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Slide 1 - Ebola Virus Microbiology 401 Fall 2007 By: Shahrzad Morim Monica Delgado Janine Gilkes
Slide 2 - Case Study- Ebola Virus VECTOR- the State Research Center for Virology and Biotechnology branch  Biosafety Level 4 Lab Designed specifically to create genetically altered viruses Bioweaponization- potentially aerosolized U.S. was considered “deep target” The former Soviet Union Program: active state-supported bio-weapon research/production (1970’s to mid 90’s) worked with incurable pathogens While working to develop a vaccine against Ebola virus, one of the scientists accidentally stuck herself with a needle contaminated with virus. She contracted Ebola hemorrhagic fever and died.
Slide 3 - Ebola Virus Brief Background Family Filoviridae Genus Ebolavirus History First emerged in 1976 Ebola River Valley, Africa Sub-types (well-known) Zaire Sudan Reston Tai (Ivory Coast) Classification Enveloped SS negative-sense RNA Structure Long, filamentous, “thread-like” structure of a filovirus “U” or “6” appearance
Slide 4 - Ebola Virus Structure-function analysis of the soluble glycoprotein, sGP, of Ebola virus Transmembrane protein, GP(1,2) GP gene encodes the soluble glycoproteins sGP and Delta-peptide. Genome has 7 genes: NP, VP35, VP40, GP, VP30, VP24, and L GP1 Viral Entry GP2 Fusion and Entry Likely pH dependent
Slide 5 - Currently Believed Animal Reservoir of Ebola Virus? Despite extensive studies, the natural and animal reservoir is unknown Seems to be the rain forests on the African continent and in the Western Pacific. Non-human primates as a source of infection for humans Carcasses of gorillas, chimpanzees and duikers during outbreaks in 2001 and 2003 High mortality from infection in these species disqualifies them from acting as reservoirs Other considered Reservoirs Plants, arthropods, and birds IRD researchers have identified bats as a potential natural reservoir of Ebola virus Of 24 plant species and 19 vertebrate species experimentally immunized with Ebola virus, only bats became infected. No clinical signs were observed in these bats This is characteristic of a reservoir species
Slide 6 - Currently Believed Animal Reservoir of Ebola Virus? Current proposal Bats Good vectors If bats are among the culprits Likely to pass virus to great apes  humans May infect humans directly Dry season More contact because of food competition Bats’ immune systems modified Virus reproduces easier In 2001-03 Survey of 1,030 animals (including 679 bats) from Ebola-affected areas Found three bat species Viral genome fragments (RNA) in the liver and spleen Evidence of immune response antibodies against virus in the serum
Slide 7 - Transmission of Ebola Virus Direct contact Blood , secretions , organs Unsterilized needles Burial ceremonies Documented human infections Handling of infected chimpanzees, gorillas, forest antelopes Airborne transmission limited evidence of human-human Incubation period 2 to 21 days Contagiousness Not during early stages As the illness progresses, bodily fluids represent an extreme biohazard
Slide 8 - Symptoms of Ebola Virus Initial Signs Fever (at least 102°F) Weakness & exhaustion Pain Severe headache Muscles & joints Abdominal pain Sore throat Nausea Dizziness Progressed Symptoms Vomiting Diarrhea Extensive bleeding Red eyes hemorrhage of sclerotic arterioles From mouth, nose, eyes, rectum & mucouse membranes Maculopapular rash Spreads over the body (often hemorrhagic) Other secondary symptoms Hypotension , Hypovolemia , Tachycardia Organ damage Internal and external bleeding
Slide 9 - Tissue Damage leads to Hemorrhaging Hemorrhagic fever syndrome late symptoms: toxic shock, hemorrhaging Direct tissue damage liver, combined with massive viremia Disseminated intravascular coagulopathy Endothelial susceptibility Subverts innate and adaptive immune responses Terminal stages diffuse bleeding, and hypotensive shock accounts for many Ebola virus fatalities
Slide 10 - Diagnostic Specialized laboratory test on blood specimens for detection of Antigens Genes of the virus Antibodies against the virus New techniques Non-invasive methods: saliva and urine samples Diagnosing ELISA Assay IgM ELISA PCR Courtesy of: http://www.ncbi.nlm.nih.gov
Slide 11 - Infection Prevention and Lab Safety Precautions Infection Prevention Isolation communication Limit direct contact Monitor those who had lose contact with infected Disinfect reusable equipment Sterilize equipment Lab Safety Precautions Education about organism Sterile environments Protective clothing Proper disposal of waste products Limit contact with contaminated medical equipment Communication
Slide 12 - Current Research On Vaccines One study found that guinea pigs were protected from Ebola virus infection by immunization with plasmids containing the viral genes for either the secreted or transmembrane forms of the viral glycoprotein (GP). This protection was correlated with antibody titer and antigen-specific T-cell responses to secreted GP or membrane GP. Another study found that harmless-Ebola-like particles (eVLPs) could confer immunological protection from Ebola virus infection.  These eVLPs were found to be immunogenic both in vitro and in vivo. Mice were vaccinated with these eVLPs, and developed high titers of Ebola virus specific antibodies, including neutralizing antibodies. Additionally, all the mice in the study were protected from Ebola virus inoculation.
Slide 13 - Bio-Warfare Biological warfare (BW) aka biological weapons, is the use of any pathogen, bacteria or virus as a weapon of war. After initial release of virus, secondary infections may occur as a result of infected individuals traveling from areas of contamination to other locations. In 1972 the Biological Weapons Convention outlawed creation and storage, but not usage, of these weapons.
