[TRANS] VIROLOGY

● The observation of a virus by electron microscopy and

OUTLINE (BOOK) the development of cell tissue culture started the
golden era of virology.
I. Introduction
● Since then a large number of viruses have been
II. General Structure of Viruses
isolated, their structure identified and their
A. Viral Nucleic Acid
replication understood, leading to the design of
B. Viral capsid
potent antiviral drugs and effective vaccines.
C. Viral Envelope
● Progress in virology over the last 50 years have been
D. Viral Receptors
considerable, leading to the eradication of smallpox
III. Virus-host cell interaction
following a worldwide vaccination programme, and
A. HIV
the likely future eradication of poliovirus.
B. Tumour viruses
● For the first time in human history an infectious
IV. Multiplication of human viruses
disease has been vanquished.
A. Attachment to the host cell
● Despite such an achieve- ment, much progress is still
B. Penetration of the viral particle
required to combat other viruses.
C. Uncoating of the viral particle
● For example, despite large financial investment and
D. Replication of viral nucleic acids and translation of
high profile studies, a cure or vaccine for HIV is still
the genome
not available.
E. Maturation or assembly of virions
● The number of commercially available antivirals is
F. Release of virions into the surrounding
also still limited, when compared to the number of
environment
antibiotics.
V. Cultivation of Human viruses
● Within the last few years, diseases caused by viruses
A. Cell Structure
in human and animals have reminded us that viral
B. The chick embryo
infections can easily spread and cause epidemics and
C. Animal Inoculation
pandemics.
VI. Control of viruses
● Recent examples are ‘swine flu’ and ‘avian flu’, both
A. Antiviral Chemotherapy
caused by influenza viruses, and also an outbreak of
B. Vaccination
severe acute respiratory syndrome (SARS), the virus
C. Viricidal effects of chemical and physical agents on
originating from Asia and spreading worldwide, and
viruses.
outbreaks of foot and mouth virus affecting cattle,
D. Control of viruses in pharmaceutical products
sheep and pigs that had important economic
VII. Viruses and gene therapy
consequences in the UK and mainland Europe.
VIII. Viruses as antimicrobials
A. Bacteriophages
B. Use of bacteriophages to treat bacterial infection II. GENERAL STRUCTURE OF VIRUSES
C. Epidemiological uses and diagnoses
IX. Prions ● Viruses are extremely diverse in size and in structure.
● The smallest virus is approximately 28 nm in size
(poliovirus), while the largest is 750nm (mimivirus).
● In simplistic terms, a virus consists of viral nucleic acid
within a protein core, the capsid (also referred to as
I. INTRODUCTION
the coat), pos- sibly surrounded by a lipidic envelope.
● In reality, there are many differences between viruses
● Viruses were first discovered at the end of the 19th
in terms of nucleic acid, capsid structure, number of
century, although the symptoms they cause were
coats and envelope composition.
identi- fied much earlier.
● Such differences account for the high diversity of
● One of the earliest pieces of evidence is the
viruses and the differences in their properties,
symptoms of poliomyelitis in an Egyptian priest
notably their resistance to antiviral drugs and viricidal
depicted on a hieroglyph.
agents.
● Virus discovery came about when the cause of an
● Viral classification is based on the physical and
infectious disease (rabies), could not be explained by
chemical properties of viruses, their structure and
the presence of bacteria.
morphology.
● Unlike bacteria, the ‘infectious materials’ were not
retained by filtration and thus viruses were then
A. Viral Nucleic acid
referred to as ‘filterable agents’ or ‘filterable viruses’.
● Until the advent of electron microscopy in the 1940s,
only the chemical nature of viruses (i.e. proteins and ● The viral genome is composed of either DNA or RNA.
nucleic acid) could be identified and their infectivity ● It can be double stranded (ds) or single-stranded (ss),
was mainly studied in animal models. linear (e.g. poliovirus) or circular (e.g. hepatitis B

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 TRANS: Chapter 5 Virology

virus), con- taining several segments (e.g. ● These subunits offer considerable economy of genetic
influenza—eight segments of minus-sense ss RNA) or information within the viral genome since only a
one molecule (e.g. poliovi- rus). small number of different subunits contribute to the
● Six groups can be distinguished depending on their formation of the capsid.
nucleic acid content. ● Viruses with an icosahedral capsid usually have
● Viruses that contain plus-sense ss RNA (e.g. capsomeres in the form of pentons and hexons.
poliovirus) can have their genome translated directly ● The number of these subunits varies considerably
by the host ribosome. between viruses: for example, adenovirus is
● The nature of the viral nucleic acid is important for constructed from 240 hexons and 12 pentons,
the effectiveness of antiviral treatments. whereas the poliovirus is composed of 20 hexons and
● For example, retroviridae such as HIV require a 12 pentons, forming a much smaller structure.
specific virus-encoded enzyme, a reverse ● Viruses with a helical capsid (e.g. influenza and
transcriptase, to convert their ss RNA into ss DNA, to mumps viruses) have their subunits symmetrically
be able to replicate within the host cell. packed in a helical array, appearing like coils of wound
● This enzyme is a primary target site of many antiviral rope under electron microscopy.
drugs. ● Although the core of such a virus is hollow, the viral
● On some occasions, the viral genome has been shown nucleic acid is embedded into ridges on the inside of
to be infectious, i.e. to cause an infection. each subunit and does not fill the hollow core.
● Such an observation is important when one considers ● Such a close association between viral nucleic acid
viricidal agents that damage the viral capsid or and capsid proteins can explain the damage caused to
envelope but not the viral nucleic acid. the nucleic acid following disaggregation of the capsid
● Furthermore, in laboratory conditions, the after chemical or physical treatments.
phenomenon of multiplicity reactivation has been
observed with poliovirus whereby random damage to Table No. 1 Structure and clinical importance of mammalian
viral capsid and nucleic acid following treatment with viruses
hypochlorite, a biocide intensively used for surface Group Virus Characteris Clinical
disinfection, resulted in complementary tics importance
reconstruction of an infectious particle by DNA viruses
hybridization of the gene pool of the inactivated Poxviruses Variola Large Variola is the
virus. particles smallpox virus, it
● This again underlines the necessity of rendering the 200-250n produces a
viral nucleic acid non-infectious following a viricidal m: systemic infection
treatment. complex with a
symmetry characteristic
B. Viral Capsid vesicular rash
affecting the face,
● The function of the capsid is to protect the viral arms and legs,
nucleic acid from detrimental chemical and physical and has a high
conditions (e.g. disinfection). mortality rate
● The capsid is composed of a number of subunits Vaccinia Vaccinia has been
named capsomeres genetically encoded by the viral derived from the
genome. cowpox virus and
● The nature and association of the capsomeres are is used to
fundamental for the virus, as they give the shape of immunize against
the capsid, but also provide the virus with resistance smallpox
to chemical and physical agents.
● The assembly of the capsomeres results in two Adenoviruses Adenoviru Icosahedral Commonly cause
different architectural styles—icosahedral and helical s particles upper respiratory
symmetries; in mammalian viruses, a more complex 80 nm in tract infections;
structure can be found, where several proteinaceous diameter tend to produce
structures envelope the viral genome core (e.g. latent infections
poxviruses, rhabdoviruses). in tonsils and
● Bacterial viruses (bacteriophages) also show a adenoids; will
complex structure consisting of a capsid head, a tail produce tumours
and tail fibres. on injection into
● The nature of the capsomeres in certain viruses hamsters, rats or
allows for the self-assembly of the capsid within the mice
host cell. Herpesviruses Herpes Enveloped, HSV1 infects oral
● The capsomeres are held together by non-covalent simplex icosahedral membranes in
intermolecular forces. virus particles children; >80%
● Such assembly also allows the release of the viral (HSV1 and 150 nm in are infected by
genome following dissociation of the non-covalently HSV2) diameter adolescence.
bonded subunits. Following the

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primary infection tropical Africa, a

the individual severe EBV
retains the HSV1 infection early in
DNA in the life predisposes
trigeminal nerve the child to
ganglion for life malignant facial
and has a 50% tumours (Burkitt’s
chance of lymphoma)
developing ‘cold Hepatitis Hepatitis B Spherical In areas such as
sores’ HSV2 is Viruses virus (HBV) envelopes South-East Asia
responsible for particle 42 and Africa, most
recurrent genital nm in children are
herpes diameter infected by
Varicella Enveloped, Causes enclosing prenatal
zoster icosahedral chickenpox in an inner transmission. In
virus (VZV) particles children; virus icosahedral the Western
150 nm in remains dormant 27 nm world the virus is
diameter in any dorsal root nucleocaps spread through
ganglion of the id contact with
CNS; release of contaminated
immune control blood or by sexual
in elderly people intercourse. There
stimulates is strong evidence
reactivation that chronic
resulting in infections with
shingles HBV can progress
Cytomegalovi Enveloped, CMV is generally to liver cancer
rus (CMV) icosahedral acquired in Papovaviruse Papilloma Naked Multiply only in
particles childhood as a s virus icosahedra epithelial cells of
150 nm in subclinical 50 nm in skin and mucous
diameter infection. About diameter membranes
50% of adults causing warts.
carry the virus in There is evidence
a dormantstate in that some types
white blood cells. are associated
The virus can with cervical
cause severe carcinoma
disease RNA viruses
(pneumonia, Myxoviruses Influenza Enveloped These viruses are
hepatitis, virus particles, capable of
encephalitis) in 100 nm in extensive
immunocomprom diameter antigenic
ised patients. with a variation,
Primary infections helically producing new
during pregnancy symmetric types against
can induce capsid; which the human
serious congenital haemagglu population does
abnormalities in tinin and not have effective
the fetus neuramini immunity. These
Epstein-Barr Enveloped, Infections occur dase spikes new antigenic
virus (EBV) icosahedral by salivary project types can cause
particles exchange. In from the pandemics of
150 nm in young children envelope influenza. In
diameter they are natural infections
commonly the virus only
asymptomatic but multiplies in the
the virus persists cells lining the
in a latent form in upper respiratory
lymphocytes. tract. The
Infection delayed constitutional
until adolescence symptoms of
often results in influenza are
glandular fever. In probably brought

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about by 70 nm in In developing
absorption of diameter countries it is
toxic breakdown responsible for
products from the about a million
dying cells on the deaths each year
respiratory Picornaviruse Poliovirus Naked One of a group of
epithelium s icosahedral enteroviruses
Paramyxoviru Mumps Enveloped Infection in particles common in the
ses virus particles children produces 28 nm in gut of humans.
variable in characteristic diameter The primary site
size, swelling of of multiplication
110-170 parotid and is the lymphoid
nm in submaxillary tissue of the
diameter, salivary glands. alimentary tract.
with helical The disease can Only rarely do
capsids have neurological they cause
complications, systemic
e.g. meningitis, infections or
especially in serious
adults neurological
Measles Enveloped Very common conditions like
virus particles childhood fever, encephalitis or
variable in immunity is poliomyelitis
size, lifelong and Rhinoviruses Naked The common cold
120-125 second attacks icosahedra viruses; there are
nm in are very rare 30 nm in over 100
diameter, diameter antigenically
helical distinct types,
capsids hence the
Rhabdoviruse Rabies Bullet-shap The virus has a difficulty in
s virus ed very wide host preparing
particles, range, infecting effective vaccines.
75-180 nm, all mammals so The virus is shed
enveloped, far tested; dogs, copiously in
helical cats and cattle are watery nasal
capsids particularly secretions
susceptible. The
incubation period Hepatitis A Naked Responsible for
of rabies is virus (HAV) icosahedra ‘infectious
extremely varied, 27 nm in hepatitis’ spread
ranging from 6 diameter by the oral–faecal
days up to 1 year. route especially in
The virus remains children. Also
localized at the associated with
wound site of sewage
entry for a while contamination of
before passing food or water
along nerve fibres supplies
to central nervous Togaviruses Rubella Spherical Causes German
system, where it particles measles in
invariably 70 nm in children. An
produces a fatal diameter; infection
encephalitis a tightly contracted in the
Reoviruses Rotavirus An inner A very common adherent early stages of
core is cause of envelope pregnancy can
surrounde gastroenteritis in surrounds induce severe
d by two infants. It is an multiple
concentric spread through icosahedral congenital
icosahedral poor water capsid abnormalities,
shells supplies and e.g. deafness,
producing when standards blindness, heart
particles of general disease and
hygiene are low.

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mental
retardation
Flaviviruses Yellow Spherical The virus is
fever virus particles spread to humans HIV Differs HIV is transmitted
40 nm in by mosquito from other from person to
diameter bites; the liver is retroviruse person via blood
with an the main target; s in that or genital
inner core necrosis of the core is secretions. The
surrounde hepatocytes leads cone-shap principal target
d by an to jaundice and ed rather for the virus is the
adherent fever than CD4+
lipid icosahedral T-lymphocyte
envelope cells. Depletion of
these cells
induces
immunodeficienc
Hepatitis C Spherical The virus is y
virus (HCV) particles spread through
40 nm in blood Hepatitis Hepatitis D An The presence of
diameter transfusions and viruses virus (HDV) RNA-contai the satellite HDV
consisting blood products. ning virus exacerbates the
of an inner Induces a that can pathogenic
core hepatitis which is only effects of HBV
surrounde usually milder replicate in producing severe
d by an than that caused cells hepatitis
adherent by HBV co-infected
lipid with HBV.
envelope The
spherical
coat of
Filoviruses Ebola virus Long The virus is HDV is
filamentou widespread composed
s rods amongst of HBV
composed populations of capsid
of a lipid monkeys. It can protein.
envelope be spread to
surroundin humans by
g a helical contact with body
nucleocaps fluids from the
id 1000 nm primates. The *CNS, central nervous system.
long, 80 resulting
nm in haemorrhagic
diameter fever has a 90%
case fatality rate
Retroviruses Human Spherical HTLV is spread
T-cell enveloped inside infected
leukemia virus 100 lymphocytes in
virus nm in blood, semen or
(HTLV-1) diameter; breast milk. Most
icosahedral infections remain
cores asymptomatic but
contain after an
two copies incubation period
of linear of 10–40 years in
RNA about 2% of
molecules cases, adult T-cell
and leukaemia can
reverse result
transcripta
se

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Hepadnavirid cytomegalovi
ae*, rus, poxvirus,
Papovavirida HBV,
e, papilloma
Adenoviridae virus,
adenovirus
Myxoviridae Bacteriophag
e T and λ
II ss DNA Bacteriophag
e M13, X174,
filamentous
bacteriophag
es
III ds RNA Reoviridae Reoviruses,
rotaviruses
IV ss RNA Picornavirida Poliovirus,
with mRNa e hepatitis A
identical in virus,
base rhinovirus
sequence
to virion
RNA
V ss RNA Paramyxoviri Influenza,
C. Viral Envelope with dae, paramyxoviru
mRNA Orthomyxovi s, measles,
● The viral capsid can be surrounded by a lipidic compleme ridae, mumps,
envelope, which originates from the host cell. ntary in FIloviridae Ebola virus
● The envelope is added during the replication process base
or following excision of the viral progeny from the sequence
host cells. to virion
● The envelope can come from the host cell nuclear RNA
membrane (e.g. herpes simplex virus) or the VI ss RNA Retroviridae HIV
cytoplasmic membrane (e.g. influenza virus). with DNA
● One characteristic of the viral envelope is that host intermedia
proteins are excluded, but proteins encoded by the te in their
viral genome are present. growth
● These viral proteins play an impor- tant serological
*Hepadnaviridae genome contains partially ds DNA and
role.
partially ss DNA.
● Enveloped viruses are generally considered to be the
most susceptible to chemical and physical conditions
and do not survive well on their own outside the host
cell (e.g. on surfaces), although they can persist
longer in organic soil (e.g. blood, exudates, faeces).
● Lipids in viruses are generally phospholipids from the
host envelope, although glycolipids, neutral fats, fatty
acids, fatty aldehydes and cholesterol can be found.

D. Viral Receptors

● In addition to these structures, glycoproteins can be

found usually protruding from the viral capsid or
embed- ded in the envelope.
● These virus-encoded structures are important for
viral infectivity as they recognize the host cell
receptor site conveying viral specificity. In
bacteriophages, these structures can take the shape
of tail fibres.
Table No. 2 Examples of types of nucleic acid in human and
bacterial viruses
Group Nucleic Family Example
acid III. Virus-host cell interactions
I ds DNA Herpesvirida Herpes
e, Poxviridae, simplex virus,

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● Viruses need to interact with a host cell as they
cannot reproduce on their own.
● They have no metabolism and cannot synthesize their
own proteins, lipids or nucleic acids.
● Thus viruses can be considered as true intracellular
parasites that grow within living cells and use their
energy and synthetic machinery to produce viral
components.
● The production and excision of viruses from the host
cell will result in cell death, although this might not
be immediate.
● Following the replication of one virus within the host
cell, hundreds of new viruses (virus progeny or
virions) can be released and infect adjacent cells
(within a tissue).
● The propagation from one infected cell to new cells,
and the subsequent destruction of tissue or cells,
provides signs of the viral disease.
● On the basis of host specificity, three major viral
groups can be distinguished: (1) viruses of bacteria
and blue-green algae, (2) plant viruses and (3) animal
(includ- ing insect) viruses.
● Viruses are usually very specific and rarely cross
species barriers, although there are some exceptions,
such as rabies and influenza that can cause diseases
in both animals and humans.
● Viruses can also be asymptomatic in certain hosts
where they do not cause an infection; the host
becomes a reservoir and can transmit the virus to a
susceptible recipient (e.g. transmission of yellow
fever to humans by mosquitoes).
● Viruses can interact with the host cell in five different
ways: (1) multiplication of the virus and destruction
of the host cell upon release of the viral progeny, (2)
multi- plication 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 (e.g. transfor- mation of a normal
cell to one having the properties of a cancerous cell)
and (5) incorporation of the viral nucleic acid in the
host cell genome without noticeable changes to the
infected cell.
● The interaction with the host cell will vary between
viruses, but will generally follow one of these five
routes.
● There is a great diversity in viral infections and viral
diseases.
● Many viral infections are asymptomatic or ‘silent’
whereby the virus replicates within the host but does
not produce symptoms of a disease.
● Other common infections produce some mild
symptoms, such as a low- grade fever and a ‘runny
nose’.
● This is the case of the common cold, caused by
rhinoviruses, from which patients make a full
recovery within a few days.
● At the other end of the spectrum, some viruses kill
their host very quickly following infection, as in the
case of haemorrhagic viruses such as the Ebola virus.
● On other occasions a range of symptoms can be
observed in different hosts.
CAJUCOM, CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● This has been the case recently with ‘swine flu’, which
produced a range of symptoms, from a slight fever to
full influenza including a high fever, vomiting, and
dizziness.
● Other problematic viruses might not cause immediate
symptoms, but following the systematic destruction
of host cells will lead to an incurable disease; e.g. HIV
and oncogenic (tumour) viruses.
A. HIV
● HIV is an enveloped virus with a cone-shaped
nucleocap- sid containing two copies of a
positive-sense ss RNA molecule and the enzyme
reverse transcriptase.
● The viral genome encodes for the following genes:
env, envelope proteins; gag, capsid proteins; pol,
enzymes involved in viral multiplication; and tat,
enzymes regulating host metabolism.
● Seventy glycoprotein spikes (gp120) project from the
envelope and interact specifically with the CD4
protein receptor on the T-lymphocyte.
● The HIV core penetrates the cell cytoplasm following
membrane fusion and is uncoated releasing the two
RNA molecules and the reverse transcriptase into the
cytoplasm.
● The RNA is copied by reverse transcriptase into ss
DNA,
which is then duplicated to form a ds DNA copy of the
original viral RNA genome. This DNA moves into the
host cell nucleus where it is integrated as a provirus
into a host cell chromosome.
● The provirus can lie dormant in the cell or can be
expressed, producing viral mRNA and proteins and
resuming the multiplication cycle producing virions.
● The viral mRNA is polycistronic, producing
polyproteins that need to be cleaved by a specific
virus-encoded protease.
● Following the assembly of viral proteins and viral
RNA, the virions bud off the infected cells.
● At present there is no prospect of any drugs that can
be developed to eliminate proviruses from infected
cells.
● Current therapy has evolved around maintaining a
high count of T4 helper lymphocytes by regulating
the viral load produced by infected cells (up to 1010
viral particles produced continuously per day).
● Indeed, the ultimate decrease in T4 helper cells
(below 200/ml of blood) seriously compromises the
host immune system and allows infection by a range
of opportunist pathogens including fungi, protozoa,
bacteria and other viruses.
● Currently, antiviral treatments (highly active
antiretrovi- ral therapy—HAART) combining a
protease inhibitor and two reverse transcriptase
inhibitors, reduce the number of HIV particles and
slow the progression of the disease by restoring and
maintaining the number of T4 helper lymphocytes.
● However, this triple therapy does not eliminate the
virus completely.
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 TRANS: Chapter 5 Virology
● Patients who stop using the drugs experience a rapid
rebound in levels of the virus in the blood and
progression of the disease.
● The inability to eliminate the virus completely calls
for lifelong therapy that is prohibitively expensive for
countries with very limited health budgets.
B. Tumour viruses
● There is a suggestion that 20% of human cancers
might have a viral origin.
● Virus infected cells change dramatically, acquiring the
characteristics of tumour cells exhibiting uncontrolled
growth.
● For example, the Epstein–Barr virus (EBV) has been
associated with the formation of lymphomas and
nasopharyngeal carcinomas, the hepatitis B and C
viruses with hepatocellular carcinoma, human
papilloma viruses with cervical cancer, human T-cell
lymphotrophic virus type 1 with adult T-cell
leukaemia/lymphoma syndrome and HIV with
Kaposi’s sarcoma.
● There are no identified single mechanisms by which
viruses induce tumours.
● The acquisition of viral genes by the host must be
followed by other events such as environmental or
dietary exposures to chemical carcinogens for cancer
to occur.
● For example, there might be an association between
EBV and malaria to trigger Burkitt’s lymphoma in
young children in Africa.
● EBV and consumption of smoked fish might trigger
nasopharyngeal carcinoma in adults in China.
● The development of liver cancer following infection
with hepatitis B virus (HVB) might be triggered by
high alcohol consumption, smoking and exposure to
fungal toxin (aflatoxins), events that damage the liver.
● It is known that the viral genome of oncogenic viruses
can integrate the host DNA.
● Indeed, viral DNA has been recovered from infected
cells.
● Following integration of the provirus, the regulation
of cell growth and division can be affected.
● There is no means of eradicating proviruses.
● However, progress in the prevention of papilloma-
virus has been made by designing an effective vaccine
combined to a successful vaccination programme that
is responsible for an important decrease in cervical
cancer cases.
IV. Virus-host cell interactions
● The objective of the replication cycle is to ensure the
multiplication of the virus with the formation of
identical viral progeny.
● Viruses differ in their replication cycle and the time to
produce and release new virions.
● The multiplication cycle of human viruses is generally
slow, from 4 to more than 40 hours.
● Bacterial viruses are generally faster and can take as
little as 20 minutes to replicate within the bacterial
host.
CAJUCOM, CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● The replication cycle can be divided into six distinct
phases that are common to all viruses, although
detail within each phase varies greatly between
viruses.
● Understanding the viral multiplication process is
crucial for the development of new antiviral drugs.
A. Attachment to the host cell
● Viral attachment to the cell surface can be divided
into three phases: (1) an initial contact mainly
dependent on brownian motion, (2) a reversible
phase during which electrostatic repulsion is reduced
and (3) irreversible changes in
virus-receptor–host-receptor configuration that
initiates viral penetration through the cell membrane.
● All viruses possess receptors on their surface, usually
in the form of glycoproteins embedded in the viral
envelope or protruding as spike from the viral capsid.
● These structures recognize and bind receptors on the
host cell and provide the virus with its high specificity
although different viruses might share the same
receptor.
● The virus–cell recognition event is similar to any
protein–protein interaction in that it occurs through a
stereospecific network of hydrogen bonds and
lipophilic associations.
● For example, the haemagglutinin receptor of
influenza virus binds the terminal glycoside residues
of gangliosides (cell surface glycolipids) of the target
cell leading directly to the virus particle adhering to
the cell.
● Similarly, the inter- action between the HIV receptor
(i.e. gp120) and the T-lymphocyte receptor (i.e. CD4)
has been intensively studied.
B. Penetration of the viral particle
● Following the irreversible attachment of the virus to
the host cell, penetration of the virus through the cell
mem- brane is initiated following two
energy-dependent mech- anisms, endocytosis or
fusion.
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 TRANS: Chapter 5 Virology
● A third mechanism has been identified in some
bacteriophages that can inject their nucleic acid
inside the bacterium.
● During endocytosis, the association between virus
receptor and host receptor triggers a number of
mechanisms that draw the cell membrane to engulf
the virus particle forming a cytosolic vacuole.
● This process is widespread among non-enveloped
viruses, but is also used with some enveloped viruses
such as influenza (orthomyxoviruses).
● Certain enveloped viruses (e.g. herpes simplex virus,
HIV) can penetrate following fusion of their envelope
with the host cell membrane, liberating the viral
capsid within the cell cytoplasm.
C. Uncoating of the viral particle
● Following penetration of the virus in a vacuole or
directly into the cell cytoplasm, the viral nucleic acid
then needs to be released from the capsid/coat(s) to
initiate viral replication. This is the uncoating process.
● For viruses that penetrate by endocytoses, the
acidification of the cytosolic vacuoles following
endosome fusion induces a conformational change in
the capsid and the release of viral nucleocapsid
(some helper proteins are associated with the viral
nucleic acid) into the cytoplasm.
● For certain viruses, such as reovirus, only a partial
uncoating is necessary for the expression of the viral
genome. The release of the nucleocapsid from
vacuoles can occur in the cytoplasm, close to the
nucleus or within the cell nucleus.
D. Replication of viral nucleic acids and
translation of the genome
● This stage of viral replication ensures that (1) the host
cell synthesis machinery is taken over by the virus,
and (2) the viral genome is replicated.
● The structure, size and nature of the viral genome are
extremely diverse and thus this stage of the viral
multiplication cycle reflects this diversity.
● Three main mechanisms are, however, common to all
viruses: the transcription of viral genes into viral
mRNA, the translation of the viral genome into
proteins, and the replication of the viral genome.
● Early transcription and translation usually occurring
immediately after the release of the nucleocapsid in
the cytoplasm is also common, and ensures the
production of early proteins such as viral
polymerases, and the hijacking of the cell synthesis
machinery.
● In addition, some viruses can encode for genes the
products of which regulate the host synthetic
processes according to the needs of the virus (e.g. tat
gene in HIV).
CAJUCOM, CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● The replication of the viral genome depends on the
type of nucleic acid carried by the virus.
● The positive strand RNA in viruses such as the
poliovirus can be used directly as mRNA following the
acquisition of a terminal sequence from the host cell.
● Negative strand RNA (e.g. in influenza virus) is
transcribed into a positive RNA complementary in
base sequence to the parent RNA using an
RNA-dependent RNA polymerase carried by the virus.
● In ds DNA viruses (e.g. adenoviruses), the nucleic acid
passes into the nucleus where it is usually transcribed
by a host DNA-dependent RNA polymerase.
● In some viruses (e.g. poxvirus), this enzyme is
contained within the virus and released during
uncoating, allowing the viral genome to be replicated
in the cell cytoplasm.
● In retroviruses (e.g. HIV), a single-stranded proviral
DNA is produced from the viral ss RNA by a viral
enzyme called reverse transcriptase.
● This unique enzyme acts both as an RNA- and
DNA-directed DNA polymerase, and has associated
RNAase activity.
● The proviral DNA can be transported to the cell
nucleus where it can be integrated within the cell
host genome by a viral integrase.
● One important difference between the host cell and
the virus is in the nature of their mRNA.
● Host cell mRNA encodes directly for functional
proteins, whereas viral mRNA is polycistronic, which
means several distinct proteins are encoded within a
single piece of mRNA. This implies that the virus
needs to use a virus-specific protease to cut at the
correct place the polyprotein produced by translation
to restore the functionality of viral proteins.
● Late protein synthesis during the replication cycle
con- cerns the production of structural components
(e.g. cap- someres) of the new virions.
E. Attachment to the host cell
● Towards the end of the multiplication process, large
amounts of viral materials accumulate within the
host.
● Viral capsid starts to form from individual structural
pro- teins.
● In certain viruses (e.g. poliovirus) the capsid self-
assembles.
● The replicated viral genome and some viral proteins
become packaged within the capsid.
● Most non- enveloped viruses accumulate within the
cytoplasm or nucleus and are only released when the
cell lyses.
● Packaging of viral components can occur within the
cyto- plasm or in the cell nucleus.
● For example, with influenza virus, the capsomeres are
transported to the cell nucleus where they combined
with the viral RNA and assemble into helical capsids.
● The envelope of enveloped viruses originates from
the host.
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 TRANS: Chapter 5 Virology
● With the influenza virus, viral proteins such as
neuraminidase and haemagglutinin migrate to the
cell membrane, displacing cell protein.
● The assembled nucleocapsids pass out from the
nucleus to the cytoplasm and as they impinge on the
altered cytoplasmic membrane they cause it to bulge
and bud off completed enveloped particles from the
cell.
● In the herpesvirus, the envelope originates from the
nucleus membrane.
● The nucleocapsid assembles into the nucleus and it
acquires its envelope as it passes through the inner
nuclear membrane.
● The complete virus is then incorporated into a vesicle
which migrates to the cell surface.
● The maturation of viruses and their assembly is not
well understood at present.
● The presence of chaperone proteins may play an
important role in the interaction between the viral
nucleic acid and the structural proteins.
F. Attachment to the host cell
● At the end of the multiplication process, the mature
virions are released from the host cell.
● This can occur in a number of different ways.
● For most non-enveloped viruses, the virus progeny
accumulates within the host cell cytoplasm and is
released following cell lysis.
● Some viruses (e.g. bacteriophages) produce a lytic
enzyme (peptide) or proteases to lyse the host
enabling the release of infectious particles, although
the host often self-disintegrates as it cannot maintain
normal housekeeping functions during a viral
infection. Enveloped viruses are usually released by a
budding process over a period of hours.
● Ultimately the host cell will die following damage to
its metabolism and housekeeping functions during
viral replication.
V. Cultivation of Human Viruses
A. Cell Culture
● support the growth of human viruses are usually
derived from humans or other primates, or from
rodents.
● may be divided into three types according to their
history:
○ Primary cell culture
➢ derived directly from an intact tissue such as
human embryo kidney or monkey kidney.
○ Secondary cell culture
CAJUCOM, CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
➢ derived from primary cultures, usually those
arising from embryonic tissue
➢ more homogenous, better characterized,
but might not be as susceptible to viral
infection as primary cell lines.
➢ a limited number of subcultures can be
performed, generally up to a maximum of
about 50 before the cells degenerate.
○ Continuous cell culture
➢ usually derived from malignant tissue
(example: HeLa cells derived from a cervical
carcinoma) and have the capacity to
multiply indefinitely in vitro.
● Primary and secondary cells are usually diploid cell
lines
● Cell culture for propagating viruses relies on the
growth of cells in a semi confluent monolayer
attached to a surface (e.g. the bottom of a flask).
● Subculture- cells are separated from the monolayer
or relevant tissue usually with trypsin to form a
suspension of single cells
● Cytopathic effect
○ Morphological change in the infected cells.
○ Usually indicates death
○ can take the form of cell shrinkage or ballooning,
or the detachment of cells from the surface of
the flask
○ usually spread to adjacent cells and will result in
the formation of a plaque that can easily be
identified following staining.
➢ Plaques -used for the enumeration of
viruses assuming ano plaque results from
infection by one virus.
● The identity of the virus is confirmed using an
appropriate viral antisera labelled with a fluorescent
dye.
● Such cells are also characterized:
○ biochemically (isoenzyme analysis)
○ immunologically (histocompatibility antigens)
○ cytogenic markers
● PERT assay (product-enhanced reverse transcriptase),
Transmission electron microscopy and infectivity
assays.
➢ Examine cells for the absence of specific
infectious agents, for the absence of
bacterial, fungal and mycoplasma
contaminants, and for retroviruses
B. The chick embryo
● Fertile chicken eggs, 9-11 days old
● used as a convenient cell system to grow a number of
human pathogenic viruses
● usefulness derives from the many types of different
tissues found in the eggs, tissues that will support the
growth of different viruses
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● EXPENSIVE to use
● Use of chicken eggs for the production of vaccines is Poliovirus Human diploid cell line,
necessarily subject to many controls. continuous cell line or
➢ eggs have to be free from specific primary, secondary or
pathogens tertiary monkey kidney cells
➢ originate from healthy flocks
● processing of the fertilized eggs must be conducted
under aseptic conditions in an area where no other
infectious agents or cells are handled at the same Rubella Human diploid cells
time.

