REVIEWER IN MICROBIOLOGY AND PARASITOLOGY

Chapter 1: Introduction to
Pharmaceutical Microbiology
Microorganisms and medicines

Modern
medicine
must
be
effective, safe and of good quality
These consists of active ingredients
and variety of other materials
(excipients) to ensure that it is
effective, stable and safe during
storage and use
Analytical pharmacists take lead
responsibility for ensuring that
components of the medicine are
present in correct physical form
and concentration
Microorganisms have the potential
to influence their efficacy and
safety

Pharmaceutical microbiology

Must encompass the subject of

sterilization
and
preservation
against microbial spoilage
Pharmacists must be responsible
for the safe, hygienic manufacture
and use of medicines
Must
know
where
the
microorganisms
arise
in
the
environment (sources of microbial
contamination and the factors that
predispose to or prevent product
spoilage)

Antibiotics

Major importance in pharmacy

Defined as a naturally occurring
substance that was produced by
one microorganism that inhibited
the growth of / killed other
microorganisms
Natural
product,
a
microbial
metabolite
Encompass
certain
synthetic
agents that are normally used
systemically (throughout the body
to treat infection)

Commercial antibiotic production

began with the discover of
penicillin in the 1940s

Notes

Understanding the physiology and

genetics of microorganisms are
important
to
produce
new
therapeutic
agents
and
understanding infections and other
diseases
Knowledge of the mechanisms
whereby microorganisms are able
to
resist
antibiotics,
colonize
medical devices and cause or
predispose
humans
to
other
disease states is essential in the
production of new drugs and in
healthcare practices

Chapter 2: Fundamental
Features of Microbiology

Microorganisms differ in shape,

size, appearance, genetic and
metabolic characteristics
These properties are used in
classifying microorganisms into
major groups (eg. Fungi, bacteria
etc.)

1.1 Viruses, viroids and prions

Viruses
-do not have cellular structure
-composed of nucleic acid surrounded
by protein
-incapable of independent duplication
-intracellular parasites
-reproduced using metabolic capacities
of the host cell
-almost all viruses are smaller than
bacteria
- Electron microscope
Viroids
-simpler than viruses

-comprising single-stranded RNA

without any associated protein (plant
pathogens)
Prions
-no nucleic acid
-atypical form of a mammalian protein
-responsible for transmissible
spongiform encephalopathies (eg.
Creutzfeldt-Jakob disease (CJD) and
bovine spongiform (BSE))
-simplest and most recently recognized
agents for infectious disease

-obligate intracellular parasites

1.2.2. Fungi
Fungi
-eukaryotes
-Non-photosynthesizing plant
- Fungus covers both yeast and moulds
Yeast
-unicellular organisms larger than
bacteria
-divide either by binary fission or budding
Budding a daughter cell arises as a
swelling or protrusion from the parent
that eventually separates that lead an
independent existence

1.2 Prokaryotes and eukaryotes

Prokaryotes
-bacteria and archaea haploid
-reproduce asexually
Eukaryotes
-All other cellular microorganisms (i.e.
Fungi, protozoa etc.)
1.2.1 Bacteria and archaea
Bacteria
-unicellular
Eubacteria
- Bacteria of interest in pharmacy and
medicine
- Rod-shaped (bacillus), Spherical (cocci),
curved or spiral cells
-divided into two groups, Gram-positive
and Gram-negative
Saprophytes
-some bacteria that contaminate or cause
spoilage of pharmaceutical materials
Parasites
-benefiting from growth on or in other
living organisms without causing
detrimental effects
Pathogens
-parasites damaging the host
Rickettsia and chlamydia

Mould
-fungi that do not form fruiting bodies
visible to the naked eye
-tangled mass of filaments
Some fungi may exhibit:
-unicellular (yeast-like)
-mycelial (mould-like)
1.2.3. Protozoa
-eukaryotic
-unicellular
-free-living motile organisms
-not normally found as contaminants of
raw materials
2 Naming of microorganisms
Organisms are known by two names:
Genus (Genera for plural) and species
Example: Staphylococcus aureus
S. aureus, E. coli
3 Microbial metabolism
Oxidation
-removal or loss of electrons
-does not invariably involve oxygen
Chemoheterotrophs
Organisms of interest in pharmacy and
medicine

