Presentation Outline

1. Some facts about rabies

2. History of the rabies virus
3. Transmission
4. Symptoms
5. Prevalence
6. Morphology and genome
7. Important viral proteins
8. Replication cycle
9. Diagnostic Techniques
10. Prevention, including pre- and post-exposure treatments
11. Conclusion
 Some Facts about Rabies
• Belongs to the family Rhabdoviridae
• Genus Lyssavirus (Greek root “lyssa”)
• Infects warm-blooded mammals, including
humans
• Causes encephalitis of the central nervous
system
• Results in death of the host
 History of Rabies Virus
• Man described the disease in 2300 B.C. in the
Eshnunna Code of Mesopotomia.
• The origin “rabhas”, meaning “to do violence”
comes from the language Sanskrit dating 3000
B.C.
• Many cultures have incorporated myths and
legends that can be connected to the rabies
virus.
 History of the Rabies Virus
•Outbreak of rabies in Europe
in 20th century spawned the
Vampire myth- linked to bats.
•Rabies epidemic in 13th
century France may have
spurred Werewolf myth as
well- linked to wolves.
•Rabies is commonly spread
by bats and wolves and other
wildlife like foxes and coyotes.
www.Davidsposters.com
 History of the Rabies Virus
• Since Roman times, man established the link
between the infectivity of a rabid dogs saliva
and the spread of the disease.
• Because there is no cure for rabies, those that
had been bitten by a rabid dog would
commonly commit suicide to avoid the painful
death that would inevitably follow.
 History of the Rabies Virus
• Louis Pasteur was the first person to diagnose
that rabies targeted the CNS.
• Also determined that nervous tissue of an
infected human or animal also contained the
virus.
• In 1890 created the rabies vaccine and saved 9
year old Joseph Meister after he had been
bitten by a rabid dog.
 Transmission of Rabies
• Is zoonotic, meaning is transmitted from
animal-animal or animal-human.
• Rabies is most commonly contracted from the
bite of an animal that is infected with the
virus.
• The virus is found in the saliva of the infected
animal.
 Transmission of Rabies
• Can also enter via any mucous membrane,
such as the eye, nose or mouth.
• Handling and skinning of infected animal
carcasses.
• Few cases of human to human transmission.
• Can be transmitted via aerosol, but that’s also
uncommon.
 Transmission of Rabies
• Inside the U.S., important reservoirs for the
transmission of the disease are wild animals such as
raccoons, skunks, foxes, bats, and coyotes.
• The time it takes for rabies to affect the animal
averages 1-3 months. Sometimes may only be a few
days, and rarely longer than a year after exposure.

www.cdc.gov/rabies.html
 Data Summaries by the United States Public Health
Service Regarding Prevalence of Rabies in the U.S.

5,540 Mammals tested

In 1955- reported by the
US Public Health Service

3,900 Animals 6,947 Animals tested

Tested in 1960 In 1994

Cockrum, 1997
 Transmission of Rabies
• The rabies infection and the symptoms that
accompany it is classified by five stages:
1. Incubation (1-3 months)
2. Prodromal, where first symptoms occur
3. Acute neurological phase
4. Coma
5. Death or recovery
 Transmission of Rabies
1. Incubation
-The animal or human is bitten by rabid animal.
-Rabies enters the body via the saliva of the infected
animal.
-The virus spreads through the nerves via retrograde
axoplasmic flow. The virus makes it’s way to the dorsal root
ganglion where it replicates.
- No symptoms are experienced at this time, and the
disease can’t be transmitted to another animal yet.
 Transmission of Rabies
2. Prodromal Stage
-Virus replicates in the dorsal root ganglion of the spinal cord
where the blood-nerve barrier is thin.
-This is where the immune system detects the virus and starts
creating antibodies.
-First symptoms are felt: fever, muscle ache, agitation,
anxiety, nausea, vomiting, ache in the bitten extremity.
-From this point the virus replicates and travels up to the
brain, and the immune system can’t keep up with fighting the
virus.
 Transmission of Rabies
3. Acute Neurological Stage
- There is rapid proliferation of the virus in the brainstem,
thalamus, basal ganglia, and spinal cord
- Brain begins to swell, encephalitis
- Fluctuation between consciousness and confusion
- Aggressive behavior, violent episodes
- Convulsions and seizures
- Hallucinations
- Hydrophobia (foaming at the mouth) induced by pharyngeal
spasms (the most common image of rabies)
 Transmission of Rabies
3. Acute Neurological Stage, continued
- Aerophobia
- Sensitivity to lights, sounds and smells
- Respiratory spasms.

