Acellular Pathogens

Viruses

Virus= Latin for poison!

Virion: Viral particle
Characteristics of Viruses:
● Infectious, acellular pathogens.
● Considered ACELLULAR due to its inability to reproduce independently
● Obligate intracellular parasites with host and cell-type specificity.
● DNA or RNA genome (never both).
● Genome is surrounded by a protein capsid and, in some cases, a phospholipid
membrane studded with viral glycoproteins.
● Lack genes for many products needed for successful reproduction, which it requires an
exploited host-cell genome to produce.

Host Range: The range of host cells a virus is capable of infecting. Often very limited.

Bacteriophage: Virus that infects bacteria.

Vector: Means by which a virus is spread from one host to another:

● Mechanical Vector: The virus is spread by physical contact with a contaminated object.
Can be a contaminated but uninfected animal, which would act less as a carrier and
more as a fomite.
● Biological Vector: Virus is directly transmitted to the new host by biting.

Viral Structures:
● Capsid: A protein coat surrounding the viral genome and a handful of reproductive
enzymes. Present in all viruses. Can be helical, polyhedral, icosahedral, or complex.
● Capsomere: Protein subunit of the capsid.
● Viral Envelope: Small portion of phospholipid membrane stolen from original host cell,
now used to protect viral capsid. Can be intracellular or cytoplasmic in origin. Only
present in enveloped viruses.
● Spikes: Protein structures that allow the virus to attach and enter a cell.
● Sheath: A shaft connecting the capsid to tail fibers and tail pins. Present in complex
bacteriophages.

Viral Genome Descriptors:

● Nonenveloped / Naked Viruses: Viruses composed of only a capsid and a genome.
● Enveloped Viruses: Viruses with a capsid surrounded by a protective viral envelope.
● ds-: Double-stranded
● ss-: Single-stranded
● +: Positive
● -: Negative
 The Viral Life Cycle

Most DNA viruses replicate in the nucleus, with the exception of some very large ones, which
replicate in the cytoplasm.

RNA viruses replicate only in the cytoplasm.

Bacteriophages, since prokaryotes lack organelles, replicate only in the cytoplasm

Virulent Phages: Viruses that kill the host cell through cell lysis.

Temperate Phages: Viruses that become part of the host cell’s chromosome and are replicated
with the cell genome until induced to make new viruses (Progeny Viruses).

Lytic Cycle: Found in virulent phages.

1. Attachment: The phage attaches to the surface of the host.
2. Penetration: The viral DNA enters the host cell.
3. Biosynthesis: Phage DNA replicates and phage proteins are made.
4. Maturation: New phage particles are assembled.
5. Lysis: The cell lyses, releasing the newly made phages.

Lysogenic Cycle: Found in temperate phages.

1. The phage infects a cell.
2. Phage DNA is incorporated into host genome.
3. Cell divides and prophage DNA is passed on to daughter cells.
4. Under stressful conditions, the prophase DNA is excised from the bacterial chromosome
and enters the Lytic Cycle, starting Biosynthesis.

Prophage: Integrated phage genome.

Lysogen: Prophage infected host.

Lysogeny: The process by which a bacterium is affected by a temperature phage. The

associated change is called lysogenic or phage conversion.

Transduction: Occurs when a bacteriophage transfers bacterial DNA from one bacterium to
another during sequential infections.
● Generalized Transduction: A piece of bacterial chromosomal DNA is transferred by the
phage during the lytic cycle.
● Specialized Transduction: Occurs at the end of the lysogenic cycle, when the
prophage is excised and bacteriophage enters the lytic cycle. Prophage can technically
be replicated as part of the host, due to being integrated into the host genome.
1. Viral Attachment and Penetration: The phage infects the cell.
 2. Integration: Phage DNA is incorporated into host genome.
3. Excision: The phage is excised from the bacterial chromosome along with a
short piece of bacterial DNA. The DNA is then packaged into newly formed
capsids.
4. Infection: Phage containing both viral and bacterial DNA infects a new host cell.
5. Recombination: Both the phage and attached bacterial DNA are incorporated
into the new host’s genome.