-These organisms obtain carbon, nitrogen

and energy by breaking down organic
compounds
Catabolic reaction
Chemical reactions by which energy is
produced by digestion of food materials.
These food materials undergo oxidation to
breakdown and release the energy
Anabolic reactions
use the produced energy to make
proteins, carbohydrates and nucleic acids
Glycolysis
Process of breaking down glucose and
release of energy
-It is a process that does not need oxygen
but he process releases only a relative
small amount the energy stored in a
sugar molecule
Fermentation
A metabolic process that releases more
energy from sugar
Note:
-Microorganisms are more versatile to the
materials that they can use as foods
Example: some bacteria can grow in the
presence of mineral salts and sugars
Example: Facultative anaerobes can
switch from respiration to fermentation if
oxygen are depleted
Metabolic products
Primary metabolites arise during the
period when a microbial culture is actually
growing (ex. Ethanol, organic acids)
Secondary metabolites produced
after cell multiplication has slowed or
stopped. These products have
commercial or therapeutic importance

Antibiotics, enzymes (biological washing

powders), toxins (Clostridium botulinum
for Botox) and carbohydrates (I.e.
Dextran)
Dextran used as a plasma expander
and for molecular separations by gel
filtration
4. Microbial cultivation
-Microorganisms can be cultivated in the
laboratory
-Most of them require relative simple
techniques and facilities
4.1 Culture media
-Also called routine laboratory media,
general purpose media and complex
media
-Mostly are readily available
-relatively inexpensive sources of protein
-Composed of different components such
as meat extracts, milk and soya, vitamins,
starch, sugars etc.
-Trypsin or other enzymes are preferred to
acids as a means of hydrolysis because
acids cause more amino acid destruction
Tryptic denotes use of enzyme
Carbohydrates- used in the form of
starch or sugars
Glucose- employed as a nutrient
-Microorganisms differ in terms of their
ability to ferment various sugars and this
can be used as an aid in the identification
-Routine culture media may be "enriched"
by the addition of milk, blood and serum
Culture media
-Can either be solid or liquid
Selective media or diagnostic media
-Restrict the growth of certain types of
microorganisms while permitting or
enhancing the growth of others

Example: antibacterial antibiotics added

to fungal media to suppress bacterial
contamination

4.2 Cultivation methods

-Most bacteria and yeast divide through
the process of binary fission
-It is where the cell enlarges or elongates
that forms a cross wall that will eventually
separate the cell into two equal
compartments each containing a copy of
the genetic material

-The bacteria more commonly grow

attached to a surface. Examples: stone,
metal, glass, epithelial surface in the
body
-Bacteria attached to a substrate are
described as sessile, and are said to
exhibit biofilm or micro-colony mode of
growth
-Planktonic cells are routinely used for
almost all the testing procedures used to
assess the activity of antimicrobial
chemicals

Yeast and Moulds

-Planktonic and biofilms exhibited

markedly different susceptibilities to
these lethal agents

Yeast multiply by budding

5 Enumeration of microorganisms

Moulds grow by the extension and

branching of hyphae to produce a
mycelium

Situations where it is necessary to

measure the number of microbial cells in
a culture, sample or specimen

Colony
-It is simply a collection of cells arising
from the multiplication of a single original
cell
-number of sufficiently large to be visible
by eye
-Colony size is limited by nutrition
availability colonies vary in shape, size
and pigmentation
that could be used for identification
Anaerobic organisms
-Can be grown on petri plates provided
that they are incubated in an aerobic jar
Catalyst consist of palladium-coated
pellets or wire
-Yeast colonies look similar to bacteria
although they may be larger and more
frequently colored
4.3 Planktonic and sessile (biofilm)
growth
Planktonic used to described freely
suspended cells
-Planktonic growth is not the normal
situation for bacteria. Example: cells
suspended in liquid medium

When measuring the levels of

microbial contamination in a raw
material or manufactured medicine
When evaluating the effects of an
antimicrobial chemical or
decontamination process
When using microorganisms in the
manufacture of therapeutic agents
When assessing the nutrient
capacity of a growth medium
Total count counting procedure
enumerating both living and dead cells
Viable count records the living cell
alone
Total viable count records all the
different species or type of
microorganisms present in a sample
Turbidity measurements most
common means of estimating the total
numbers of bacteria present in a sample
Traditional methods of viable counting all
suffer from the same limitations:

