Reoviruses

Gastroenteritis

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Key points
1.The Reoviridae comprise four genera which infect humans; they are characterized by the possession
of 10–12 segments of double-stranded RNA encased within an unenveloped capsid of icosahedral
symmetry.
2.When viewed under the electron microscope, the characteristic wheel-shaped virus particles measure
70–75 nm in diameter.
3.Group A rotaviruses are the major cause of acute gastroenteritis in infants and young children
worldwide.
4.A unique feature of rotavirus replication is the transient acquisition of an envelope during the
process of budding into the endoplasmic reticulum.
5.Rotaviruses produce NSP4, the first described viral enterotoxin.
6.Diagnosis of rotavirus gastroenteritis is usually made by the detection of viral antigen (VP6) using
enzyme-linked immunosorbent and immunochromatographic assays.
7.A distinct winter seasonality of rotavirus infection is evident in temperate countries; infection is year-
round in tropical countries.
8.Rehydration and restoration of electrolyte balance are the primary aims of treatment of acute rotavirus
gastroenteritis.
9.Two live oral rotavirus vaccines are entering childhood immunization schedules and promise to
substantially reduce the global rotavirus disease burden.
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Classification

Four of the nine genera of the Reoviridae family infect

humans:
• Orthoreovirus – reovirus types 1, 2 and 3
• Orbivirus and Coltivirus – various serogroups
• Rotavirus – of seven species, formerly called serogroups
(A–G), Rotavirus A–Rotavirus C have been detected in
humans. Multiple genotypes exist within Rotavirus A.

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Rotaviruses
Characteristic
• None-enveloped, genome is composed of 11 segments of double-
stranded RNA
• Also found in other mammals and birds, causing diarrhoea.
• In human divided into major groups:
Group A infections are most common
Group B has been associated with outbreaks in adults in China
Group C is responsible for sporadic cases of diarrhea in infants around the
world

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 The rotavirus replication cycle

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Pathology
• Account for 50-80% of all cases of viral gastroenteritis.
• Usually endemic, but responsible for occasional outbreaks.
• Causes disease in all age groups but most severe symptoms in neonates and
young children. Symptomatic infections again common in people over 60.
• Asymptomatic infections common in adults and older children. 24-48 hr
incubation period followed by an abrupt onset of vomiting and diarrhoea; a
low-grade fever may be present.
• Up to 30% mortality rate in malnourished children, responsible for up to half
a million deaths per year.
• 80% of the population have antibody against rotavirus by the age of 3.

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Clinical Features
Incubation period - thought to be <4 days
Fever (can be high grade (>39C in 30%))
Vomiting, nausea precede diarrhea
Diarrhea
- usually watery (no blood or leukocytes)
- lasts 3-9 days
- longer in malnourished and immune deficient individuals.
- Hemorrhagic gastroenteritis (GE) or necrotizing enterocolitis (NEC)  seen in neonates
Dehydration is the main contributor to mortality.
Secondary malabsorption of lactose and fat, and chronic diarrhea are possible
Watery diarrhea due to net secretion of intestinal fluid
Activation of the enteric nervous system -possible role of enterotoxin
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Transmission INDIA RIVER GANGA 

• More frequent during the winter.

• Faecal-oral spread.
• Fomites
• Food and water-borne spread is possible
• Spread via respiratory route is speculated
• Only 10 to 100 infectious virus particles are needed to cause infection.
Diagnosis
• Electron microscopy or by the detection of rotavirus antigens in
faeces by ELISA or other assays.
Treatment & Prevention
• Supportive - oral, IV rehydration
• Live attenuated vaccines now available for use in children.
• Large numbers of viruses are shed in fecal matter, from 100 to 1000 particles per milliliter.
• Hand-washing and sanitation does not work due to the resistant nature of the virus. Similar
hospitalisation rates in countries with high and poor sanitation
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 Retroviruses
Acquired immune deficiency syndrome; HTLV-1.

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Key points
1.Some retroviruses, including human T-cell lymphotropic virus (HTLV)-I, the cause of
human T-cell leukaemia, cause tumors in natural hosts.
2.HIV-1 is the cause of the acquired immune deficiency syndrome (AIDS), a persistent
infection leading to loss of CD4+ T cells, immunodeficiency and many opportunistic
infections.
3.Disease status can be measured by sequential changes in CD4 count and viral load in
plasma (HIV RNA copy number).
4.HIV has a global distribution; it is spread by sexual intercourse, mother-to-child
transmission and via blood and blood products. The greatest incidence is in sub-Saharan
Africa and South-East Asia.
5.HIV replication can be inhibited by drugs that block co-receptor binding, membrane
fusion, reverse transcription, integration and protein cleavage during maturation.
6.Combination antiretroviral therapy (cART) reduces the appearance of drug resistance and,
combined with control of opportunistic infections, substantially improves survival.

