1. The Ebola Virus Mario Rodriguez Solivan 200-90-3863

2. What is Ebola?  First discovered in 1976  Severe often fatal viral disease  Human and non-human primates  Lytic life cycle  Does not integrate in host genome  Causes viral hemorrhagic fever  With an incubation period of 2 to 21 days  Internal bleeding

3. Taxonomy  Group: Group V  Order: Mononegavirales  Monos = single(strand)  Negare = negative  ssRNA (-)  Family: Filoviridae  Genus: Ebolavirus  Species: Zaire, Reston, Ivory Coast, Sudan, Bundibugyo

4. Morphology  Pleomorphic  Filamentous  800-14,000 nm in length  80-100 nm diameter  970nm smallest found that can cause infection

5. Genes  7 main segments app. 19kb  NP: structural nucleoprotein  N terminal interacts with the RNA to contact the rest of the proteins in matrix and envelope  VP35: plays a strong roll in mRNA synthesis and replication of the negative strands  VP40: MAIN TARGET for research as of 2012  Bridges the lipid envelope with the nucleocapsid  Mutations or removal of the protein have shown to attenuate the virus

6. Genes  GP/SGP: glycoprotein and secreted glycoprotein  Non-structural can confound the immune system  VP30: important for budding out of membrane  VP24: Inhibits INF-alpha/beta and INF-gamma signaling  L: L-RNA polymerase  Copies the negative strand to make the positive transcripts

7. Gene Map

8. Origin  New serological studies  Fruit bats (Pteropodidae)  Contain specific antibodies to Ebola  Probable Natural reservoir  Chimpanzee, Gorillas, smaller primates  Carriers of the virus  Some strains and species will affect them

9. Ecological Cycle

10. Different Species  Zaire  Congo region of Africa (Zaire)  Most lethal of all species  Up to 90% mortality rate  Sudan  Region of Sudan and Uganda  High mortality rate  App. 53%  Bundibugyo  Bundibugyo District of Uganda  Province Orientale Democratic Republic of Congo  2007 – 2012 App. 36.3% moratality rate

11. Different Species  Reston  Named after Reston, Virginia  First Discovered in crab-eating macaques  Mutation from other Ebola Virus  Mostly non-pathogenic towards humans  Philippines caused human infection  Less aggressive than African Ebola  Extremely hazardous to monkeys

12. Different Species  Côte d'Ivoire  AKA Taï Forest and Ivory Coast  First found in Taï Forest of the Côte d'Ivoire in Africa  Highly infectuous to chimpanzees  Swiss ethologist infected during necropsy on the chimps  “Dengue-like” symptoms a week later  Transported to Switzerland for treatment  2 weeks later she was released from hospital  Did not fully recover for 6 weeks after release

13. Viral Life Cycle 1. Virion entrance  Endocytosis 2. Formation of endosome 3. Endosome binds with lysosome  Formation of endolysosome 4. Acidification of endolysosome  Release genetic material 5. Replication of ssRNA(-)  Form (+) strand 6. Transcription of newly formed ssRNA(+)  Assisted by VP30, VP35 and L protein 7. Translation  mRNA encoding for GP 8. Travel to ER  Where GP is synthesized 9. GP is further modified in Golgi  Delivered to plasma membrane in secretory vesicles 10. All viral proteins now assemble with the membrane associated proteins  Virions bud from cell surface  GP is also secreted

14. Pathogenesis  Hemorrhagic fever  Multisystem syndrome  Damaged vascular system  Internal bleeding  Body regulations abnormal

15. Mode of Attack  Endothelial cells  Walls of vascular system  Cytokines  Inflammation  Hepatic Cells  Swelling  Cytokines  TNF-alpha, IL-6, IL-8  Hemorrhages all over

16. Symptoms  First signs  Headache  Fever  Joint and muscle aches  Weakness  Vomiting  Diarrhea  Fatigue  Advanced signs  Internal bleeding  Orificial bleeding  Cutaneous ruptures  Hepatic inflamation  Cytokines  Renal disfunction