Relatively labor intensive

Not easy to automate
Slow, because they require an
incubation period for colonies to
develop or liquid cultures to
become turbid

Fast methods for viable counting

Epifluorescent technique uses

fluorescent dyes that exhibit
different colors in living and dead
cells
Enzyme assays detects living
cells that generate adenosine
triphosphate (ATP)
Resistance, capacitance or
impedance of a culture medium
changes as a result of bacteria or
yeast growth and metabolism
Manometric techniquesappropriate for monitoring the
growth of organisms that consume
or produce significant quantities of
gas during their metabolism

6 Microbial genetics
-The nature of the genetic material
possessed by a microbial cell and the
manner in which that genetic material
may be transferred to other cells depends
largely upon whether the organism is a
prokaryote or a eukaryote.
6.1 Bacteria
-genes essential for growth and
metabolism of bacteria are contained on
a chromosome of double-stranded DNA
-Plasmids replicate independently of the
chromosome within the cell
6.2 Eukaryotes
-microorganisms possess a nucleus that
normally contains one or more pairs of
linear chromosomes in which the ds DNA
is complexed with protein
-cells may divide asexually
-nucleus undergoes mitosis

-have potential for sexual reproduction

-possess plasmids
6.3 Genetic variation and gene
expression
Genotype
Genetic composition of an organism
-refers to the genes that the organism
possesses
-Microorganisms may also use an
alternative strategy of genetic
adaptation, by which they acquire new
gens either by mutation or by conjugation
-In bacteria, mutation is an important
mechanism by which resistance to
antibiotics and other antimicrobial
chemicals is achieved
7 Pharmaceutical importance of the
major categories of microorganisms
-Viral infections are among the most
dangerous and difficult to cure
Virus
-are not free-living
- Incapable of growing on manufactured
medicines or raw materials so they do not
cause product spoilage
-easy to destroy by heat, radiation or
toxic chemicals
-Penicillin and cephalosporin antibiotic are
produced by fungal species
-bacterial spores have the ability to
survive not only in dry conditions but in
other adverse environments
Protozoa
-do not possess cell wall
-do not survive drying well
8 Preservation of microorganisms
-Microorganisms are employed in a
variety of tests and assays, particularly
those to measure the activity of antimicrobial chemicals

-Organisms that readily form spores and

most fungi can be stored for months or
even years in this way provided that the
culture medium does not evaporate to
dryness
-Reference cultures can be obtained in a
freeze-dried form from internationally
accessible culture collections

Chapter 3: Bacteria
BACTERIA
-smallest free-living organism
-a prokaryote that lacks a true nuclear
membrane
-represent a large and diverse group of
microorganisms that can exist as single
cells or as cell clusters
BACTERIAL DIVERSITY AND UBIQUITY
-diversity can be seen in terms of
morphology (variation in cell size and
shape), adaptation to environmental
extremes, survival strategies and
metabolic capabilities
Refer to Table 3.1 pg. 25
CELL SIZE AND SHAPE
-smallest free-living organisms
-size measured in microns (micrometers)
-vary in size as small as 0.1-0.2 m in
diameter to those that are >5 m in
diameter
-majority of the bacteria are 1-5 m long
to 1-2 m in diameter
-divided into major groups on purely
morphological grounds
Shape:
Cocci (round)
Ovoid
Rod or bacillus (cylindrical)
Vibrios (curved)
Spirochaetes (long rigid curved with
multiple spirals)
Cell wall
-essential for the maintenance of the
shape and integrity of the bacterial cell

-target for antibiotics that attack and kill

bacteria
-provide strong, rigid structural
component to withstand osmotic
pressures
-most bacterial cell walls have murein
(peptidoglycan layer)
-peptidoglycan is composed of N-acetyl
muramic acid (NAM) and N-acetyl
glucosamine (NAG)
CELLULAR COMPONENTS
Gram stain
-treating a film of bacteria dried on a
microscope slide with a solution of crystal
violet, followed by a solution of iodine;
these are then washed with an alcohol
solution
Gram positive cell wall
-contains teichoic acids and lipoteichoic
acids
-during an infection, lipoteichoic acids
molecules trigger an inflammatory
response
Gram negative cell wall
-compose of proteins, lipoproteins,
phospholipids and lipopolysaccharides
(LPS) that is unique to gram negative
bacteria
LPS
-determines the antigenicity of the gram
negative cell
-made up of lipid A, ore polysaccharides
and O-specific polysaccharide
-Lipid A is responsible for the toxic
properties of gram negative bacteria
Cytoplasmic membrane
-involved in various transport and enzyme
functions associated with the membrane
-transports of nutrients, energy
generation and electron transports
-selective barrier between the cytoplasm
and the cell environment

