100 are known pathogenic to humans Zoonotic (except urban yellow fever and dengue) 3 syndromes: 1. 2. 3. Fevers of undifferentiated type with or without a maculopapular rash and benign Encephalitis(inflammation of brain) Hemorrhagic fevers (usually sever and fatal)
ARBOVIRUSES
TOGAVIRUS
Genus Alphavirus
Spherical 70 nm diameter 42 capsomeres (nucleocapsid ) Genome: 11-12kb positive-sense, single stranded RNA Enveloped 3 or 4 major structural polypeptides (2 glycosylated) Assembly: budding through host cell membranes
FLAVIVIRIDAE
Genus Flavivirus Spherical 40-60 nm diameter Genome: 11kb,Single stranded , positive-sense RNA (INFECTIOUS!) Enveloped 3 structural polypeptides, (2 glycosylated) Assembly: within endoplasmic reticulum
A L P H AV I RU S
Genomic Length
TRANSCRIPTION
Subgenomic (26S)
Viral Replication
Genomelength mRNA
FLAVIVIRUS REPLICATION
Replication: Cytoplasm
Particle Assembly: intracellular vesicles
ANTIGENIC
Alphavirus
Cross-section in immunotechniques (antigenically related) HI,ELISA,IF test define 8 serogroups ID specific virus: Nt tests
Flavivirus
Antigenically related Nt test : ID 8 complexes
PATHOGENESIS
myeloid and lymphoid cells or vascular endothelium CNS (depends on ability of virus)
CLINICAL FINDINGS
Incubation: 4 to 21 days Inapparent infections (common) Mild flu-like illness & encephalitis Sudden onset of severe headache, chills and fever, nausea and vomiting, generalized pain, and malaise. Drowsiness and stuporous (24-48 hrs.) Severe: mental confusion, tremors, convulsions and coma 4-10 days (fever)
LABORATORY DIAGNOSIS
Direct Detection & Recovery
Specimens: blood (early infection), CSF, tissue specimen Common cell lines: Vero, BHK, HeLa, & MRC-5 Mosquito Cell Lines Intracerebral inoculation of suckling mice or hamsters Appropriate biosafety precautions!
SEROLOGY
Neutralizing & hemagglutination-inhibiting antibodies HI test (simplest diagnostic test) ELISA (most sensitive) Serum 1st sample (after onset) 2nd sample (2-3 weeks later)
Cross reactivity must be considered! (Alphavirus & Flavivirus )
Immunity
-permanent after single infection
-immunity may be modified prior exposure to another same group of viruses
Epidemiology
Eastern equine Encephalitis Most severe (hisghest case-fatality rate) Culex tarsalis and birds St. Louis Encephalitis
T I C K B O R N E F L AV I V I R U S A N D M O S Q U I T O - B O R N E F L AV I V I R U S C Y C L E
YELLOW FEVER
Flaviviridae Family Mosquito-borne Local lymph nodes, spleen, & heart,(degenerative changes) liver and kidney(necrotic lesion) , bone marrow
myocardium(shock) [days]
Blood(early onset)
LABORATORY DIAGNOSIS
Virus Detection/Isolation Blood ( first 4 days), tissue specimen (immunochemistry),postmortem tissue (intracerebral inoculation of mice), cell lines Viral antigen/nucleic acid ELISA or PCR
SEROLOGY
IgM (1st week) ELISA (presumptive) Fourfold or greater rise of titer (confirmation) Immunity Neutralizing antibodies (complete protection)
Treatment, Prevention, Control NO antiviral drug therapy Vaccination (most effective treatment) 17D strain and virulent Asibi strain (vaccine) Good antibody response (95%)
TRANSMISSION CYCLE
DENGUE
Breakbone fever Mosquito-borne (Aedes aegypti & Aedes albopictus )
CLINICAL MANIFESTATIONS
4-7 days ( range 3-14 days) Onset fever (2-7 days), malaise, chills, & haedache Pain in back, joints, muscles, eyeballs saddleback form Rash (3rd or 4th day ) lasts up to 1-5 days Enlargement of lymph nodes Mild febrile illness
Dengue hemorrhagic fever/dengue shock syndrome Increased vascular permeability with plasma leakage Pre-existing dengue antibody
LABORATORY DIAGNOSIS
Reverse transcriptase-PCR-based methods(rapid ID) serum Inoculation of mosquito cell line with px serum with
nucleic acid
E/M viral protein-specific capture IgM or IgG ELISA HI test
Immunity
Nt test and molecularbased assays (4 serotypes ) Infection (lifelong protection) Reinfection of virus of different serotype (dengue hemorrhagic fever)
