocytosis
b. fusion
d. b and c
Concept Questions
13. Clear patches in cell cultures that indicate sites of virus infection are
called
a. plaques
c. colonies
b. pocks
d. prions
185
16. Label the parts of these viruses by indicating the capsid, nucleic acid,
envelope, and other features. Can you identify them as to capsid type
and group?
Concept Questions
These questions are suggested as a writing-to-learn experience. For each
question, compose a one- or two-paragraph answer that includes the factual
information needed to completely address the question.
1. a. Describe 10 unique characteristics of viruses (can include
structure, behavior, multiplication). (159, 160)
b. After consulting table 6.1, what additional statements can
you make about viruses, especially as compared with
cells? (160163)
2. a. Explain what it means to be an obligate intracellular
parasite. (160)
b. What is another way to describe the sort of parasitism exhibited by
viruses? (161)
3. a. Characterize viruses according to size range. (161, 162)
b. What does it mean to say that they are ultramicroscopic? (161)
c. That they are filterable? (161)
4. a.
b.
c.
d.
e.
f.
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Critical-Thinking Questions
Critical thinking is the ability to reason and solve problems using facts and
concepts. These questions can be approached from a number of angles, and
in most cases, they do not have a single correct answer.
1. a. What characteristics of viruses could be used to characterize them
as life forms?
b. What makes them more similar to lifeless molecules?
2. a. Comment on the possible origin of viruses. Is it not curious that
the human cell welcomes a virus in and hospitably removes its
coat as if it were an old acquaintance?
b. How do spikes play a part in the action of the host cell?
3. a. If viruses that normally form envelopes were prevented from
budding, would they still be infectious? Why or why not?
b. If the RNA of an influenza virus were injected into a cell by itself,
could it cause a lytic infection?
4. The end result of most viral infections is death of the host cell.
a. If this is the case, how can we account for such differences in the
damage that viruses do (compare the effects of the cold virus with
those of the rabies virus)?
b. Describe the adaptation of viruses that does not immediately kill
the host cell and explain what its function might be.
5. a. Given that DNA viruses can actually be carried in the DNA of the
host cells chromosomes, comment on what this phenomenon
means in terms of inheritance in the offspring.