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RhabdovirusesRhabdoviruses

Hugh B. FackrellHugh B. Fackrell

Fackrel@Uwindsor.caFackrel@Uwindsor.ca

Rhabdo.pptRhabdo.ppt

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RhabdovirusesRhabdoviruses

StructureStructureClassificationClassificationMultiplicationMultiplicationClinical manifestationsClinical manifestationsEpidemiologyEpidemiologyDiagnosisDiagnosisControlControl

Baron’s Web Site

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StructureStructure

Anti-sense ssRNAAnti-sense ssRNAgenome codes for five proteinsgenome codes for five proteinsbullet shaped capsid (60-180 nm)bullet shaped capsid (60-180 nm)lipid envelopelipid envelopeglycoprotein peplomersglycoprotein peplomers

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Rabies VirusRabies Virus

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Rhabdovirus proteinsRhabdovirus proteins

Protein L -RNA dependent RNA Protein L -RNA dependent RNA polymerase polymerase

Protein G- surface antigenProtein G- surface antigenProtein N -RNA binding proteinProtein N -RNA binding proteinProtein NS- phosphoproteinProtein NS- phosphoproteinProtein M-membrane/matrix proteinProtein M-membrane/matrix protein

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Rhabdovirus G proteinRhabdovirus G protein

Glycoprotein in peplomerGlycoprotein in peplomer 64-68,000 MW64-68,000 MW

Induces protective virus neutralizing Induces protective virus neutralizing antibodyantibody

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Rabies virus genomeRabies virus genome

ssRNA 12 kbasessRNA 12 kbaseantisense antisense leader at 3’ endleader at 3’ endintergenic region between each geneintergenic region between each gene

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MultiplicationMultiplication

1903. Adelchi 1903. Adelchi Negri, an Italian Negri, an Italian physician found physician found negri bodies.negri bodies.

in cytoplasm of in cytoplasm of CNSCNS

Negri Body

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MultiplicationMultiplication

Attaches to host via G proteinAttaches to host via G proteinpenetrationpenetrationuncoated in cytoplasmuncoated in cytoplasmProtein synthesisProtein synthesis

5 complementary mRNA developed5 complementary mRNA developed RNA dependent RNA polymeraseRNA dependent RNA polymerase

Positive strand of RNAPositive strand of RNA template for antisense RNAtemplate for antisense RNA

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Rhabdo Virus ReplicationRhabdo Virus Replication

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+ mRNA

+ ssRNA

- ss RNA

RNA polymerase

Cytoplasm

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TransmissionTransmission

Bite of infected animalBite of infected animalEating infected meat ( wild animals)Eating infected meat ( wild animals)Airborne transmission-Airborne transmission-

bat caves 2 cases in USA (1950-88)bat caves 2 cases in USA (1950-88) lab workers - 2 cases in USA( 1950-1988)lab workers - 2 cases in USA( 1950-1988)

Corneal transplants - 6 cases Corneal transplants - 6 cases no known exposure -22%no known exposure -22%

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Infections from Trauma BitesInfections from Trauma Bites

Zoonosis: Zoonosis: Reservoir in wild animalsReservoir in wild animalsSkunks, Bats, Racoons, FoxesSkunks, Bats, Racoons, Foxes

Transmissible to man and domestic animals Transmissible to man and domestic animals by bitesby bites

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DistributionDistribution

GlobalGlobal all mammals all mammals Endemic in dogs in Asia, Africa,Endemic in dogs in Asia, Africa,

India 17,000 deaths/year, 3,000,000 vaccinesIndia 17,000 deaths/year, 3,000,000 vaccines Philppines canine rabies 25,000 /yr Philppines canine rabies 25,000 /yr

Mexico, Central and south America,Mexico, Central and south America,Canada, U.S.A., western Europe Canada, U.S.A., western Europe

Wildlife rabies -increasingWildlife rabies -increasing canine rabies controlled.canine rabies controlled.