Treatment or Control NO antiviral drug NO vaccines (difficult to develop) DHF: fluid replacement therapy control: breeding places & insecticides
BUNYAVIRIDAE
Spherical 50-300 nm Genome: double-segmented, negative sense and ambisense, singlestranded RNA, 10-14 kb Enveloped 3 or 4 major structural polypeptides, 2 gycosylated Replication: cytoplasm Assembly: budding through host cell membranes
BUNYAVIRIDAE
transovarial transmission :Woodland mosquito(Aedes triseriatus) & sandflies Hosts: squirrels,chipmunks, & rabbits Inactivated: heat, detergents, formaldehyde, and low pH
SANDFLY FEVER
Genus Phlebovirus Bordering Mediterranean sea Female sandfly (Phlebotomus papatasii) Small (pass through nets) and feeds at night Incubation: 3-6 days
small, itching papules (5 days) Headache, malaise, nausea, fever, photophobia, stiffness of neck & back, abdominal pain, and leukopenia. NO specific treatment Insect repellents at night and insecticides
COLORADO TICK FEVER Mountain fever or tick fever Genus Coltivirus (tick) mild, febrile disease without rash 4-6 incubation days Sudden fever and myalgia Headache, muscle & joint pain, lethargy, and nausea & vomiting Self-limited
RODENT-BORNE VIRUSES
Intra-species or interspecies Rodents or bats (reservoir) Contact with body fluids or excretions Examples: Lassa Fever Hantavirus infections South American hemorrhagic fevers
BUNYAVIRUS
Genus Hantavirus (Hantaviruses)
HFRS and HPS Transmissions: inhaling aerosols rodent excreta (urine,feces,saliva)
MANIFESTATIONS:
Fever, headache, and myalgia Progressive pulmonary edema, severe respiratory compromise No hemorrhage
LABORATORY DIAGNOSIS:
Detection of viral nucleic acid (reverse transcriptase-PCR) Immunohistochemistry of viral Ag ( fixed tissues)
THERAPY
Maintain adequate oxygenation of hemodynamic functioning Ribavirin (HPS)
Prevention:
rodent control Avoid inhalation of aerosolized dried excreta
Spherical ARENAVIRUS 50-30nm Genome: double-segmented,negative sense and ambisense; single stranded RNA; 10-14kb Virion has transcriptase 4 major polypeptides Enveloped Replication: cytoplasm Assembly: incorporate ribosomes and bud from plasma membrane
ARENAVIRUS
Old world and New world viruses NW: Grp. A (Pichinde virus) Grp. B (Machupo Virus) Chronic infections (rodents)
LASSA FEVER
Highly virulent Rodent Reservoir: house rat (Mastomys natalensis) Incubation: 1-3 weeks High fever, mouth ulcers, severe muscle aches, skin rashes with hemorrhages, pneumonia, and heart & kidney damage; deafness Fetal death
Standard precautions(hospital)
Avaoid contact contaminated body fluids and blood
LYMPHOCYTIC CHORIOMENINGITIS
Wild house mouse (Mus musculus) Via mouse droppings acute disease Aspetic meningitis or mild systemic inlfuenza-like illness Subclinical infections Serology diagnosis: ELISA (IgM & IgG Ab) Immunochemistry Viral culture (Viro cells) Reverse transcriptase-PCR (viral nucleic acid)
FILOVIRIDAE
Long filaments, 80 nm (varying length) Marburg 665nm Ebola 805nm Genome: negative-sense, Non-segmented single-stranded RNA ;19kb 7 genes Enveloped(glycoprotein) encoded in two reading frames transcriptional editing Virions released by budding (plasma membrane)
Zaire
Sudan Reston Ivory Coast
HIGHLY VIRULENT! (Biosafety Level 4) Destroyed by heating (30 mins.) at 60oC Ultraviolet Gamma-radiation Lipid solvents Bleach Phenolic disinfectants
Marburg virus 1967 lab. Workers exposed to tissues of African green monkeys (Cercopithecus aethiops) Patients to medical personnel transmissions Ebola Virus 1976 close, prolonged contact with patients, blood and excreta -subtypes: Zaire and Sudan (Virulent!) 2008- pigs (Philippines)
LABORATORY DIAGNOSIS:
ELISA for viral antigens (serum) Reverse transcriptase-PCR (clinical specimens) Fresh Virus isolates (Vero and MA-104) Under maximum biologic containment
Reservoir
Rodent or Bat
Human (accidental) Human to Human (direct contact)
PREVENTION:
Isolation facilities Strict barrier techniques Therapy NO antiviral therapy Maintaining renal function & electrolyte balance No vaccine