Slide 14 - Benefits and Hazards of U.S Aid Benefits of US Aid   - Russian Allies - Collaborative efforts in finding a cure for Ebola Decreased Fear A cut of the profits Hazards of US Aid - VECTOR removed from biowarfare threat list; however, 4 other weapons labs exist with no U.S. inspection - Difficult to verify whether former Soviet Scientists are using the American supported research for peaceful purposes - Lack of Accountability
Slide 15 - Which is more dangerous As a Potential Bio-Weapon? Ebola Virus CDC classifies it as: Category A bioterrorism agent Sudden, severe onset of symptoms Believed to hide out in animals No approved vaccine or treatment Short incubation period May kill victim before transmitted to others (high death rate) Outbreaks seem to burn out quickly Smallpox Virus CDC classifies as Category A bioterrorism agent Incubation period between contraction and the first obvious symptoms of the disease is usu. 12-14 days Initially vague, flu-like symptoms, turn into severe symptoms (fever, severe pain, characteristic rash) Humans are the only natural host Historic death rate of 1 in 3 Vaccination in 1967 Effective if administered up to 4 days after viral exposure and before rash appearance No effective treatment In 1979, WHO declared eradication
Slide 16 - Smallpox (Variola Virus) Genus: orthopoxvirus DS DNA: one of the largest viral genomes Relatively stable If aerosolized, infective for at least several hours (without sunlight or UV exposure) Acute contagious disease Two main forms Variola major (30% fatal) Variola minor (<1% fatal) Two rare forms (almost always fatal) Hemorrhagic Malignant
Slide 17 - Which is more dangerous As a Potential Bio-Weapon? CDC: an agent must have the following properties to be used for a maximum credible event Be highly lethal Be easily produced in large quantities Be communicable from person to person Have no treatment or vaccine Smallpox Extremely contagious Transmission through air Can be produced in large scale Can spread in any climate or season Contaminates in a short time Preventive measures/treatments Routine smallpox vaccination was discontinued in the U.S. in 1972 Limited vaccine with questionable potency Vaccine complications
Slide 18 - References  1) Adamcek, K., Eanes, M., Shaw, S., Virology Project: Ebola Virus. Retrieved: 11/8/07 http://www.biosci.ohiou.edu/virology/Ebola 2) Miller, J. Russian Scientist Dies in Ebola Accident at Former Weapons Lab. The New York Times. Published: 5/25/2004. Retrieved: 11/12/2007 http://query.nytimes.com 3) Pappalardo, J. From Russia, with bugs [US subsidizing Russian Biological Warfare Lab]. Dallas Observer. Published: 6/22/2000. Retrieved: 11/14/07 http://freepublic.com 4) “Biological Warfare.” Wikipedia: The Free Encyclopedia. Retrieved: 11/12/07 www.wikipedia.org 5) Emerging Infectious Diseases. National Center for Infectious Diseases. Vol. 5, No.4, July-Aug 1999. Retrieved 11/8/07 http://www.cdc.gov/ncidod/eid/vol5no4/pdf/v5n4.pdf 6) Tanna, J.H, Preventing “Dark Winter”-The Public Health System’s Role in Strengthening National Security. Vol. 1, No.4, Spring 2002. Retrieved 11/8/07 http://www.carnegie.org/reporter/04/preventing/index.html 7) Leroy, E., Fruit Bats a Reservoir for Ebola Virus. Indigo Base, IRD. November 2005. Retrieved 11/8/07 www.ird.fr/us/actualities/fiches/2005/fiche231.htm 8) World Health Organization. Ebola Haemorrhagic Fever. Retrieved 11/8/07
Slide 19 - Refrences http://www.who.int/mediacentre/factsheets/fs103/en/ 9) Hoenen et al. Ebola Virus: Unravelling Pathogenesis to Combat a Deadly Disease. Trends Mol. Med. May 2006, 12(5): 206-215 10) Leroy, EM, Kimulugui, B, Pourrut, X et al. Fruit Bats as Reservoirs of Ebola Virus. Nature. 2005. 438:575–576 11) Pourrut, X, Kumulungui, B, Wittmann, T et al. The Natural History of Ebola Virus in Africa. Microbes and Infection. 2005. 7:1005–1014 12) Retrieved 11/11/07 http://www.nytimes.com/packages/html/health/20030506_Ebola_COMPARE/sci_Ebola_COMPARE_01.html# 13) World Health Organization. Smallpox. Retrieved 11/8/07 http://www.who.int/mediacentre/factsheets/smallpox/en/ 14) September 2003, p. 9733-9737, Vol. 77, No. 18 0022-538 15) BMC Microbiology 2003 3:6   doi:10.1186/1471-2180-3-6 16) www.biomedcentral.com/1471-2180/3/6/figure/F3 www.biosci.ohiou.edu/virology/Ebola/Dectec3.jpg (Courtesy of http://www.ncbi.nlm.nih.gov/ICTVdb/Images/Murphy/ebola_cell.htm 17) World Health Organization  http://www.who.int/mediacentre/factsheets/fs103/en/ Provisional Revision: 2007 18) Leroy, Eric M., et al. Multibple Ebola Virus Transmission Events and Rapid Decline of Central African Wildlife. Science 303: 387 -389. 2004 19) Facts sheet: Community Health Administration http://edcp.org/factsheets/ebola.html Journal of Virology
Slide 20 - Thank you!