Baculovirus (recombinant Insect vector

vaccine)
Virus Cultivation

Hepatitis A virus Human diploid cell

C. Animal Inoculation

● Some animals (e.g. rodents, primates) have to be

Influenza virus Fertilized hen’s used to culture certain viruses in order to study
eggs—membrane bounding
antiviral and vaccine effectiveness.
the amniotic cavity
● use of animals follows strict ethical guidelines and is
extremely expensive
● used to grow viruses or to test an antiviral or vaccine,
Smallpox Fertilized hen’s eggs— growth of the virus is indicated by signs of disease or
chorioallantoic membrane death.

VI. Control of Viruses

Measles and mumps Chick embryo cells

Human herpesvirus 3
(shingles, herpes zoster)
Rabies virus
Varicella zoster virus
Tick-borne encephalitis virus
Yellow fever virus
A. Antiviral Chemotherapy
Human diploid cells
● important for persons at high risk, notably
immunocompromised patients
● Most antivirals are prodrugs that need to be
Chick embryo cells or in activated within the cell, usually by a kinase, and
human diploid cells other cellular enzymes.
● Can act at different stages of the viral replication
cycle
● A number of targets are being investigated to
Human diploid cells prevent viral attachment to the host cell:
➢ competition for CD4 receptors using a
pentapeptide identical in sequence to the
terminal amino acids of HIV gp120
Chick embryo cells ➢ inhibition of herpes simplex virus (HSV)
ribonucleotide reductase
➢ Competition for the cell receptor using a
Chick embryos hexapeptide fusion sequence at the
N-terminus of the influenza haemagglutinin
viral receptor

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● Proteases are particularly important for the uncoating

process preventing the release of viral nucleocapsid
and for the cleavage of viral polypeptide gene
products 1. Antiviral therapy HIV
➢ e.g. indinavir sulphate
● The replication of viral DNA is also a well-exploited ● no cure for HIV infections
target with the use of nucleoside analogues ● Role of antivirals is to slow or halt disease
(idoxiuridine is incorporated into viral and cellular progression
DNA instead of thymidine),non- nucleoside analogues ● Drugs for HIV called antiretrovirals have considerably
(nevirapine and foscarnet) and oligonucleotides. prolonged the life expectancy of patients, although
● Problems of antiviral therapy: not without some important side effects
○ viral diseases only become apparent after ● Antiretroviral treatments
extensive viral multiplication and tissue damage ○ aim to reduce HIV plasma levels for as much and
have occurred, delaying treatments. as long as possible; prolonged the life expectancy
○ Many antivirals are toxic (nucleoside analogues) of patients.
➢ Since viral replication often depends on the ○ usually given together or as a combo to avoid
use of host cell enzymes. emerging viral resistance.
○ antiviral monotherapy often leads to the ● Initiation of HIV treatment (HAART- highly active
development of virus resistance antiretroviral therapy)- complex and involves two
● The use of prodrugs has improved drug adsorption nucleoside reverse transcriptase inhibitors and two
● Oligonucleotide therapies: non- nucleoside reverse transcriptase inhibitors.
● Use for prophylaxis after exposure is also possible,
where a patient has been exposed to
METHODS PRINCIPLES EFFECT
HIV-contaminated materials
➢ Example: needle injury
Antisense Production of an Triggers the ● Adverse Effects:
nucleotides RNA sequence formation of ○ immune reconstitution syndrome
complementary to double ○ lipodystrophy syndrome - fat redistribution,
single-stranded viral stranded insulin resistance, hyperglycemia and
RNA duplex, dyslipidemia
inhibiting viral ○ damaging to liver function
RNA replication ○ osteonecrosis following long-term combination
treatments side effects
● Side effects:
○ gastrointestinal disturbance, anorexia,
Antigen methods Formation of triple Inhibit pancreatitis, liver damage, dyspnea, cough,
helix of DNA transcription headache, insomnia, dizziness, fatigue, blood
disorders, myalgia, arthralgia, rash, urticaria and
fever.

Decoy methods Production of Inhibit

synthetic decoys transcription 2. Antiviral therapy Herpesvirus infections
corresponding to a
specific nucleic acid ● Family of viruses that include:
sequence which ○ the herpes simplex virus
binds virally ○ chickenpox (varicella)
encoded regulatory ○ shingles (herpes zoster)
proteins ○ cytomegalovirus.
● Mild herpes simplex virus infections in healthy
individuals are treated with a topical antiviral drug
(e.g. treatment of cold sores)
Ribozymes Production of RNA Cleave other ● A systemic antiviral drug is used for severe infections
molecules (oligo RNA sequences (e.g. neonatal herpes infection, infection in
(ribo)nucleotides) at specific sites immunocompromised patients)

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● Antiviral treatments for chickenpox are
recommended in patients at risk and in neonates to
reduce risks of severe diseases.
● In healthy adults, chickenpox treatment taken within
24 hours of the appearance of a rash may decrease
the duration and severity of symptoms.
● In shingles treatment is taken within 72 hours of the
onset of rash
● Antivirals for herpes are also associated with a
number of side effects which vary depending on the
drug, but may include nausea, vomiting, stomach
pain, head- ache, fatigue, rash, and increase in serum
and urine uric acid.
● Antivirals for the treatment of cytomegalovirus are
usually given to immunocompromised patients and
they tend to be more toxic with notable
nephrotoxicity (e.g.cidofovir) and a number of
documented side effects (e.g.ganciclovir, foscarnet)
● DNA polymerase inhibitor:
○ Acyclovir
○ Valaciclovir
○ Valganciclovir
○ Vidarabine
○ Famciclovir
○ Ganciclovir
○ Cidofovir
● Ribavirin-- nucleoside analogue
3. Antiviral therapy Viral Hepatitis
● Hepatitis B and C are major causes of viral chronic
hepatitis
● Initial treatment for acute hepatitis B:
○ interferons (peginterferon alfa-2a) which may
reduce the risk of chronic infection; Choice of
antivirals depends upon the initial response to
peginterferon-alfa, emerging viral resistance, and
co- infection with HIV.
● Treatment of chronic hepatitis C:
○ a combination of ribavirin and peginterferon-alfa
is recommended, although the choice and
duration of treatment depends upon the viral
genotypes and viral load.
● Side effects:
○ Nausea
○ Vomiting
○ Abdominal pain
○ Diarrhea
● Hepatitis A- caused by contaminated food and water
4. Antiviral therapy Influenza
CAJUCOM, CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● Oseltamivir was extensively used for the prevention
and control of the swine flu outbreak in the UK in
2009
● Zanamivir
● Limitations in the usefulness of the drug:
○ The drug needs to be taken within a few hours
of the onset of symptoms, which proved very
difficult with a number of symptoms from mild
‘cold-like’ to severe ‘flu-like’ symptoms reported.
○ the side effects, especially in young children
and adolescents, have been very severe,
prompting many parents to stop the medication
5. Antiviral therapy Respiratory syncytial virus
● responsible for severe bronchiolitis notably in
infants.
● A monoclonal antibody (palivizumab) or an antiviral
drug (ribavirin) is indicated for the treatment of RSV
B. Vaccination
● most successful measure against microbial infections,
and particularly viral infections
● Vaccines
○ are preparations containing antigens that elicit a
specific and active immunity against an infecting
agent
○ Can induce the innate and the adaptive (cel-
lular, humoral) parts of the immune system
● The success of vaccination relies on the prevention
of a disease from occurring.
● Vaccination is particularly indicated:
○ to protect persons at risk (e.g. hepatitis B,
varicella zoster vaccines for healthcare workers)
○ prior to travelling (hepatitis A virus, Japanese
encephalitis, yellow fever, tick- borne
encephalitis virus)
○ to prevent cancer (human papillomavirus
vaccine preventing cervical cancer)
○ given to protect from a viral outbreak (e.g.
measles, mumps and rubella vaccine—MMR)
○ Exposure to rabies
● Viral vaccines are prepared using different methods:
○ MMR vaccine and live (oral) poliomyelitis
vaccines are based on the use of live attenuated
viruses, which are not as virulent as the original
virus.
○ Hepatitis A virus and influenza vaccines rely on
chemically inactivated (formaldehyde,
propiolactone) virus particles or components
(surface antigens).
○ Hepatitis B virus vaccine is a recombinant
vaccine where the viral DNA encoding for a virus
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 TRANS: Chapter 5 Virology
surface antigen (HBsAg) is expressed in yeast
(Saccharomyces cerevisiae) or mammalian cells
(Chinese hamster ovary cells or other suitable
cell lines). The surface antigen is then purified.
○ Human papillomavirus (HPV) vaccine contains
virus-like particles and recombinant capsid
protein expressed in yeast or using a baculovirus
as an expression system.
● Viruses or their components can be prepared from
animals, fertilized hen’s eggs, in suitable cell cultures
or in suitable tissues, or by culture of genetically
engineered cells.
● It should be noted that most viral vaccines contain
one or more adjuvants, e.g. aluminium salts (antigens
are absorbed to the aluminium salts)
● Immunoglobulin plays a role in the protection of
patients with a compromised immunity against viral
infections.
○ Human normal immunoglobulin (HNIG) is
prepared from a pool of donated human plasma
that has been checked to be non-reactive for
hepatitis B surface antigen, hepatitis C virus and
HIV (types 1 and 2).
○ Intramuscular normal immunoglobulin - protect
against hepatitis A virus in immunocompromised
patients where the disease is highly endemic and
to protect against or attenuate measles infection
in immunocompromised patients.
● Disease-specific immunoglobulin pool of plasma
obtained from specific human donors who have
high-levels of the specific antibody required.
○ Disease-specific hepatitis B immunoglobulin is
used following accidental inoculation by a risk
material (e.g. needlestick injury) or for infants
born from mothers infected with the virus.
○ Disease-specific rabies immunoglobulin is
available following the bite of an animal
suspected of carrying the disease or originating
from an area where the disease is endemic
○ Disease-specific varicella zoster immunoglobulin
is indicated for individuals who are at a high risk
such as neonates whose mothers develop
chickenpox, or for those exposed to the virus
while requiring intensive care or prolonged
special care, and for immunocompromised
individuals.
C. Viricidal Effects of chemical and physical agents on
viruses
● use of viricidal disinfectants on hard surfaces and
viricidal antiseptics on skin is important
● Chemical and Physical agents
○ often associated with organic materials, such as
secretions from the host, blood, faeces; which
enable them to persist on surfaces for longer
CAJUCOM, CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
periods of time and to survive better viricidal
challenges
○ Virus are not particularly resistant to chemical or
physical agents
○ small non-enveloped viruses (e.g. poliovirus) are
more resistant than large enveloped viruses (e.g.
HIV, influenza)
○ lipid-rich envelope being damaged easily by
chemical and physical agents.
● The viricidal activity of biocides (antiseptics,
disinfectants) varies and not all biocides show a
strong viricidal activity against non-enveloped viruses
(e.g. cationic biocides, phenolics and alcohols).
● Chemical Agents
○ Biocidal activity depends upon a number of
factors, such as concentration, contact time,
presence of soiling, and formulation.
○ Viruses present only a few target sites to
biocides, mainly the envelope (when present),
glycoproteins, the capsid and viral nucleic acid.
○ Destruction or alteration of the viral nucleic acid
would ensure complete viral inactivation.
○ Capsids are shown to be severely damaged in
the presence of highly reactive biocides such as
aldehydes (e.g. glutaraldehyde) and oxidizing
agents (e.g. peracetic acid, hydrogen peroxide).
○ Less reactive biocides, such as quaternary
ammonium compounds (QACs) and biguanides
(e.g. chlorhexidine) have been shown to damage
viral capsid to a lesser extent, explaining the
limited activity of these agents against
non-enveloped viruses.
○ Viral genome- infectious part of the virus and is
the ideal target for biocides.
○ Helical capsid structures are more susceptible
since the destruction/alteration of capsid is more
likely to cause damage to the viral nucleic acid
● Physical Agents
○ heat and irradiation are viricidal
○ play an important role in the control of viral
contaminants in pharmaceutical products
○ susceptible to exposure to temperatures above
60°C for 30 minutes
➢ used for the inactivation of viral
contaminants, such as HIV, in blood
products.
○ Other viruses such as the hepatitis B virus are
less susceptible and appropriate assurance of the
absence of such a virus is needed.
○ survive well at low temperatures and they can be
routinely stored at −40°C to −70 °C
○ UV irradiation and ionizing radiation (-ray and
accelerated electrons) are viricidal - destruction
of the viral nucleic acid
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○ Ionizing irradiation and thermal processes are ○ Minimize the risk of generating replicating
used for terminal sterilization processes applied viruses or to eliminate helper viruses when used
to medical and pharmaceutical products during production.
D. Control of viruses in pharmaceutical products ● For retroviridae-derived vectors, genetic modification
ensure that the recombinant retroviruses are
● The presence of certain viruses needs to be rendered replication-incompetent.
controlled in pharmaceutical products derived from ● Adeno-associated virus vectors (rAAV) are deficient
human and animal origin. adenovirus in which certain genes necessary for viral
○ Blood products such as human plasma for replication have been replaced
fractionation intended for: ● Poxviruses- family of viruses has an enveloped DNA
➢ manufacture of human antithrombin III
➢ human coagulation factor VII, VIII, IX, XI
➢ dried prothrombin complex
➢ dried fibrinogen
➢ normal immunoglobulin
➢ human α1- proteinase inhibitor
➢ human von Willebrand factor, for which
tests for the presence of antibodies against
HIV-1 and HIV-2
➢ hepatitis B surface antigen (HBsAg)
➢ antibodies against hepatitis C virus
○ For the urine-derived urofollitropin (obtained
from post- menopausal women), tests for
hepatitis virus antigens and HIV antigen are
needed.
● stringent controls are applied to the raw materials, in
process samples and to the final product.

● The type of inactivation measures used must be

validated against a range of representative viruses
(i.e. enveloped, non-enveloped, DNA, and RNA
VIII.
viruses)
● For the preparation of vaccines, the inactivation
process must ensure that it does not affect the
antigenicity while killing the virus and other potential
contaminants such as mycoplasmas
VII. Viruses and Gene Therapy
● Certain viruses or virus components are now used as
vectors for the delivery of genes to targeted cells
● Gene Therapy:
○ viruses are being used in gene transfer medicinal
products (GTMP) and these include:
➢ adenoviruses (AAV),
➢ poxviruses,
➢ retroviruses,
➢ lentiviruses,
➢ adeno- associated viruses
➢ Herpesviruses.
○ Genetically modified to transfer genetic material
to human somatic cells in vivo (i.e. injected
directly into the patient’s body) or ex vivo (i.e.
transferred into host cells before administration).
Viruses as antimicrobials
A. Bacteriophages
● viruses that infect only bacteria
● first described at the end of the 19th century
● typically 20–200nm in size and are highly diverse in
their structure and host range
● all bacterial species can be infected by a phage
○ Phages
➢ are extremely specific in their host range
and some will only infect a specific bacterial
strain
➢ proved to be very useful genetic tools over
the years, since they are easy to propagate
to high concentration and easy to study.
➢ used to elucidate the viral multiplication
cycle and their study has led to many
discoveries
● T-phage
○ referred to as ‘tadpole-shaped’
○ head (often icosahedral) that contains the viral
genome

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○ a tail which functions to recognize the host

receptor, attach and subsequently serve as a
nucleic acid injection device.
● One of the main differences between such phages
and common mammalian viruses is that these phages
inject their viral genome inside the host cell.
● Phage replication cycles
○ two scenarios have emerged
○ one resulting in the lysis of the bacterial host--
the lytic cycle
○ other resulting in the viral nucleic acid being
integrated into the host genome—the lysogenic
cycle
● Lytic cycle
○ involves the reproduction of viruses using a host
cell to manufacture more viruses; the viruses
● virulent phage- Infection with a lytic phage
then burst out of the cell.
● Transduction
● Lysogenic cycle
○ process where genes carried on the prophage
○ The viral nucleic acid which has integrated the
can then be expressed in the new host.
host genome is prophage, and the host cell that
○ responsible for gene transfer between bacteria.
contains the viral genome is lysogenic.
○ genetic factors transferred by transduction
○ incorporation of the viral genome into the host
encode for antibiotic resistant determinants
cell genome, infecting it from within.
and/or virulence factors such as toxins.
○ Use of lysogenic phages is best confined to the
genetic engineering of bacteria.
● Lysogeny is an extremely common phenomenon and
through evolution most bacteria will host several
prophages
● sequencing of the whole bacterial genome often
indicates the presence of prophages

B. Use of bacteriophages to treat bacterial

infection

● ● combat bacteria responsible for dysentery outbreaks

● first against against Vibrio cholera (work of Hankin)
and then Shigella Shiga (work of d’Herelle)
● Phages targeting bacteria causing a number of
diseases such as anthrax, scarlet fever, cholera and
diphtheria were quickly isolated
● The introduction of antibiotics in the early 1940s
resulted in the end of phage therapy in the West
● use of phages to combat Listeria monocytogenes in
ready- to-eat meat and poultry ( US)-2006,
cheese (EU)2009
● combat Pseudomonas aeruginosa in ear infection is
now in phase III clinical trials.

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● Phage preparations are also successfully employed in ● devoid of nucleic acid and are extremely resistant to
aquaculture against Lactococcus garvieae, the cause heating and ultraviolet irradiation
of a serious fish disease. ● fail to produce an immune response in the host.
● Ways to prepare a phage product : ● Clinical symptoms take a long time to develop, up to
○ ‘Natural’ phages from the environment 20 years in humans, but the disease has an inevitable
non-replicating phages genetically modified progression to paralysis, dementia and death.
phages lysogenic phages ● variant or vCJD - attacked young adults with an
○ Non-replicating phages have been used with average age of 30 rather than the 60-year-olds who
some degree of success, but in this case the lytic typically succumb to classical sporadic C
effect is short lived. ● Scrapie- prion found in sheep
○ Genetically modified phages have also been
used, whereby detrimental genes can be
removed and phage virulence genes added; VIRUS RECAP & COVID
patents might be easier to file
○ lysogenic phages can be used following FAMILY PAPOVAVIRIDAE
Characteristic dsDNA genome; icosahedral capsid, no
treatment to remove their lysogenic property.
s envelope
● Phage Therapy
Viruses and Human papilloma Skin and genital
○ use of phages for surface disinfection and diseases virus (HPV) warts, benign head
antisepsis or for the treatment of a bacterial and neck tumors,
infection anogenital warts
○ unlikely to supplant anti- biotic therapy in the Polyomavirus (BK Mild or
future and JC viruses asymptomatic
○ Advantages: Highly specific and Easy to isolate infects humans) primary infection,
virus remains
and propagate
dormant in
○ Disadvantages: Phage therapy could result in kidneys;
toxic shock from bacterial lysis; i.e. release of reactivation in
bacterial components such as lipopolysaccharide immunocompromis
wall material ed patients causes
C. Epidemiological uses and diagnosis hemorrhagic
cystitis (BKV) or
progressive
● high specificity of phage against their host leads to
multifocal
other applications leukoencephalopat
● Phage typing hy (JCV)
○ differentiates distinct strains of the same
bacterial species on the basis of their
susceptibility to phages. ● Papillomavirus
○ relevant with bacteria such as Salmonella ○ dsDNA
enterica serovar Typhi and to some extent ○ Over 100 types of human (HPV)
○ Infect skin and mucous membranes
Staphylococcus aureus that can be ‘typed’ during
○ Causes:
an outbreak ■ Warts
● Molecular techniques based on genetic differences ■ Cervical cancer
are now preferred for S. aureus. ■ Laryngeal carcinoma
● Application for phage is in developing rapid ○ No culture is performed, but molecular
diagnostics against slow-growing bacteria such as assays are used for diagnosis
Mycobacterium tuberculosis.

FAMILY PARVOVIRIDAE
Characteristics ssDNA virus; icosahedral capsid;
IX. Prions
no envelope
Virus Parvovirus B-19
● distinct class of infectious agents
Diseases Erythema infectiosum (fifth
● have unique and disturbing properties disease), aplastic crises in
● can be recovered from the brains of infected patients with chronic hemolytic
individuals as rod-like structures which are oli- anemias, and fetal infection and
gomers of a 30kDa glycoprotein. stillbirth

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FAMILY POXVIRIDAE Replicate in the cytoplasm

Characteristics Largest and most complex of all except orthomyxoviridae and
viruses; brick-shaped virion with retroviridae they replicate in
non conforming symmetry the nucleus
referred to a scomplex; dsDNA Smallest RNA virus:
genome Picornaviridae
Virus Smallpox, molluscum Largest RNA virus:
contagiosum and orf viruses Paramyxoviridae
Diseases smallpox is a generalized Arbovirus (arthropod-borne):
infection with pustular rash; Bunyaviridae, Flaviviridae and
molluscum manifests as benign Togaviridae
nodules of the skin; orf
manifests as localized papules/
vesicles of the skin FAMILY ARENAVIRIDAE
Characteristics Enveloped, irregular-shaped
● Poxviruses capsid containing
○ dsDNA, replicate in cytoplasm of the cell two-segmented ssRNA genome
○ Largest viruses Virus Lymphocytic choriomeningitis
○ Complex construction (LCM), Lassa Fever Virus,
MAchupo virus, Junin virus,
○ Brick shape, lipid envelope
Sabia virus
○ Resistant to environmental factors
Diseases LCM causes asymptomatic to
● Smallpox: variola influenza-like to aseptic
○ Eradicated by the world health organization meningitis-type disease; Lassa
○ Good vaccine and no animal reservoir fever virus causes influenza-like
● Vaccinia disease to severe hemorrhagic
○ Used to immunize against smallpox fever

FAMILY BUNYAVIRIDAE
Characteristics Segmented, ssRNA genome;
spherical or pleomorphic capsid
with envelope
Virus Arboviruses including the
california encephalitis group
containing LaCrosse virus, and
non arthropod-borne virus
including hantaviruses
(containing sin nombre virus);
Rift Valley fever virus, Crimean
Congo hemorrhagic fever virus
Diseases Encephalitis for arboviruses;
pneumonia or hemorrhagic
fever for hantaviruses

FAMILY IRIDOVIRIDAE
Characteristics dsDNA, icosahedral capsid FAMILY CALICIVIRIDAE
Virus Iridovirus Characteristics non-enveloped , icosahedral
Diseases African swine fever virus capsid surrounding ssRNA
genome
RNA VIRUSES In general: single stranded, Virus Norwalk and hepatitis E viruses
helical capsid, enveloped, Diseases Nausea, vomiting and diarrhea
reprilate in the cytoplasm (Norwalk), hepatitis similar to
Single-stranded RNA virus that caused by hepatitis A virus
except Reoviridae (dsRNA) except for extraordinarily high
Icosahedral capsid: case fatality rate (10-20%)
picornaviridae, caliciviridae, among pregnant women
reoviridae, retroviridae,
togaviridae and flaviviridae FAMILY CORONAVIRIDAE
Naked (not enveloped): Characteristics ssRNA genome; helical capsid
picornaviridae, caliciviridae and with envelope
reoviridae

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Virus Coronavirus
Diseases Common cold; possible
gastroenteritis, especially in
children
Coronavirus
○ ssRNA, eveloped
○ Pleomorphic, spherical capsid
○ Large club-shaped spikes on surface gives
“corona” effect
○ Coronaviruses are pleomorphic, roughly
spherical, medium-sized, enveloped RNA
viruses
■ Prefix corona- results from the viral
structure and the crown-like
surface projections on the external
surface of the virus that can be
seen with electron microscopy
■ Human respiratory coronaviruses
cause colds, and occasionally
pneumonia in adults
■ Viral transmission is person to
person via contaminated
respiratory secretions or aerosols
■ Virus is present in the highest
concentration in the nasal
passages, where it infects the nasal
epithelial cells
● In november, 2002, SARS (SARS CoV-1) was identified
as the cause of a worldwide outbreak. It first
emerged in the Guangdong province in china.
○ Because of its sensitivity and specificity,
molecular testing by RT-PCR remains the
recommended method for laboratory
diagnosis
○ Although nucleic acid testing by RT-PCR is
the most useful diagnostic test available, the
CAJUCOM, CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
virus is capable of growth in cell nature
using the Vero-E6 cell line
● Severe acute respiratory syndrome coronavirus 2
(SARS-CoV-2) is the etiologic agent of the current
rapidly growing outbreak of coronavirus diseases
(COVID-19), originating from the city of Wuhan,
Hubei province china.
○ Coronavirus disease 2019 (COVID-19) is
cause by a new coronavirus first identified in
WUha, China, in December 2019
○ Although most people who have COVID-19
have mild symptoms, COVID-19 can also
cause severe illness and even death
○ Some groups, including older adults and
people who have certain underlying medical
conditions, are at increased risk of severe
illness
● On february 11, 2020, the world health organization
announced an official name for the disease that is
causing the 2019 novel coronavirus outbreak.
○ The new name of this diseases is
coronavirus disease 2019, abbreviated as
COVID-19
○ In COVID-19, CO stands for corona, Vi for
virus and D for disease. Formerly, this
disease was referred to as 2019 novel
coronavirus or 2019-nCoV
● The most common symptoms of COVID-19 are:
○ Fever
○ Dry cough
○ Fatigue
● Other symptoms that are less common and may
affect some patients include:
○ Loss of taste or smell
○ Nasal congestion
○ Conjunctivitis (also known as red eyes)
○ Sore throat
○ Headache
○ Muscle or joint pain
○ Different types of skin rash (usually toes)
○ Nausea or vomiting
○ Diarrhea
○ Chills or dizziness
● Symptoms of severe COVID-19 disease include:
○ Shortness of breath
○ Loss of appetite
○ Confusion
○ Persistent pain or pressure in the chest
○ High temperature (above 38 C)
● Other less common symptoms:
○ Irritability
○ Confusion
○ Reduced consciousness (sometimes
associated with seizures)
○ Anxiety
○ Depression
○ Sleep disorders
○ More severe and rare neurological
complications such as strokes, brain
inflammation, delirium and nerve damage
MAYO CLINIC INSIGHTS: WHAT IS COVID-19 VARIANT STRAIN
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● Variant- mutation that occurs in the virus overtime.
So just with natural evolution overtime. The virus
infects people it has the opportunity to replicate its
genome, and everytime it replicates there are
chances for mutations to occur
● Infected millions of people and it replicated its
genome billions of times and airs have been
incorporated these results in mutation
● Variants were expected
● 3 primary variants of COVID-19
○ United kingdom variant
■ Higher potential for transmission
■ Spread faster and easier between
people
■ Doesn’t cause more severe disease
or lead to a higher rate of
hospitalizations
○ South african variant
■ Higher potential for transmission
○ Brazil variant
■ Still new, more to learn
● Why do variants have a geographical name?
○ Media and scientific reports are referring to
these variants where they were first
identified
○ By the time we learn about them they are
spread through many countries if not
throughout the world, so it’s more
important to understand that this occurs
naturally, we expect this to happen
● Do the current vaccines offer protection against these
variants?
○ I think that it is safe to say that the vaccines
will offer protection to the currents strains
of COVID-19
○ It is important to think about the efficacy or
effectiveness of the vaccine on a continuum
rather than a binary yes and no
■ UK variant appears that the pfizer
and moderna vaccine will offer
strong protection against that
strain
■ South african variant, there are
some evidence that the efficacy
may be reduced (may be from 95%
efficacy to 85% efficacy against
preventing severe COVID disease)
● How do we stop new strains from emerging?
○ Viruses can’t mutate if they don’t replicate,
so:
■ Vaccination
■ Masking
■ Physical distancing
CAJUCOM, CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
○ Are going to be really important to drive
down the infections, and that will slow the
emergence of new variants in mutating
IN-DEPTH: WHY COVID VARIANTS JUST GOT NEW NAMES
● The world health organization wants to stop using
names such as the UK and India variants the health
agency just ruled out a new naming system
● UK variant = alpha variant
● South african variant = beta variant
● Brazil variant = gamma variant
● India variant = delta variant
● The new naming system is based on the greek
alphabet, will be easier, more practical, and less
stigmatizing than the location based
● If we don’t change the names and the terminologies
the, the racism will be sustained
● Hate crime
● Sop AAPI hate documented 6,600 targeting asian
american
● There is an increase in asian american hate after
former american president Donald Trump tweeted
“chinese virus”
● The asian americans are attacked because of the
terminologies
● The challenge in the greek alphabet is it only has 24
letters and the WHO used 10 already
The time from exposure to COVID-19 to the moment when
symptoms begin is, on average, 5-6 days and can range from
1-14 days
● This is why people who have been exposed to the
virus are advised to stay remain at home and stay
away from others, for 14 days, in order to prevent the
spread of the virus, especially where the testing is not
easily available
Both isolation and quarantine are methods of preventing the
spread of COVID-19. Quarantine is used for anyone who is in
contact with someone infected with the SARS-CoV-2 virus,
which causes COVID-19, whether the infected person has
symptoms or not.
● Quarantine means that you remain separated from
others because you have been exposed to the virus
and you may be infected and can take place in a
designated facility or at home
● For COVID-19, this means staying in the facility or at
home for 14 days
Isolation is used for people with COVID019 symptoms or who
have tested positive for the virus
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● Being in isolation means being separated from other ● Extend the respirator under your chin, protecting
people, ideally in a medical facility where you can both your mouth and nose
receive clinical care ● Pull the top strap over your head placing it on the
● If isolation in a medical facility is not possible and you crown of your head
are not in a high risk group of developing severe ● Pull the bottom strap over your head placing it on the
disease, isolation can take place at home base of the neck
● If you have symptoms, you should remain in isolation ● Lastly, perform a user seal check, do this by using
for at least 10 days plus an additional 3 days without your hands to cover the surface of the respirator and
symptoms gently exhale checking if the respirator bulges slightly
● If you are infected and do not develop symptoms, you ● Then take a deep breath checking if the facepiece
should remain in isolation for 10 days from the time collapses slightly
you tested positive ● If air escapes through the edges, re-adjust the
○ respirator and perform another seal check. Do this
every time you put the respirator on
HAND-WASHING STEPS USING THE WORLD HEALTH ● If a respirator is not available, put on a face mask
ORGANIZATION (WHO) TECHNIQUE ● Extend the facemask to your chin, protecting your
mouth and nose, if the mask has loops put them
● Wet hands with water around your ears. If it has ties, secure them at the
● Apply enough soap to cover all hand surfaces base of your neck and at the crown of your head
● Rub hands palm to palm ● Put on a face shield or goggles
● Right palm over the left dorsum with interlaced ● Lastly, put on your gloves
fingers and vice versa ● Pull down the gloves so that it covers the wrist of the
● Palm to palm with fingers interlaced gown
● Back of fingers to opposing palms with fingers
interlocked DEMONSTRATION OF DOFFING (TAKING OFF) PERSONAL
● Rotational rubbing of left thumb clasped in right palm PROTECTIVE EQUIPMENT (PPE)
and vice versa
● Rotational rubbing backwards and forwards with ● Before exiting the patient’s room
clasped fingers of right hand in left palm and vice ● Remove and discard your gloves
versa ○ Glove in glove
● Rinse hands with water ■ Pinch the outside of the glove
● Dry hands thoroughly with a single use towel ■ Peel downwards pulling the glove
● Use the towel to turn off the faucet inside out
■ With your ungloved hand slide
DEMONSTRATION OF DONNING (PUTTING ON) PERSONAL your finger under the wrist of your
PROTECTIVE EQUIPMENT (PPE) glove
■ Peel downwards turning the glove
● Identify and gather the proper PPE to put on inside out
○ Appropriately sheeted isolation gown ■ Discard the gloves
○ N95 filtering facepiece respirator or higher ○ Beaking method
level respiratory protection ■ Pinch the outside of your glove
○ If a respirator is not available, face mask near the wrist
○ A shield or goggles ■ Using your finger pull the glove
○ And a pair of disposable patient inside out and over the fingers
examination gloves forming a beak
● Perform hand hygiene by using alcohol based ■ With the beak hand pinch the
sanitizer or washing your hands with soap and water opposite glove at the wrist and pull
for at least 20 seconds downwards turning the glove
● Put on the isolation gown inside out
● Tie all ties or snaps all snaps ■ With the ungloved hand pull the
● You may need assistance with another health care beaked glove off touching only the
provider inside of the glove
● Put on the N95 respirator ● Remove your gown. Untie all ties or unsnap all snaps
● Fit it to your nose with both hands. Do not bend the some gowns can be broken rather than untied in that
respirator