-Mesosomes (invaginations)
Cytoplasm
-consists of 80% water and contains
enzymes that generates ATP
-compose of the ribosomes, nucleoid and
inclusion granules
Nucleoid
-singular, closed circular molecule of
double-stranded DNA
Plasmids
-small, circular pieces of double-stranded
extrachromosomal DNA
Ribosomes
-site of protein synthesis
-Two subunits: 30S and 50S
Inclusion granules
-serves as the storage material for
carbon, nitrogen and sulfur or phosphorus
Flagella
a. Monotrichous a single polar
flagellum
b. Lophotrichous two or more
flagella at one pole of the cell
c. Amphitrichous single/tuft of
flagella at each end of the cell
d. Peritrichous flagella distributed
over the entire cell
Pili and Fimbriae
a. Pili (pilus) join bacterial cell in
preparation of DNA transfer from
one cell to another
b. Fimbriae for adherence of cells to
one another and to environmental
surfaces
Glycocalyx (Slime Layer and Capsule)
-general substances that surround cells
-gelatinous polymer of polysaccharide,
polypeptide, or both

a. Capsule substance is organized

and firmly attached to cell wall
b. Slime layer unorganized and
loosely attached to the cell wall
-functions as protective barrier
Biofilms
-any surface for microbial habitat
-usually contains more than one species
of bacteria which exist and cooperate
together
-biofilm formation begins with attaching
to surface and form a cement cells to the
surface to protect the bacteria from
hazardous materials cells to the surface
to protect the bacteria from hazardous
materials
Bacterial sporulation
-not part of the reproductive cycle but
enables the bacteria to survive in adverse
environmental conditions
-all sterilization process for
pharmaceutical products have been
designed to destroy bacterial spore
Bacterial toxin
-Normal flora bacteria that are not
pathogenic but can be opportunistic
-Pathogenic bacteria that is not a
normal flora and cause diseases
-Toxin products of bacteria that produce
immediate host cell damage
Endotoxin
-the lipid A component of LPS
-ability to induce fever and activate B
lymphocytes
-generally released from lysed or
damaged cell
Exotoxin
A-B toxin contains B subunit that binds
to host cell receptor and bound to the A
subunit that mediates enzyme activity for
toxicity
Cytolytic toxis does not have A and B
subunit but work enzymatically to attack
cell that causes cell lysis
Superantigen toxin stimulate large
number of immune response cell resulting
to massive inflammatory response

Multiplication and division cycle

-bacteria multiply in number by binary
fission
-each daughter cell contains same
material of the mother cell
-DNA replication is dependent on
temperature
Traditional cultivation
Cultivation
-a process of growing microorganisms in
culture
-meet its nutritional and environmental
requirements
Phases of culture media
1. Broth (liquid) nutrients dissolve in
water
2. Agar (solid) with solidifying agent
3. Biphasic with solid and liquid
phase
Traditional cultivation
Types of bacteria by nutrient
requirements
I.
Fastidious
complex or extensive nutritional
requirements
-culture media should be
supplemented
II.

Non-fastidious
-Basic nutritional requirements

Media classifications and functions

1. Supportive media
support growth of most nonfastidious organisms
Beef extract, peptone, water agar
Nutrient agar
2. Enriched media (non-selective
isolation media)
- Nutrient supplement added
to supportive media for
growth of fastidious
microbes
- SBA and CAP
3. Selective media
-permits growth of certain bacteria
while inhibitory to other
HE agar plate and S agar plate
4. Differential media
-provides a distinct cultural
appearance of microorganism