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Retroviruses
Single stranded, positive sense, enveloped, icosahedral.
Distinguished from all other RNA viruses by presence of an unusual enzyme,
reverse transcriptase.
Retro = reversal
RNA is serving as a template for DNA synthesis.

Two genera of human interest

LentivirusLentus = slow Human immunodeficiency viruses 1 & 2 (HIV-1 & -2)
Human T-cell lymphotropic virus-bovine leukemia virus group (HTLV-BLV)
Human T-cell leukemia viruses 1 & 2 (HTLV-1 & -2)

Acquired immunodeficiency syndrome (AIDS) was first reported in US in 1981.

By 1984, AIDS was recognized as an infectious disease caused by a virus.

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Structure
The viral envelope formed from host cell
membrane; contains 72 spiked knobs. These
consist of a transmembrane protein TM (gp
41), which is linked to surface protein SU (gp
120) that binds to a cell receptor during
infection. The virion has cone-shaped,
icosahedral core, containing the major capsid
protein CA (p24). Between capsid and
envelope is an outer matrix protein, MA
(p17).

Two identical copies of positive sense ssRNA genome (retroviruses are diploid).
Enzymes: reverse transcriptase, integrase and protease.

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 HIV Replication
The first phase of viral entry, reverse transcription and integration into host genome is
accomplished by viral proteins.

The second phase of synthesis and processing of viral genomes, mRNAs and structural
proteins, uses host cell machinery.

• Attachment to specific cell surface receptor: gp120 binds CD4 molecule on the helper
T cells, monocytes and dendritic cells
• Viral entery.
• Reverse transcription of viral RNA into DNA. The resulting double stranded DNA is
called provirus.
• Integration of provirus into host cell DNA. The viral integrase cleaves the
chromosomal DNA and covalently inserts the provirus. The insertion site is random.
• Transcription and translation of viral DNA sequences. The provirus is transcribed
into a full length mRNA by the cell RNA polymerase II.
• Assembly and maturation of progeny virus.
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Medical Microbilogy 2019 - NNU 14
Transmission
Transfusions: whole blood, plasma, clotting factors or cellular
fractions of blood.
Contaminated needles: accidentally or sharing needles by drug
users.
Sexual contact: HIV is present in semen and vaginal secretions;
either homoxesual or heterosexual contact
Perinatal: Transplacental, during delivery or via breast milk.

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Pathogenesis and clinical significance (1)
• Initial infection:
– genital tract macrophages
– HIV disseminates via blood
– Dendritic cells in lymphoid tissue
– CD4+ lymphocytes

• Acute phase viremia: several weeks after the initial infection, 1/3 – 2/3 of individuals
experience an acute disease syndrome similar to infectious mononucleosis. Circulating
antibody appears in 1 – 10 weeks after the initial infection (seroconversion).
• Latent period: lasts from months to many years (average 10 years). During this period,
90% of HIV proviruses are transcriptionally silent. Although there is continuous loss of
CD4+ cells in which HIV is replicating, active replacement through stem cell
multiplication is occurring. The infection remains clinically asymptomatic as long as the
immune system is functional.

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Pathogenesis and clinical significance (2)
• Clinical complications during the latent period: there are multiple non-specific
conditions such as persistent generalized lymadenopathy, diarrhea, chronic fevers,
night sweats and weight loss. The more common opportunistic infections such as
herpes zoster and candidiasis may occur repeatedly during this period. The CD4+ cell
count remains normal or gradually declines but is greater than 200 / ul. Progression
from asymptomatic infection to AIDS is not sudden.

• AIDS: Coinfection with HHV-6 can transactivate transcription from the silent HIV
provirus, increasing HIV replication. Any stimulation of an immune response causing
activation of resting T cells also activates HIV replication. Appearance of HIV
mutants with altered antigenic specificity which are not recognized by the existing
humoral antibody or cytotoxic T lymphocytes; also contributes to progression with
CD4+ count falling below 200 / ul and appearance of serious diseases and
opportunistic infections.
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Opportunistic Infections

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Laboratory Diagnosis
Antigen / antibody detection
ELISA, serum
HIV-1 & -2 antibodies, HIV-1 CA (p24) antigen
Screening of blood donors
Western Blotting

PCR
Viral RNA or DNA provirus
Blood or tissue specimens
Quantitative PCR (viral load): to determine disease stage and
treatment follow up.