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instance break the gown gently avoiding a forceful hepatocellular

movement carcinoma
● Reach up to the shoulders carefully pull, roll the gown
down and away from the body
● Flaviviruses
● Dispose the used gown
○ sssRNA, enveloped
● You may now exit the patient’s room ○ Icosahedral symmetry
● Perform hand hygiene by using alcohol based ○ St. louis encephalitis, west nile virus, yellow
sanitizer or washing your hands with soap and water fever, dengue, hepatitis C
for at least 20 seconds. Be sure to clean your wrist ○ Not isolated by culture
where the edge of the glove was located ● Hepatitis C virus
● Carefully remove the face shield or goggles by ○ Blood or sexual contact
○ No other vector
carefully removing the straps upwards and away from
○ Chronic liver infection
your head. DO NOT TOUCH THE FRONT OF THE FACE ○ Serology most common diagnosis
SHIELD OR GOGGLES ○ Also detected in blood by molecular assays
● Remover and discard the respirator
● If respirator, remove the bottom strap, by grabbing FAMILY ORTHOMYXOVIRIDAE
only the strap carefully over your head Characteristic Segmented, ssRNA genome; helical capsid
● Grasp the top strap and bring it carefully over your s with envelope; three major antigenic
types, influenza A, B, and C; types A and B
head
cause nearly all human disease
● Pull the respirator away from your face without
Viruses and Influenza A Influenza (malaise,
touching the front of the respirator diseases headache, myalgia,
● If facemask, carefully untie the straps or unhook cough); primary
them from your ears influenza
● Pull the mask away from your face without touching pneumonia; in
the front of the facemask children,
bronchitis, croup,
● Lastly, perform hand hygiene
otitis media
Influenza B Similar to mild
influenza
● Orthomyxoviridae
RNA VIRUS
○ ssRNA; eight segments
○ Helical symmetry capsid, enveloped
FAMILY FILOVIRIDAE ○ Replicate in cytoplasm
Characteristics Enveloped, long, filamentous ● Influenza viruses
and irregular capsid forms with ○ Two important surface Ag
ssRNA ■ Neuraminidase
Family Ebola (aka ebola-reston) and ■ Hemagglutinin
marburg viruses ○ Segmented genomes: heavy reassortment
Diseases Severe hemorrhages and liver ■ Antigenic shift
necrosis; mortality as high as ● Phenomenon of slight
90% antigenic change seen in
influenza viruses over
time as a result of minor
FAMILY FLAVIVIRIDAE mutations in the ssRNA
Characteristic ssRNA genome surrounded by spherical ■ antigenic drift
s and icosahedral capsid ● Phenomenon whereby an
Viruses and Arboviruses St. louis and west often unexpected change
diseases including yellow nile encephalitis, occurs in influenza virus
fever, west nile, dengue and yellow strains. This antigenic
japanese fever change is often so drastic
encephalitis and St. that it triggers pandemics
Louis encephalitis ○ Severely in elderly, immunocompromised
viruses ○ Pandemics
Hepatitis C virus Acute and chronic ■ 1918 (spanish flu)
hepatitis; strong ■ Avian flu or swine flu
correlation ○ Can be isolated in culture
between chronic
HCV infection and FAMILY PARAMYXOVIRIDAE

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Characteristic ssRNA genome; helical capsid with Polivirus Coxsacke A:

s envelope; no segmented genome like herpangina,
orthomyxoviruses Coxsackievirus hand-foot-mouth
Viruses and Measle virus Measles, atypical group A disease
diseases measles (occurs in
those with waning Coxsackievirus Coxsackie B:
vaccine immunity), group B pleurodynia,
and subacute - Echovirus pericarditis and
sclerosing - enterovir myocarditis
panencephalitis uses
Enterovirus 70:
Hallmark rash of acute hemorrhagic
measles infection is conjunctivitis
referred to as Hepatitis A virus Hepatitis with short
Koplik’s sports, (enterovirus type incubation, abrupt
which are bluish 72) onset, and low
white sports with a mortality; no
red halo located on carrier state
the buccal or labial Rhinovirus Common cold
mucosa (common cold
Mumps Mumps viruses)
Parainfluenza virus Adults: upper ● Picornavirus
respiratory, rarely ○ ssRNA
pneumonia ○ Very small: approximately 27 nm in
diameter
Children: ○ No envelope, icosahedral symmetry
respiratory ○ Replicate in cytoplasm of cell
including croup, ● Enteroviruses
bronchiolitis, and ○ Fecal-oral transmission
pneumonia ■ Polioviruses: 3 types
Respiratory Infants: ■ Coxsackie A: 24 types
syncytial virus bronchiolitis, ■ Coxsackie B: 6 types
(RSV) pneumonia and ■ Echoviruses: 34 types
croup ○ Many cultivable
● Rhinoviruses
Children: upper ○ Common cold virus
respiratory ○ More than 100 serotypes
○ Can be isolated in culture
○ Acid-sensitive
○ Limited to upper respiratory tract
● Hepatitis A virus
○ 25% of hepatitis
○ Usually fecal-oral transmission
○ No growth in cell culture
○ Serology for diagnosis

FAMILY REOVIRIDAE
Characteristics Segmented, dsRNA genome;
icosahedral capsid with no
● Koplik’s spots envelope
○ Little spots inside the mouth that are highly Family Rotavirus
characteristic of the early phase of measles Diseases Gastroenteritis in infants and
(rubeola) children 6 months to 2 years
● Subacute sclerosing panencephalitis (SSPE) ● Reovirus
○ Progressive, disabling, and deadly brain ○ Respiratory enteric orphan
disorder related to measles (rubeola) ○ dsRNA, 60 to 80 nm
infection ○ nonenveloped , icosahedral symmetry
○ Replicates in the cytoplasm
FAMILY PICORNAVIRIDAE ● Rotaviruses
Characteristic ssRNA genome; icosahedral capsid with ○ Fecal-oral
s no envelope ○ Causes infantile diarrhea
Viruses and Enteroviruses: Poliovirus: Polio ○ Not isolated in laboratory, ELISA
diseases

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○ At least six serotypes ○ Travels up sensory nerves to the central

nervous system
FAMILY RETROVIRIDAE ○ Incubation of 2 to 16 weeks
CharacteristicssRNA genome; Icosahedral capsid with ○ 100% fatal if untreated
s envelope; reverse transcriptase converts ○ Negri bodies
genomic RNA into DNA
Virus Human T-cell leukemia and FAMILY TOGAVIRIDAE
T-lymphotropic lymphoma, and Characteristic ssRNA genome; Icosahedral capsid with
viruses (HTLV-1 and tropical spastic s envelope; family contains arboviruses and
HTLV-2) paraparesis for non arthropod-borne rubella virus
HTLV-1; no known Virus and Rubella virus Rubella (mild
disease associated Disease exanthematous
for HTLV-2 disease)
Disease Human Most diseases in Arboviruses Eastern, western
immunodeficiency humans caused by including and venezuelan
virus types 1 and 2 HIV-1; infected alphaviruses equine encephalitis
(HIV-1 and HIV-2( cells include CD4 ● Togaviruses
(helper) T ○ ssRNA
lymphocytes, ○ Enveloped, icosahedral symmetry
monocytes and ● Arboviruses
some cells of the ○ Eastern and western equine encephalitis
central nervous ■ Bird reservoir
system; ■ Mosquito vectors
asymptomatic ○ Symptoms include fever, encephalitis, rash
infection, acute ○ Cell culture possible, but serology most
flu-like disease, commonly used
AIDS related ● Rubella virus
complex, and AIDS, ○ Transmitted by droplets
AIDS associated ○ Infection in children mild
infections and ○ Congenital disease serious
malignancies ○ Mother contracts rubella in first trimester
● Retroviruses ○ Vaccine developed for children because of
○ ssRNA enveloped, icosahedral symmetry congenital disease
○ All have reverse transcriptase ○ Cell culture possible but serology most
○ Many can cause tumors in animals common
○ Replicate in nucleus and cytoplasm
○ HIV 1 and 2, human T-lymphotropic virus 1
and 2
● HIV REFERENCES
○ Can lead to AIDS
○ Does not form tumors Notes from the discussion by: Ms. Marilyn Ngo
○ Infects CD4 + cells - the CD4 should not be
below 200 microliter University of Santo Tomas powerpoint presentation: Ms.
■ If lower 200 microliter = AIDS Marilyn Ngo
○ Destroys immune system
○ Characteristic secondary diseases: Course book: Hugo & Russell’s Pharmaceutical Microbiology
pneumocystis pneumonia, CMV, kaposi’s (2011) 8th ed.
sarcoma
○ Diagnosis: ELISA and western blot analysis

FAMILY RHABDOVIRIDAE Edit History Log

Characteristics ssRNA genome; helical capsid Date Remarks
with envelope, bullet-shaped Date Here Version 1 completed by DIMAPANAT,
Virus Rabies virus VICEDO
Diseases Rabies Date Here Edits made by
● Rhabdoviruses Date Here Evaluated by XXX
○ ssRNA, bullet-shaped, enveloped Date Here Compiled to Master File
○ Replicate in cytoplasm
● Rabies virus
○ Encephalitis
○ Bite of infected animal

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TRANS: VIROLOGY

OUTLINE (VIDEO DISCUSSION)

I. Virology
II. Viral Ultrastructure
III. Viral Replication
IV. Viral Transport
V. Sample Sites and Associated Viral Agents
VI. Viral Identification
VII. RNA Viruses

I. Virology

● Virology – study of viruses

● Viruses – are obligate intracellular parasites unable to
self-replicate
● Once inside living cells, they induce the host cell to
synthesize virus particles.
● the genome is either DNA or RNA (single or double
stranded)
● Do not have a system to produce ATP.
● Range in size from 25 to 270 nm
● Classification is based on nucleic acid type, size and
shape of virion, and presence or absence of an
envelope.
○ Insert description here
➢ Insert description here III. Viral Replication

Steps in Viral Replication

II. Viral Ultrastructure
1. Attachment and absorption
● Virion is the entire viral particle. 2. Penetration
● Capsid is the protein coat that encloses the genetic 3. Uncoating
material. 4. Early viral mRNA synthesis
● Capsomer is the protein subunit that makes up the 5. Early viral protein synthesis
capsid.
● Nucleocapsid is composed of the capsid and genetic
material.
● The envelope is the outer coating composed of a
phospholipid bilayer, which is composed of
viral-encoded glycoproteins and sometimes viral
encoded matrix proteins

6. Viral genome replication

7. Late viral mRNA synthesis
8. Late viral protein synthesis
9. Assembly
10. Release

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IV. Viral Transport

● Samples for viral culture must be placed into a viral
transport medium (VTM)
● VTM contains:
a. Buffered Saline
b. Protein Stabilizers
c. Antimicrobials that inhibit bacterial and fungal
growth
● Samples for viral cultures can be refrigerated in VTM
for about 48 hours, but they should never be frozen
at -20C
● Samples can be stored at -70C; however, infectivity
will be diminished

V. SAMPLE SITES AND ASSOCIATED VIRAL AGENTS:

RESPIRATORY INFECTIONS
• Upper respiratory tract infections - are commonly
caused by viruses, including rhinovirus, influenza
virus, parainfluenza virus, respiratory syncytial virus
(RSV), Epstein-Barr virus (EBV), and coronavirus.
• Croup and bronchitis - can be caused by influenza
virus, parainfluenza virus, RSV, and adenovirus.
• Pneumonia- in children can be caused by RSV,
parainfluenza virus, adenovirus, and varicella-zoster
virus (VZV).
• Pneumonia in adults - can be caused by influenza
virus, VZV, cytomegalovirus (CMV), and RSV.

VIRAL MENINGITIS
● Caused by enterovirus. echovirus, herpes simplex
virus type 1 (HSV-I), HSV-2. and VZV 2.
● Viral meningitis is often less severe than bacterial
meningitis. Aseptic meningitis is an older term
referring to meningitis not caused by easily cultured
bacteria. The term has become synonymous with viral
meningitis

NEONATAL INFECTIONS
● are acquired in Utero, during childbirth, or soon after
SPECIMEN PROCESSING FOR DIAGNOSING VIRAL DISEASES
childbirth.
Viruses are in highest concentrations during the first
● The infections can be caused by HSV, CMV, and
several days following onset of symptoms. Therefore,
rubella virus
samples should be collected early in the disease
course. Samples should generally come from the
ENCEPHALITIS
infected site.
● Encephalitis typically has a viral etiology.
● Skin infections: Rash site and, depending on the
● Infections are caused by Hsv, vzv, and arboviruses.
virus, serum and urine
Arboviruses are genetically unrelated viruses
● Respiratory infections: Sputum or throat swabs
transmitted by arthropods (e.g., mosquitoes); they
● Central nervous system (e.g., meningitis and
include the families Togaviridae (eastern and western
encephalitis): For diagnosis of meningitis,
equine viruses) and Flaviviridae (St. Louis encephalitis
cerebrospinal fluid (CSF) and serum, as well as stool
virus).
or throat swabs, can be collected because viruses are
● A number of animals, including birds and horses,
sometimes shed into these sites. In cases of
serve as reservoirs for these viruses. Rabies virus is
encephalitis, brain biopsy material and sometimes
also an uncommon cause of encephalitis.
serum are used.
● Encephalitis is an infection of the brain or spinal cord
● Urogenital infections: Needle aspirates and
and is much more severe than viral meningitis.
endocervical and urethral swabs
● Gastrointestinal tract: Stool samples and rectal swabs
● Eye infections: Eye swabs and corneal scrapings

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CUTANEOUS INFECTIONS ○ RNA viruses produce cytoplasmic inclusions

- Caused by HSV-1, HSV-2, VZV, echovirus, measles (assembled in the cytoplasm)
virus, rubella virus, enterovirus, molluscum ○ HSV and VZV cause intranuclear inclusions
contagiosum vims, and parvovirus B-19 ○ CMV induces enlarged (cytomegalic) cells
- Cutaneous infections often result in a rash that, with a basophilic intranuclear inclusion
depending on the virus, can have a variety of referred to as “owl eye” inclusion
presentations

VIRAL ISOLATION
● Cell cultures
○ Primary cell cultures
GENITAL INFECTIONS ○ Established cell lines
● Frequently caused by HSV-2 and human ● Ebmbryonated eggs
papillomavirus ● Animal models
● Genital tract infections are typically sexually
transmitted

GASTROENTERITIS
● Caused by a number of viruses including rotaviruses,
Norwalk viruses, adenoviruses, and caliciviruses
● The symptoms can range from mild self-limiting
diarrhea to severe diarrhea with dehydration,
particularly with rotavirus infections in young children

ELECTRON MICROSCOPY
● Due to size, most individual virions can only be seen
by electron microscopy. However, the poxyviruses are
about the size of some small bacteria
● Electron microscopy is sometimes used to identify
Norwalk viruses, astrovirus, calicivirus, and
coronavirus
EYE INFECTIONS ● Electron microscopy is expensive, requires expertise,
● Caused by HSV, adenovirus, and VZV and is usually not very sensitive
● Viruses can cause conjunctivitis if it is severe

VI. VIRAL IDENTIFICATION

CELLULAR INCLUSIONS
● are diagnostic for many viruses
○ Because most DNA viruses replicate in the
nucleus, they often produce nuclear
inclusions. However, some DNA viruses are
assembled elsewhere in the cell

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OTHER METHODS OF IDENTIFICATION

● Detection of host antibodies directed against specific
viruses
● Direct detection of viral antigens in clinical specimens
● Viral gene probes and nucleic acid amplification (e.g.
polymerase chain reaction [PCR])
●

VIRUS SIZES AND ILLUSTRATIONS

VII. DNA VIRUSES

HERPESVIRUSES
● Herpesviruses are icosahedral shaped, have an
envelope, and range in size from 90-100 nm
● They are members of the family Herpesviridae
● Except for neonates, infections are more severe in
adults than in children
● All herpesviruses produce latent infections
● Sites of latency include leukocytes and peripheral
nerves
● Reactivation may result from physiological stress, the
symptoms are milder than primary infection. The
● Smallest: bacteriophage (24nm) exception is shingles, which is a reactivation of VZV
● Poliovirus (30 nm) ● Herpesviruses include VZV, HSV-1 and -2, CMV, EBV,
● Rhinovirus (30 nm) and human herpes viruses 6, 7, and 8
○ HSV-1: oral herpes
○ HSV-2: genital herpes

HERPES SIMPLEX VIRUS TYPE 1 (HSV 1)

● Infections associated
○ Mouth lesions and fever blisters (cold sores)
○ Include mild fever and general malaise
● Diagnosis
○ Clinical symptoms
○ Immunologic assays that detect viral
antigens
○ Viral isolation

● HSV will grow in continuous cell line (e.g. HEp-2 and

A549) and established cell lines (e.g. MRC-5)
● In A549 cells, syncytia are sometimes seen
● MRC-5 infected cells develop cytoplasmic granules
that become large, round, and refractive (BALLOON
CELLS) clusters (foci) of infected cells appear in a few
days
DNA AND RNA VIRUSES
● HSV are smart enough to deliver large transgenes to
neurons
○ Packaging capacity: up to 150kb
○ Transduction efficiency: ⅗
○ Immune response: 5/5

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○ Infection: most dividing/non-dividing cells

(ideal for neuronal cells)
○ Expression: transient
○ Integrating: No (but may replicate
separately from the host)
○ Genetic material: double stranded linear
DNA

HERPES SIMPLEX VIRUS TYPE 2 (HSV 2)

● Infections associated:
○ Genital herpes: a common sexually
transmitted disease (STD)
○ Lesions appear on the penis, cervix, and SHINGLES
vagina
○ Has been linked to cervical carcinoma ● Reactivation of VZV in the peripheral or cranial nerves
○ Infant infection acquired during childbirth leads to shingles and occurs mainly in the elderly
can cause severe eye infections and central ● Characterized by skin vesicles, often on one side of
nervous system (CNS) damage the body, and severe pain around the skin lesions
● Complications include CNS disorders, eye problems,
● Diagnosis
adn facial paralysis
○ Clinical symptoms
● Diagnosis is often based on clinical symptoms
○ Immunologic assays that detect viral
antigens
○ Viral isolation
○ serology

CYTOMEGALOVIRUS (CMV)

● Notes:
VARICELLA-ZOSTER VIRUS (VSV) ○ MOT is through contact with saliva or blood
○ CMV results in persistent infections in
● Notes: humans, including endothelial cells and
○ MOT is via respiratory aerosis from vesicular leukocytes. The tubular cells of the human
skin lesions of infected individuals kidney shed CMV for prolonged periods into
● Infections associated the urine
○ Chickenpox ● Infections associated
■ Primarily a childhood illness; ○ Pneumonia and encephalitis
however, symptoms are more ○ Congenital infections are severe and cause
severe in adults developmental problems for the newborn
● Incubation period is from 1 to 2 weeks ● Diagnosis
● Symptoms include a rash and fever ○ Serologic testing
● Individuals are contagious 48 hours before the rash ○ Viral isolation from blood, respiratory
and will remain contagious until scabbing of all secretions, or urine
lesions ● The virus grows best in human fibroblast cells (e.g.
● Prevention: routine use of vaccine has greatly WI-38 and MRC-5), where it will produce
reduced the incidence of chickenpox characteristic intranuclear inclusions previously
described

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EPSTEIN-BARR VIRUS (EBV)

● notes :
○ The virus is transmitted in saliva
○ The incubation period lasts 1-2 months
● Infections associated:
○ Infectious mononucleosis HUMAN HERPESVIRUS-7
○ Burkitt lymphoma
○ Hodgkin’s disease
● Notes:
○ Nasopharyngeal carcinoma
○ HHV-7 is also very common virus, with
●
serologic prevalence rates in healthy adults
● Diagnosis
about 90%
○ Signs and symptoms
○ The virus is mostly transmitted via saliva and
■ Symptoms include fever, enlarged
infects lymphocytes
lymph nodes, and swollen tonsils
● Infections associated:
○ Hematologic abnormalities include
○ In immunocompetent individuals, infections
lymphocytosis and the presence of reactive
are mild or asymptomatic
(atypical) lymphocytes
○ HHV-7 causes about 5% of all cases of
○ Serology
Roseola; neurologic involvement is rare

HUMAN
HUMAN HERPESVIRUS-6 HERPESVIRUS-8

● HHV-6 is a very common virus and is acquired by ● Notes

respiratory secretions ○ The virus is probably transmitted via oral
● It infects T lymphocytes seretions
● Infections associated: ○ Little is known about primary HHV-8
○ Infections are generally mild or subclinical in infections
immunocompetent individuals ○ In latent infections, viral DNA has been
○ HHV-6 causes Exanthem Subitum, also found in B cells and peripheral blood
known as Roseola or Sixth disease mononuclear cells
○ Sixth disease is a childhood disease ● Infections associated:
characterized by fever, rash, and sore throat; ○ Kaposi sarcoma in immunosuppressed
neurologic involvement is rare patients (e.g. acquired immunodeficiency
syndrome [AIDS])
○ HHV-8 is also known as Kaposi Sarcoma
herpes virus (KSHV)

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● Variola virus is considered a potential bioterrorism

agent
● Infections associated
HUMAN PAPILLOMA VIRUS (HPV) ○ Variola major causes smallpox
○ Variola minor strains produced milder
● Notes: infections with a fatality rate of less that 1%
○ HPV has an icosahedral shaped, enveloped ● Due to a worldwide vaccination program, the World
virion, the viruses range in size from 40-50 Health Organization was able to declare the world
nm smallpox free in 1979
○ Member of the family Papillomarividae ● The vaccine virus is an attenuated vaccine that
● Infections associated: prevents variola infection
○ Plantar wars, genital warts, flat warts
○ Some HPVs are associated with cervical
cancer
● Genital warts:
○ Most common STD in the U.s. and HPV is
the most common cause of cervical cancer
○ A vaccine was approved for use in females.
The vaccine, designed to prevent cervical
cancer, contains the strains of HPV most
often associated with this disease

OTHER POXVIRUSES

● Molluscipoxvirus
○ Causes molluscum Contagiosum: a skin
infection that occurs worldwide
○ Characteristic inclusion:
Henderson-Patterson Bodies
● MonkeyPoxvirus
○ Causes Zoonosis
○ Found primarily in Africa
○ An outbreak recently occurred in the U.s.
○ Infections were traced back to rodents
- imported from Africa that transmitted the
POXVIRUSES virus to Prairie dogs
● Poxviruses are large, ranging in size from 220 to 450
nm
● Virions also contain a DNA polymerase for DNA
replication and an RNA polymerase system for
transcription of viral genes
● Poxviruses replicate entirely within the cytoplasm
● Poxviruses belong to the family Poxviridae

ADENOVIRUSES
● Notes
○ Adenoviruses are naked icosahedral virions
○ They range in size from 70 to 90 nm
VARIOLA VIRUS ○ Adenoviruses belong to the family
Adenoviridae and have been isolated from
humans and animals

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● Infections associated immune responses. As more T cells are destroyed,

○ Respiratory tract infections, especially in immune function deteriorates. CD4 is the primary
young children receptor for the virus
○ Urinary tract and gastrointestinal infections, ● Infection in most dividing/non-dividing cells
and pharyngitis ● Infection: stable
○ Eye infections in newborns, ● Integrating: yes
immunocompromised patients, and military ● Genetic material: RNA
recruits (because of close living conditions)
are common
○ Keratoconjunctivitis
● Sample collection:
○ Done through throat swabs, eye samples,
and stool samples
● Packaging capacity: 8 kb
● Transduction efficiency: 4/5
● Immune response: 5/5
● Infections: most dividing/non-dividing cells, with high
transduction rate towards primary cells
● Expression: transient HIV INFECTION IN TARGET T CELLS
● Integrating: No
● Genetic material: double stranded linear DNA

VII. RNA VIRUSES

RETROVIRUSES
● Retroviruses have an icosahedral shaped, enveloped
virion. They range in size from 80 to 130 nm
● Retroviruses contain an RNA-dependent DNA
polymerase (reverse transcriptase) for replication
● Reverse transcriptase uses viral RNA as a template to
make double-stranded DNA that then moves into the
nucleus where it is integrated into the host
chromosome
● This stage is referred to as a provirus. The genome is
transcribed into mRNA by host RNA polymerase ● Virus replication occurs at a high rate in lymphoid
● Only infectious to dividing cells tissue, but the patient remains asymptomatic for
● Packaging capacity: 8 kb many years. The host is able to replace infected T
● Transduction efficiency: 3/ 5 cells as fast as they are destroyed. This condition is
● Immune response: 4/5 referred to as “clinical latency”
● Infection: dividing cells ● Eventually, the virus begins to destroy T cells faster
● Expression: stable than they can be replaced. As immune function is
● Integrating: yes compromised, the patient presents with chronic and
● Genetic material: RNA recurrent infections, including Pneumocystis
pneumonia, CMV infections, mycobacteriosis,
cryptosporidosis, candidiasis, and toxoplasmosis. This
LENTIVIRUSES stage is sometimes referred to as AID-related complex
● As immune function continues to deteriorate, the
● Notes:
infections become more severe and life threatening.
○ Spread of the virus by sexual contact with
This stage is referred to as AIDs or full-blown AIDS
infected individuals (homosexual or
● HIV has also been associated with malignant
heterosexual), intravenous drug use,
conditions such as Kaposi sarcoma, cancer, and B cell
congenital transmission, or contaminated
lymphomas
blood products
● Diagnosis is by clinical history, serology, and detection
○ The human immunodeficiency virus (HIV-1)
of viral antigens or RNA
and HIV-2 are members of the genus
Lentivirus
WESTERN BLOT
● Disease associated
○ HIV is the causative agent of AIDS ● Confirmatory test for determining true HIV infection
○ HIV-1 causes a more severe infection and is
much more prevented than HIV-2

● The virus initially infects macrophages and dendritic

cells, then the host’s CD4 positive T cells. These cells
are key to both humoral-mediated and cell-mediated