MacConkey
Bacterial Growth Curve 4 Phases
1. LAG phase little or no cell
division
-intense metabolic activity
2. LOG (Logarithmic ) phase AKA
Exponential growth phase
-cell begins to divide
-active cellular reproduction with
constant minimum generation time
-cells are at their most active state
3. STATIONARY phase growth rate
slows down (no. of cells = no. of
microbial deaths = population
stabilizes) period of equilibrium
4. DEATH phase logarithmic
decline
-number of deaths exceeds the
number of new cells formed

Chapter 4: Fungi
Fungi
Can be yeasts or molds
Yeast
Reproduce by budding
Unicellular
Molds
Multicellular
Woolly or fuzzy appearance in culture
Made up of mycelium
- Intertwining structures called
hyphae
Hyphae
- Tube-like structures
- Fundamental units of fungi
Parts of Hyphae
Aerial (reproductive)
- Above the surface
Vegetative (thallus)
- Inside the medium
- Absorbs water and nutrients
Types of hyphae

 Septate
- With frequent crosswalls
Sparsely septate
- Aseptate
- Few cross walls
Structures of hyphae
Conidiophore/Sporangiophore
- Stalks for conidia/sporangium
Conidia/Sporangia
- Asexual structures that form at
the hyphae of a conidiophore or a
sporangiophore
- Spores are in a sac (sporangia)
- Spores are not in a sac (conidia)
Vesicles
- Enlarged structure at the tip of
the conidiophore
Columella
- Dome shaped structure at the tip
of sporangiophore
Other hyphae
Spiral
- Coiled hyphae
Racquet
Favic chandelier
- Antler hyphae
Hyaline
Non-pigmented fungi
Dematiaceous
Darkly-pigmented (melanin)
Dimorphism
Dimorphic fungi
Ability to exist in two forms
Yeast or Spherule phase - 37C w/ CO2
Mold - 25C w/ ambient air

Asexual
- Results in the formation of
Conidia
a. Conidia - spores that form on the
hyphae or conidiophore
b. Macroconidia - large, multicellular
c. Microconidia - small, unicellular
- Spores develop from vegetative
mycelium
d. Blastoconidia (Blastospores) hyphae or pseudo hyphae, daughter cell
that buds from mother cell
e. Chlamydoconidia
(Chlamydiospores) -formed from
"rounding up" an enlargement of hyphal
segments; Terminal (tip); Sessiles (sides);
Intercalary (within)
f. Arthroconidia (Arthrospores) fragmentation of the hyphae into barrel or
rectangular shaped spores
- Spores contained in sacs (sporangia)
g. Sporangiospores - produced at the
tip of sporangiophore or aseptate hyphae;
Zygomycetes

Chapter 5: Virus
1 INTRODUCTION
Viruses first discovered at the end of
the 19th century, although they were
identified much earlier.
- It all started when rabies (an
infectious disease) could not be
explained by the presence of
bacteria.
- They were then referred to as
filterable agents/filterable viruses
- Electron Microscopy identification
of the chemical nature of viruses
start of the golden era of virology
led to the design of potent antiviral
drugs and effective vaccines

Polymorphism (Polymorphic Fungi)

Have both Yeast and mold form in
culture

2 GENERAL STRUCTURES OF VIRUSES

Reproduction

Viruses Diverse in size and structure.

Smallest = 28nm (poliovirus), Largest =
750nm (Mimi virus).

- Viral classification is based on the

physical and chemical properties of
viruses, their structure and morphology

- Bacterial Viruses: complex structure that

consists of a capsid head, a tail and tail
fibers

VIRUS COMPOSITION:
Viral nucleic acid within a protein core
(the capsid/coat), possibly surrounded by
a lipidic envelope

2.3 Viral envelope

- Viral capsid can be surrounded by a
lipidic envelope (can come from either
the host cells nuclear membrane or
the cytoplasmic membrane)
- Host proteins in the viral envelope are
excluded, but proteins encoded by the
viral genome are present
- Enveloped viruses are considered
the most susceptible to chemical
and physical conditions, and do
not survive well on their own
outside the host cell

2.1 Viral Nucleic Acid

- Viral genome is made up of either
DNA or RNA.
- Can either be double stranded or
single stranded, linear or circular,
containing several segments or one
molecule.
2.2 Viral Capsid
-

protects the viral nucleic acid

from detrimental chemical and
physical conditions
composed of a number of subunits
called capsomeres (genetically
encoded by the viral genome)
they give the shape of the capsid
and provide the virus the
resistance to chemical and physical
agents
capsomeres are bonded together
by non-covalent intermolecular
forces