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Treatment
Anti-retroviral drugs
Nucleoside analog reverse transcriptase inhibitors
Act by serving as a chain terminator (Zidovudine (AZT), Didanosine (ddi), Lamivudine (3TC))
Non-nucleoside reverse transcriptase inhibitors
Act by targeting the enzyme itself (Efavirenz Delaviridine Nevirapine)
Protease inhibitors
Interfere with the processing of polyproteins in the budding virion, resulting in non-infectious
particle. (Ritonavir, Amprenavir, Indinavir, Lopinavir)
Multidrug therapy
RT has no proofreading activity, resulting in production of many errors during viral DNA
synthesis which leads to mutations in all HIV genes and accumulation of mutant viral strains.
In presence of an antiviral drug, there is strong selection for mutations that confer resistance
to that drug.
Early therapy
Viral load is a prognostic indicator of the rate of progression to AIDS. Infection should be treated
as aggressively and as early as possible to minimize initial spread of the virus and give a
lower chance for mutants to arise.

Vaccine: not yet available/ Blood supply screening/ Perinatal transmission: AZT therapy
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HTLV
HTLV-1 and -2 have 65% nucleotide sequence homology
Genetically and biologically similar

HTLV-1
Adult T-cell leukemia
HTLV-associated myelopathy/tropical spastic paraparesis

HTLV-2
Hairy cell leukemia

Transmission:
Vertical transmission, Sexual, Blood products

Laboratory diagnosis
Screening of blood donors using ELISA
Hairy cell leukemia
Confirmation by western blotting
a rare lymphocytic leukemia, of B cell origin; caused by
PCR HTLV-2. it is characterized by malignant cells that look
ciliated.

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Caliciviruses and Astroviruses
Diarrhoeal disease

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Key points
1.Caliciviruses (noroviruses and sapoviruses) and astroviruses are common
causes of diarrhoeal disease transmitted worldwide by the faecal–oral route and
via fomites. Diagnosis is established by RT-PCR, enzyme immune assays or
electron microscopy.
2.They cause diarrhoea and vomiting with an incubation period of 12–72 h (3–4
days for astroviruses). Illness lasts for 1–4 days, and virus may be shed for up
to 2 weeks.
3.The very young and elderly are most at risk and may require rehydration.
Prolonged excretion occurs in some immunocompromised patients.
4.Cold foods are an important source, as are shellfish harvested from
contaminated seawater.

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Norovirus, Sapovirus
Family Caliciviridae
Non-enveloped RNA viruses
with ss [+] sense RNA
27-35 nm in size
Contains single capsid protein
Resistant to environmental pressure: detergents, drying, and acid.
Transmitted by fecal-oral route in contaminated water and food.
Cause outbreaks of gastroenteritis.
Disease resolves after 48 hours, without serious consequences

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ASTROVIRUS
Human serotypes: HuAstV 1-8
Non-enveloped small ss RNA virus, 27-32nm in size
Round with an unbroken, smooth surface
EM appearance of a 5 or 6 pointed star within smooth edge
Infants and children <7 years are most often affected, elderly and immune
compromised persons also can be infected.
Short incubation period 1-4 days
Nausea, vomiting, abdominal cramping and watery diarrhea
Constitutional symptoms-fever, malaise, headache
Endemic worldwide
Transmission- person-to-person [fecal-oral]
Outbreaks due to fecal contamination of sea-food or water

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Coronaviruses

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Key points
1.Coronaviruses are widespread among mammals and birds, affecting many
organ systems and causing a range of diseases.
2.Human coronaviruses 229E and OC43 are major causes of the ‘common cold’.
These, as well as the newly discovered HCoV NL-63 and HKU1, can cause both
upper respiratory tract infection and sometimes lead to lower respiratory tract
infections in all age groups.
3.SARS CoV emerged from bats, adapted in other small wild mammals (e.g.
civet cats) and acquired efficient human transmission leading to a global outbreak
of a novel disease. However, unusual features of its
pathophysiology allowed public health measures to
interrupt virus transmission in humans.
4.No vaccines or antivirals are in routine clinical
use for any HCoV.
civet cats
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Coronavirues
Enveloped
Replicates in cytoplasm of animal cells
Single-strand 30 kb RNA genome
With 5’ cap & poly-A tail
Respiratory, enteric, hepatic, neurological
Cause 30% respiratory infections
Can acquire genes by horizontal transfer and co-infection
Three main classes

SARS-CoV -2 responsible for the death of more than two and a half millions
patient all over the world