Life Cycle of Viruses With Animal Hosts:

1. Virus attaches to target cell.
2. Cell engulfs the virus by endocytosis.
3. Fusion and uncoating: viral genome released.
4. Viral genome enters the nucleus where it is replicated by viral RNA or DNA polymerase.
5. Viral mRNA is used to make viral proteins.
6. New particles are made and released into the extracellular fluid. The cell, which is not
killed in the process, continues to produce viral particles.
-ssRNA viruses cannot be translated by host ribosomes until --ssRNA is replicated into
+ssRNA by viral RNA-dependent RNA polymerase (RdRP).

Life Cycle of Viruses With Plant Hosts: Rarely lethal to the host. Usually RNA genome,
+ssRNA specifically.

Retrovirus: A type of +ssRNA virus that uses reverse transcriptase within the capsid to make
a dsDNA copy of its genome to insert into the host. The inserted viral genome is called a
provirus.

Latent Virus: Viruses capable of remaining dormant inside the cell either inside or outside the
host genome.

Viral Growth Curve:

1. Inoculation: Inoculum of virus binds to cells. Number of infectious virions briefly drops.
2. Eclipse: Virions penetrate cells. Number of infectious virions stays at a constant low.
3. Burst: Host cells release many viral particles. Number of infectious virions rises sharply.
4. Burst Size: Total number of virions released per bacterium. Number of infectious virions
plateaus.

6.3. Isolation, Culture, and Identification of Viruses

● Define “filterable agent” and explain why this term was used to describe viruses

o Infected host cells are cultured and grown, and then the growth medium can
be harvested as a source of virus.
o Virions in the liquid medium can be separated from the host cells by either
centrifugation or filtration.
 o Filters can physically remove anything present in the solution that is larger
than the virions; the viruses can then be collected in the filtrate.

● Very briefly describe the cultivation of viruses

o Grown in vivo (within a whole living organism, plant or animal) or in vitro

(outside a living organism in cells in an artificial environment)
o Animal viruses require cells within a host animal or tissue-culture cells derived
from an animal
o Bacteriophages can be grown in the presence of a dense layer of bacteria
called a bacteria lawn grown in a 0.7% soft agar in a dish or flask
o In vivo host sources can be a developing embryo in an embryonated birds egg
§ Serves as an incubator for viral replication, target sites include the
amniotic cavity, the chorioallantois membrane, or the yolk sac
o In Vitro studies, a primary culture is freshly prepared from animal organs or
tissues, cells are extracted by mechanical scraping or mincing to release cells
or by enzymatic method using trypsin or collagenase to break up tissue and
release single cells into suspension
§ Primary cultures usually have limited life span, contact inhibition (cell
density increases and cell contact triggers mitosis to stop) prevents
the density of the cells from becoming too high.
§ To prevent contact inhibition cells must be transferred to a secondary
cell culture and periodically cells density must be reduced by pouring
off some cells and adding fresh medium to provide space and
nutrients to maintain cell growth
o Continuous cell lines, derived from transformed cells and tumors, are often sub
cultured many times or even grown indefinitely. May not exhibit anchorage
dependency (will grow in suspension) and may have lost their contact
inhibition, result continuous cell lines can grow in piles or lumps resembling
small tumor growths.

● Explain how viruses may be identified

o Samples can be observed under a brightfield, electron, or fluorescent

microscope
o Cytopathic effects (CPEs) are distinct observable cell abnormalities due to viral
infection
§ Loss of adherence to surface of container, changes in cell shape from
flat to round, shrinkage of the nucleus, vacuoles in cytoplasm, fusion
of cytoplasmic membranes and formation of multinucleated syncytia,
inclusion bodies in the nucleus or cytoplasm, and complete cell lysis.
 Viroids, Virusoids, and Prions

Viroid (virus-like) consists only of a short strand of circular RNA capable of self-replication.
Unlike viruses, viroids do not have a protein coat to protect their genetic information.

Virusoids are subviral particles best described as non--self-replicating ssRNAs. Unlike viroids,
they require that the cell also be infected with a specific “helper” virus (which are all from the
family of Sobemoviruses).

Prion is a misfolded rogue form of a normal protein found in the cell. It can be infectious by
stimulating other endogenous normal proteins to become misfolded therefore forming plaques.
Cannot be treated by antivirals. Potentially spread by bites and consumption of necrotic tissues?

Animal Viruses: Viruses that infect macroorganisms.