Result of the assembly of the capsomeres

is either of the two:
1. Icosahedral Symmetry capsomeres in the form of pentons
and hexons
2. Helical Symmetry subunits are
symmetrically packed in a helical
array (coils of wound rope when
viewed under an electron
microscope)
- core is hollow, viral nucleic acid is
embedded into ridges on the inside
of each sub unit
- In mammalian viruses: more complex
structure several proteinaceous
structures envelope the viral genome
core

2.4 Viral receptors

- they recognize the host cell receptor
site conveying viral specificity
- Viruses possess receptors on their
surface in the form of glycoproteins
usually embedded in the viral
envelope OR protruding as a spike
from the viral capsid
- They recognize and bind receptors
on the host cell
3 VIRUS-HOST CELL INTERACTIONS
- viruses need to interact with their host
cell because they cannot reproduce on
their own
- they are considered as intracellular
parasites
- they have no metabolism and cannot
synthesize their own proteins, lipids or
nucleic acids
- production and excision of viruses can
result in cell death
Three major groups based on host
specificity: 1) viruses of bacteria and
blue-green algae, 2) plant viruses, and 3)
animal (including insect) viruses
Viruses are very specific, and rarely
cross species barriers (exception:
rabies and influenza can cause
diseases in both animals and
humans)

- Viruses can also be asymptomatic in

certain hosts (the host can become a
reservoir, and transmit the virus to a
susceptible recipient)
5 ways in which the virus can
interact with the host cell:
1. Multiplication of the virus and
destruction of the host cell upon release
of the viral progeny
2. Multiplication of the virus and release
of the virions without the immediate
destruction of the host cell
3. Survival of the virus in a latent stage
without noticeable changes to the
infected cell
4. Survival of the infected cell in a
dramatically altered or transformed state
5. Incorporation of the viral nucleic acid in
the host cell genome without noticeable
changes to the infected cell
4. MULTIPLICATION OF HUMAN
VIRUSES
4.1 Attachment to the host cell
Three phases: 1) an initial contact mainly
dependent on Brownian motion, 2)
reversible phase wherein the electrostatic
repulsion is reduced and 3) irreversible
changes in virus-receptor-host-receptor
configuration that initiates viral
penetration through the cell membrane
-

Virus-cell recognition is similar to any

protein-protein interaction they both
occur through a stereospecific network
of hydrogen bonds and lipophilic
associations

4.2 Penetration of the viral particle

- starts with the irreversible attachment
of the virus to the host cell

Two energy-dependent mechanisms;

endocytosis or fusion
Followed by penetration of the virus
through the cell membrane
Third mechanism is identified in some
bacteriophages that can inject their
nucleic acid inside the bacterium

4.3 Uncoating of the viral particle

- After penetration in a vacuole or
directly in the cytoplasm, the viral
nucleic acid has to be released from
the capsid/coat in order to initiate viral
replication uncoating process
4.4 Replication of viral nucleic acids
and translation of the genome
- the viral replication stage ensures that
1) the host cell synthesis machinery is
taken over by the virus, and 2) the
viral genome is replicated
Three main mechanisms common to
all viruses:
1. Transcription of viral genes into viral
mRNA
2. Translation of the viral genome into
proteins
3. Replication of the viral genome
- The common early transcription and
translation that occurs immediately after
the release of the nucleocapsid ensures
the production of early proteins (viral
polymerases, etc), and the hijacking of
the cell synthesis machinery
- Replication depends on the type of
nucleic acid carried by the virus
- Positive strand RNA: can be used
directly as mRNA following the acquisition
of a terminal sequence from the host cell
- Negative strand RNA: first transcribed
into positive RNA
- An important difference between the
host cell and the virus in the nature of
their mRNA
- Host cell mRNA encodes directly for
functional proteins