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● Interpretation of results
○ No bands, negative
○ In order to be interpreted as positive a
minimum of 3 bands directed against the
following antigens must be present: p24,
p31, gp41, or gp120/160
○ CDC criteria require 2 bands of the
following: p24, gp41, or gp120/160
HUMAN T-CELL LYMPHOTROPIC VIRUS (HTLV)
● This group of viruses includes HTLV-1 and HTLV-2
● These viruses are transmitted via sexual contact,
mother to child by breastfeeding, and parenteral drug
use
● Infections associated:
○ HTLV-1 has been linked to adult T-cell
leukemia and HTLV-1 associated
myeiopathy/tropical spastic paraparesis
(HAM/TSP)
○ Although HTLV-2 has not been associated
with malignancies, it has been linked to a
neurologic disease resembling HAM/TSP
ORTHOMYXOVIRUSES
● Virions contain a segmented RNA genome and have a
helical-shaped virion with an envelope
● They range in size from 75-125 nm
● The influenza viruses A, B, and C are the only
members of the family orthomyxoviridae
● Orthomyxoviruses have hemagglutinin (HA) and
neuraminidase (NA) on their surface. These
molecules are immunogenic, and antibodies to these
molecules confer protection
○ HA allows the viruses to attach to the
surface of respiratory epithelial cells and
also agglutinate red blood cells. NA has
enzymatic activity, cleaving budding viruses
from infected cells
● ○ An infected cell can cause fusion with
● Antigenic drift:
○ occurs when point mutations occur in the
viral genes encoding the HA and NA spikes.
○ Antigenic drift can occur within any of the
three influenza viruses
● Antigenic shift:
○ occurs following a major change
(reassortment) of the RNA genome when a
CAJUCOM, CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
single host cell is infected with two different
influenza viruses.
○ Among the influenza viruses, antigenic shift
only occurs in influenza A viruses.
○ Antigenic shift results in a new combination
of viral surface glycoproteins (e.g. from
H1N1 to H2N1)
● Influenza can infect other animals, including birds and
pigs
○ In these animals, the virus often undergoes
recombination events, resulting in new
strains.
○ Epidemics and pandemics are generally due
to antigenic shifts.
○ Other viruses such as HIV, can also undergo
antigenic shift
● An influenza trivalent vaccine is available: each year
the formulation of the vaccine can vary as the centers
for Disease Control and Prevention tries to predict
which influenza strains will predominate in the
upcoming flu season
● Diagnosis is based on clinical symptoms, serology,
direct antigen detection, and viral isolation
PARAMYXOVIRUSES
● Paramyxoviruses have helical-shaped enveloped
virions. They range in size from 150 to 300 nm.
● The family Pammyxoviridae contains:
○ paramyxoviruses,
○ morbilliviruses,
○ pneumoviruses, and
○ megamyxoviruses
● Parainfluenza virus causes childhood croup, which is a
respiratory infection characterized by fever and a
hoarse cough.
● There are four human parainfluenza viruses: 1–4.
● Morbillivirus causes rubeola or measles, typically a
childhood illness.
○ Necrotic vesicles with a white center
surrounded by erythema on the oral
mucosa, referred to as Koplik spots, are a
characteristic of measles
○ Vaccination programs have nearly
eliminated measles in developed countries.
Respiratory Syncytial Virus (RSV)
● member of the genus Pneumovirus
● RSV causes respiratory and ear infections that are
most common in newborns and young children.
● RSV is characterized by the formation of syncytia.
adjacent cells, producing giant
multinucleated cells. The virus can be grown
in human heteroploid cells such as Hep-2,
HeLa, and A549
○ Infections associated:
○ Bronchitis, pneumonia in infants and
children
Mumps Virus
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● Infections associated:
● Mumps is an infection of the parotid glands, causing
swelling and difficulty in swallowing.
○ Rare in developed countries because of
widespread use of a vaccine.
● Aspirin is never used in the treatment of fever (due to
a virus infection) because it causes/triggers Reye’s
Syndrome. Use paracetamol instead
○ Reye’s Syndrome is a rare but serious
condition that causes swelling in the liver
and brain.
PICORNAVIRUSES
● Picornaviruses have a naked virion ranging in size
from 20 to 30 nm.
● The family Picornaviridae includes a number of
viruses such as the:
○ enterovirus,
○ hepatitis A virus, and
○ the rhinoviruses.
Enterovirus
● Members of the genus Enterovirus (e.g., poliovirus,
coxsackie viruses, and echoviruses) are a common
cause of a variety of human infections worldwide
● Infections associated:
○ Acute nonspecific febrile syndrome
▪ Infections of the respiratory and
gastrointestinal tracts
▪ Diagnosis is generally made by
nucleic acid amplification tests of
clinical specimens:
● Serum
● CSF
● Throat swabs
● Rectal swabs, etc.
Poliovirus
● Poliovirus is transmitted by the fecal-oral route
● The virus initially infects the gastrointestinal tract but
spreads to the CNS
● Infections associated:
○ Paralytic polio and meningitis
● Vaccines
○ The Salk vaccine is a formalin-inactivated
vaccine
▪ Routinely used by most countries
○ The Sabin vaccine is an attenuated vaccine
▪ Produces a stronger immune
response
▪ It (attenuated virus)
sometimes produce severe
infections
○ Because of vaccination programs, the
current risk for polio is extremely small.
Coxsackie Virus
● Infections associated:
○ Coxsackie A viruses
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▪ Hand, foot, and mouth disease of
humans
▪ Conjunctivitis
○ Coxsackie B viruses:
▪ Myocarditis, meningitis
Rhinovirus
● The rhinoviruses grow better at temperatures just
below core body temperature (e.g., 33°C). This is
near the typical temperature of the nasal passage.
● Infections associated:
○ Common colds
▪ Other viruses, including
coronaviruses, are also associated
with colds
▪ Over 100 serotypes are known,
and immunity to one does not
provide immunity to the others.
▪ Infection prevention:
handwashing and avoiding
hand-to-nose contact
ROTAVIRUSES
● Have a double-stranded RNA genome
● The virion is about 70 nm in diameter and has a
wheel-like (spokes) appearance
● Rotaviruses belong to the family Reoviridae
● Infections associated:
○ Gastrointestinal infections in children less
than 2 years of age (during winter months)
○ Diagnosis:
▪ Antigen detection via latex
agglutination or ELISA and, less
commonly, immunoelectron
microscopy.
CALICIVIRUSES
● The family Caliciviridae contains four genera:
○ norovirus,
○ sapovirus,
○ lagovirus, and
○ vesivirus
● They are small, naked viruses
● The noroviruses and Norwalk viruses (members of
the genus Norovirus) are highly contagious and are
important causes of gastroenteritis
TOGAVIRUSES
can ● The virions are about 70 nm in diameter and contain
an envelope
● The family Togaviridae contains two genera
○ Rubivirus: Rubella virus is the only member
of this genus. The virus causes mild
infection.
○ Alphavirus: This genus contains about 25
viruses, all of which are transmitted by
anthropods
● Infection associated:
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○ German measles HEPATITIS VIRUSES

○ However, it can produce severe congenital
infections if women are infected in the early
Incubation Method of Diagnostic
Disease Pathogen Symptoms
stage of pregnancy, therefore, pregnant Period Transmission Test
Fever,
women and women of childbearing age are Hepatitis HAV, headache, IgM
2–6 weeks Ingestion
often tested for immunity. Rubella is rare in A Picornaviridae malaise,
jaundice
antibodies

developed countries because of an effective Severe,

liver
vaccine. Hepatitis HBV, damage, Parenteral, IgM
3–26 weeks
B Hepadnaviridae chronic sexual contact antibodies
disease
occurs
Same as
Hepatitis PCR of viral
FLAVIVIRUSES C
HCV, Flaviviridae HBV, more
chronic
2–33 weeks Parenteral
RNA

● Many viruses belonging to the family Flaviviridae are Severe liver

Parenteral,
damage,
arboviruses Hepatitis
HDV, Deltaviridae high 6–26 weeks
when IgM
D co-infected antibodies
● Important members of this family include: mortality
rate
with HBV

○ West Nile Virus (WNV), Pregnant

women
▪ First reported in the US in 1999 in may be at
IgM
high risk
New York Hepatitis
HEV, Caliciviridae and show 2–6 weeks Ingestion
antibodies,
E PCR of viral
▪ Birds are primary reservoir for high
RNA
mortality,
WNV, and mosquitoes are the not chronic
disease
vectors
▪ WNV typically produces mild or
asymptomatic infections in many Hepatitis A Virus (HAV)
otherwise healthy individuals.
However, the most serious
● HAV contains RNA, and the naked virion has an
complication of WNV infection is
icosahedreal shape
fatal encephalitis (inflammation of
● have ranges in size from 24 to 30 nm
the brain).
● It is a member of the family Picornaviridae and the
▪ Laboratory diagnosis: Can be made
genus Hepatovirus
by ELISA antigen capture RT-PCR
● Infections are spread by fecal-oral route and are
○ St. Louis encephalitis virus,
generally due to poor sanitation and hygiene. Food
○ Yellow fever virus, and
handling transmission is common.
○ Dengue virus
● Humans can also acquire the infection from
contaminated shellfish, including shrimp, oysters,
scallops, etc. Vaccines are available.
RHABDOVIRUSES ● Clinical Characteristics
● Have a bullet-shaped, enveloped capsid ranging in o The incubation period is 15–40 days
size from 150 to 350 nm, a member of the family o Liver involvement (jaundice) nausea,
Rhabdoviridae and the genus Lyssavirus anorexia, and malaise
● The rabies virus gains entry into humans by animal o Mortality rate is less than 1%
(e.g., cat, dog, or raccoon) bites, as well as with bats ● Diagnosis
● Infections caused: o Clinical symptoms and liver enzymes,
○ Rabies particularly alanine aminotransferase, are
▪ The virus first infects the muscle elevated
tissue but preferentially infects o Serology
neurons. The virus migrates along ▪ Anti-HAV IgM is positive in acute
the peripheral nerves to the CNS. infections
The disease progresses to produce ▪ Anti-HAV IgG (positive) and
convulsions, coma, and fatal anti-HAV IgM (negative) indicate a
encephalitis. past HAV infection
● Diagnosis is through medical history of animal bites ▪ General serology testing also
and a positive direct fluorescent-antibody test. includes ruling out hepatitis B virus
● Detection of Negri bodies inclusions in infected brain
cells has a low sensitivity and is not recommended Hepatitis B Virus (HBV)
● A rabies vaccine is available to prevent infection. It is
only administered to those at risk for exposure to the ● HBV contains partially double-stranded DNA
rabies virus. ● The complete virion has an envelope, ranges in size
● Post exposure treatment is effective if administered between 42 and 47 nm, and is sometimes referred to
within 72 hours. Without rapid treatment, the as Dane particles
infection is essentially 100% fatal. ● The virus is unusual in that an RNA intermediate is
required for replication of the genome. The virus
needs a viral-encoded reverse transcriptase for the
replications

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● Member of the family Hepadnaviridae ○ Acute HCV infection is often mild or

● Infections are spread by contaminated body fluids,
including blood. HBV can be sexually transmitted
● Infections are associated with contaminated blood
products, needle sticks, tattoos, body piercing,
intravenous drug abuse, and renal dialysis
● HBV vaccines are available and are recommended for
all healthcare workers
● Clinical Characteristics:
○ The incubation period is 50–180 days
○ Acute infections produce symptoms
resembling HAV infections
○ Chronic infections are common and can
result in cirrhosis and hepatocellular
● Diagnosis:
○ Clinical symptoms and elevated liver
enzymes
○ Serology
asymptomatic and is rarely diagnosed in this
phase. HCV is more likely to cause chronic
hepatitis, resulting in cirrhosis, than HBV.
HCV infections are one of the most common
reasons for liver transplant in the US.
● Diagnosis
○ Elevated liver enzymes
○ Serologic indicators (anti-HCV and HCV
antigen) and nucleic acid amplification
○ The virus has not been grown in cell
structure
Hepatitis D Virus (HDV)
● Contains RNA, and the naked viruses range in size
from 35 to 37 nm. HDV is also called the delta virus
● HDV requires but does not encode for HBsAg;
therefore, it only replicates in cells also infected with
HBV
o Coinfection occurs when an individual
acquires both HDV and HBV at the same
time. A superinfection is when a patient
with an HBV infection is exposed to HDV.
Superinfections are more severe than
coinfections
● Diagnosis:
o Detection of anti-HDV and HDV RNA
o Serologic markers for HBV will also be
positive; in particular, HBsAg

● Hepatitis B Panel Other Human Hepatitis Viruses

HBsAg Anti-HBc Anti-HBs ● Hepatitis E Virus (HEV)

No HBV ○ HEV contains RNA, and virions range in size
- - -
infection from 32 to 34 nm
Early HBV ○ HEV is spread by the fecal-oral route, often
+ - -
infection in contaminated water. It is the most
Acute HBV common cause of hepatitis in some
+ + -
infection countries with poor sanitation.
Window ○ Diagnosis: Serology
- + - ● Hepatitis G Virus (HGV)
period
Past infection - + + ○ HGV contains RNA and has an envelope
Immunization - - + ○ It is in the same family Flaviviridae, as HCV
○ Although HGV is most commonly
transmitted by contact with blood, it can
Hepatitis C Virus (HCV) also be sexually transmitted and transmitted
from mother to children. Infection seems
● HCV contains RNA and has a lipid envelope relatively common worldwide, but HGV is
● The virus is a member of the family Flaviviridae and believed to be nonpathogenic
the genus Hepacivirus
● Infections:
○ HCV is the most common cause of non-A, VIRUSES
non-B (NANB) hepatitis. It is common ● Smallest infectious agents (ranging from about 20 to
worldwide. 300 nm in diameter) and contain only one kind of
Spread through contaminated blood products, organ nucleic acid (RNA or DNA) as their genome
transplants, renal dialysis, and intravenous drug o Nucleic acid is encased in a protein shell,
abuse which may be surrounded by a
No vaccine currently exists for HCV lipid-containing membrane.
● Clinical Characteristics o The entire infectious unit is termed a VIRION
○ Incubation period is 2–25 weeks

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VIRION
● Mature virus containing a nucleic acid core 4. Size and Shape
surrounded by a protein coat, with or without an ○ Can be visualized directly by electron
envelope microscopy
1. Nucleic acid - each virus contains nucleic acid ○ Size ranges from 20 to 300 nm
core, the genome, either single-stranded or
double-stranded
○ DNA - deoxyribonucleic acid
○ RNA - ribonucleic acid

2. Nucleocapsid - each nucleocapsid contains a

nucleic acid core surrounded by a protein coat,
the capsid, which consists of many subunits
named capsomeres; the capsid symmetry is
either:
○ Helical - rod shaped
○ Icosahedral - cube-like

REPLICATION OF VIRUS PARTICLES

1. Attachment
o Specific cell receptor
o Responsible for varying cell tropism
2. Penetration
o Virus passing through cell membrane
o May take cell membrane as protection
3. Uncoating
o Removes all part of the capsid
3. Enveloped or Naked
o Exposes the nucleic acid
○ Some viruses have envelopes from the
4. Biosynthesis
host cell membrane or nuclear
o Proteins, nucleic acids, and other
membrane
components
○ Some viruses have no envelopes
o Some made in tremendous excess
(naked)
5. Morphogenesis
o Components assembled
o Often uses enzymes encoded by virus
6. Release
o Budding through membrane
o Lysis of membrane

CLASSIFICATION

1. NUCLEIC ACID COMPOSITION

● DNA or RNA
● Single or double stranded
2. CAPSID
● Helical or Icosahedral
3. ENVELOPES

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● Presence or Absence o Immunofluorescence, enzyme-linked

● Acquired from nuclear cytoplasmic membrane immunosorbent assay (ELISA)
4. SITE OF VIRION ASSEMBLY 2. Electron microscopy
● Nucleus or cytoplasm o Morphology of virus particles
3. Light microscopy
o Histologic appearance
SPECIMEN COLLECTION, TRANSPORT, AND STORAGE o Demonstration of inclusion bodies
● Storage: 4०C
● Transport: 70०C Antigen Detection
1. Throat swabs or washings 1. Enzyme Immunoassay (EIA)
2. Rectal swabs ○ Plate-based assays
3. Skin scrapings or lesion swabs ○ Lateral flow technology
4. Urine 2. Nasopharyngeal aspirate
5. Body fluid or vesicle ○ Respiratory syncytial virus (RSV)
6. Tissue biopsies ○ Influenza
3. Stool
CLINICAL VIROLOGY: CULTURE METHOD ○ Rotavirus
● 1937: Propagated yellow fever virus in chicken ○ Adenovirus
embryos ➢ Advantages and Disadvantages
● Growth of viruses ● Advantages
1. Chicken embryos: Historical ○ Result available quickly, point-of-care testing
2. Tissue explants: Research use only ● Potential Problems
3. Cell culture ○ Reduced sensitivity compared to cell culture
o Primary cell culture or PCR: 40% to 80%
o Diploid cell lines ○ Labor intensive
o Continuous cell lines ○ Not well suited a core laboratory

Nucleic Acid Amplification

CULTURE METHOD ○ Allows the amplification of specific target DNA
sequences by a factor
1. Primary cell cultures ○ PCR is the most common method but other variations
● Derived directly from donor (animal or are being developed and used
human) ○ Detection of the PCR product usually has been by
● Most common are kidney cells agarose gel electrophoresis, probe hybridization, or
● Rhesus monkey, rabbit kidney DNA sequencing
2. Diploid cell line ○ Real-time PCR has streamlined the amplification and
● Prepared from animal tissues the detection processes
● Usually fibroblasts from lung or foreskin ○ Advantages of PCR
3. Continuous cell lines ○ Extremely high sensitivity
● Single cell type that can propagated ○ Fast turnaround time for real-time PCR
indefinitely ○ Nucleic Acid amplification
● Derived from tumors or mutagenic ○ Disadvantages of PCR
treatment of primary culture cell ○ Extremely liable to contamination
○ High degree of operator skill required
HeLa cells: HENRIETTA LACKS ○ Not easy to set up quantitative assay
Among the important scientific discoveries of the last century ○ A positive result may be difficult to interpret
was the first immortal human cell line known as “HeLa” – a
remarkably durable and prolific line of cells obtained during Serologic Tests for Viral Infections
the treatment of Henrietta’s cancer by John Hopkins ○ Classic Techniques
researcher Dr. George Gey in 1951. o Complement fixation tests
o Hemagglutination inhibition tests
Over the past several decades, this cell line has contributed to o Immunofluorescence techniques
many medical breakthroughs, from research on the effects of o Counterimmunoelectrophoresis
zero gravity in outer space and the development of the polio ○ Older Techniques
vaccine, to the study of leukemia, the AIDS virus, and cancer ○ Radioimmunoassay
worldwide. ○ Newer Techniques
○ Enzyme-linked immunoassay (EIA)
○ Particle agglutination
NON-CULTURE METHODS ○ Western Blot
○ Recombinant immunoblot assay
○ Lateral flow “rapid” devices
Direct Examination
1. Antigen detection

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COVID-19 TESTING serve as a possible treatment for

those who are seriously ill from
● Crucial to guiding our next steps in the fight against COVID-19
COVID-19
DNA VIRUS
TWO MAIN CATEGORIES OF TESTS
IN GENERAL:
1. Diagnostic Tests DNA DOUBLE STRANDED, ICOSAHEDRAL CAPSID,
VIRUSES REPLICATE IN THE NUCLEUS
● Can tell you if you currently have an
•Double stranded DNA viruses except Parvovirus
infection
(ssDNA)
● Molecular Tests •Icosahedral capsid except Poxvirus (complex)
○ Work by looking for the virus’s •Enveloped (ether-sensitive/labile):
genetic material in a sample from Herpesviridae, Hepadnaviridae, Poxviridae
you •Naked (ether-sensitive/stable): Adenoviridae,
○ The sample is usually analyzed in a Papovaviridae, Parvoviridae
lab; some can be analyzed at point •Replicate in the nucleus except Poxviridae
of care (cytoplasm)
○ FDA-authorized tests are very •Smallest DNA virus: Parvoviridae
•Largest DNA virus: Poxviridae (largest virus)
accurate
○ Results are returned anywhere
FAMILY ADENOVIRIDAE
from a minute to several days
CHARACTERISTICS dsDNA genome; icosahedral
depending on the Test capsid, no envelope
● Antigen Tests VIRUS Adenovirus
○ See if there are viral proteins in a DISEASES Pharyngitis,
sample taken from inside your pharyngoconjunctival fever,
nose with a swab keratoconjunctivitis,
○ Simpler and may provide results pneumonia, hemorrhagic
cystitis, disseminated
quicker than any molecular tests;
diseases, and gastroenteritis
sometimes within minutes in a in children
doctor’s office
○ Can be less accurate and may need FAMILY HEPADNAVIRIDAE
confirmation with an additional CHARACTERISTICS Partly dsDNA genome;
diagnostic test icosahedral capsid with
○ Can be made more available envelope; virion also called
because they are easy and simple Dane particle; surface
antigen termed Australia
2. Antibody Tests (Serology test)
antigen
● Antibodies are produced by your body when VIRUS Hepatitis B virus
you are infected by a virus and help your
DISEASES Acute infection with
immune system fight of the infection
resolution (90%); fulminant
● Detects the antibodies to the virus using a
hepatitis most infected with
blood sample
delta virus (1%); chronic
● If the antibody test finds antibodies in the
hepatitis, persistence of
blood it likely means the person has been
HBsAg (9%) followed by
previously infected with the virus
resolution (disappearance of
● Antibodies DO NOT show if you have a
HBsAg), asymptomatic
current infection and SHOULD NOT be used
carrier state, chronic
to diagnose a current infection
persistent (systemic disease
● Results can help us better understand
without progressive liver
questions about exposure to COVD-19 by
disease), or chronic active
○ Helping identify who has been
live disease (progressive
infected and has developed
liver damage)
antibodies
○ If antibodies may provide
protection from future infection
HEPADNAVIRUS
○ Who may still be at risk
○ Who may be eligible to donate a
part of their blood—called ○ DsDNA with a short single-stranded region
convalescent plasma—which may ○ Icosahedral core with envelope

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○ Hepatitis B HERPESVIRUS
Two important antigens
1. Surface Ag ○ dsDNA, replicate in nucleus
2. Core Ag ○ Icosahedral nucleocapsid, envelope
○ Virus not isolated in culture ○ Most prominent feature: Latency
○ Serologic test ○ Herpes simplex virus
Transmission through blood ○ Two types
1. Transfusion 1. Cold sores
2. Intravenous (IV) drug abuser 2. Genital
3. Sexual contact ○ Skin and mucous membrane infections
Vaccine ○ Encephalitis
○ Engineered ○ Isolated easily in culture
○ Infants at birth, health care workers

FAMILY HERPESVIRIDAE VARICELLA-ZOSTER VIRUS

CHARACTERISTICS dsDNA genome; icosahedral capsid
with envelope; at least 8 human
○ Chickenpox and shingles
herpes viruses known
○ Vaccine important in controlling outbreaks
VIRUS AND Herpes simplex Gingivostomatitis,
○ Tzanck stain: Giant cells
DISEASES virus type 1 pharyngitis, herpes
○ Isolation more difficult than HSV
(HSV-1) labialis (cold sores),
conjunctivitis,
CYSTOMEGALOVIRUS
keratitis,
encephalitis
Herpes simplex Genital infection ○ Isolated from blood
virus type 2 ○ In adults: Syndrome similar to mononucleosis, many
(HSV-2) infect kidney (shed in urine)
Varicella-zoster Chicken pox Laboratory Tests
virus (VZV) (varicella), shingles ○ Shell vial culture
(zoster) ○ Serology
Epstein-Barr Infectious
virus (EBV) mononucleosis,
progressive EPSTEIN-BARR VIRUS
lymphoreticular
disease, oral hairy ○ Heterophile-positive infectious mononucleosis (85%)
leukoplakia in ○ Can produce tumors
HIV-infected ○ Not isolated in culture
patients ○ Serologic diagnosis
Cytomegaloviru Asymptomatic
s (CMV) infection, congenital
disease of the HUMAN HERPESVIRUS 6, 7, AND 8
newborn,
symptomatic Human Herpesvirus-6
disease of
○ Exanthema subitum/roseola infantum
immunocompromis
○ Sixth disease
ed host,
○ Spread by respiratory route
heterophile-negativ
○ Molecular assays used for detection
e infectious
mononucleosis
Human Herpesvirus-7
Human Roseola, (exanthem ○ Cause small percentage of roseola
herpesvirus 6 subitum), fever,
and 7 (HHV-6 malaise, rash, Human Herpesvirus-8
and HHV-7) leukopenia, and ○ Found in Kaposi’s sarcoma
interstitial
pneumonitis in Kaposi’s sarcoma is a type of cancer that forms in the lining of
organ transplant in blood and lymph vessels
patients ○ The tumors (lesions) of Kaposi’s sarcoma typically
Human Kaposi’s sarcoma appear as painless purplish spots on the legs, feet, or
herpesvirus-8 face
(HHV) ○ Lesions can also appear in genital area, mouth, or
lymph nodes

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○ In severe Kaposi’s sarcoma, lesions may develop in

the digestive tract and lungs
People infected with human immunodeficiency virus (HIV) –
the virus that causes AIDS – have the highest risk of Kaposi’s
sarcoma.

REFERENCES

Notes from the discussion by Ma’am Marilyn Ngo

University of Santo Tomas powerpoint presentation:

Course book: Hugo and Russel’s Pharmaceutical

Microbiology 8th Edition

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Date Remarks

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 [TRANS] UNIT 6: PROTOZOA
● Most parasites have a life cycle that often involves
OUTLINE several hosts; this means that survival and transmission
between different hosts requires the parasite to exhibit
I. Introduction more than one physiologically distinct form.
A Parasitism
B Habitats
HABITATS
C Physiology of parasitic protozoa
II. Blood and tissue parasites
● Parasites inhabit a wide range of habitats within their
A Malaria
hosts.
B Trypanosomatids
● Some parasites will inhabit only one site through- out
C Toxoplasma gondii their life cycle, but many move to various sites within
the body.
III. Intestinal Parasites ● Such movement may require the formation of motile
A Subtopic A cellular forms, and it will produce a significant change in
B Subtopic B the physiology and morphology of the parasite as a
C Cryptosporidium parvum result of environmental change.
● Parasites moving from the gut to other tissues, for
IV. Trichomonas and free-living amoebas example, will encounter higher levels of oxygen, changes
D Trichomonas vaginalis in pH and significant exposure to the host immune
E Free-living opportunist amoebas response.
● When life cycles involve more than one host organism
V. Host response to infection these changes are greater.
F Immune response ● The reasons why parasites move to various sites in with
G Immune pathology the host is driven by evasion of host immune attack and
H Immune evasion to aid transmission.

VI. Detection of parasites PHYSIOLOGY OF PARASITIC PROTOZOA

I Methods of detection
i Antibody-based technologies ● Parasitic protozoa, like their free-living counterparts, are
ii DNA-based technologies single-celled eukaryotic organisms that utilize flagella,
iii Alternative methods cilia or amoeboid movement for motility.
J Analysis of samples ● The complexity of some parasite life cycles means that
some species may exhibit, at different times, more than
i Clinical samples
one form of motility.
ii Environmental samples
● All pathogenic protozoa are heterotrophs, using car-
bohydrates or amino acids as their major source of
VII. Control of protozoan parasites
carbon and energy.
K Chemotherapy ● Some parasitic protozoa utilize oxygen to generate
i Mechanisms of action and selective toxicity energy through oxidative phosphorylation, but many
ii Drug resistance protozoan parasites lack functional or ‘typical’
L Other approaches to control mitochondria, or have mitochondria that do not function
i Biological control like those in mammalian cells.
ii Vaccination ● As a result of this adaptation many parasites exhibit a
fermentative metabolism that functions even in the
I. INTRODUCTION presence of oxygen.
● The reason for the utilization of less efficient
PARASITISM fermentative pathways is not clear, but it is presumably
due in part to the fact that such parasites survive in
environments where oxygen is only present occasionally
● Parasitism is a specific type of interaction between two
or at low levels.
organisms that has many features in common with other
● For some parasites oxygen is toxic, and they appear to
infectious processes, but host–parasite interactions
utilize it possibly in an effort to remove it and thus main-
often operate over a longer timescale than those seen
tain an anaerobic metabolism.
with other pathogens.
● The metabolism of parasites is highly adapted, with
● This extended process results in significant host–parasite
many possessing unique organelles such as kinetoplasts
interaction at the cellular and organismal level.
and hydrogenosomes.
● It is known, for example, that some parasites alter the
● Many synthetic pathways that are found in other
behaviour of the host, while others, such as Giardia
eukaryotes are absent because many metabolic
lamblia, induce biochemical change in the host cells at
intermediates or precursors such as lipids, amino acids
the site of infection (the duodenal epithelium).
and nucleotides are actively scavenged from their
environment.