17. Diagnostics  Virus isolation  Monoclonal antibodies that bind to NP  Determine the species  Reverse transcription- PCR  Primers designed specifically for NP region  Real-time quantitative RT-PCR  Green Dye  Primer set used to amplify the L-protein genes  Antigen-capture ELISA  Monoclonal antibodies  NP, VP40, GP  Immunizing mice with rNP  React with epitopes in carboxyl groups at NP terminals  AWESOME FACT!  The rNP of the Reston EBOV could only detect Reston NP… but the Zaire rNP could detect NP from all 5 species!

18. Prevention NO VACCINES AVAILABLE  Avoid traveling to areas of known outbreaks  Check CDC before traveling  Wash your hands frequently  Like any other infectious disease  Avoid bush meat  Wild animal meat in developing countries markets  Avoid contact with infected people  Body fluids, exposed tissues, blood, semen  Follow infection-control procedures  Gloves, masks, eye shields, careful disinfections  Don't handle remains  Dead bodies are still contagious

19. TIM-1 T-cell immunoglobulin and mucin domain 1  This human protein binds to the EBOV GP  Recent Study  Reduction of this receptor reduces infection ARD5  Monoclonal Antibody  Blocked EBOV binding and infection  Anti-TIM-1  New antiviral approach

20. Statistics

21. 1976-2012

22. 2014  http://www.euronew s.com/2014/04/12/w est-africa-doctors- fight-to-contain- ebola-outbreak/  https://www.youtube.com/watch?v=5BRJ PCsUM2w

23. Questions  What kind of genetic material does Ebola have?  A. dsRNA+  B. ssDNA-  C. ssRNA-  D. dsDNA-  E. ssRNA+  What GROUP does EBOV belong to?  A. V  B. III  C. I  D. II  E. IV  In what organelle is GP (glycoprotein) synthesized?  A. Mitochondria  B. Nucleus  C. Golgi  D. ER  E. Lysosome  Why is it suspected that the fruit bat is the natural reservoir for EBOV?...  ANSWER OUT LOUD

24. Citations  http://jvi.asm.org/content/77/3/1793  http://en.wikibooks.org/wiki/Structural_Biochemistry/Glyco proteins/Ebola_Virus  http://edusanjalbiochemist.blogspot.com/2014/04/the- deadly-ebola-virus-frequently-asked.html  http://www.iflscience.com/health-and-medicine/new- antiviral-shows-potential-against-ebola  http://www.elmundo.es/salud/2014/04/16/534d2762ca474 14d4e8b456d.html?cid=SMBOSO25301&s_kw=twitter  http://www.virology.ws/2012/01/18/how-lethal-is- ebolavirus/  https://www.google.com.pr/search?q=province+orientale +&oq=province+orientale+&aqs=chrome..69i57j0l5.5777j0j7 &sourceid=chrome&essm=93&ie=UTF-8  http://www.microbiologytext.com/index.php?module=Bo ok&func=displayarticle&art_id=494  http://www.stanford.edu/group/virus/filo/eboci.html  http://www.pnas.org/content/108/20/8426  http://www.cell.com/cell/abstract/S0092-8674(13)00886-6  http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3485981/  http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3100998/fi gure/fig01/  http://www.cdc.gov/vhf/ebola/outbreaks/guinea/  http://wwwnc.cdc.gov/travel/notices/alert/ebola-guinea  http://www.who.int/csr/don/2014_04_ebola/en/  http://wwwnc.cdc.gov/eid/article/3/1/97-0107_article.htm  http://www.microbiologytext.com/index.php?module=Bo ok&func=displayarticle&art_id=494  http://www.ncbi.nlm.nih.gov/pubmed/24283270  http://www.infectioncontroltoday.com/news/2014/05/ebo la-virus-disease-in-west-africa-226-cases-149-deaths.aspx  http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1459631/  http://www.euronews.com/2014/04/12/west-africa- doctors-fight-to-contain-ebola-outbreak/