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Symptoms and Diagnostic Tests for SARS-CoV 1 and 2
Initial Symptoms:
fever of 38о C or higher, headaces, body aches, and malaise.
Week Later:
dry cough, difficulty breathing and severe diarrhea are seen in patients.
Recovery: starts after 5 to 6 days
Early Diagnosis:
patient is given antibiotics, antiviral, and steroids used for atypical pneumonia.
Patient is are quarantined in specially ventilated rooms.
Laboratory tests:
RT-PCR (reverse transcription-polymerase chain reaction) assay
Detection of SARS-CoV RNA
EIA (enzyme immunoassay)
Detection of serum antibody to SARS-CoV RNA

Enzyme-linked immunosorbent assays (ELISA)

Detects antibodies against the virus produced in response to infection

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Medical Microbilogy 2019 - NNU 31
RT-PCR

Antigen Testing

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Medical Microbilogy 2019 - NNU 33
 Palestine

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Rhabdoviruses

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Key points
1.Rabies has the highest case-fatality (100%) rate of any infectious disease once
clinical symptoms are observed. There are at least 55 000 cases annually worldwide.
2.Over 99% of human rabies infections are due to bites from a rabid dog. Most of
these occur in developing countries and the majority in children under the age of 15.
3.Pre- and post-exposure vaccination is available and both are extremely effective.
4.A paradigm shift is required to focus rabies control efforts towards the elimination
of rabies in domestic dogs.
5.Reducing the risk of rabies transmission to humans means implementation of
vaccination schemes for both domestic animals and wildlife.
6.The number of reported human rabies cases due to bat bites is on the increase
worldwide.
7.Vaccination of people in high-risk groups (laboratory workers, animal handlers and
those travelling to rabies endemic areas) is recommended.

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Rhabdovirs
Structure
Anti-sense ssRNA
genome codes for five proteins
bullet shaped capsid (60-180 nm)
lipid envelope A peplomer is a glycoprotein spike on a viral capsid or viral
envelope. These protrusions will only bind to certain receptors on
glycoprotein peplomers
the host cell: they are essential for both host specificity and viral
infectivity.
Transmission
• Bite of infected animal
• Eating infected meat ( wild animals)
• Airborne transmission-
– bat caves 2 cases in USA (1950-88)
– lab workers - 2 cases in USA( 1950-1988)
• Corneal transplants - 6 cases
• no known exposure -22%
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 Distribution
• Global all mammals
• Endemic in dogs in Asia, Africa,
– India 17,000 deaths/year, 3,000,000 vaccines
– Philppines canine rabies 25,000 /yr
• Mexico, Central and south America,
• Canada, U.S.A., western Europe
– Wildlife rabies -increasing
– canine rabies controlled.

• Described 5 century B.C. and 4 century B.C. by ancient

th th

Greeks

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Pathology of Rabies
Virus enters local tissue through bites
Travels along sensory nerves to CNS
Virus multiplies in CNS neurons
Hippocampus and cerebellum
Incubation Period
viremia usually 4-6 weeks
Infects other organs severe head or neck bites -2 weeks
range 5 days- 2 years
Clinical Manifestations
Fever, Malaise, Headache,
Sensory disturbances,
Respiratory muscle spasms ,
Swallowing muscle spasms Medical Microbilogy 2019 - NNU 39
Paralytic Rabies
‘Dumb’ rabies
Flaccid paralysis including respiratory muscles
Coma and Death

‘Furious’ rabies
Excitability- CNS disturbances
Recurrent spasms of muscles involed in swallowing
Hydrophobia
Choking panic
Delerium, convulsions

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Treatment of Suspected Rabies
Wash bite - soap, water and disinfect .
Rabies antiserum around skin of bite area
Vaccinate with HDCV immediately
Intramuscular
Deltoid in adults
Thigh in children
Days 0,3,7
Booster day30,90
Tetanus antiserum & antibiotics

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 Prion diseases
(transmissible spongiform encephalopathies)
Creutzfeldt–Jakob disease; Gerstmann–Sträussler–Scheinker syndrome; fatal
familial insomnia; iatrogenic Creutzfeldt–Jakob disease; kuru; variant Creutzfeldt–
Jakob disease; bovine spongiform encephalopathy; scrapie