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 TRANS: PROTOZOA
● This minimizes energy expenditure, which is finely
balanced in parasites and means that the membrane of
parasitic protozoa is rich in transporters.
● Secretion of haemolysins, cytolysins, proteolytic
enzymes, toxins, antigenic and immunomodulatory
molecules that reduce host immune response also
occurs in pathogenic protozoa.
● Survival of parasites is partly due to their high rate of
reproduction, which may be either sexual or asexual;
some organisms such as Plasmodium exhibit both forms
of reproduction in their life cycle.
● Simple fission is characteristic of many amoeba, but
some species also undergo nuclear division in the cystic
state (cysts are forms required for survival outside the
host) with each nucleus giving rise to new trophozoites
(the growing, motile and pathogenic form).
II. BLOOD AND TISSUE PARASITES
● This section considers the life cycles, disease and pathol-
ogy of some blood and tissue parasites; this is not an
exhaustive list but covers some of the most important
species.
● These diseases are commonly associated with travel to
tropical and subtropical countries, but diseases such as
leishmaniasis are frequently seen in southern Spain and
France.
● It should also be noted that climate change is altering
the geographical distribution of many parasitic diseases.
A. MALARIA
● Malaria has been a major disease of humankind for
thousands of years.
● Despite the availability of drugs for treatment, malaria is
still one of the most important infectious diseases of
humans, with approximately 200–500 million new cases
and 1–2.5 million deaths each year.
● Protozoa of the genus Plasmodium cause malaria and
four species are responsible for the disease in humans:
P. falciparum, P. vivax, P. ovale and P. malariae. P.
falciparum and P. vivax account for the vast majority of
cases, although P. falciparum causes the most severe
disease.
● Other species of plasmodia infect reptiles, birds and
other mammals.
● Malaria is spread to humans by the bite of female
mosquitoes of the genus Anopheles but transmission by
inoculation of infected blood and through congenital
routes is also seen.
● These mosquitoes feed at night and their breeding sites
are primarily in rural areas.
DISEASE
● The most common symptom of malaria is fever,
although chills, headache, myalgia and nausea are
frequently seen and other symptoms such as vomiting,
diarrhoea, abdominal pain and cough occasionally
appear.
● In all types of malaria, the periodic febrile response
(fever) is caused by rupture of mature schizonts (one of
the cell forms arising as part of the life cycle).
● In P. vivax and P. ovale malaria fever occurs every 24–48
hours, whereas in P. malariae, maturation occurs every
72 hours.
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● In falciparum malaria fever may occur every 48 hours,
but is usually irregular, showing no distinct periodicity.
● Apart from anaemia, most physical findings in malaria
are often non-specific and offer little aid in diagnosis,
although enlargement of some organs may be seen after
prolonged infection.
● If the diagnosis of malaria is missed or delayed,
especially with P. falciparum infection, potentially fatal
complicated malaria may develop.
● The most frequent and serious complications of malaria
are cerebral malaria and severe anaemia.
LIFE CYCLE
● Plasmodia have a complex life cycle involving a number
of life cycle stages and two hosts.
● The human infective stage comprises the sporozoites
(c.1–7 μm), which are produced by sexual reproduction
in the midgut of the mosquito (vector) and migrate to its
salivary gland.
● When an infected Anopheles mosquito bites a human,
sporozoites are injected into the bloodstream and are
thought to enter liver parenchymal cells within 30
minutes of inoculation.
● In these cells the parasite differentiates into a spherical,
multinucleate schizont which may contain 2000–40 000
uninucleate merozoites.
● This process of growth and development is termed
exoerythrocytic schizogony.
● This exoerythrocytic phase usually takes between 5 and
21 days, depending on the species of Plasmodium;
however, in P. vivax and P. ovale the maturation of
schizonts may be delayed for up to 1–2 years.
● These ‘quiescent’ parasites are called hypnozoites.
● Clinical illness is caused by the erythrocytic stage of the
parasite life cycle; no disease is associated with
sporozoites, the developing liver stage of the parasite,
the merozoites released from the liver, or gametocytes.
● The common symptoms of malaria are due to the
rupture of erythrocytes when erythrocytic schizonts
mature.
● This release of parasite material triggers a host immune
response, which in turn induces the formation of
inflammatory cytokines, reactive oxygen intermediates
and other cellular products.
● These proinflammatory molecules play a prominent role
in pathogenesis, and are probably responsible for the
fever, chills, sweats, weakness and other systemic symp-
toms associated with malaria.
● In P. falciparum malaria, infected erythrocytes adhere to
the endothelium of capillaries and postcapillary venules,
leading to obstruction of the microcirculation and
localized anoxia.
● The pathogenesis of anaemia appears to involve
haemolysis or phagocytosis of parasitized erythrocytes
and ineffective erythropoiesis.
B. TRYPANOSOMATIDS
● The family Trypanosomatidae consists of two genera,
Trypanosoma and Leishmania.
● These are important pathogens of humans and domestic
animals and the diseases they cause constitute serious
medical and economic problems.
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 TRANS: PROTOZOA
● Because these protozoans have a requirement for
haematin obtained from blood, they are called haemo-
flagellates.
● The life cycles of both genera involve insect and
vertebrate hosts and have up to eight life cycle stages,
which differ in the placement and origin of the
flagellum.
● Trypanosomatids have a unique organelle called the
kinetoplast.
● This appears to be a special part of the mitochondrion
and is rich in DNA.
● Two types of DNA have been found in the kinetoplast:
maxicircles that encode many mitochondrial enzymes,
and minicircles, which serve a function in the process of
RNA editing.
● Replication of trypanosomatids occurs by single or
multiple fission, involving first the kinetoplast, then the
nucleus, and finally the cytoplasm.
● There are four major diseases associated with this
group: Chagas disease is caused by Trypanosoma cruzi;
sleeping sickness (African trypano- somiasis) is
associated with T. brucei; cutaneous and mucocutaneous
leishmaniasis are caused by a range of species including
Leishmania tropica, L. major, L. mexicana, L.
amazonensis and L. braziliensis; and visceral leish-
maniasis, which is also known as kala-azar, is typically
caused by L. donovani.
AMERICAN TRYPANOSOMIASIS (Chagas disease)
● Chagas disease begins as a localized infection that is fol-
lowed by parasitaemia and colonization of internal
organs and tissues.
● Infection may first be evidenced by a small tumour
(chagoma) on the skin.
● Symptoms of the disease include fever, oedema and
myocarditis (infection of the heart muscle) with or
without heart enlargement, and meningoencephalitis in
children.
● The acute disease is frequently subclinical and patients
may become asymptomatic carriers; this chronic phase
may result, after 10–20 years, in cardiopathy.
● Chagas disease is transmitted by several genera of
triatomine bugs (Triatoma, Rhodnius and Panstrongylus)
and in nature the disease exists among wild mammals
and their associ- ated triatomines.
● Human trypanosomiasis is seen in almost all countries of
the Americas, including the southern USA, but the main
foci are in poor rural areas of Latin America.
● T. cruzi exhibits two cell types in vertebrate hosts, a
blood form termed a trypomastigote, and in the tissues
(mainly heart, skeletal and smooth muscle, and reticu-
loendothelial cells) the parasite occurs as an amastigote.
● Trypomastigotes ingested when the insect takes a blood
meal from an infected host transform into epimastigotes
in the intestine.
● Active reproduction occurs and in 8–10 days metacyclic
trypomastigote forms appear which are flushed out of
the gut with the faeces of the insect.
● These organisms are able to penetrate the verte- brate
host only through the mucosa or abrasions of the skin;
hence, transmission does not necessarily occur at every
blood meal.
● Within the vertebrate the trypomastig- otes transform
into amastigotes, which, after a period of intracellular
multiplication at the portal of entry, are released into
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
the blood as trypanosomes; these invade other cells or
tissues, becoming amastigotes again.
● The pathology of the infection is associated with
inflammatory reactions in infected tissues.
● These can lead to destruction of the infected tissue and
if it involves heart tissue this can cause acute
myocarditis.
● Parasite enzymes may also cause cell and tissue damage.
● In the absence of parasites, an autoimmune pathological
process seems to be mediated by T lymphocytes (CD4+)
and by the production of certain cytokines; these induce
a polyclonal activation of B-lymphocytes and the
secretion of large quantities of autoantibodies.
● Figure 6.1 The malaria parasite life cycle involves two
hosts. During a blood meal, a malaria-infected female
Anopheles mosquito inoculates sporozoites into the
human host (1). Sporozoites infect liver cells (2) and
mature into schizonts (3), which rupture and release
merozoites (4). (Of note, in P. vivax and P. ovale a
dormant stage (hypnozoites) can persist in the liver and
cause relapses by invading the bloodstream weeks, or
even years later.) After this initial replication in the liver
(exo-erythrocytic schizogony, A), the parasites undergo
asexual multiplication in the erythrocytes (erythrocytic
schizogony, B). Merozoites infect red blood cells (5). The
ring stage trophozoites mature into schizonts, which
rupture releasing merozoites (6). Some parasites
differentiate into sexual erythrocytic stages
(gametocytes) (7). Blood-stage parasites are responsible
for the clinical manifestations of the disease. The
gametocytes, male (microgametocytes) and female
(macrogametocytes), are ingested by an Anopheles
mosquito during a blood meal (8). The parasite’s
multiplication in the mosquito is known as the
sporogonic cycle (C). While in the mosquito’s stomach,
the microgametes penetrate the macrogametes,
generating zygotes (9). The zygotes in turn become
motile and elongated (ookinetes) (10), which invade the
midgut wall of the mosquito where they develop into
oocysts (11). The oocysts grow, rupture and release
sporozoites (12), which make their way to the
mosquito’s salivary glands. Inoculation of the
sporozoites into a new human host perpetuates the
malaria life cycle (1).
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 TRANS: PROTOZOA
● Figure 6.2 (a) Mature schizonts of Plasmodium vivax. P.
vivax schizonts are large, have 12–24 merozoites, and
may fill the red blood cell (RBC). RBCs are enlarged 1.5–2
times and may be distorted. Under optimal conditions
Schüffner’s dots may be seen. (b) Leishmania tropica
amastigotes within an intact macrophage. (c)
Toxoplasma gondii trophozoites in the bronchial
secretions from an HIV-infected patient. (d) Trophozoites
of Giardia intestinalis. Each cell has two nuclei and is
10–20 mm in length. (e) Trophozoites of Entamoeba
histolytica with ingested erythrocytes, which appear as
dark inclusions. (f) Oocysts of Cryptosporidium parvum
(upper left) and cysts of Giardia intestinalis (lower right)
labelled with immunofluorescent antibodies. (g)
Trophozoites of Trichomonas vaginalis.
● Figure 6.3 An infected triatomine insect vector (or
‘kissing’ bug) takes a blood meal and releases
trypomastigotes in its faeces near the site of the bite
wound. Trypomastigotes enter the host through the
wound or through intact mucosal membranes, such as
the conjunctiva (1). Common triatomine vector species
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
for trypanosomiasis belong to the genera Triatoma,
Rhodinius and Panstrongylus. Inside the host, the
trypomastigotes invade cells, where they differentiate
into intracellular amastigotes (2). The amastigotes
multiply by binary fission (3) and differentiate into
trypomastigotes, and then are released into the
circulation as bloodstream trypomastigotes (4).
Trypomastigotes infect cells from a variety of tissues and
transform into intracellular amastigotes in new infection
sites. Clinical manifestations can result from this
infective cycle. The bloodstream trypomastigotes do not
replicate (different from the African trypanosomes).
Replication resumes only when the parasites enter
another cell or are ingested by another vector. The
‘kissing’ bug becomes infected by feeding on human or
animal blood that contains circulating parasites (5). The
ingested trypomastigotes transform into epimastigotes
in the vector’s midgut (6). The parasites multiply and
differentiate in the midgut (7) and differentiate into
infective metacyclic trypomastigotes in the hindgut (8).
Trypanosoma cruzi can also be transmitted through
blood transfusions, organ transplantation,
transplacentally, and in laboratory accidents.
AFRICAN TRYPANOSOMIASIS (sleeping sickness)
● Sleeping sickness (African trypanosomiasis) is caused by
Trypanosoma brucei, of which there are two
morphologically indistinguishable subspecies: T. brucei
rhodesiense and T. brucei gambiense.
● After infection the parasite undergoes a period of local
multiplication then enters the general circulation via the
lymphatics.
● Recurrent fever, headache, lymphadenopathy and
splenomegaly may occur.
● Later, signs of meningoencephalitis appear, followed by
somnolence (sleeping sickness), coma and death.
● T. brucei, unlike T. cruzi, multiplies in the blood or
cerebrospinal fluid.
● Trypanosomes ingested by a feeding fly must reach the
salivary glands within a few days, where they reproduce
actively as epimastigotes attached to the microvilli of
the salivary gland where they transform into metacyclic
trypomastigotes, which are found free in the lumen.
● Around 15–35 days after infection the fly becomes
infective through its bite.
● The pathology of the infection is due to inflammatory
changes associated with an induced autoimmune demy-
elination of nerve cells.
● Interestingly, the immunosuppressive action of
components of the parasite’s membrane is probably
responsible for frequent secondary infections such as
pneumonia.
● Liberation of common surface antigens (the mechanism
involved in immune evasion) in every trypanolytic crisis
(episode of trypanosome lysis) leads to antibody and
cell-mediated hypersensitivity reactions.
● It is believed that some cytotoxic and pathological
processes are the result of biochemical and immune
mechanisms.
CUTANEOUS AND MUCOCUTANEOUS LEISHMANIASIS
● Leishmaniasis is the term used for diseases caused by
species of the genus Leishmania that are transmitted by
the bite of infected sand flies.
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● The lesions of cutaneous and mucocutaneous
leishmaniasis are localized to the skin and mucous
membranes.
● Visceral leishmaniasis is a much more severe disease,
which involves the entire reticuloendothelial system,
and is discussed in section 2.2.4 of this chapter.
● Cutaneous leishmaniasis appears 2–3 weeks after the
bite of an infected sand fly as a small cutaneous papule;
this slowly develops and often becomes ulcerated and
develops secondary infections.
● Secondary or diffuse lesions may develop.
● The disease is usually chronic but may occasionally be
self-limiting.
● Leishmaniasis from a primary skin lesion may involve the
oral and nasopha- ryngeal mucosa.
● Leishmania species that infect humans are all
morphologically similar and only exhibit one form, the
intracellular amastigotes (3–6μm long and 1.5–3μm in
diameter).
● Promastigotes are found in the sand fly.
● In mammalian hosts amastigotes are phagocytosed by
macrophages, but resist digestion and divide actively in
the phagolysosome.
● The female sand fly ingests parasites in the blood meal
from an infected person or animal and these pass into
the stomach where they transform into promastigotes,
and multiply actively.
● The parasites attach to the walls of the oesophagus,
midgut and hindgut of the fly, and some eventually
reach the proboscis and are inoculated into a new host.
● The obvious symptoms of this infection are caused by
the uptake of parasites by local macrophages.
● Host response to infection produces tubercle-like
structures designed to limit the spread of infected cells.
● Some lesions may resolve spontaneously after a few
months but other types of lesion may become chronic,
sometimes with lymphatic and bloodstream
dissemination.
● In infections due to L. braziliensis there is a highly
destructive spread of infected macrophages to the oral
or nasal mucosa.
● In L. mexicana, L. amazonensis and L. aethiopica
infections the disease becomes more disseminated.
● The immunological response of the host plays an
important factor in determining the precise pathology of
the disease and this is apparent from the more severe
type of infection seen in individuals with HIV.
● In Europe and Africa several rodents may act as
reservoirs of the disease, but in countries such as India,
transmission can occur in a human–sand fly– human
cycle without rodent intervention.
● In rural semi-arid zones of Latin America, both wild and
domestic dogs enter the epidemiological chain and the
vector is a common sand fly, Lutzomyia longipalpis,
abundant in and around houses.
● The disease is more common in children in both Latin
America and the Mediterranean area.
VISCERAL LEISHMANIASIS (kala-azar)
● Like cutaneous leishmaniasis, visceral leishmaniasis
begins with the formation of a nodule at the site of
inoculation but this lesion rarely ulcerates and usually
disappears in a few weeks.
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● However, symptoms and signs of systemic disease such
as undulating fever, malaise, diarrhoea, organ
enlargement and anaemia subsequently develop.
● In more serious cases of visceral leishmaniasis the
parasites, which can resist the internal body tempera-
ture, invade internal organs (liver, spleen, bone marrow
and lymph nodes), where they occupy the reticuloen-
dothelial cells.
● The pathogenic mechanisms of the disease are not fully
understood, but enlargement occurs in those organs
that exhibit marked cellular alteration such as
hyperplasia.
● Parasitized macrophages replace tissue in the bone
marrow.
● Patients with advanced disease are prone to
superinfection with other organisms which are pre-
dominantly bacterial.
C. TOXOPLASMA GONDII
● The term coccidia describes a group of protozoa that
contains the genus Cryptosporidium (see intestinal
parasites) as well as a number of important veterinary
parasites.
● Toxoplasma gondii is an intestinal coccidian but the
major pathology of infection is associated with other
tissues and organs.
● T. gondii infects members of the cat family as definitive
hosts and has a wide range of intermediate hosts.
Infection is common in many warm-blooded animals,
including humans.
● In most cases infection is asymptomatic, but devastating
disease can occur congenitally in children as a result of
infection during pregnancy.
● T. gondii infection in humans is a worldwide problem,
although the rates of human infection vary from country
to country.
● The reasons for these variations include environmental
factors, cultural habits and the presence of domestic and
native animal species.
● The frequency of postnatal toxoplasmosis acquired by
eating raw meat and by ingesting food contaminated by
oocysts from cat faeces (oocyst formation is greatest in
the domestic cat) is not well established but is thought
to be significant.
● Widespread natural infection is possible because
infected animals may excrete millions of resistant
oocysts, which can survive in the environment for
prolonged periods (months–years).
● Mature oocysts are approximately 12 μm in diameter
and contain eight infective sporozoites.
● T. gondii infection in most animals including humans is
asymptomatic.
● Severe disease in humans is observed only in
congenitally infected children and in immuno-
suppressed individuals.
● The most common symptom associated with postnatal
infection in humans is lymphadenitis which may be
accompanied by fever, malaise, fatigue, muscle pains,
sore throat and headache (flu-like symptoms).
● Typically infection resolves spontaneously in weeks or
months, but in immunosuppressed individuals, a fatal
encephalitis may occur producing symptoms such as
headache, disorientation, drowsiness, hemiparesis,
reflex changes and convulsions.
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● Prenatal T. gondii infections often target the brain and
retina and can cause a wide spectrum of clinical disease.
● Mild disease may consist of impaired vision, whereas
severely diseased children may exhibit a ‘classic tetrad’
of signs: retinochoroiditis, hydrocephalus, convulsions
and intracerebral calcifications.
● Hydrocephalus is the least common but most dramatic
lesion of congenital toxoplasmosis.
● The life cycle of T. gondii was only fully described in the
early 1970s when felines including domestic cats were
identified as the definitive host and various warm-
blooded animals were identified as intermediate hosts.
● T. gondii is transmitted by three mechanisms:
congenitally, through the consumption of uncooked
infected meat and via faecal matter contamination.
● Cats acquire Toxoplasma by ingesting any of three
infectious stages of the organism: the rapidly multiplying
forms, tachyzoites, the dormant bradyzoites (cysts) in
infected tissue and the oocysts shed in faeces.
● The probability of infection and the time between infec-
tion and the shedding of oocysts varies with the stage of
T. gondii ingested.
● Fewer than 50% of cats shed oocysts after ingesting
tachyzoites or oocysts, whereas nearly all cats shed
oocysts after ingesting bradyzoites.
● When a cat ingests tissue cysts, the cyst wall is dissolved
by intestinal and gut proteolytic enzymes, which causes
the release of bradyzoites.
● These enter the epithelial cells of the small intestine and
initiate the formation of numerous asexual generations
before the sexual cycle begins.
● At the same time that some bradyzoites invade the
surface epithelia, other bradyzoites penetrate the lamina
propria and begin to multiply as tachyzoites
(trophozoites).
● Within a few hours, tachyzoites may disseminate to
other tissues through the lymph and blood.
● Tachyzoites can enter almost any type of host cell and
multiply until the cell becomes packed with parasites.
● The host cell then lyses and releases more tachyzoites to
enter new host cells.
● The host usually controls this phase of infection, and as a
result the parasite enters the ‘resting’ stage in which
bradyzoites are isolated in tissue cysts.
● Tissue cysts are formed most commonly in the brain,
liver and muscles.
● These cysts usually cause no host reaction and may
remain dormant for the life of the host.
● In intermediate hosts, such as humans, the
extraintestinal cycle of T. gondii is similar to the cycle in
cats except that there is no sexual stage.
● Most cases of toxoplasmosis in humans are probably
acquired by the ingestion of either tissue cysts in
infected meat or oocysts in food contaminated with cat
faeces.
● Bradyzoites from the tissue cysts or sporozoites released
from oocysts invade intestinal epithelia and multiply.
● T. gondii may spread both locally to mesenteric lymph
nodes and to distant organs by invading the lymphatic
and blood systems.
● Focal areas of necrosis (caused by localized cell lysis)
may develop in many organs.
● The extent of the disease is usually determined by the
extent of injury to infected organs, especially to vital and
vulnerable organs such as the eye, heart and adrenals.
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● Opportunist toxoplasmosis in immunosuppressed
patients usually represents reactivation of chronic infec-
tion.
● The predominant lesion of toxoplasmosis— encephalitis
in these patients—is necrosis, which often results in
multiple abscesses, some as large as a tennis ball.
● Figure 6.4 Leishmaniasis is transmitted by the bite of
female phlebotomine sandflies. The sandflies inject the
infective stage, promastigotes, during blood meals (1).
Promastigotes that reach the puncture wound are
phagocytosed by macrophages (2) and transform into
amastigotes (3). Amastigotes multiply in infected cells
and affect different tissues, depending in part on the
Leishmania species (4). This originates the clinical
manifestations of leishmaniasis. Sandflies become
infected during blood meals on an infected host when
they ingest macrophages infected with amastigotes (5,
6). In the sandfly’s midgut, the parasites differentiate
into promastigotes (7), which multiply and migrate to
the proboscis (8).
III. INTESTINAL PARASITES
● Gut protozoan parasites include Entamoeba histolytica,
Giardia lamblia, Dientamoeba fragilis, Balantidium sp.,
Isospora sp. and Cryptosporidium parvum.
● All these organisms are transmitted by the faecal–oral
route and most of them are cosmopolitan in their
distribution.
● A good example of this is Giardia, which is found in
nearly all countries of the world.
● In many developed countries, including the UK and USA,
it is one of the most commonly identified waterborne
infectious organisms.
● Cryptosporidium, like Toxoplasma, has a complex life
cycle utilizing both sexual and asexual reproduction.
● In contrast, Giardia and Entamoeba have simple life
cycles utilizing only asexual reproduction.
● These latter organisms are members of a small group of
eukaryotes that do not have mitochondria.
● It had long been assumed that they never had
mitochondria, but recent studies showing the presence
of mitochondrial-like enzymes and structural proteins
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suggest that it is more likely that this organelle was lost
as a result of metabolic/physiological adaptation.
A. GIARDIA LAMBLIA (sy. Intestinalis, duodenalis)
● Giardia duodenalis (syn lamblia and intestinalis) is the
causative agent of giardiasis, a severe diarrhoeal
disease.
● The incidence of Giardia infection worldwide ranges
from 1.5% to 20% but is probably significantly higher in
countries where standards of hygiene are poor.
● The most common route of spread is via the faecal–oral
route, although spread can also occur through ingestion
of contaminated water and these modes of transmission
are particularly prevalent in institutions, nurseries and
daycare centres.
● Recent outbreaks and epidemics in the UK, USA and
eastern Europe have been caused by drinking
contaminated water from community water supplies or
directly from rivers and streams. Many animals harbour
Giardia species that are indistinguishable from the
human infective types.
● There is now clear evidence from genotyping studies
that the G. duodenalis species is made up of a number
of genetically distinct groups which may represent
species.
● This has raised the question of the existence of animal
reservoirs of Giardia.
● Recent findings of Giardia-infected animals in
watersheds from which humans acquired giardiasis, and
the successful interspecies transfer of these organisms,
suggests that human giardiasis can be acquired by
zoonotic transfer, however it is not clear if the major
route of transfer is from animal to human or from
human to animal.
● More recently, it has been recognized that Giardia
infection may be transmitted by sexual activity,
particularly among homosexual men.
● This organism exhibits only two life cycle forms: the
vegetative binucleate trophozoite (10–20 μm long × 2 –
3 μ m w i d e ) and transmissible quadrinucleate cyst
( 1 0 – 1 2 μ m l o n g × 1 – 3 μ m w i d e ).
● Trophozoites have four pairs of flagella and an adhesive
disc, which is thought to help adhesion to the intestinal
epithelium.
● Division in trophozoites is by longitudinal fission.
● It was long believed that Giardia was a non-pathogenic
commensal.
● However, we now know that Giardia can produce
disease ranging from a self-limiting diarrhoea to a severe
chronic syndrome.
● Immune-competent individuals with giardiasis may
exhibit some or all of the following signs and symptoms:
diarrhoea or loose, foul-smelling stools; steatorrhoea
(fatty diarrhoea); malaise; abdominal cramps; excessive
flatulence; fatigue and weight loss.
● Infected individuals with an immune deficiency or
protein-calorie malnutrition may develop a more severe
disease and will exhibit symptoms such as interference
with the absorption of fat and fat-soluble vitamins,
retarded growth, weight loss, or a coeliac disease-like
syndrome.
● Giardia infection is initiated by ingestion of viable cysts,
the infective dose of which can be as low as one cyst,
although infection initiated by 10–100 viable cysts is
more likely.
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● As the cysts pass through the stomach the low pH and
elevated CO2 induce excystation (cyst–trophozoite
transformation).
● From each cyst two complete trophozoites emerge and
these rapidly undergo division then attach to the
duodenal and jejunal epithelium.
● Once attached, they will undergo division, and 4–7 days
later they will detach and begin to round up and form
cysts (encystment).
● This process is thought to be induced in response to bile.
● The first cysts are found in faeces after 7–10 days.
● The underlying pathology of giardiasis is not fully
understood.
● The trophozoites do not invade the mucosa, and
although their presence may have some physical effects
on the surface it is more likely that some of the
pathology is caused by inflammation of the mucosal cells
of the small intestine causing an increased turnover rate
of intestinal mucosal epithelium.
● The immature replacement cells have less functional
surface area and less digestive and absorptive ability.
● This would account for the microscopic changes seen in
infected epithelia.
● It has been suggested that other mechanisms may exist,
e.g. toxin production, but to date no such molecule has
been observed.
● Figure 6.5 Members of the cat family (Felidae) are the
only known definitive hosts for the sexual stages of
Toxoplasma gondii and thus are the main reservoirs of
infection. Cats become infected with T. gondii by
carnivorism (1). After tissue cysts or oocysts are ingested
by the cat, viable organisms are released and invade
epithelial cells of the small intestine where they undergo
an asexual cycle followed by a sexual cycle and then
form oocysts, which are then excreted. The
unsporulated oocyst takes 1–5 days after excretion to
sporulate (become infective). Although cats shed oocysts
for only 1–2 weeks, large numbers may be shed. Oocysts
can survive in the environment for several months and
are remarkably resistant to disinfectants, freezing and
drying, but are killed by heating to 70 °C for 10 minutes.
Human infection may be acquired in several ways: (A)
ingestion of undercooked infected meat containing
Toxoplasma cysts (2); (B) ingestion of the oocyst from
faecally contaminated hands or food (3); (C) organ
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transplantation or blood transfusion; (D) transplacental ● The initial superficial ulcer may deepen into the
transmission; (E) accidental inoculation of tachyzoites. submucosa and become chronic.
The parasites form tissue cysts, most commonly in ● Spread may occur by direct extension, by undermining
skeletal muscle, myocardium and brain; these cysts may of the surrounding mucosa until it sloughs, or by
remain throughout the life of the host. penetration that can lead to perforation.
● If the trophozoites gain access to the vascular or
B. ENTAMOEBA HISTOLYTICA lymphatic circulation, metastases may occur first to the
● Entamoeba histolytica is the causative agent of amoebic liver and then by direct extension or further metastasis
dysentery, another infection transmitted via the faecal– to other organs, including the brain.
oral route.
● The severity of this and related pathologies caused by
this organism can vary from diarrhoea associated with
the intestinal infection to extraintestinal amoebiasis
producing hepatic and often lung infection.
● The prevalence of amoebiasis in developing countries
reflects the lack of adequate sanitary systems.
● It had long been known that most infections associated
with E. histolytica are asymptomatic or exhibit minimal
symptomology, but in the late 1980s a separate, but
morphologically and biochemically similar species, E.
dispar, was identified.
● This organism exhibits limited pathogenicity and in many
cases produces no symptoms but is commonly
misidentified as E. histolytica.
● This species is the most likely cause of ‘asymptomatic
Entamoeba infection’.
● E. histolytica has a relatively simple life cycle and, like
Giardia, exhibits only two morphological forms: the tro-
phozoite and cyst stages.
● Trophozoites vary in size from 10 to 60 μm and are
actively motile.
● The cyst is spherical, 10–20 μm in diameter, with a thin
transparent wall. Fully mature cysts contain four nuclei.
● Symptoms of amoebic dysentery are associated with ● Figure 6.6 Cysts are resistant forms and are responsible
mucosal invasion and ulceration. for transmission of giardiasis. Both cysts and
● Mucosal erosion causes diarrhoea, the severity of which trophozoites can be found in the faeces (diagnostic
increases with the level of invasion and colonization. stages) (1). The cysts are hardy, and can survive several
● Symptoms can also be affected by the site of the months in cold water. Infection occurs by the ingestion
infection. of cysts in contaminated water, food or by the
● Peritonitis as a result of perforation has been reported in faecal–oral route (hands or fomites) (2). In the small
connection with severe amoebic infection. intestine, excystation releases trophozoites (each cyst
● Extraintestinal amoebiasis is usually associated with liver produces two trophozoites) (3). Trophozoites multiply by
infection, causing abscesses and/or enlargement. longitudinal binary fission, remaining in the lumen of the
● The abscess appears as a slowly enlarging liver mass and proximal small bowel where they can be free or attached
will cause noticeable pain. to the mucosa by a ventral sucking disc (4). Encystation
● Jaundice may also occur due to blockage of the bile. occurs as the parasites transit toward the colon. The cyst
● Pleural, pulmonary, and pericardial infection results from is the stage found most commonly in non-diarrhoeal
metastatic spread from the liver, but can also manifest in faeces (5). Because the cysts are infectious when passed
other parts of the viscera or give rise to a brain abscess. in the stool or shortly afterwards, person-to-person
● However, these complications are uncommon. transmission is possible. Although animals are infected
● The life cycle of E. histolytica is simple, but the ability of with Giardia, their importance as a reservoir is unclear.
trophozoites to infect sites other than the intestine make
it more complex than that of Giardia.
● Infection is initiated by ingestion of mature cysts, and Cryptosporidium parvum
again, excystation occurs during transit through the gut.
● After this, trophozoites rapidly divide by simple fission to
● ubiquitous coccidian parasite that causes
produce four amoebic cells which undergo a second divi-
cryptosporidiosis in humans
sion; thus each cyst yields eight trophozoites.
● Survival outside the host depends on the resistant cyst ● other species are known to cause infection in
form. immunocompromised patients and in total this genus
● The pathology of the disease is only partially under- comprises 19 – 20 distinct species
stood. ● Recently there has been a renaming of the major
● The process of tissue invasion has been well studied and species in this group to reflect the host specificity of
involves binding and killing of the host cells by spe- cific the particular species or genotype
adhesin molecules and the action of a pore-forming ● The life cycle of the parasite is complex but is
protein, amoebapore. completed in a single host.