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EpidemiologyEpidemiology

Described 5Described 5 th th century B.C. and 4century B.C. and 4 th th century century B.C. by ancient GreeksB.C. by ancient Greeks

Rabid dogsRabid dogs

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Pasteur Pasteur

evidence of infections agent in saliva, CNS, evidence of infections agent in saliva, CNS, peripheral nerves.peripheral nerves.

He attenuated the agentHe attenuated the agent used it to protect against rabies Joesph used it to protect against rabies Joesph

Meister. Meister.

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Pathology of RabiesPathology of Rabies

Virus enters local tissue through bitesVirus enters local tissue through bitesTravels along sensory nerves to CNSTravels along sensory nerves to CNS Virus multiplies in CNS neuronesVirus multiplies in CNS neurones Hippocampus and cerebellumHippocampus and cerebellumviremiaviremiaInfects other organsInfects other organs

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Clinical ManifestationsClinical Manifestations

Fever, Malaise, Headache, Fever, Malaise, Headache, Sensory disturbances, Sensory disturbances, Respiratory muscle spasms , Respiratory muscle spasms , Swallowing muscle spasmsSwallowing muscle spasms

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Paralytic RabiesParalytic Rabies

‘‘Dumb’ rabiesDumb’ rabiesFlaccid paralysis including respiratory Flaccid paralysis including respiratory

musclesmusclesComa and DeathComa and Death

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‘Furious’ rabies‘Furious’ rabies

excitability- CNS disturbancesexcitability- CNS disturbancesrecurrent spasms of muscles involed in recurrent spasms of muscles involed in

swallowingswallowing 17-50%17-50% ‘‘Hydrophobia”Hydrophobia” choking panicchoking panic

delerium, convulsionsdelerium, convulsions

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Incubation Period Incubation Period

usually 4-6 weeksusually 4-6 weekssevere head or neck bites -2 weekssevere head or neck bites -2 weeks range 5 days- 2 yearsrange 5 days- 2 years

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Canine RabiesCanine Rabies

Dog acts as if it had a sore throat or something Dog acts as if it had a sore throat or something caught in its throat.caught in its throat.

Paralysed - “Dumb” rabiesParalysed - “Dumb” rabiesagitated or aggressive furious rabiesagitated or aggressive furious rabiesthroat muscle spasms - droolingthroat muscle spasms - droolingDog has difficulty swallowingDog has difficulty swallowing appears to be foaming at the mouth, eventually appears to be foaming at the mouth, eventually

becomes staperous and dies.becomes staperous and dies.

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Wild life rabiesWild life rabies

ONTARIO - Foxes, Racoons, Skunks - ONTARIO - Foxes, Racoons, Skunks - 59% 59%

Bats/cave dust 14%Bats/cave dust 14%

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DiagnosisDiagnosis

Clinical symptomsClinical symptomsCan be detected without symptoms.Can be detected without symptoms. Fluorescent antibodies -developed 1958.Fluorescent antibodies -developed 1958.Isolate animalIsolate animal observe 5 daysobserve 5 days

if symptoms disappear - not rabies -if symptoms disappear - not rabies -autopsy animal - Negri bodies - Brain autopsy animal - Negri bodies - Brain

cellscells

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Laboratory DiagnosisLaboratory Diagnosis

Antibodies in serial serum specimensAntibodies in serial serum specimensvirus cultured from salivavirus cultured from saliva virus antigenvirus antigen

skin biopsyskin biopsy detected by fluorescent antibodydetected by fluorescent antibody

mouse testmouse test inject saliva into mouseinject saliva into mouse isolate virusisolate virus

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Treatment of Suspected Rabies Treatment of Suspected Rabies

Wash bite - soap, waterWash bite - soap, waterDisinfect Disinfect

quaternary ammoniumquaternary ammonium 0.1% benzalkonium chloride 0.1% benzalkonium chloride Iodine or 70% ethanol.Iodine or 70% ethanol.