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Key points
1.Prion diseases are fatal neurodegenerative disorders with very lengthy incubation
periods caused by unconventional agents. They are transmitted by inoculation or
ingestion; the intracerebral route of transmission results in the shortest incubation
period.
2.Prion diseases occur in mammals, including scrapie in sheep, bovine spongiform
encephalopathy (BSE) in cattle and Creutzfeldt–Jakob disease (CJD) in man.
3.The infectious agents are known as prions, which are thought to lack nucleic acid
and appear to consist entirely of a modified host-encoded protein, the prion protein.
4.Prion diseases do not elicit conventionally detectable immune responses.
Asymptomatic or subclinical infections are very difficult to detect.
5.Human prion diseases occur in sporadic, familial and acquired forms, all of which
are rare diseases. In 1996, a new form of human prion disease known as variant CJD
was identified, which results from infection with the BSE agent, probably via the oral
route.
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PrPc (for cellular)
The normal protein.
A transmembrane glycoprotein normally found at the surface of certain cells (e.g., neural
and hematopoietic stem cells).
Has its secondary structure dominated by alpha helices (probably 3 of them).
Easily soluble.
Easily digested by proteases.
Encoded by a gene designated (in humans) PRNP located on chromosome 20.

PrPsc(for scrapie)
Abnormal, disease-producing protein (infectious).
Has the same amino acid sequence and number as the normal protein; that is, their
primary structures are identical but:
- its secondary structure is dominated by beta conformation
- Insoluble in all solvents.
- Highly resistant to digestion by proteases.
- When PrPsc comes in contact with PrPc, it converts the PrPc into PrPsc (even in the test
tube). Medical Microbilogy 2019 - NNU 44
Creutzfeldt-jakob disease (CJD) - Sporadic
• Accounts for »85% of all CJD cases.
• Estimated incidence of one case/1 million population per year with equal sex ratio.
• A peak age of onset between 55 and 75 yrs (mean:61.5yrs).
Clinical Features
• Dementia: rapidly progressive, visual abnormalities.
• Ataxia: cerebellar dysfunction, including muscle incoordination,gait and speech
abnormalities.
• Most patients develop pyramidal and extrapyramidal dysfunction with abnormal
reflexes, spasticity, tremors, and rigidity.
• Psychaitric manifestations:some patients may also show behavioral changes with
agitation, depression, or confusion.
• Myoclonus: most constant physical sign,present in > 90% pts.
• Invariably fatal, with a median illness duration of 4 months & death occurs within 12
months of illness onset in 85–90% of pts.
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The transmissible nature of CJD was first described in 1968, after
intracerebral inoculation of a brain biopsy tissue from a CJD patient into
a chimpanzee.

Person-to-person transmission of the CJD agent was reported in:

1.Corneal transplant
2.Contaminated EEG depth electrodes
3.Neurosurgical instruments
4.Cadaveric pituitary-derived gonadotropin
5.Human growth hormone (hGH)
6.Dura mater grafts
7.Blood transfusion?

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Creutzfeldt-jakob disease - familial
It accounts for 5–15% of CJD patients.
Autosomal dominant inheritance pattern with a family history
of CJD.
Most patients presented with personality changes followed by
progressive dementia and a Parkinsonian syndrome.
The mean age at onset was 44.8 years, and the mean duration
of illness was 4.2 years.
Spongiform changes, neuronal loss, and mild gliosis were
predominantly seen in frontal and temporal lobes .

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Gerstmann-str¨aussler-scheinker syndrome (GSS)
A familial disease with autosomal dominant inheritance,
First described in1936 by Gerstmann, Str¨aussler, & Scheinker.
It occurs at an estimated annual incidence of 5 cases/100 million
population.
Neurologic signs and symptoms: cerebellar ataxia, gait
abnormalities, dementia, dysarthria, ocular dysmetria, and
hyporeflexia or areflexia in the lower extremities with infrequent
myoclonus and diagnostic EEG, and a neuropathologic feature of
numerous amyloid plaques.
GSS is considered a variant of the familial form of CJD, but it is
primarily associated with mutations at codon 102 of the prion
protein gene. Medical Microbilogy 2019 - NNU 48
Fatal familial insomnia (FFI)
A familial disease with autosomal dominant inheritance, have been
reported since 1939.
Patients with severe dementia and bilateral symmetrical degeneration
of the thalamus
Progressive insomnia and autonomic dysfunction, followed by
dysarthria, tremor, and myoclonus.
Neuropathologic examination showed neuronal degeneration and
reactive astrocytosis confined to the anterior and dorsomedial nuclei of
the thalamus; no spongiform changes or inflammatory infiltrates were
noted.
Rare sporadic cases of FFI were reported.
FFI is primarily associated with a mutation at codon 178 of the prion
protein gene.
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Kuru
Kuru was once found among the Fore tribe in Papua New Guinea in 1957.
Ingesting brain tissue of dead relatives for religious reasons was the route of
transmission whose rituals included eating the brain tissue of their recently
deceased members of the tribe.
Since this practice (ritualistic cannibalism ) was halted, the disease has
disappeared.
Clinically, the disease resembles CJD.

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