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 TRANS: PROTOZOA

● Infection follows the ingestion of oocysts associated

with contaminated water or food
● According to the World Health Organization (WHO)
the health significance of C. parvum is high due to the
persistence of the organism in the environment.
○ Cattle represent the most important reservoir of
C. parvum but other mammals, domestic and
wild, can be infected and act as carriers of the
disease, even if asymptomatic
● genetically distinct subspecies exist that can be
divided into two groups:
○ Group I organisms infect humans only (soon to
be reclassified as C. hominis)
○ Group II organisms infect a wider range of hosts
➢ several large outbreaks of this infection in
the UK and USA in which water was
identified as the initial vehicle for
transmission
● Members of this genus are intracellular parasites
infecting the intestinal mucosal epithelium.
● The two major life cycle forms are the oval oocyst and Figure 6.2
the sporozoite. a. Mature schizonts of Plasmodium vivax. P. vivax
● C. parvum infections are often asymptomatic, but schizonts are large, have 12– 2 4 merozoites, and may
symptoms such as profuse watery diarrhoea, stomach fill the red blood cell (RBC). RBCs are enlarged 1.5 – 2
cramps, nausea, vomiting and fever are typical times and may be distorted. Under optimal
○ symptoms can last from several days to a few conditions Schüffner’s dots may be seen.
weeks in immunocompetent individuals b. Leishmania tropica amastigotes within an intact
○ Immunocompromised patients' infection can macrophage
become chronic, lasting months or even years. c. Toxoplasma gondii trophozoites in the bronchial
➢ mean infective dose for immunocompetent secretions from an HIV - infected patient.
people is dependent on the strain of C. d. Trophozoites of Giardia intestinalis . Each cell has two
parvum it is considered to be approximately nuclei and is 10 – 20 mm in length.
100 cells, and infants are more vulnerable to e. Trophozoites of Entamoeba histolytica with ingested
infection erythrocytes, which appear as dark inclusions.
● Diarrhoea is a major cause of childhood mortality and f. Oocysts of Cryptosporidium parvum (upper left) and
morbidity as well as malnutrition in developing cysts of Giardia intestinalis (lower right) labelled with
countries. immunofluorescence antibodies.
○ Cryptosporidium is the third most common g. Trophozoites of Trichomonas vaginalis.
cause of infective diarrhoea in children in such
countries, and consequently it plays a role in the ● During infection a variety of changes are seen such as
incidence of childhood malnutrition. partial villous atrophy, crypt lengthening and
● Cryptosporidium infection has a higher nutritional inflammation
impact in boys than girls, because of the need for ○ responses are probably due in part to cell
micronutrients in boys to build up larger muscle damage that occurs during the growth of
mass. the intracellular forms
○ breast - feeding does offer some protection ● Cryptosporidiosis is resolved by the immune system
against infection. in healthy patients normally within 3 weeks
● In immunocompromised individuals Cryptosporidium
infection causes a severe gastroenteritis, and often
the parasites infect other epithelial tissues causing
pneumonia
○ The mortality rate due to C. parvum in AIDS
patients is between 50% and 70%.
● Infection occurs when oocysts (Figure 6.2 f) excyst
following environmental stimuli (typical intestinal
conditions) and parasitize the epithelial cells which
line the intestine wall (Figure 6.8 ). After several
further stages of the cycle, two forms of oocyst are
produced; soft - walled oocysts reinitiate infection of
neighbouring enterocytes while hard-walled cysts are
expelled in the faeces.

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Figure 6.8: Life Cycle of Cryptosporidium
IV. Trichomonas and free-living amoebas
Trichomonas vaginalis
● common sexually transmitted parasite.
● Infection rates vary from 10% to 50%, with the
highest reported rates found in the USA.
● Infections are usually asymptomatic or mild although
symptomatic infection is most common in women.
● Trichomonads are all anaerobes and contain
hydrogenosomes.
● found in very few other anaerobic eukaryotes and is
often termed the ‘ anerobic mitochondrion ’
● A number of functions have been assigned to it, and
it has been shown to function in the generation of
ATP.
● does not exhibit a life cycle as only the motile
(flagellated/amoeboid) trophozoite (Figure 6.2 g) has
been seen and division is by binary fission.
● Trichomonads have a pear - shaped body 7 – 15 μ m
long, a single nucleus, three to five forward-directed
flagella, and a single posterior flagellum that forms
the outer border of an undulating membrane
● Trichomoniasis in women is frequently chronic and is
characterized by vaginal discharge and dysuria
○ inflammation of the vagina is usually diffuse and
is characterized by reddening of the vaginal wall
and migration of polymorphonuclear leukocytes
into the vaginal lumen (these form part of the
vaginal discharge).
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● Because there is no resistant cyst, transmission from
host to host must be direct.
● The inflammatory response in trichomoniasis is the
major pathology associated with this organism;
however, the mechanisms of induction are not
known.
● It is likely that mechanical irritation resulting from
contact between the parasite and vaginal epithelium
is a major cause of this response but the organism
produces high concentrations of acidic end - products
and polyamines, both of which would also irritate
local tissues.
Free-living opportunist amoebas
● often forgotten group of protozoans.
● The two major groups, Naegleria and Acanthamoeba,
infect humans and both can cause fatal encephalitis.
● Both types of infections are rare, with less than 200
cases of Naegleria fowleri infection recorded
worldwide and approximately 100 – 200 cases of
Acanthamoeba ulcerative keratitis per year
● commonly associated with contact lens use and it is
thought that infection is caused by a combination of
corneal trauma and dirty contact lenses
● Both types of amoeba produce resistant cysts and
Naegleria also exhibits a flagellate form.
● Both Acanthamoeba and Naegleria are free - living
inhabitants of fresh water and soil
○ Naegleria fowleri (the human pathogen)
reproduces faster in warm waters up to 46 ° C.
○ Treatment of water by chlorination or ozonolysis
does not entirely eliminate cysts and both
amoebae have been isolated from air
conditioning units.
1. Naegleria fowleri
● is the causative agent of primary amoebic
meningoencephalitis, a rapidly fatal disease that
usually affects children and young adults.
● In all cases, contact with amoebae occurs as a result
of swimming in infected fresh water.
● The organisms enter the brain via the olfactory tract
after amoebae are inhaled or splashed into the
olfactory epithelium.
● The incubation period ranges from 2 to 15 days and
depends both on the size of the inoculum and the
virulence of the strain.
● The disease appears with the sudden onset of severe
frontal headache, fever, nausea, vomiting and stiff
neck.
● Symptoms develop rapidly to lethargy, confusion and
coma and in all cases to date the patient died within
48 – 72 hours.
2. Acanthamoeba castellanii, A. culbertsoni and other
pathogenic Acanthamoeba species
● can cause opportunist lung and skin infections in
immunocompromised individuals
● Where amoebae spread from such lesions to the
brain, they can cause a slowly progressive and usually
fatal encephalitis.
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● Acanthamoeba can cause an ulcerating keratitis in
healthy individuals, usually in association with
improperly sterilized contact lenses.
● The presence of cysts and trophozoites in alveoli or in
multiple nodules or ulcerations of the skin
characterizes acanthamoebic pneumonitis and
dermatitis.
● Spread of amoebae to the brain produces an
encephalitis, characterized by neurological changes,
drowsiness, personality changes and seizures in the
early stages of infection, which progress to altered
mental status, lethargy and cerebellar ataxia.
● The end point of infection is usually coma followed by
death of the patient.
● Acanthamoeba keratitis is characterized by painful
corneal ulcerations that fail to respond to the usual
anti - infective treatments.
● The infected and damaged corneal tissue may show a
characteristic annular infiltrate and congested
conjunctiva.
● If not successfully treated, the disease progresses to
corneal perforation and loss of the eye or to a
vascularized scar over thinned cornea, with impaired
vision.
V. Host response to infection
● Mechanisms to control parasitic protozoa are similar
to those utilized for other infectious agents
● divided into non - specific mechanism(s) and specific
mechanism(s) involving the immune system
1. non - specific mechanisms include those that
affect the entry of parasites into the red blood
cell
a. sickle cell haemoglobin trait and lack of the
Duffy factor on the erythrocyte surface make
the red cell more resistant to invasion by
Plasmodium
b. These traits are commonly found in
populations from malaria - endemic regions.
c. A second example of a non - specific factor is
the presence of trypanolytic factors in the
serum of humans which confer resistance to
T. brucei
d. Non - specific factors can play a key role in
resistance, usually they work in conjunction
with the host’s immune system.
Immune response
● Unlike most other types of infection, protozoan
diseases are often chronic, lasting for months to
years. When associated with a strong host immune
response, this type of long - term infection is apt to
result in a high incidence of immunopathology.
● The impact of HIV infection on many parasitic
diseases has highlighted this relationship.
● Different parasites elicit different humoral and/or
cellular immune responses (Chapter 9 ).
● In malaria and trypanosome infections, antibodies
appear to play a major role in immunity, although it
would seem that for many organisms both humoral
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
and cellular immunity are required for killing of
parasites.
● Cellular immunity is believed to be the most
important mechanism in the killing of Leishmania and
Toxoplasma
● Cytokines are involved in the control of both the
immune response and also the pathology of many
parasitic diseases.
● Helper (h) and cytotoxic (c) T cells play major roles in
the induction/control of the response.
● The various subsets of these produce different
profiles of cytokines.
● For example, the Th1 subset produces gamma
interferon (IFN - γ ) and interleukin - 2 (IL - 2) and is
involved in cell - mediated immunity.
● In contrast, the Th2 subset produces IL - 4 and IL - 6,
and is responsible for antibody - mediated immunity.
● The induction of the correct T - cell response is key to
recovery.
● The Th1 subset and increased IFN - γ are important
for the control of Leishmania , T. cruzi and
Toxoplasma infections, whereas the Th2 response is
more important in parasitic infections in which
antibody is a major factor.
● It is important to recognize that the cytokines
produced by one T - cell subset can up - or down -
regulate the response of other T - cell subsets; IL - 4
will down - regulate Th1 cells for example.
● The cytokines produced by T and other cell types do
not act directly on the parasites but induce changes
in the metabolism of glucose, fatty acid and protein in
other host cells.
● Cytokines can also stimulate cell division and,
therefore, clonal expansion of T - and B - cell subsets.
● This can lead to increased antibody production
and/or cytotoxic T - cell numbers.
● The list of cytokines and their functions is growing
rapidly, and it would appear that these chemical
messages influence all phases of the immune
response.
● They are also clearly involved in the multitude of
physiological responses (fever, decreased food intake,
etc.) observed in an animal ’ s response to a
pathogen, and in the pathology that results.
Immune pathology
● The protozoa can elicit humoral responses in which
antigen – antibody complexes are formed and these
can trigger coagulation and complement systems.
● Immune complexes have been found circulating in
serum and deposited in the kidneys where they may
contribute to conditions such as glomerulonephritis.
● In other tissues these complexes can also induce
localized hypersensitivities.
● It is thought that this type of immediate
hypersensitivity is responsible for various clinical
syndromes including blood hyperviscosity, oedema
and hypotension.
● Another important form of antibody - mediated
pathology is autoimmunity.
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 TRANS: PROTOZOA
● Autoantibodies to a number of different host
antigens (e.g. red blood cells, laminin, collagen and
DNA) have been demonstrated.
● These autoantibodies may play a role in the
pathology of parasitic diseases by exerting a direct
cytotoxic effect on the host cells, e.g. autoantibodies
that coat red blood cells produce haemolytic
anaemia; they may also cause damage through a
build - up of antigen – antibody complexes.
● Many parasites can elicit the symptoms of disease
through the action of their surface molecules such as
the pore - forming proteins of E. histolytica that
induce contact - dependent cell lysis, and
trypanosome glycoproteins that can fix and activate
complement resulting in the production of
biologically active and toxic complement fragments.
● A range of parasite - derived enzymes such as
proteases and phospholipases can cause cell
destruction, inflammatory responses and gross tissue
pathology.
Immune evasion
● Parasites exhibit a number of mechanisms that allow
them to evade host immune response.
● Two such mechanisms are displayed by
trypanosomes, which are able to exhibit both
antigenic masking and antigenic variation.
● Masking means the parasite becomes coated with
host components and therefore is not recognized as
foreign.
● In addition parasites can undergo antigenic variation
which results in surface antigens being changed
during the course of an infection.
● The ultimate goal of these is that the host's immune
response is evaded.
● Parasites can also suppress the host ’ s immune
response either to the parasite specifically or to
foreign antigens in general.
● This, however, can cause a number of problems, as
general immune suppression may make the individual
more susceptible to secondary infection.
VI. Detection of parasites
● The detection of parasites in the host and
environment (including foods) is vital for proper
treatment and also for understanding mechanisms of
transmission.
● In addition the use of molecular typing is of major
benefit in studying epidemiology and for some of the
zoonotic potential of parasites.
Methods of detection
● detection of these organisms in samples requires the
use of methods such as microscopy and DNA
amplification.
● The most commonly used method involves
microscopy and this can be applied to clinical as well
as environmental samples.
● For some organisms this approach remains the
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● ‘ gold standard ’ for detection and identification.
● The advantages of microscopy are speed, cost and
availability.
● fluorescent - labelled antibodies raised to
● species - specific antigens can be utilized to help
identify organisms to species or subspecies level, and
other stains can be used to help determine the
viability of cells.
● Examples of such stains include fluorescein diacetate
● which is cleaved by esterases in viable cells, releasing
fluorescein (gives a green fluorescence) and
propidium iodide which is excluded from viable cells
but taken up by cells with damaged membranes
(gives red fluorescence).
● However, there are a number of obvious limitations
including the requirement for well - trained staff to
perform the microscopy, limits of sensitivity of the
method and, for some parasites, difficulty in
differentiating species based on morphology.
● the parasite may not be present in easily available
samples. Thus a variety of other approaches are used
to help in detection.
Antibody-based technologies
● For the major protozoan parasites a number of
immunology - based methods exist to detect the
presence of organisms in clinical samples.
● These include the use of agglutination, complement
fixation and enzyme - linked immunosorbent assay
(ELISA).
● The most commonly used method is ELISA and this
can be used to detect the presence of antigens in
samples (direct assay) or antigen - specific antibodies
in patients ’ serum (indirect).
● The ELISA method has a number of advantages in that
it can be automated and has good specificity and
sensitivity.
● Although a number of ELISA - based detection kits are
available their cost can be a limitation to use
especially in those developing countries where
parasitic infections are endemic.
● The use of indirect ELISA methods for determining
the infection status of the patient can also be difficult
because previous exposure to the parasite can cause
problems and the immune status of the patients can
also impact on antibody production.
DNA-based technologies
● Nucleic acids, DNA or RNA, are found in all types of
microorganism and the sequences of these molecules
can be used to help in the identification of species
and individuals.
● The robust nature of DNA has allowed researchers to
use these sequences in a variety of applications
including forensics, archaeology (sequences from
animal tissues over 100 years old have routinely been
determined)and epidemiology.
● The advent of highly sensitive DNA amplification
technologies such as the polymerase chain reaction
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 TRANS: PROTOZOA
(PCR) has allowed the development of molecular
tools for the identification and detection of parasites.
● Initial work on these techniques focused on their
ability to help define species and many of these
studies utilized the ribosomal DNA genes and were
very successful.
● Using these genes it was possible to discriminate
many genera and species, but the genes lacked the
level of sequence variation required for separation
below the level of species (important for some
pathogenic organisms and for epidemiological
studies).
● In addition ribosomal DNA sequencing was also not
always appropriate for identifying species that are
highly diverse.
● Other DNA markers that have been used included
genes that encode for metabolic enzymes and
structural proteins.
● For example Cryptosporidium can be speciated by
using a combination of target genes including the
Cryptosporidium oocyst wall protein (COWP), heat
shock protein (hsp70), dihydrofolate reductase
(DHFR) and 18S ribosomal DNA.
● Complete genomes are now available for the major
protozoan parasites and this has helped in the
development of improved methods for species
identification.
● Other DNA - based techniques are now available such
as the multiplex PCR (amplification of more than one
gene at a time) and quantitative real - time PCR
(qPCR), and these have improved specificity and
sensitivity in detection and increased the usefulness
of DNA technology for the detection of parasites in
environmental as well as clinical samples.
● It is also possible to assess the viability of protozoan
parasites using reverse transcription PCR (RT - PCR).
● This is due to the fact that mRNAs degrade quickly
once parasites are killed.
● However, it should be noted that increasing evidence
exists to show that mRNA can survive in some cells
for up to 24 hours
● A recent method, DNA microarray, is a fast
developing technology.
● This is a ‘ chip ’ based system that measures the
binding of target DNA sequences (parasite) from
samples (fluorescently labelled using PCR) to
complementary sequences bound to the chip surface.
● The binding of parasite sequence DNA and the
amount that is bound can be detected using
fluorescence (Figure 6.9 ).
● In theory this technique allows the detection of
multiple parasite species at the same time in one
sample.
Alternative methods
● Alternative methods are being developed for the
detection of parasites; these include mass
spectrometry (MS) and biosensors.
● MS can be used by identifying known surface
characteristics of appropriate life cycle forms.
● This type of MS uses low - energy electrons to
generate a fingerprint of the cells.
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● Other mass spectrometry methods can generate
fingerprints based on parasite - derived peptides and
metabolites.
● These can be detected easily with a high level of
sensitivity and can be used in complex mixtures that
would be present in clinical and environmental
samples.
● Over the past 10 years the production of kits for near
- patient testing has driven the development of
detection technology and this has generated interest
in the use of biosensors.
● These sensors detect the presence of ‘ marker
molecules ’ that are specific to a particular pathogen
and utilize mechanical, electrochemical or
piezoelectric methods to generate a signal that can
be detected.
● Such sensors can be combined onto ‘ chips ’ to detect
a range of pathogens.
Analysis of samples
Clinical samples
● Diagnosis of parasitic infection is dependent on the
demonstration of the parasite in appropriate
samples.
● The type of samples can vary from blood where
preparation can be minimal (e.g. sample of smears
for microscopy) to faeces or intestinal aspirates.
● Faecal samples require more processing: for example,
fresh or preserved stools can be concentrated to
increase the yield of the parasites by sedimentation
using the formol – ether or formol – ethyl acetate
techniques or by faecal parasite flotation method
using copper sulphate.
● These concentrates can be stained for microscopy.
● For the extraction of DNA from samples, a number of
methods exist; however, there are various resin -
based kits that will separate DNA from complex
biological samples.
Environmental samples
● Many parasites have life cycle forms that can survive
in the environment.
● These act to initiate infections in susceptible hosts;
for some parasites this only requires ingestion of no
more than 10 cells or cysts, thus the ability to detect
them in the environment is vital.
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● This section focuses on water and foods as they are
often major routes of transmission for parasites such
as Giardia and Cryptosporidium which are included in
the monitoring standards for potable waters (drinking
- waters) by regulatory bodies in both the European
Union and the USA.
● In addition, food and pharmaceutical companies that
utilize water in their processes are required to test for
a number of pathogens, including waterborne
parasites.
● Foods can also be a source of infection and methods
for the detection of pathogens in foods are as
important as those for water samples.
● A number of methods can be used to detect and to
identify parasites but the performance of these
techniques will often be impaired when used on real
samples.
○ This is due to a number of factors that include:
➢ the levels of parasites present
➢ volume of samples required for analysis
➢ the presence of other microorganisms
➢ the presence of compounds that interfere
with detection
➢ turbidity of samples (water)
➢ high levels of protein, lipids and
carbohydrates in samples (foods).
○ Often these factors combine to make detection
difficult.
○ For example, if levels of a parasite in water are
low ( < 1/100 ml) several litres must be collected
and analysed.
○ This is possible, but would require the sample
volume to be reduced for analysis. This can be
achieved either by filtration or centrifugation.
○ If the sample contains sediment, both of these
approaches are difficult; however, methods such
as tangential flow filtration (fluid passed parallel
to the filter) can be used.
● Waters can also contain organic acids that can inhibit
the PCR process so these must be removed from the
sample before analysis.
● DNA - based technology may also fail to detect the
presence of a parasite if the DNA present in the
sample is degraded, e.g. by other organisms.
● In foods, the process can be even more complex and
separation of parasites from the foodstuff is often the
factor that impacts most on the limits (sensitivity) of
the method
VII. Control of protozoan parasites
● It is now clear that the best approach for the
successful control of parasites requires the
integration of a number of methods which draw upon
our increasing understanding of the parasites ’ life
cycle, epidemiology and host response to infection
Chemotherapy
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● The origins of chemotherapy are closely linked to the
development of antiparasitic agents, but there has
been slow progress in the development of new and
novel antiprotozoal agents over the past 30 years.
● Recently, with the support of the WHO and
government - sponsored research, new antiparasitic
drugs are slowly coming into the market.
● Interestingly there are still a number of protozoan
parasite infections such as cryptosporidiosis for which
there is no effective treatment.
Mechanisms of action and selective toxicity
● For many of the commonly used antiprotozoal drugs
the modes of action and mechanisms of selective
toxicity are well understood, although for some the
precise mechanism remains unclear.
● The most common antiprotozoal drugs and their
modes of action are shown in Table 6.1.
● Considering the drugs in relation to modes of action,
dapsone and the sulphonamides block the
biosynthesis of tetrahydrofolate by inhibiting
dihydropteroate synthetase, while the 2,4 - diamino -
pyrimidines (proguanil and pyrimethamine) block the
same pathway but at a later step catalysed by
dihydrofolate reductase.
● The drugs that interfere with nucleic acid synthesis
include those that bind to the DNA and intercalate
with it such as chloroquine, mefloquine and quinine,
and also pentamidine, which is unable to intercalate
but probably interacts ionically.
● Other compounds such as benznidazole and
metronidazole may alkylate DNA through activation
of nitro groups via a one - electron reduction step.
● Several of these compounds, however, including
chloroquine, mefloquine, quinine and metronidazole,
have more than one potential mode of action.
● Chloroquine, for example, inhibits the enzyme haem
polymerase, which functions to detoxify the cytotoxic
molecule haem that is generated during the
degradation of haemoglobin.
● Metronidazole is reduced in the parasite cell and
forms a number of cytotoxic intermediates, which can
cause damage not only to DNA but also to
membranes and proteins.
● Tetracycline targets protein synthesis in Plasmodium
via a similar mechanism to that seen in bacteria:
inhibition of chain elongation and peptide bond
formation.
● Eflornithine interferes with the metabolism of the
amino acid ornithine in T. brucei gambiense by acting
as a suicide substrate for the enzyme ornithine
decarboxylase.
● Albendazole has recently been shown to have
significant antigiardial activity, although its mode of
action is unclear.
● In Leishmania , amphotericin B binds to ergosterol
in the membrane making it leaky to ions and small
molecules (e.g. amino acids), while the antiprotozoal
drugs atovaquone and primaquine bind to the
cytochrome bc 1 complex and inhibit electron flow.
● The antitrypanosomal drug melarsaprol is most likely
to act by blocking glycolytic kinases, especially the
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 TRANS: PROTOZOA

cytoplasmic pyruvate kinase, although it may also

disrupt the reduction of trypanothione. ● The early success in malaria control can be attributed
to the use of professional spray teams who treated
Table No 6.1 Common antiprotozoal drugs and their modes of the inside of huts with DDT, without any direct
action involvement of the infected population.
Drug Mode of Mechanism Target ● However, the problem with pesticides like DDT is that
action of selectivity organism(s) they lack specificity, and as application is not always
(if known) well directed there is often destruction of a wide
Dapsone Cofactor Unique Plasmodium range of insects, which may have undesirable side
synthesis target spp. effects.
Proguanil Cofactor Differences Plasmodium ● Further problems include the accumulation of
synthesis in the target spp. pesticide residues in the food chain and pesticide
Pyrimethamine Cofactor Differences Plasmodium resistance in the target organism.
synthesis in the target spp. ● Window screens and bed nets do prevent mosquito
Sulphonamides Cofactor Differences Plasmodium bites, however, and there has been a lot of interest in
synthesis in the target spp. using bed nets impregnated with insecticide.
Benznidazole Nucleic acid Activation in Trypanosoma ● Environmental control was a major strategy used
synthesis the parasite spp. before the development of modern insecticides.
Chloroquine Nucleic acid Differential Plasmodium ● A good example of this is mosquito control through
synthesis uptake spp. the removal of breeding sites by drainage, land
Mefloquine Nucleic acid Differential Plasmodium reclamation projects, removal of vegetation
synthesis uptake spp.
overhanging water, speeding up water flow in canals,
Metronidazole Nucleic acid Activation in Giardia, T
and periodic drainage and drying out of canals. Life
synthesis the parasite richomonas,
cycle forms that enter the water system, such as cysts
Entamoeba
and oocysts of Giardia and Cryptosporidium , can
Pentamidine Nucleic acid Differential Leishmania
present a major public health problem.
synthesis uptake spp.
● These forms are often resistant to common
Quinine Nucleic acid Differential Plasmodium
disinfection methods and require physical removal
synthesis uptake spp.
from waters.
Eflornithine Protein Differences T. brucei
function in the target gambiense ● Cysts and oocysts can be destroyed by use of proper
Tetracycline Protein Differential Plasmodium
sewage treatments such as anaerobic digestion, but
function uptake spp. these systems require regular maintenance in order
Benzimidazoles Microtubule Differences Giardia , to remain effective.
function in the target Trichomonas
Amphotericin Membrane Differences Leishmania Biological control
B function in the target spp.
Atovaquone Energy Differences Plasmodium ● Biological control is an active but developing area.
metabolism in the target spp. ● Genetic control of insect vectors, particularly the use
Melarsoprol Energy Target T. brucei of irradiated sterile males, has been widely
metabolism pathway gambiense publicized, and the release of chemically sterile males
in the more has been attempted to control anopheline
parasite important mosquitoes.
Primaquine Energy Differences Trypanosoma ● Other similar methods include the release of closely
metabolism in the target spp. related species within the environment in order to
produce sterile hybrids.
Drug resistance ● Genetically modified mosquitoes are currently being
developed that are resistant to Plasmodium infection,
● As with bacteria, drug resistance in some parasites and larvivorous fish have also been employed for
such as Plasmodium is a major problem and tends to mosquito control; other organisms considered for the
appear where chemotherapy has been used same purpose include bacteria, fungi, nematodes and
extensively. predatory insects.
● This problem is exacerbated by the fact there are so ● One of the best - studied agents is the bacterium
few drugs available for the control of some parasites, Bacillus thuringiensis ; the spore or the isolated toxin
which utilize the same five basic resistance from this species can be used as a very effective and
mechanisms that are displayed by bacteria: specific insecticide.
(1) metabolic inactivation of the drug;
(2) use of efflux pumps Vaccination
(3) use of alternative metabolic pathways
(4) alteration of the target ● Where exposure to infection is likely to occur, killing
(5) elevation of the amount of target enzyme the parasite as it enters the host is a sensible
approach to control.
Other approaches to control ● There are two options available, chemoprophylaxis or
vaccination.