Rabies antiserum around skin of bite areaRabies antiserum around skin of bite areaVaccinate with HDCV immediatelyVaccinate with HDCV immediatelyTetanus antiserum & antibioticsTetanus antiserum & antibiotics

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Vaccine Vaccine

Duck embryo vaccine has side effectsDuck embryo vaccine has side effectsHuman diploid cell vaccine Human diploid cell vaccine

Merieux InstituteMerieux Institute

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Human Diploid Cell VaccineHuman Diploid Cell Vaccine

HDCV =Merieux vaccineHDCV =Merieux vaccine1% allergic encephalitis1% allergic encephalitis

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Pre-exposure vaccinePre-exposure vaccine

Merieux vaccine (HDCV)Merieux vaccine (HDCV)lyophilized dose in syringelyophilized dose in syringe

intradermal injection intradermal injection 0.1 ml0.1 ml0,7,21,28 days0,7,21,28 days

Booster every 2 yearsBooster every 2 years

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High risk PopulationsHigh risk Populations

Rabies research lab workersRabies research lab workersRabies diagnostic labsRabies diagnostic labsSpelunkersSpelunkersVeterinariansVeterinariansAnimal control workersAnimal control workersInternational travelersInternational travelers

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Postexposure ProphylaxisPostexposure Prophylaxis

IntramuscularIntramuscular Deltoid in adultsDeltoid in adults Thigh in childrenThigh in children Days 0,3,7Days 0,3,7 Booster day30,90 Booster day30,90

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Reactions to HDCVReactions to HDCV

LocalLocal<10% redness, induration, 24-48 hours<10% redness, induration, 24-48 hours

GeneralGeneral slight fever, malaiseslight fever, malaise1/10,000 type I hypersensitivity1/10,000 type I hypersensitivity1/1,000 type III 2-21 days1/1,000 type III 2-21 days

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Single verified case of transient neuroparalytic illness after

HDCV

Single verified case of transient neuroparalytic illness after

HDCV

Bernard et al 1982, JAMA 248:3136-3138Bernard et al 1982, JAMA 248:3136-3138

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ControlControl

Domestic AnimalDomestic Animal intradermal vaccinationintradermal vaccination

Wild animalsWild animals oral vaccines meatballsoral vaccines meatballs

Humans Vaccinations Humans Vaccinations high risk populationshigh risk populations

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Rabies free countries Rabies free countries

England, Australia, Japan,Sweden, Spain.England, Australia, Japan,Sweden, Spain.Due to vaccination Due to vaccination strict import regulations.strict import regulations.6 months quarantine. 6 months quarantine.

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RABIESRABIES RhabdovirusRhabdovirus

ssRNAssRNA

zoonosis: Reservoir in wild animalszoonosis: Reservoir in wild animals

e.g. Skunks, Bats, Raccoons, Foxese.g. Skunks, Bats, Raccoons, Foxes

Transmissible to man and domestic animalsTransmissible to man and domestic animals

to man by bitesto man by bites

Infections from Trauma bitesInfections from Trauma bites

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EpidemiologyEpidemiology

Described 5th century B.C. and 4th century Described 5th century B.C. and 4th century B.C. by ancient GreeksB.C. by ancient Greeks

Pasteur - found evidence of infections agent Pasteur - found evidence of infections agent in saliva, CNS, peripheral nerves.in saliva, CNS, peripheral nerves.

He attenuated the agent and used it to He attenuated the agent and used it to protect against rabies. protect against rabies.

1903. Adelchi Negri, an Italian physician 1903. Adelchi Negri, an Italian physician found negri bodies.found negri bodies.