- Viral mRNA is polycistronic, several

distinct proteins are encoded within a
single piece of mRNA
- The virus needs to use a virusspecific protease to cut the
polyprotein produced in order to
restore the functionality of the viral
proteins
- Late protein synthesis during the
replication cycle is important in the
production of structural components of
the new virions
4.5 Maturation or assembly of virions
- large amounts of viral materials
accumulate within the host towards the
end of the multiplication process
- viral capsid starts to form from
individual structural proteins
- packaging of viral components can
occur within the cytoplasm or in the cell
nucleus
- chaperones are important in the
interaction between the viral nucleic acid
and the structural proteins
4.6 Release of virions into the
surrounding environment
- Mature virions are released from the
host cell at the end of the multiplication
process
- Most non-enveloped viruses: virus
progeny accumulates within the host cell
cytoplasm and is released following cell
lysis
- Some viruses: produce a lytic enzyme
(peptide) or proteases to lyse the host
which enables the release of infectious
particles - the host often self-disintegrates
due to the lack of maintenance
- Enveloped viruses: usually released by a
budding process over a period of hours
5. CULTIVATION OF HUMAN VIRUSES
5.1 Cell culture

- Support for human virus growth is

usually derived from humans, other
primates or rodents
Cell culture according to history:
1. Primary diploid cell line, derived
directly from intact tissue (human embryo
kidney, monkey kidney)
2. Secondary diploid cell line, derived
from primary cultures, more homogenous
and characterized, but not as susceptible
to viral infection
3. Continuous from malignant tissues,
and have the power to multiply in vitro
- cell culture for propagating greatly relies
on the growth of cells in a semiconfluent
monolayer attached to a surface
- to sub-culture, cells are separated from
monolayer or relevant tissue with trypsin
in order to form a suspension of single
cells suspended cells are used to seed a
new flask
- the media used to grow consists of basic
nutrients and salts supplemented with
serum
- Cytopathic effect is an indication of cell
death cell shrinkage, ballooning or
detachment of cells from the surface of
the flask
- cytopathic effect can spread to adjacent
cells and result to the formation of a
plaque (can easily be identified following
staining) can be used for the
enumeration of viruses assuming one
plaque results from infection by one virus
5.2 The chick embryo
- useful because of the different tissues
found in the eggs that can support the
growth of different viruses
5.3 Animal inoculation
- some animals are used in order to study
antiviral and vaccine effectiveness, and
can also be a source of cell lines for cell
cultures

- growth of virus is indicated by signs of

disease or death
6. CONTROL OF VIRUSES
- vaccination is the most successful
against microbial infections
7 VIRUSES AND GENE THREAPY
- some viruses/virus components are used
as vectors for the delivery of genes to
targeted cells
- Some viruses used in gene transfer
medicinal products (GTMP): adenoviruses,
poxviruses, retroviruses, lentiviruses,
adeno-associated viruses, and
herpesviruses
8 VIRUSES AS ANTIMICROBIALS
8.1 Bacteriophages
- Bacteriophages are viruses that only
infect bacteria
- 20-200nm in size and are highly diverse
in their structure and host range
- all bacteria species can most likely be
infected by a phage
- complex phages are tadpole-shaped, it
consists of a head (contains the viral
genome), a tail (recognizes the host
receptor, attaches, and serves as the
nucleic acid injection device)
- Phages inject their viral genome inside
the host cell
Two phage replication cycles:
1. Lytic cycle results in the lysis of the
bacterial host
2. Lysogenic cycle viral nucleic acid
being integrated into the host genome
- Lytic phage infection aka virulent phage
results in the replication (within the
susceptible bacteria) and release of
infectious phage progeny from the host
cell following cell lysis

- Transduction when the genes carried

by the prophage can be expressed in the
new host, it is responsible for gene
transfer between the bacteria
8.2 Use of bacteriophages to treat
bacterial infection
- the discovery of the lytic properties of
phages led to their use as potential
bactericidal agents
8.3 Epidemiological uses and
diagnosis
- Phage typing differentiates distinct
stains of bacteria based on their
susceptibility to phages
- further development of rapid diagnostics
9 PRIONS
- causative agents of the
neurodegenerative diseases bovine
spongiform encephalopathy, scrapie in
sheep, and Creutzfeldt-Jakob disease in
humans
- distinct class of infectious agents that
have unique and disturbing properties
- rod-like structures that can be recovered
from the infected brain of the patient
- lacks a nucleic acid and are resistant to
heating and UV radiation, they also
cannot produce an immune response in
the host