4. Coma
- Spasms decrease because paralysis occurs
- Heart arrhythmias and irregular heartbeat persist
- Cerebral encephalitis causes major organs to collapse
- Fixed pupils that are unresponsive to light
 Transmission of Rabies
5. Death or Recovery
- Death results from encephalitis of the brain.
- Heart failure along with major organ failure are the main
causes of death.
- Only four known cases of human recovery, and they were
permanently brain damaged.
 Morphology of rabies virus
•Negative single-stranded RNA genome
•Rod shaped with a flexible nucleocapsid tapering at
one end
– Nucleocapsid contains important viral proteins

180nm\75nm

http://www.ncbi.nlm.nih.gov
N-protein
makes up most of the nucleocapsid
catalyzes replication process
G-protein
glycoprotein, come together to form the spikes of the viral envelope
mediates fusion
its presence triggers immune response of host
determines pathogenic ability of the virus

L-protein
Small part of nucleocapsid
Codes for RNA polymerase
Functions in polymerizing activity
P-protein
Also makes up a small portion of the nucleocapsid
Contributes to the ability of the L-protein to both bind and move down template
strand during the attachment of nucleotides
Voyles 1993
• A membranous envelope surrounds the nucleocapsid
– Envelope contains both viral and glycoproteins
– Derived from the lipids of the host cell membrane
– Viral proteins of envelope form spikes that aid in viral
attachment

http://www.microbe.org1microbes/virus_or_bacterium.asp
 Diagnostic Techniques
1. Histological examination for Negri bodies
sections/impresssion smears of brain –Sellers
technique. -negri bodies are intracytoplasmic masses
round/oval eosinoplilic with basophilic inner granules (3-
27µm size) -of viral nucleocapsids found in the brain tissue
-problem is that negri bodies are only present in 50-80% of
rabies cases
2. Direct fluorescent antibody test
-Antemortem: In salivary, corneal, conjunctival smears or
biopsy from nape of neck.
-PM: in impression smears of the cut surface of salivary
gland, brain stem, cerebellum, hipocampus.
Jogai 2002
 Negri body

http://www.med.sc.edu:85/virol/negri-bris.jpg
 3. Immunohistochemical technique
-confers the presence of viral antigens in organs
outside the NS (GI-tract, heart, etc.)
-biopsies are stained with immunoperoxidase, to
expose antigens, and treated w/ labeled rabies-
specific antibodies to detect antigens
-benefits
▪ reduces risk to technician because tissue sample are
embedded in formalin-fixed paraffin

▪ can examine the spread of the virus in organs outside of

the nervous system

Jogai 2000
 4. Mouse inoculation test
-brain material from the patient in question is inoculated into
mice to see if it leads to fatality
-this procedure takes at least a week

5. RT-PCR
-Reverse Transcriptase-Polymerase Chain Reaction
-can make a DNA copy of the viral genome and use PCR, with
a primer specific to the rabies genome, to determine its
presence
Detection of rabies antibodies: by ELISA in serum and CSF.

Meslin 1996
 Post-exposure Prophylaxis
1. Wash bite wound thoroughly with soap and water/QACs
2. Isolate the animal if possible
3. Human Rabies immunoglobulin 20IU/kg or equine antirabies serum 40
IU/kg; ½ IM –gluteal and other ½ infiltrated around the bite wound.
4. Vaccines generally confer immunity after 2 weeks; so bites closer to CNS
( like face, neck etc must be given Ig)
5. Cell culture vaccine (HDCV)- 1ml should be given IM on days 0,3,7,14,30
and 90 in the deltoid
6. Give TT prophylaxis as needed
7. Treatment may be discont if animal remains helathy after 10 days of
observation; or if it is found –ve for rabies on lab PM examination.
8. Pre-exposure immunization: can be offered to persons at high isk of
exposure such as lab staff working with virus/samples, vets, animal
handlers etc. 3 injections of HDC on days 0, 7 and 28. Boosters may be
given at 1-3 y intervals.
 Vaccine:
• Neural vaccines:
– Pasteur vaccine
– Fermi vaccine
– Semple vaccine
– Beta propionolactone vaccine
– Suckling mouse brain vaccine
• Non – neural vaccines:
– Duck egg vaccine
– Cell culture vaccines:
• First generation cell culture –HDCV
• Second generation cell culture - purified chick embryo cell vaccine ( PCEC) and Purified Vero
cell rabies vaccine (PVRV)
• Third generation rabies vaccine: clinical trials.
Thank You!!!!!!