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Pathology of RabiesPathology of Rabies

Bites/ Virus enters local tissue.Bites/ Virus enters local tissue.Travels along sensory nerves to CNSTravels along sensory nerves to CNS Virus multiplies in CNS neuronsVirus multiplies in CNS neuronsHippocampus and cerebellumHippocampus and cerebellumviremiaviremiaInfects other organsInfects other organs

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RabiesRabies

Clinical Course: Clinical Course: Fever, Malaise, Headache, Fever, Malaise, Headache, Sensory disturbances, Sensory disturbances, Respiratory muscle spasms , Respiratory muscle spasms , Swallowing muscle spasmsSwallowing muscle spasmsChoking panic “ Hydrophobia”Choking panic “ Hydrophobia”CNS disturbances - excitability CNS disturbances - excitability Delirium, ConvulsionsDelirium, Convulsions

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Rabies: Terminal SymptomsRabies: Terminal Symptoms

Flaccid paralysis including respiratory musclesFlaccid paralysis including respiratory musclesComa and DeathComa and DeathIncubation: 4-6 weeksIncubation: 4-6 weeksif severe head or neck bites -2 weeksif severe head or neck bites -2 weeksIsolate animalIsolate animal observe 5 daysobserve 5 daysif symptoms disappear - not rabies -if symptoms disappear - not rabies - use fluorescent antibodyuse fluorescent antibodyIf symptoms get worse, kill animal - Negri bodies - Brain If symptoms get worse, kill animal - Negri bodies - Brain

cellscells

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Treatment of Suspected RabiesTreatment of Suspected Rabies

1.Wash bite - soap, water, alcohol, lavage with 1.Wash bite - soap, water, alcohol, lavage with Iodine.Iodine.

2. Inject hyper immune serum around skin of bite area2. Inject hyper immune serum around skin of bite area3. Vaccinate immediately - Human tissue culture 3. Vaccinate immediately - Human tissue culture

Vaccine - Vaccine - Duck embryo vaccine has side effectsDuck embryo vaccine has side effects 1% allergic encephalitis1% allergic encephalitis

4. If symptoms develop. Death very likely.4. If symptoms develop. Death very likely.

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Rabies cont’dRabies cont’d

1958 - Fluorescent antibodies used to detect rabies.1958 - Fluorescent antibodies used to detect rabies.Can be detected without symptoms.Can be detected without symptoms.

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Rabies: IncidenceRabies: Incidence

Rabies - World wide - all mammals Rabies Rabies - World wide - all mammals Rabies endemic in dogs in Asia, Africa, Mexico, endemic in dogs in Asia, Africa, Mexico, Central and south America,Central and south America,

In Canada, U.S.A., western Europe - Rabies in In Canada, U.S.A., western Europe - Rabies in dogs controlled.dogs controlled.

Wild life rabiesWild life rabiesONTARIO - Foxes, Racoons, Skunks - 59% ONTARIO - Foxes, Racoons, Skunks - 59% Bats/cave dust 14% Bats/cave dust 14%

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Rabies Rabies

Control: Pets vaccinatedControl: Pets vaccinated Cats, Dogs, Horses, CattleCats, Dogs, Horses, Cattle

Humans at risk: Veterinarians, Vaccinations - Humans at risk: Veterinarians, Vaccinations - originally 14 injections - agonising.originally 14 injections - agonising. Now intramuscular.Now intramuscular.

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Rabies in dogs:Rabies in dogs:

Dog acts as if it had a sore throat or Dog acts as if it had a sore throat or something caught in its throat.something caught in its throat.

Paralyzed - “Dumb” rabiesParalyzed - “Dumb” rabiesagitated or aggressive furious rabiesagitated or aggressive furious rabiesthroat muscle spasms - droolingthroat muscle spasms - drooling

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RabiesRabies

Dog has difficulty swallowingDog has difficulty swallowing appears to be foaming at the mouth, eventually appears to be foaming at the mouth, eventually

becomes staperous and dies.becomes staperous and dies.Rabies free countries include: England, Australia, Rabies free countries include: England, Australia,

Japan,Japan,Sweden, Spain.Sweden, Spain.Due to vaccination and strict import regulations.Due to vaccination and strict import regulations.6 months quarantine.6 months quarantine.