15
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 TRANS: PROTOZOA
● Unfortunately long - term chemotherapy can have
adverse side effects, and in the absence of symptoms
members of the at - risk population may fail to take
the treatment.
● Vaccination would seem to be the ideal method of
parasite control, as lifelong resistance may result
from just a single treatment.
● Despite a huge amount of effort, the only successful
parasite vaccines are those for the control of
veterinary parasites.
● However, there has been significant success with the
development of recombinant vaccines for the control
of malaria.
● The recent development of DNA vaccines may be of
use in the control of parasites.
● In this method the DNA encoding an important
parasite protein is injected into host cells and the
foreign ‘ vaccinating ’ protein is synthesized in or on
the surface of the cell.
● This intracellular foreign protein enters the cell ’ s
major histocompatibility complex (MHC) class 1
pathway resulting in a cell - mediated immune
response.
● In contrast, a protein that is extracellular enters the
MHC class 2 pathway, which results primarily in an
antibody or humoral response.
● Despite advances in technology, few commercially
available parasite vaccines exist and there are none
for use in humans. Several vaccines are being
evaluated in clinical trials and most of these are
against malaria.
● In addition there are vaccines being evaluated against
other parasites, for example a Leishmania vaccine co -
administered with BCG (bacillus of Calmette and
Guerin) is under trial.
● Recently, DNA and viral vector - based vaccines have
been tested in clinical trials including one utilizing an
adenovirus human serotype 35 - based vector
encoding the malarial circumsporozoite protein.
PPT
I.
PARASITOLOGY
Association Between Parasite and Host
● A parasite is a living organism, which takes its
nourishment and other needs from a host.
● Host is an organism which supports the parasite. The
parasites included in medical parasitology are
protozoa, helminthes, and some arthropods.
○ Understanding parasites which do not ordinarily
produce disease in healthy (immunocompetent)
individuals but do cause illness in individuals
with impaired defense mechanism
(opportunistic parasites) is becoming of
paramount importance because of the
increasing prevalence of HIV/AIDS in our
country.
● There is a dynamic equilibrium which exists in the
interaction of organisms.
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● Any organism that spends a portion or all of its life
cycle intimately associated with another organism of
a different species is considered as symbiont
(symbiote) and this relationship is called symbiosis
(symbiotic relationships).
● Three common symbiotic relationships between two
organisms:
○ Mutualism
○ Commensalism
○ Parasitism
BASIC CONCEPTS OF PARASITOLOGY
Different Kinds of Parasites
● Ectoparasite – a parasitic organism that lives on the
outer surface of its host, e.g. lice, ticks, mites etc.
● Endoparasites – parasites that live inside the body of
their host, e.g. Entamoeba histolytica.
● Obligate Parasite - This parasite is completely
dependent on the host during a segment or all of its
life cycle, e.g. Plasmodium spp.
● Facultative parasite – an organism that exhibits both
parasitic and non-parasitic modes of living and hence
does not absolutely depend on the parasitic way of
life, but is capable of adapting to it if placed on a
host. E.g. Naegleria fowleri
● Accidental parasite – when a parasite attacks an
unnatural host and survives.
E.g. Hymenolepis diminuta (rat tapeworm).
● Erratic parasite - is one that wanders in to an organ in
which it is not usually found.
E.g. Entamoeba histolytica in the liver or lung of
humans
Different Kinds of Hosts
● Definitive host – a host that harbors a parasite in the
adult stage or where the parasite undergoes a sexual
method of reproduction.
● Intermediate host - harbors the larval stages of the
parasite or an asexual cycle of development takes
place. In some cases, larval development is
completed in two different intermediate hosts,
referred to as first and second intermediate hosts.
● Paratenic host – a host that serves as a temporary
refuge and vehicle for reaching an obligatory host,
usually the definitive host, i.e. it is not necessary for
the completion of the parasite's life cycle.
● Reservoir host – a host that makes the parasite
available for the transmission to another host and is
usually not affected by the infection.
● Natural host – a host that is naturally infected with
certain species of parasite.
● Accidental host – a host that is under normal
circumstances not infected with the parasite.
LAB DIAGNOSES
● Depending on the nature of the parasitic infections,
the following specimens are selected for laboratory
diagnosis:
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 TRANS: PROTOZOA
○ Blood – in those parasitic infections where the
parasite itself in any stage of its development
circulates in the blood stream, examination of
blood film forms one of the main procedures for
specific diagnosis
Example:
฀ In Malaria the parasites are found inside
the red blood cells
฀ In Bancroftian and Malayan filariasis,
microfilariae are found in the blood
plasma
○ Stool – examination of the stool forms an
important part in the diagnosis of intestinal
parasitic infections and also for those helminthic
parasites that localize in the biliary tract and
discharge their eggs into the intestine.
○ Urine – when the parasite localizes in the
urinary tract, examination of the urine will be of
help in establishing the parasitological
diagnosis.
Example:
฀ In urinary Schistosomiasis, eggs of
Schistosoma haematobium are found in
the urine.
฀ In cases of chyluria caused by Wuchereria
bancrofti, microfilariae are found in the
urine
○ Biopsy material - varies with different parasitic
infections.
Example:
฀ spleen punctures in cases of kala-azar,
฀ muscle biopsy in cases of Cysticercosis,
Trichinelliasis, and Chagas’ disease,
฀ skin snip for Onchocerciasis.
○ Urethral or vaginal discharge – for Trichomonas
vaginalis
○ Sputum – examination of the sputum is useful in
the following:
Example:
฀ In cases where the habitat of the parasite is
in the respiratory tract, as in Paragonimiasis,
the eggs of Paragonimus westermani are
found.
INDIRECT EVIDENCES
● Cytological changes in the blood:
○ Example:
฀ eosiniphilia often gives an indication of
tissue invasion by helminthes, a reduction
in white blood cell count is an indication of
kala-azar
฀ anemia is a feature of hookworm
infestation and malaria
● Serological tests – are carried out only in laboratories
where special antigens are available.
TREATMENT
● Many parasitic infections can be cured by specific
chemotherapy. The greatest advances have been
made in the treatment of protozoal diseases.
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
● For the treatment of intestinal helminthiasis, drugs
are given orally for direct action on the helminthes.
To obtain maximum parasiticidal effect, it is desirable
that the drugs administered should not be absorbed
and the drugs should also have minimum toxic effect
on the host.
PREVENTION
● Measures may be taken against every parasite
infecting humans. Preventive measures designed to
break the transmission cycle are crucial to successful
parasitic eradication. Such measures include:
○ Reduction of the source of infection- the
parasite is attacked within the host, thereby
preventing the dissemination of the infecting
agent. Therefore, a prompt diagnosis and
treatment of parasitic diseases is an important
component in the prevention of dissemination.
○ Sanitary control of drinking water and food.
○ Proper waste disposal – through establishing
safe sewage systems, use of screened latrines,
and treatment of night soil.
○ The use of insecticides and other chemicals
used to control the vector population
○ Protective clothing that would prevent vectors
from resting in the surface of the body and
inoculate pathogens during their blood meal.
○ Good personal hygiene.
○ Avoidance of unprotected sexual practices.
SPECIMEN COLLECTION AND PROCESSING
● Diagnosis of parasite infections often depends on
observing parasite forms that include protozoa, ova,
larva, or adult forms. Specimen types include stools
(most common), tissue, urine, sputum, and blood.
○ Stool samples should be free of antimicrobial
agents or other substances that inhibit parasite
growth. Barium (from enemas) can obscure
parasites during microscopic examination.
฀ At least 3 grams of fecal sample on three
consecutive days are required for most
parasite analyses.
฀ Because urea and acidic pH inhibit some
parasites and distort their morphology,
stool should be free of urine.
฀ Liquid stools are best to detect
trophozoites, whereas formed stools are
best to detect ova and cysts.
TERMINOLOGIES
Table No. Terminologies
TERMINOLOGY DEFINITION
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An asymptomatic host that harbors a 5. Less toxic preservatives generally substitute zinc
Carrier parasite and is capable of transmitting it sulfate for mercury. Compared to PVA, these
to others preservatives do not provide the quality of
Cestode Tapeworm preservation of intestinal protozoa.
Ciliate Protozoa motile by means of cilia
Symbiotic relationship beneficial to one
Commensalism member and harmless Optimal Detection of Parasites Often Requires Concentration
to another of Specimen
Thick-walled stage of protozoa resistant 1. Gross examination of stool may detect adult forms,
Cyst
to adverse conditions particularly helminths (worms).
Host supporting the adult or sexual 2. Concentration procedures for feces remove debris
Definitive Host
phase of a parasitic life cycle that could obscure parasites. Barium is not removed
Ectoparasite Parasite found on the surface of a host during concentration procedures.
Endoparasite Parasite found inside a host 3. Fecal concentration methods
Filariae Blood or tissue roundworms a. Formalin-ethyl acetate sedimentation
Flagellate Protozoa motile by means of flagella b. Zinc sulfate flotation
Gravid Containing ova c. Sheather sugar flotation
Worms that include nematodes 4. Blood concentration methods
Helminths (roundworms), cestodes (tapeworms), a. The Knott method uses low-speed
and trematodes (flukes) centrifugation to concentrate blood samples
Hermaphroditic Organism capable of self-fertilization suspected of containing minimal numbers of
Living organism that harbors another parasites.
Host b. Buffy coat slides are used for Leishmania or
organism
Hydatid cyst Larval stage of Echinococcus granulosus. Trypanosoma detection
Intermediate Host containing the asexual phase of a
host parasite MEDICALLY-IMPORTANT PARASITES
Larva Juvenile stage of a parasite
Immature Schizont:
Early stage of asexual sporozoa INTESTINAL PROTOZOA
Schizont Mature Schizont: ● General Characteristics:
Developed stage of asexual sporozoa ○ Pseudopods are extensions of cytoplasm
trophozoite providing motility unique to amebae.
Symbiotic relationship beneficial to both ○ Trophozoite and cyst stages are part of the
Mutualism amebae life cycle.
species
Nematode Roundworm ○ Most amebic infections are spread to humans
through contaminated water.
Oocyst Encysted form of an egg
○ Cyst is the infective stage, whereas the
An organism that obtains its nutrients
trophozoite is the active reproduction stage
Parasite from another organism (the host) while
destroyed by stomach acid.
harming the host
○ Laboratory identification:
Symbiotic relationship in which one
฀ Microscopic identification of cysts (in
Parasitism member benefits at the expense of
formed stools) and trophozoites (in liquid
another member (the host)
stools) based on size, nuclear
An association between two or more characteristics, and inclusions
Symbiosis
organisms of different species ○ Size is one of the most important criteria for
identification.

MORPHOLOGIC TERMS ASSOCIATED WITH PROTOZOA

STOOL PRESERVATIVES

1. Stool specimens should not be frozen, and Table No. Morphologic terms associated with protozoa
unpreserved specimens should not be stored at room TERMINOLOGY DEFINITION
temperature longer than a couple of hours. Karyosome Area of chromatin within the nucleus
2. Formalin (5 or 10%) is an all-purpose preservative to Peripheral Nucleic acid combined with protein
preserve stool specimens for concentration chromatin found along the nuclear membrane
procedures. Excystation Development of a cyst into a trophozoite
3. Polyvinyl alcohol (PVA) is a mercury-containing
Encystation Development of a trophozoite into a cyst
preservative for preparing permanent stained smears.
Chromatoid bar Rod-shaped, RNA containing structure
4. Sodium acetate formalin (SAF) is a mercury-free
preservative that can be used to preserve stool
samples for both concentration and permanent
stained smears INTESTINAL AMOEBAE

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Entamoeba histolytica
● The only amoeba pathogenic for the gastrointestinal
tract.
● Cysts are infective when ingested. Excystation occurs
in the small intestines. Infective cysts are passed in
stools and are resistant to environmental stress.
● Infections caused:
○ Amoebic colitis
฀ Amoebic colitis is characterized by
abdominal cramping, anorexia, fatigue,
and diarrhea. It can also cause ulcers and
dysentery.
○ Liver abscess
● Morphology:
○ Cyst characteristics:
฀ Cysts range in size from 8-22 um, and they
are spherical.
฀ It contains 1-4 nuclei; peripheral
chromatin is fine and uniformly
distributed.
฀ Karyosome is centrally located
฀ Cytoplasm is finely granular with
chromatoid bars with round ends.

○ Trophozoite characteristics:
฀ Trophozoites range in size from 5-70 um
and they are motile by means of
pseudopods
฀ E. histolytica trophozoites contain one
nucleus, and they resemble those found in
the cyst
฀ Cytoplasm is finely granular and may
contain red blood cell (RBC) inclusions.
฀ The presence of intracellular RBCs in
intestinal amoebae is considered
diagnostic of E. histolytica

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Morphologically, E. histolytica is identical to the nonpathogen

E. dispar. These two species can be differentiated by
immunologic assays detecting surface antigens

● Laboratory Diagnosis:
○ Intestinal Amoebiasis
฀ Examination of a fresh dysenteric fecal
specimen or rectal scraping for trophozoite
stage.
฀ Examination of formed or semiformed
feces for cyst stage.
○ Extraintestinal Amoebiasis
฀ Diagnosed by the use of scanning
procedures for liver and other organs.
฀ Specific serologic tests, together with
microscopic examination of the abscess
material, can confirm the diagnosis.
● Treatment:
○ Acute, fulminating amebiasis is treated with
metronidazole followed by iodoquinol
○ Asymptomatic carriage can be eradicated with
iodoquinol, diloxanide furoate, or paromomycin

○ Metronidazole, chloroquine, and diloxanide

furoate can be used for the treatment of extra
intestinal amoebiasis

Entamoeba coli
● Notes:
○ E. coli is generally nonpathogenic but may cause
intestinal problems in immunosuppressed
patients.
○ If found in a stool specimen, E. coli can indicate ○ Trophozoite characteristics:
the presence of pathogenic organisms. ฀ Trophozoites range in size from 10 to 60
○ Needs to be differentiated from E. histolytica for um, and they are motile by means of
purposes of treatment short/blunt pseudopods.
● Morphology: ฀ Contain single nucleus with coarse,
○ Cyst Characteristics: unevenly distributed chromatin, and they
฀ Cysts range in size from 8 to 40 um, and resemble those found in the cyst.
they are spherical. ฀ Cytoplasm is coarse and vacuolated, with
฀ It contains 1-8 nuclei; the peripheral bacterial inclusions.
chromatin is coarse and unevenly
distributed. Young cysts may contain a
large central glycogen mass.
฀ Karyosome is eccentric and large
฀ Cytoplasm is coarse with thin chromatoid
bars with pointed ends

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○ Microscopic examination of stool for

trophozoites

Blastocystis hominis
● Notes:
○ Currently classified as an amoeba but rRNA
analysis indicates it is related to algae and water
moulds
● MOT:
○ Contaminated food and water
● Infections caused:
○ DIARRHEA and ABDOMINAL PAIN
● Morphology: Giardia lamblia
○ The classic form varies in diameter from 4 to 60 ● Taxonomy: G. duodenalis and G. intestinalis are
um and contains a large central body that fills synonyms
about 90% of the cell volume ● Infection caused:
○ There is an outer ring of cytoplasm with several ○ Giardiasis (“traveler’s diarrhea”)
nuclei around the central body. ● MOT:
● Lab diagnosis: ○ Exposure to contaminated water and food;
○ Microscopic examination of stool sample campers and hunters are prone to infection
○ The organism may be detected in wet mounts or after drinking untreated water from streams
trichome –stained smears of fecal specimens ● Cysts are the infective stage.
● Treatment: ○ Cysts pass through the stomach and excyst in
○ Treatment with iodoquinol or metronidazole the duodenum.
○ Trophozoites attach to the duodenum mucosa. ü
○ Encystation occurs in the large intestines, and
the cysts will pass in the stool
● Morphology:
○ Cyst characteristics:
฀ G. Lamblia cysts are oval-shaped, and the
average size ranges from 12 um to 8 um
wide
฀ Cysts contain 4 nuclei with no peripheral
chromatin
฀ Cytoplasm is retracted from the cyst wall
and may contain 2 to 4 comma-shaped,
median bodies

FLAGELLATES
● General characteristics:
○ Flagellates are a subclass of protozoa that have
one or more flagellum that provide motility.
○ All flagellates have a trophozoite stage, but
several lack the cyst stage.
○ Many flagellates live in the small intestines.
○ Giardia lamblia is the only pathogenic flagellate;
it causes mild to moderate diarrhea. Severe
infections can lead to malabsorption.
● Diagnosis:

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○ Trophozoite characteristics:
฀ G. lamblia trophozoites have an average
size of 15 um wide
฀ Motile, pear-shaped with bilateral
symmetry and two large nuclei on each
side of a central axostyle
฀ “OLD MAN with EYEGLASSES”
฀ Contain 2 oval-shaped nuclei without
peripheral chromatin
฀ Possess 4 pairs of flagella
฀ 2 median bodies, 2 axonemes, and a
sucking disk are present
฀ Diagnoses:
● Microscopic examination of stool
samples for trophozoites and cysts ü
● Other diagnostic tests (EnteroTest®)
and antigen detection immunologic
assays
● Lab diagnosis:
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
○ Examination of diarrheal stool- trophozoite or
cyst, or both may be recovered in wet
preparation.
○ In examinations of formed stool (e.g. in
asymptomatic carriers) only cysts are seen.
○ Giardia species may occur in “showers”, i.e.
many organisms may be present in the stool on
a given day and few or none may be detected
the next day.
○ Therefore, one stool specimen per day for 3
days is important
● Treatment:
○ For asymptomatic carriers and diseased
patients, the drug of choice is quinacrine
hydrochloride or metronidazole
● Prevention:
○ Asymptomatic reservoirs of infection should be
identified & treated.
○ Avoidance of contaminated food and water.
○ Drinking water from lakesand streams should be
boiled, filtered and/or iodine treated.
○ Proper waste disposal and use of latrine
Chilomastix masnili
● Notes:
○ MOT: contaminated food or water containing
the cyst stage (infective)
○ Generally nonpathogenic but has been
associated with disease in immunosuppressed
patients
● Diagnoses:
○ Microscopic examination of stool sample
● Cyst characteristics:
○ The cyst ranges in size from 5 to 10 um in length
and is oval-shaped
○ C. mesnili contains a single nucleus without
peripheral chromatin
○ The karyosome is large and centrally located
○ The cytostome is well defined
● Trophozoite characteristics:
○ Size ranges from 5 to 25 um in length and 5 to
10 um in width, they are pear-shaped and
motile
○ Single nucleus without peripheral chromatin
○ Karyosome: eccentric and small
○ Flagella: Three anterior and one posterior
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○ Cytostome is very large and a spiral groove is ● Prevention:

present ○ The reservoir for this flagellate and lifecycle is
○ It is considered to be a non-pathogenic, and no unknown. Thus, specific recommendation for
treatment is recommended prevention is difficult. However, infection can be
avoided by maintenance of adequate sanitary
conditions.

CILIATES
● General characteristics:
○ Motile by cilia
○ Trophozoites and cysts are part of the life cycle
○ Balantidium coli is the only species pathogenic
for human

Dientamoeba fragilis

● Notes:
○ Many cases of diarrhea caused by D. fragilis
occur in individuals living in close quarters such
as inmates, college students, and military
recruits
○ Infects the mucosal lining of the large intestines. Balantidium coli
○ There is no cyst stage, and the life cycle is not ● MOT:
well-defined ○ Contaminated (feces) or food
● Infections associated: ● Causes Balantidiasis, characterized by diarrhea to
○ Diarrhea dysentery
○ Abdominal pain ● Diagnosis:
○ Anal pruritus (itching) ● Microscopic examination of stool for cysts or
● Diagnoses: trophozoites
○ Microscopic examination of trophozoites in the ● Morphology:
stool ○ Cyst characteristics:
● Trophozoite characteristics: ฀ Ranges in size from 43 to 65 um and is
○ Size ranges from 5 to 19 um; they are motile by round in shape
means of hyaline pseudopods and are ฀ Contains 2 nuclei; one, the macronucleus
round-shaped is kidney -shaped and very large, the
○ Most cells contain two nuclei without peripheral micronucleus is round and much smaller
chromatin but with clumps of nuclear chromatin (rarely seen)
○ The cytoplasm is vacuolated with bacterial ฀ Has a double cyst cell wall with numerous
inclusions cilia between the two cell walls
● Treatment:
○ The therapeutic agent of choice for this
infection is iodoquinol, with tetracycline and
parmomycine as acceptable alternatives

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○ Oval oocyst ranges in size from 4 to 6um

○ Oocysts contain 4 sporozoites enclosed within a
thick cell wall
○ The cytoplasm may contain several dark
granules
● Laboratory diagnosis:
○ Diagnosis is made by finding oocysts in fecal
smears when using a modified Kinyoun acid–
fast stain.
○ Serological tests are not available.
● THERE IS NO EFFECTIVE DRUG THERAPY

○ Trophozoites characteristics:
฀ Range in size up to 100 um in length and
70 um in width
฀ Like the cyst, trophozoites contain 2 nuclei
฀ Has one or two contractile vacuoles with
cilia around the cell

Cyclospora cayetanensis
● MOT:
○ Through ingestion of basils, snowpeas, mesclun,
strawberries
● Humans are the only host for C. cayetanensis
● Infections caused
○ Nonbloody diarrhea
● Diagnosis:
○ Examination of stained fecal smears; the oocysts
will stain with the modified Kinyoun’s acid-fast
stain and are 8 to 10 um in diameter
● Treatment:
○ The treatment of choice is trimethoprim
● Treatment: sulfamethoxazole.
○ The drug of choice is tetracycline; iodoquinol
and metronidazole are alternative agents.

B. coli: fig. a. trophozoite, fig. b. cyst

INTESTINAL SPOROZOA

Cryptosporidium parvum
● MOT:
○ Through contaminated food or water (rodent,
cow, pig, or chicken feces) of infected oocyst
● Infections caused:
○ Cryptosporidiosis, moderate to severe diarrhea
○ In patients with AIDS, Cryptosporidium
infections are an important cause of death due
to dehydration
● Oocyst characteristics:

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Isospora belli [Cytoisospora belli appears on the web]

● MOT:
○ Ingestion of the infective oocyst in
contaminated food and water
● Humans are the definitive host; there are no
intermediate hosts
● Infections caused:
○ Isosporiasis, characterized by mild diarrhea to
severe dysentery
● Oocyst characteristics:
○ The oval oocyst ranges in size from 25 to 40 um
in length
○ The cytoplasm is granular and contains two
sporoblasts that contain 4 sporozoites each
● Laboratory diagnosis:
○ Diagnosis is made by finding the typical oocysts
in fecal specimens. Serologic tests are not
available.
● Treatment: Plasmodium falciparum (P. falciparum)
○ The treatment of choice is
trimethoprim-sulfamethoxazole
Table No. Malignant Tertian Malaria
NOTES INFORMATION
Cytoplasm: small, fine, pale blue ring
Young Chromatin: 1-2 small red dots, Applique
trophozoite or Acole form
Size: 1/5 to 1/3 diameter of RBC
Cytoplasm: thing, blue ring or
Mature
comma-shape
trophozoite
Chromatin: 1 or 2 medium sized red dots
Merozoites: 8-24
Schizont
Dark brownish-black pigment
Shape: banana/sickle/sausage/crescent
Color: blue (male), dense blue (female)
Gametocyte Nucleus: reddish pink
Pigment: few blue black granules in the
center of the cytoplasm or scattered
RBC Normal in size
Pigments Maurer’s dots & Christopher’s dots
Stages in
peripheral Ring forms and gametocytes
blood
Asexual cycle 48 hours
● Treatment:
○ Because chloroquine-resistant stains of P.
falciparum are present in many parts of the
world, infection of P. falciparum may be treated
with other agents including mefloquine,
quinine, guanidine, pyrimethamine-sulfadoxine,
and doxycycline

EXTRAINTESTINAL PROTOZOA
Plasmodium

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Plasmodium vivax (P. vivax) Pigment: Large black granules in the

cytoplasm
RBC Normal
Table No. Benign Tertian Malaria Pigments Ziemann’s dots
NOTES INFORMATION Stages in
Cytoplasm: irregular blue, thick ring peripheral All
Young
Chromatin: large red dots blood
trophozoite
Size: 1/4 to 2/3 diameter of RBC Asexual cycle 72 hours
Mature Cytoplasm: large, blue, irregular, small ● Treatment:
trophozoite particles of brownish orange pigment ○ The treatment regimen, including the use of
Merozoites: 12-24 primaquine to prevent relapse from latent liver
Schizont Large, compact red granules against the stages is similar to that used for P. vivax
pale blue cytoplasm infection
Female: Oval or rounded, dense blue
with dense red triangular nucleus; many
particles of orange pigment in cytoplasm
Gametocyte
Male: Rounded, pale blue with a round
central pale nucleus; some particles of
orange pigment in cytoplasm
RBC Enlarged, pale
Pigments Schuffner’s dots
Stages in
peripheral All
blood
Asexual cycle 48 hours Plasmodium malariae (P. malariae)
● Treatment:
○ Chloroquine is the drug of choice for the
suppression and therapeutic treatment of Table No. Quartan
P.vivax, followed by primaquine for radical cure NOTES INFORMATION
and elimination of gamatocytes Cytoplasm: thick, dense blue ring with
Young some granules of black pigments
trophozoite Chromatin: 1 large red dot Size: 1/4 to
2/3 diameter of RBC
Cytoplasm: either band form or round,
Mature compact, dark blue with many black
trophozoite particles of pigment Chromatin: a round
dot or a red band
Merozoites: 8-10 each one a large red
Schizont
spot enclosed by pale cytoplasm
Shape: Large, oval, or round
Color: male (pale blue), female (dense
blue)
Gametocyte
Nucleus: 1 round spot of red chromatin
Plasmodium ovale (P. Ovale) Pigment: Large black granules in the
cytoplasm
RBC Normal
Table No. Pigments Ziemann’s dots
NOTES INFORMATION Stages in
Cytoplasm: regular, dense blue ring peripheral All
Young
Chromatin: medium-sized red dots blood
trophozoite
Size: 1/4 to 2/3 diameter of RBC Asexual cycle 72 hours
Cytoplasm: either band form or round, ● Treatment:
Mature compact, dark blue with many black ○ Chloroquine is also the drug of choice for the
trophozoite particles of pigment suppression and therapeutic treatment of P.
Chromatin: a round dot or a red band malariae, followed by primaquine for radical
Merozoites: 8-10 each one a large red cure and elimination of gamatocytes
Schizont
spot enclosed by pale cytoplasm
Shape: Large, oval, or round
Color: male (pale blue), female (dense
Gametocyte
blue)
Nucleus: 1 round spot of red chromatin

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Babesia microti
● Notes:
○ B. microti is an erythrocytic intracellular
parasite, like Plasmodium spp., that can also
cause hemolytic anemia. Babesiosis is a
self-limiting infection; death is a rare outcome.
○ B. microti causes babesiosis, which can affect
the spleen, liver, and kidneys.
○ The infective sporozoite is transmitted to
humans by a tick bite (Ixodes scapularis).
● Diagnosis:
○ Made by blood smear examination and
serologic testing. It is difficult to differentiate
Babesia spp. from Plasmodium spp.
● Ring form characteristics:
○ Size ranges from 3 to 5 um.
○ Cytoplasm: Minimal with two or more
chromatin dots. Two to four rings per RBC are
often seen, sometimes appearing like a
"Maltese cross."
Naegleria fowleri
Toxoplasma gondii
● Infections caused:
○ Toxoplasmosis is characterized by a broad
spectrum of symptoms depending on the
individual's state of health. T. gondii has a
predilection for central nervous system (CNS)
infections.
➢ In healthy individuals, toxoplasmosis often
resembles infectious mononucleosis and
produces fatigue, swollen lymph glands,
fever, and myalgia. The disease can
become chronic and affect the heart and
liver.
CONCEPCION, DIMAPANAT, GARCIA, HERNANDEZ, VICEDO | 2H PH
○ Congenital toxoplasmosis occurs in premature
or antibody-deficient infants, where symptoms
include splenomegaly, jaundice, and fever
● Mode of Transmission:
○ The sexual stage of reproduction occurs in the
intestinal tract of house cats. The infective form
(oocysts) of the parasite is passed in the stool,
and the ingestion of cat feces-contaminated
food and water can produce infection.
○ Ingestion of undercooked meat (lamb and pork)
containing viable tissue cysts
○ Transplacental transmission from the infected
mother to the fetus
● Morphology:
○ Because tachyzoites and bradyzoites are small
and no single organ is typically involved, it is
difficult to diagnose infection by microscopic
examination of tissue samples.
○ Tachyzoites (trophozoites) range in size from 1
to 3 um and are crescent to round in shape.
○ Cysts contain many bradyzoites.
● Diagnosis:
○ Serologic testing for Toxoplasma antibody
Toxoplasma gondii
○ Microscopic examination of Giemsa–stained
preparations shows crescent–shaped
trophozoite. Cysts may be seen in the tissue.
● Treatment:
○ Treatment is with a combination of sulfadiazine
and pyrimethamine.
● Notes:
○ Causes amebic meningoencephalitis, which is
often fatal within 3-6 days
○ N. fowleri is found in lakes, ponds, and
swimming pools where the water is warm.
○ Life cycle: Trophozoites are the infective stage.
○ The amebae are contracted from contaminated
water, where trophozoites enter the body
through the nasal mucosa and migrate along the
olfactory nerve to the brain
● Morphology:
○ Cyst characteristics
➢ The round cyst ranges in size from 10 to 13
um.
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➢ N. fowleri cysts contain a single nucleus,

without peripheral chromatin.
➢ The karyosome is centrally located and
large.
○ Trophozoite characteristics
➢ Size ranges from 10 to 23 um, and they are
motile by means of blunt pseudopods.
➢ Trophozoites contain a single nucleus,
without peripheral chromatin.
➢ The cytoplasm is granular and vacuolated.
● Flagellate characteristics:
○ Flagellates range in size from 7 to 15 um and are
pear shaped. They are motile by means of two
flagella.
○ The single nucleus is indented.
○ Karyosome is centrally located and large.
○ The cytoplasm is granular and vacuolated
● Diagnosis:
○ Made by finding the organism (ameba) in
cerebrospinal fluid (CSF) or brain biopsies.
● Treatment of free-living amoebic infections is largely
ineffective.
Pathogenic Free-Living Amebae

Naegleria fowleri

● Primary amebic meningoencephalitis (PAM)

Acanthamoeba spp.
● Granulomatous amebic encephalitis (GAE)
● Granulomatous skin and lung lesions (primarily
immunocompromised)
● Amebic keratitis

Balamuthia mandrillaris
● GAE + granulomatous skin and lung lesions (primarily
healthy)

Acanthamoeba
● Extraintestinal protozoa
● Causes amebic encephalitis and amebic keratitis
(cornea infection)
● The life cycle is not well characterized.
● The eye is directly invaded by trophozoites, producing
keratitis.
● Skin, respiratory tract, and CNS infections are caused by
the cyst or trophozoite stage (unknown entry route).
● Treatment of free-living amoebic infections is largely
ineffective.
● Diagnosis is made by finding the cyst or trophozoite
stages in CSF.
Trichomonas vaginalis
● Causes vaginitis in women, whereas men are generally
asymptomatic carriers
● T. vaginalis is a sexually transmitted disease and can
infect neonates (aspiration pneumonia) during delivery.
Trophozoites are the infective stage and infect the
epithelial or mucosal lining of the vagina, urethra, and
prostate gland.
● T. vaginalis does not have a cyst stage.
● Diagnosis: Trophozoites are usually detected during a
microscopic urinalysis.
● Treatment: Metronidazole is the drug of choice. If
resistant cases occur, re-treatment with higher doses is
required
Trophozoite characteristics
● Trophozoites average about 30 um in length. They are
motile, with an undulating membrane, and are pear
shaped.
● Single prominent nucleus
● Flagella: Three to five anterior and one posterior
● Large axostyle with cytoplasmic granules

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Laboratory diagnosis ● Pathogenic species

● In females, T. vaginalis may be found in urine sediment, ● Xenodiagnosis - which consists of allowing an
wet preparations of vaginal secretions or vaginal uninfected, laboratory-raised reduviid bug to feed on the
scrapings. patient and, after several weeks, examining the
● In males it may be found in urine, wet preparations of intestinal contents of the bug for the organism.
prostatic secretions or following massage of the prostate
gland.
● Contamination of the specimen with feces may confuse
T.vaginalis with T.hominis

Trypanosoma brucei

● T. brucei causes African trypanosomiasis or sleeping

Extraintestinal Protozoa: HEMOFLAGELLATES
● Hemoflagellates inhabit the blood and tissues of
humans.
● Four stages of development: amastigote, promastigote,
epimastigote, and trypomastigote
○ Amastigote: Non Flagellated oval form, found
in tissue
○ Promastigote: Flagellated stage found in the
vector, rarely seen in the blood
○ Epimastigote: Long, slender flagellated form
found in arthropod vectors
○ Trypomastigote: Has an undulating membrane
running the length of the body; found both in
the vector and bloodstream of humans
● Transmission to humans is by arthropod bites
sickness, and infection affects the lymphatic system and
CNS.
● Swollen lymph nodes at the posterior base of the neck
(Winterbottom's sign) are sometimes present.
● Subspecies gambiense and rhodesiense are named
according to their geographic location.
● For the acute stages of the disease the drug of choice is
suramin with pentamidine as an alternative.
● In chronic disease with CNS involvement, the drug of
choice is melarsoprol.
● Alternatives include tryparsamide combined with
suramin.
Trypanosoma cruzi
● Causes Chagas disease or American trypanosomiasis,
which is characterized by lesion formation (chagoma),
conjunctivitis, edema of the face and legs, and heart
muscle involvement leading to myocarditis.
● Mostly found in South
● The drug of choice is nifurtimox.
● Alternative agents include allopurinol & benzimidazole

Trypanosoma

● Diagnosis is made by microscopic examination of blood

and CSF and serologic testing.
● Trypanosomiasis occurs mainly in Africa and South Leishmania
America.

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sticky side of the tape is pressed to

● Human leishmaniasis
● Cutaneous leishmaniasis is characterized by skin and
mucous membrane ulcers.
● Disseminated leishmaniasis: Liver, spleen, and
reticuloendothelial involvement
● Diagnosis: Finding the amastigote in the blood or tissue
and serologic testing
● Mainly a disease of Africa, Eastern Europe, and
South/Central America
● The drug of choice is sodium stibogluconate, a
pentavalent antimonial compound. Alternative
approaches include the addition of allopurinol and the
use of pentamidine or amphotericin B.
the perianal skin.
■ Because the gravid female deposits
eggs in the perianal folds during the
late evening hours, yield is best in
children if the specimen is collected
in the morning
● Egg morphology
○ Eggs range in size from 50 to 60 um in length
and 30 um in width. They are oval in shape,
and one side is flat
○ The shell is thick and double walled
○ The ovum contains a developing embryo

Nematodes: Roundworms
● General characteristics:
○ Nematodes (roundworms) are a class of
helminths
● Diagnosis: based on adult, larvae, or egg morphology
● MOT: ingestion of eggs, transmitted by larvae which gain
entry through the skin to the intestines
● Adult nematodes have a tapered, cylindrical body with
an esophagus and longitudinal muscles
● Nematodes cause diseases associated with the
intestines and the skin, including diarrhea, vomiting, and
skin lesions
● Nematodes are placed into one of two groups: the
intestinal nematodes and intestinal-tissue nematodes.
○ Intestinal nematodes
■ Enterobius vermicularis
■ Trichuris thrichiura
■ Ascaris lumbricoides
■ Strongyloides stercoralis
■ Necator americanus
■ Ancylostoma duodenale
○ Intestinal-tissue nematodes
■ Trichinella spiralis
■ Dracunculus medinensis

Enterobius vermicularis

● AKA: pinworm, seatworm

● Infections: enterobiasis (itching and inflammation of the Trichuris trichiura
anus)
● Common especially in school-age children ● AKA: whipworm
● Diagnosis: ● Infections: RECTAL PROLAPSE, colitis, abdominal pain
○ SCOTCH-TAPE METHOD and bloody diarrhea Notes: Eggs from human feces are
■ Based on the identification of ova infective. After ingestion of the ova, the larvae develop
from a tape preparation in which the into adults in the intestines
● Egg morphology

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○ Eggs range in size from 50 to 60 um in length
and 25 um in width and are football-shaped
with clear plugs at each end
○ shell is smooth and yellow to brown
○ Egg contains a developing embryo
● Adult
morphology
○ Infertile ova are oval and measure up to 90 um
in length.
○ Fertile eggs are round and range in size up to
75 um in length and 50 um in width. The shell
is thick and contains a developing embryo

○ The female adult measures 35-50 mm long,

and the male is 30-45 mm long. Adults are
● Adult morphology
rarely found in the stool
○ Females are 20-30 cm long
○ The anterior end resembles a whip handle
○ Males are 15-31 cm long with a curved
○ The posterior end resembles a whip and is gray
posterior end
in color

Strongyloides stercoralis
Ascaris lumbricoides
● AKA: threadworm
● AKA: giant intestinal roundworm Infection: Ascariasis
● MOT: skin contact with contaminated soil (filariform
(bowel obstruction)
larvae)
● can also migrate to the lungs where they cause
● After penetration, larvae migrate through the skin and
pulmonary disorders and to other body sites
repeated exposure to larvae can produce an allergic
● Eggs are passed in human feces Diagnosis: Examination
reaction commonly referred to as larva migrans
of stool for eggs and adult forms
● Diagnosis: Stool examination for rhabditiform larvae
● Egg morphology
● Morphology of rhabditiform larvae

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○ Larvae range in size up to 700 um in length ○ The buccal cavity of N. americanus contains a
○ The tail is notched pair of cutting plates; whereas A. duodenale
has teeth
adult

egg

Hookworms
● Necator americanus (New World hookworm) and
Ancylostoma duodenale (Old World hookworm) cause
human hookworm infections.
● MOT: Humans acquire the filariform larvae through skin
penetration
● The larvae migrate to the lungs and then to the
intestines
● Diagnosis: Identification of larvae in sputum or ova from
stool specimens
● Egg morphology
○ Eggs of two species are identical
○ Eggs are oval and measure 56-75 um long by
36-40 um wide
○ Eggs are thin-shelled and a developing embryo
can sometimes be seen inside
● Adult morphology
Trichinella spiralis
○ Adult worms firmly attach to the intestinal
mucosa and are rarely seen ● Parasite is found worldwide; pigs are important
○ Adults range in size from 7 to 11 mm in length; reservoirs
Ancylostoma worms tend to be slightly larger ● MOT: eating contaminated undercooked pork
than Necator worms ● Infection: Trichinosis
○ (symptoms: diarrhea, blurred vision, muscle
edema, and coughing)
● Diagnosis: examining muscle tissue for encysted larvae

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● Lab tests reveal eosinophilia, leukocytosis, and increased

creatine kinase and lactate dehydrogenase
● Morphology
○ Larvae range in size up to 125 um in length
and 7 um in width. Many encysted forms are
found in striated muscle tissue
○ Adults range up to 1 mm in length and reside
in the intestinal tract

Filariae

● General Characteristics of the Filarial Parasites

Dracunculus medinensis
● Causes guinea worm infection; symptoms include
allergic reactions and painful ulcers
● Most infections occur in Africa, India, and Asia.
● Humans acquire the infection by the ingestion of
infected copepods (water fleas) carrying the larvae.
● Diagnosis: Observing worms emerging from ulcerated
areas of the body
● Morphology of adults
○ Adult females range in size up to 1 m in length
and 2 mm in width.
○ Males are about 2 cm in length
● Filarial parasites are an order of nematodes consisting of
adult threadlike worms.
● Filariae inhabit the circulatory and lymphatic system and
are also found in muscle, connective tissue, and serous
cavities.
● Four primary species cause disease in humans:
○ Wuchereria bancrofti
○ Brugia malayi
○ Loa loa
○ Onchocerca volvulus.
● Transmission of the filariae parasites occurs after the
bite of mosquitoes or other arthropods.
● Diagnosis is by microscopic examination of blood or
tissue for microfilariae. Some species migrate in the
daytime, whereas others migrate at night (nocturnal
periodicity). It is therefore important to draw blood in
the morning (between 10 A.M. and noon) and in the
evening (between 10 P.M. and midnight).
● Most filarial disease occurs in Africa and South America

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Loa loa
● MOT: through the bite of the Chrysops (deer) fly
● Causes subcutaneous tissue infections and infections of
the conjunctival lining of the eye
● Most infections occur in Africa.
● Diagnosis: Blood smear examination for microfilariae
● Microfilariae characteristics
○ Size ranges from 250 to 300 um in length.
○ Contains a row of nuclei that extends to the
tail of the parasite
○ A sheath is present

Wuchereria bancrofti
● causes Bancroftian filariasis, a condition that produces
lesions in the lymphatics. Elephantiasis may result.
● Disease occurs in the tropics and subtropics.
● Vectors for the disease are the Culex, Aedes, and
Anopheles mosquitoes.
Brugia malayi ● Diagnosis: Examination of blood smears for microfilariae
● Microfilariae characteristics
● MOT: transmitted by the Anopheles or Aedes mosquito.
○ Ranges in size from 250 to 300 um in length
● B. malayi causes Malayan filariasis, a condition that
○ No nuclei are found in the tail.
produces lesions in the lymphatics, and elephantiasis
○ A sheath is present.
may result.
● Most infections occur in the Far East, Japan, and China.
● Diagnosis is by microscopic examination of blood smears
for microfilariae.
● Samples collected at night offer the largest yield.
● Microfilariae characteristics
○ Size ranges from 200 to 300 um in length
○ Two nuclei are located in the tail
○ A sheath is present.

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Cestodes: Tapeworms

Onchocerca volvulus ● Class Cestoda: General Characteristics

● O. volvulus causes river blindness; eye infections may ● consist of a rounded head, called a scolex, and long
lead to blindness. strobila or chain of proglottids (multiple segments) of
● Most infections occur in Africa, South America, and varying stages of maturity.
Mexico.
● Transmission of the infective microfilariae is by the bite
of the Simulium (blackfly).
● Diagnosis: Tissue or ophthalmologic analysis for
microfilariae
● Microfilariae characteristics
○ Size ranges from 150 to 360 um in length.
○ No nuclei are located in the tail.
○ A sheath is present

● They have no digestive tract of its own at any point in its

life cycle.
● The scolex has specialized means of attaching to the
intestinal wall, namely suckers, hooks, or sucking
grooves.
● All species are parasitic during both the adult and larval
developmental stages.
● All cestodes have stages of oncosphere in the life cycle.

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● In humans the tapeworm can reach a length of 10

● Scolex (pl: Scolices)
○ Suckers, Hooks and Bothria (with slit-like
groove with weak suction powers and usually
two in number)
● Neck
○ Undifferentiated stem cells that give rise to
proglottids in strobila.
● Unique structure: Proglottids
○ Contains both male and female organs.
○ Essentially a whole reproductive package in
one segment of the strobila.
● Integument
○ Microtriches (like Microvilli of vertebrate small
intestine), on surface of proglottid;
○ Absorb nutrients.
meters (>30 feet) (it’s the longest of the tapeworms) and
produce over a million eggs per day!
● Disease: diphyllobothriasis
● Distribution: Scandinavia, northern Russia, Japan,
Canada, USA.
● Morphology:
○ Diphyllobothrium latum can be indentified by
its scolex with 2 elongated sucking grooves by
which the worm attaches to the intestinal wall.
○ The proglottids are wider than they are long,
and the gravid uterus is in the form of a
rosette.
○ Larva is called plerocercoid .
● Life cycle of Diphyllobothrium latum
○ Definitive hosts: humans Intermediate hosts:
■ copepod crustacea
■ freshwater fish.
○ Humans are infected by eating raw or
under-cooked fish containing plerocercoids.
○ In the small intestine, the larvae attach to the
gut wall and develop into adult worms.
○ Gravid proglottids release fertilized eggs.
○ The immature eggs must be deposited in fresh
water for the life cycle to continue.
● Transmission: fecal-oral Invasive stage: plerocercoid
● Clinical disease:
○ infection by D. latum causes little damage in
the small intestine.
○ In some individuals, megaloblastic anemia
occurs as a result of vitamin B12 deficiency
caused by preferential uptake of the
asymptomatic, vitamin by the worm.
○ Most but abdominal patients are discomfort
and diarrhea can occur.
● Diagnosis depends on finding the typical eggs, oval,
yellow-brown eggs with an operculum (lidlike opening)
at one end, in the stools.
● Prevention involves adequate cooking of fish and proper
disposal of human feces.

Diphyllobothrium latum
● the fish tapeworm
● Important parasites of man.
Taenia solium
● Definitive hosts can be humans, dogs, foxes, cats, mink,
● the pork tapeworm
bears, and seals.
● Larvae cause cysticercosis.
● Localisation: small intestine.
● Distribution: worldwide, but is endemic in areas of Asia,
South America, Eastern Europe.

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● Morphology: can be indentified by its scolex with 4
suckers and circle of hooks and by its gravid proglottids,
which have 7-12 primary uterine branches.
● Larva is called cysticercus. A cysticercus consists of a pea
sized fluid-filled bladder with an invaginated scolex.
● Life cycle of Taenia solium
○ Transmission: fecal-oral;
○ Definitive host: humans; Intermediate host:
pigs;
○ Humans can be infected by eating raw or
undercooked pork containing the larvae
cysticercus.
○ In the small intestine, the larvae attach to the
gut wall and take about 3 months to grow into
adult worms.
○ The gravid terminal proglottids detach daily,
are passed in the feces.
● When in the brain, it may cause severe central nervous
system dysfunction. - Cerebral cysticercosis
○ Most common and distinct symptom is sudden
onset epilepsy.
○ Brain imaging can now spot cysticercus in the
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Taenia saginata
● beef tapeworm
● Most common tapeworm in humans.
● Morphology: large species reaching up to 20 m can be
indentified by its scolex with 4 suckers without hooklets.
Its gravid proglottids have 17-35 primary uterine
branches. No hooks on scolex.
● Causes taeniasis saginata.
● Distribution: worldwide but is endemic in areas of Asia,
South America, Eastern Europe.
● Larva of T. saginata is called cysticercus..
● Life cycle of Taenia saginata
○ Definitive host: humans
■ Intermediate host: cattle
○ Humans can be infected by eating raw or
undercooked beef containing larvae.
○ In the small intestine, the larvae attach to the
gut wall. The gravid terminal proglottids
detach, are passed in the feces, and are eaten
by cattle.
○ Laboratory diagnosis: gravid proglottids (with
17-35 uterine branches) may be found in the
stools.
○ Prevention of taeniasis saginata involves
cooking beef adequately and preventing cattle
from ingesting human feces by disposing of
waste properly
● do not cause a serious disease.
● Usually asymptomatic but may cause dizziness,
abdominal pain, diarrhea, headache and nausea.
● Proglottids are obvious in feces.
● Clinical manifestation of teniasis soleum and teniasis
saginata: abdominal pain, nausea, diarrhea, weight loss,
infection may be asymptomatic
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Echinococcus granulosus
Hymenolepis nana
● dwarf tapeworm
● Distribution: worldwide, commonly in the tropics.
● Morphology: it is only 2-3 cm in length.
○ Scolex has round form and contains suckers
and hooks.
○ The neck is very long and thick.
○ Strobila has 200 proglottides.
○ The uterus has an excretory ostium. Eggs are
released from it into the feces.
● Transmission: fecal-oral (by the ingestion of eggs from
contaminated food or water).
● Invasive stage: egg
● Clinical disease: asymptomatic, but diarrhea and
abdominal cramps may be present.
● Diagnosis can be proved by observing eggs in stool.
● Prevention consists of good personal hygiene and
avoidance of fecal contamination of food and water.
● Distribution: founded primarily in shepherds living in the
Mediterranean region, the Middle East, and Australian,
USA (western states).
● Morphology: worms are up to 3-5 mm.
○ Scolex has suckers and hooks.
○ The neck is short.
○ Strobila has 3-5 proglottides.
○ Posterior segment (mature) is the largest and
contains the uterus with the haustrums,
genital pore situated in the back of the
proglottid.
● Life cycle of Echinococcus granulosus
○ Transmission: fecal-oral.
○ Invasive stage: egg.
○ Definitive hosts: dogs, other carnivorous
mammals.
○ intermediate hosts: sheep, other herbivorous
mammals, humans.
○ Worms in the dog’s intestine liberate
thousands of eggs, which are ingested by
sheep (or humans).
● The oncosphere embryos emerge in the small intestine
and migrate to the liver also to the lungs, bones, and
brain. The embryos develop into large fluid filled hydatid
cysts.
● The life cycle is completed when the entails of
slaughtered sheep are eaten by dogs.
● Diagnosis: made by clinical manifestations.
○ Asymptomatic, but liver cysts may cause
hepatic dysfunction
○ Cysts in the lungs can erode into a bronchus,
causing bloody sputum, end cerebral cysts can
cause headache and focal neurologic signs.
● Diagnosis: made by routine X - ray, observation of
eosinophilia, serologic tests.
● Prevention of human disease involves not feeding the
entrails of slaughtered sheep to dogs.

Echinococcus
multilocularis
● alveolar
hydatid tapeworm
or the small fox
tapeworm

● Distribution: Northern Europe, Siberia, Canada, the USA.

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● Morphology: a small tapeworm that is 3- 6 mm long. The

segmented worm contains a scolex with suckers and
hooks.
● Localisation in a definitive host: small intestine.
● Many of the features of this organism are the same as
those of E. granulosus, but the definitive hosts are
mainly foxes and the intermediate hosts are various
rodents.
● Humans are infected by accidental ingestion of food
contaminated with fox faeces.
● Life cycle of Echinococcus multilocularis
○ Definitive hosts: foxes, wolves, jackals,
coyotes, dogs.
○ Intermediate hosts: various rodents.
● Invasive stage for humans: egg.
● Transmission: fecal-oral.
● Disease occurs primarily in hunters and trappers. Within Schistosoma japonicum
the human liver, the larvae form multiloculated cysts
with few protoscoleces. ● Life cycle involves alternating parasitic stages in
● The cysts continue to proliferate, producing a mammalian hosts and free living stages
honeycomb effect of hundreds of small vesicles (without ○ Egg and miracidium
fluid). ○ First stage (mother) sporocyst
● The clinical picture usually involves jaundice and weight ○ Second stage (daughter) sporocyst
loss. ○ cercaria
● Diagnosis: serological and imaging tests. ○ Schistosomulum
○ Adult schistosome
● Primarily parasites of the portal vein and its branches
● Each female fluke deposits 500-2000 immature eggs/day
● Embryonation takes place within 10-12 days
● Eggs escape through ulcerations in the intestinal lumen
and are passed out with the feces
● Schistosoma japonicum in eternal copula
○ Males have a gynecophoral canal which
receives the female during copulation

Trematode: Flukes
● BLOOD FLUKES
● Schistosomes
○ Schistosoma japonicum : Oriental blood fluke
○ Schistosoma haematobium: Vesical blood
fLuke
○ Schistosoma mansoni: Manson’s blood fluke

Schistosoma mansoni
● Schistosoma mansoni male and female
○ Female inside the gynecophoral canal of
male

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● Distended belly is one of the symptoms of

Schistosomiasis

Blood flukes: Diagnosis

● Schistosomiasis
○ Eggs may not be demonstrable in the feces
○ Infections where there is scarring prevent
passage of eggs into the intestinal lumen
○ Stool examination technique
Schistosoma Haematobium ■ Merthiolate-iodine formalin
● Schistosoma Haematobium adult concentration technique (MIFC)
● Sensitive for moderate and
heavy infections
● Not adequate for light
infections (less than 10
eggs/ grams of stool)
■ Kato Katz Technique
● For enumeration of eggs
● Most commonly used for
evaluating epidemiology,
effect of control measures,
drug trials
■ Immunodiagnosis
● Intradermal tests for
● Schistosoma Haematobium ova immediate cutaneous
○ Note the presence of terminal spine hypersensitivity using adult
worm extracts
● Indirect hemagglutination
using adult worm and egg
antigens
● Circumoval precipitin test
● Enzyme-Linked
Immunosorbent Assay
(ELISA) using soluble
antigens of adults and eggs

Blood flukes: Treatment

● Praziquantel (heterocyclic prazinoisaquinolone
compound)
● Single dose of 40- 50 mg/kg
● 25 mg/kg in two doses
● 20 mg/kg in three doses
Blood flukes: Pathogenesis and clinical manifestations
● Schistosomiasis Lung flukes
● Host granulomatous reaction to eggs ● Paragonimus westermani
● Pneumonitis due to schistosomula in the lungs ● Oriental Lung Fluke
● Hepatosplenic disease ● Paragonimus philippinensis
● Colonic schistosomiasis ● Paragonimus siamensis (cats)
● Cerebral schistosomiasis
Lung flukes: Paragonimus westermani
Blood flukes: Schistosomiasis ● Paragonimus westermani adult
○ Hermaphroditic

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○ Body covered with spines

○ Reddish brown
○ Measures 4-6 mm in width and 3.5-5 mm in
thickness
○ Resembles a coffee bean
○ Adult worms are found in pairs or in threes in
fibrotic capsules or cysts in the lungs

Lung Flukes: Epidemiology of Paragonimiasis

● First Intermediate Host
○ Brotia asperata (snail)
○ Where miracidium develops into 1 sporocyst
and 2 redial stages of development
● Second Intermediate Host
○ Sundathelpusa philippina or Parathelpusa
grapsoides (former name)
○ Harbors the metacercaria that is infective to
man

Lung Flukes: Paragonimus westermani

● Man gets infected after ingestion of raw or insufficiently
cooked crabs harboring the metacercariae

● Paragonimus westermani ova

○ Yellowish brown
○ Thick-shelled
○ Operculated with a thickened abopercular egg
○ May be seen in the sputum or in feces if the
sputum is swallowed

Lung Flukes: Pathogenesis and clinical manifestations

● Paragonimiasis
○ Cough
○ Hemoptysis
○ Symptoms consistent with pulmonary
tuberculosis
○ Misdiagnosed as PTB

Lung Flukes: Paragonimus westermani life cycle

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● It becomes sexually mature in the bile ducts

Liver Flukes Fasciola Hepatica

● Adult worm
○ Large, broad, flat body
○ Anterior end forms a prominent cephalic cone
○ Small oral and ventral suckers
○ Long and highly branched intestinal caeca

Lung Flukes: Diagnosis of Paragonimiasis

● Radiographs aid in diagnosis
● Definitive diagnosis is based on the finding of ova in the
sputum, stool or less frequently in aspirated material
from abscesses or pleural effusions
● Multi-dot ELISA

Liver Flukes Fasciola Gigantica

● Adult worm
○ Larger
○ More lanceolate
○ Less developed shoulders (shorter cephalic
cone)
○ Larger ventral sucker

Lung Flukes:Treatment of Paragonimiasis

● Praziquantel
○ Drug of choice
○ 25 mg/kg body weight 3x a day for three day
● Bithionol
○ 15 – 25 mg/kg / day on alternate days for a
total of 10-15 days

Liver Flukes

LIver Flukes: Fasciola species

● Found in the liver and biliary passages of humans and
Liver Flukes Fasciola Gigantica and Hepatica
ruminants
● Fasciola hepatica
○ Sheep liver fluke
○ Temperate liver fluke
● Fasciola gigantica
○ Giant liver fluke
○ Tropical liver fluke
● Mode of transmission is by ingestion of metacercariae
found in edible aquatic plants or by drinking water with
floating metacercariae
● Metacercariae exists in the duodenum or jejunum and
liberate the juvenile fluke
● Juvenile fluke penetrates the intestinal wall and reaches
the liver capsule
● The parasite burrows into the liver parenchyma where it
grows and develops

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● First intermediate host: ○ Stimulates inflammation in the biliary

○ Lymnea philippinensis epithelium leading to fibrosis
● Second Intermediate Host
○ Watercress Liver Flukes Fasciola species diagnosis
○ Grass ● Microscopy
● Serologic test
Liver Flukes Fasciola hepatica ○ Low specificity because of cross reactivity with
● Fasciola hepatica Ova antigens of other parasites
○ Large ● RFLP
○ Ovoid ○ PCR restriction Fragment Length
○ Operculated Polymorphism
○ Bile stained
○ Unsegmented Liver Flukes Fasciola species treatment
○ Larger but very similar to Fasciola hepatica ova ● Bithionol
○ 20-50 mg/kg body weight on alternate days to
complete 10 to 5 doses
● Triclabendazole
○ Also a recommended drug of choice due to:
■ Efficacy
■ Safety
■ Ease of use

Liver Flukes Fasciola species Epidemiology

● Worldwide distribution
● Economic importance livestock raising
● In the Philippines, the dominant species is Fasciola
gigantica affecting cattle and water buffalos
● Few human cases are reported locally

Liver Flukes Clonorchis sinensis

● Chinese liver fluke
● Oriental Liver Fluke
● Distome of China
Liver Flukes Fasciola species
● First intermediate Host:
○ Bulimus fuchsiana (snail not found in the
Philippines)
● Second Intermediate Host:
○ Ctenopharyngodon idellus (fish)
● Adult Worm
○ Narrow, oblong, flatworm
○ Oral sucker is slightly larger than the ventral
sucker
○ Blind intestinal caeca are simple and extend to
the caudal region
○ Life span is 20-30 years

Liver Flukes Fasciola species Pathogenesis and Clinical

Manifestations
● Fascioliasis
● Asymptomatic
● Can produce fever
● Right upper quadrant abdominal pain Hypereosinophilia
● Acute or invasive phase
○ Migration from intestine to liver
○ Traumatic and necrotic lesions in liver
parenchyma
● Chronic or latent phase
○ Asymptomatic
○ Parasite has reached the bile ducts Adult Worm
○ Obstruction

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● Ova ○ 25 mg/kg three times a day for two days

○ Bile stained ○ 60 mg/kg in three doses for one day
○ Flask-shaped ○ May be used together with albendazole for
○ Operculated light and moderate infections
○ Contains a miracidium when oviposited
○ Does not hatch in water but is ingested with a Liver Flukes Clonorchis sinensis epidemiology
molluscan host ● Transmission is due to consumption of raw, undercooked
○ Has a terminal spine fish and salted and dried fish harboring the
○ Electric bulb in shape metacercariae
○ Infective to snails only ● Over 30 million people are infected in Southeastern Asia
● No reported cases in children below 10 years old
Ova ● Endemic in:
○ China
○ Japan
○ Korea
○ Vietnam

TREMATODES

Liver Flukes epidemiology of Paragonimiasis

● Has a global distribution
● In the Philippines
○ Leyte
○ Sorsogon
○ Mindoro
Camarines
○ Samar
Liver Flukes Clonorchis sinensis Life cycle ○ Davao
○ Cotabato
○ Basilan

INTESTINAL FLUKES

Intestinal Flukes: Fasciolopsis buski

● Giant intestinal fluke of man
● Parasite of the intestines of humans and pigs
● Mode of transmission is by ingestion of encysted
metacercariae on aquatic plants
● The viable metacercariae exist in the duodenum and
becomes mature in about three months
● Elongated
● Oval
● 20 – 75 mm in length and 8 -20 mm in width
● Covered with spines
Liver Flukes Clonorchis sinensis Pathogenesis and Clinical ● No cephalic cone
Manifestations ● Unbranched intestina caeca which reach up to the
● Clonorchiasis posterior end
● Provokes intense proliferation of intestinal epithelium
● Acute stage (less than 1 month of infection)
○ Chills
○ Fever
● Chronic stage
○ Cirrhosis
○ Portal hypertension

Liver Flukes Clonorchis sinensis diagnosis

● Detection of parasite egg in stool
● Clonorchis, Opisthorchis and Heterophyid ova may not
be differentiated under ordinary light microscope
● ELISA with crude Clonorchis sinensis antigen
● Enzyme immunoassay (EIA)
● Polymerase Chain Reactions Intestinal Flukes: Fasciolopsis buski life cycle

Liver Flukes Clonorchis sinensis treatment

● Praziquantel

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○ Korea
○ India
● Endemicity in the Philippines has not been
demonstrated yet
● Fasciolopsiasis in Filipinos are probably imported cases

Intestinal Flukes: Pathogenesis of Fasciolopsis buski

● Fasciolopsiasis
● Pathological changes caused are:
● Traumatic
○ Inflammation and ulceration
● Obstructive Intestinal Flukes: Echinostoma ilocanum
○ Intestinal obstruction due to heavy infection ● Garrison’s fluke
● Toxic ● Echinostomid
○ Due to absorption of worm metabolites by the ● There are several species that infect man
host ● There are 2 identified echinostomids that infect man in
Intestinal Flukes: Diagnosis of Fasciolopsis buski the Philippines:
● Detection of parasite eggs in stool ○ Echinostoma ilocanum
○ Provided with an operculum ○ Artyfechinostomum malayanum
○ Large ● Adult
○ Unembryonated when laid ○ Reddish gray
● Resemble Fasciola eggs ○ Horse-shape collar of spines (circum oral disk)
● Provided with an operculum around the oral suckers
○ 49-51 collar spines
○ Integument is covered by plaque like scales
○ Simple intestinal caeca

Adult

Intestinal Flukes: Treatment of Fasciolopsiasis

● Praziquantel Echinostoma ilocanum showing circum-oral disk
○ 25 mg/kg for 3 doses for one day
○ Side effects:
■ Dizziness
■ Drowsiness
■ Epigastric pain

Intestinal Flukes: Epidemiology of Fasciolopsiasis

● Endemic in
○ Southeast Asia
○ China

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Life Cycle

Intestinal Flukes Treatment Echinostoma ilocanum

● Praziquantel
○ 25 mg/kg for 3 doses for one day
● Echinostoma ilocanum Ova ○ No alcohol; no fats must be taken 24 hours
○ Straw-colored before and after treatment
○ Operculated ○ Only water must be taken within 3 hours of
○ Ovoid medication
○ Similar to Fasciola and Fasciolopsis buski ova
Intestinal Flukes Epidemiology Echinostoma ilocanum
Ova ● Echinostoma ilocanum is endemic in:
○ Northern Luzon
○ Leyte
○ Samar
○ Mindanao provinces
● Artyfechinostomum malayanum
○ First reported in 1987
○ Northern and Central Luzon

Intestinal Flukes Heterophyids

● Mode of transmission is by ingestion of metacercariae
encysted in fish
● Metacercariae in the abdomen excysts, liberating a larva
that attaches to the intestinal wall
● Heterophyes heterophyes Adult
Intestinal Flukes: Pathogenesis and Clinical Manifestations ○ Elongated
Echinostoma ilocanum ○ Oval or pyriform
● Man gets infected when metacercariae in the second ○ Measures less than 2 mm in length
intermediate hosts are ingested ○ Integument has fine scale-like spines
● Inflammation at the site of attachment of adults ○ Some species have gonotyl or genital sucker
● Ulceration
● Diarrhea (sometimes bloody) heterophyes Adult
● Abdominal pains
● General intoxication

Intestinal Flukes: Diagnosis Echinostoma ilocanum

● Detection of characteristic eggs in the stool

● Heterophyes heterophyes Ova

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○ Light brown in color

○ Ovoid in shape
○ Operculated
○ A fully developed symmetrical miracidium is
already present
○ Operculum fits into the egg smoothly
○ No abopercular protuberance like that of
Clonorchis sinensis ovum

Ova

Intestinal Flukes Heterophyids life cycle

REFERENCES

Notes from the discussion by Ma’am Marilyn Ngo

University of Santo Tomas powerpoint presentation: PPT

Intestinal Flukes Heterophyes heterophyes Pathogenesis and
from Ma’am Marilyn Ngo
Clinical Manifestations
● Heterophyiasis Course book: Hugo & Russell's Pharmaceutical Microbiology
● Inflammation at the site of attachment
8th edition
● Manifestations are consistent with peptic ulcer
(observed among infected individuals in Compostela
Valley)
○ Upper abdominal discomfort
○ Gurgling abdomen
Intestinal Flukes Diagnosis Pathogenesis and Clinical Edit History Log
Manifestations Date Remarks
● Detection of eggs in the stool using Kato Katz method Date Here Version 1 completed by CONCEPCION,
● Care must be taken to distinguish them from Clonorchis DIMAPANAT, GARCIA, HERNANDEZ.
and Opisthorchis ova VICEDO
Intestinal Flukes Treatment Pathogenesis and Clinical Date Here Edits made by VICEDO
Manifestations Date Here Evaluated by VICEDO
● Praziquantel Date Here Compiled to Master File
○ 25 mg/kg in 3 doses for 1day

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