the second conference of virology

Egyptian Society of Virology The 6th International Conference of Virology

VIRAL RISKS AND ENVIRONMENTAL CHANGES The Role of National & International Organizations November 29 – December 2, 2016 Hurghada, Egypt

Conference Chairman Prof. Dr. Mohamed A. Shalaby President, Egyptian Society of Virology, Faculty of Veterinary Medicine, Cairo University

Conference Co-Chairman Prof. Dr. Khalid A. El-Dougdoug Faculty of Agriculture, Ain Shams University Prof. Dr. Ali M. Zaki Faculty of Medicine, Ain Shams University

General Secretary Prof. Dr. Aboulata E. Aboulata Plant Pathology Institute, ARC, Giza

Conference Rapporteur General Prof. Dr. Ahmed A. El-Sanousi Faculty of Veterinary Medicine, Cairo University

Organizing Committee 

Prof. Dr. Sayed H. Salem



Dr. Ahmed Kamal



Dr. Ayman H. M. El-Deeb



Dr. Allam Arafat Megahed



Dr. Basem Mohamed A. Ahmed

Scientific Committee 

Prof. Dr. Mostafa A. El-Kady



Prof. Dr. Badawy Othman



Prof. Dr. Ahmed El-Kafrawy



Prof. Dr. Seham Elzeedy



Prof. Dr. Mansour M. Hashem



Prof. Dr. Hussein A. Hussein



Prof. Dr. Ausama A. Yousif



Dr. Ahmed Abd El Wahed

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Sponsors The organizing committee would like to thank the following sponsors of the conference through either advertisement, cash donations, donations in kind, or assisting participants:

1.

Food and Agriculture Organization of the United Nations (FAO)

10. United BioMed (MEVAC)

2.

General Organization for Veterinary Services (GOVS), Egypt

11. Al-Madar co

3.

Animal Health Research Institute (AHRI), NLQP, Dokki, Egypt

12. MSD co

4.

Academy of Scientific Research & Technology (ASRT), Egypt

13. Life Tech co

5.

Al-Hesn Vet Quarantine

14. Pharmachem international

6.

IFT Corporation

)‫الشركة الدولية االولى (المحجر الصحى االقليمى بجيبوتى‬15.

7.

Egyptain Veterinarian Syndicate

16. ALBORAQ Group

8.

Ismailia Misr for Poultry

17. AL-TAREQ Media

9.

Ceva Sante Animale

18. EIC. Comp

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Information about the Egyptian Society of Virology (ESV) Egyptian Society of Virology (ESV) was established in 2000. It is located at Dept. of Microbiology. Faculty of Agriculture, Ain Shams University, P.O. Box 68, Hadaiyk Shaubra, 11241 Cairo, Egypt (The secretary office of ESV is located at the Department of Virology, Faculty of Veterinary Medicine, Cairo University, 12211-Giza, Egypt.), Email: [email protected]. The ESV Website is www.esveg.org, which promotes the exchange of information and stimulates collaboration among scientists active in all aspects of virology. ESV organizes annual meeting, provides representation on national and international council, and organizes Journal of Virological Sciences (JVS). ESV coordinates scientific research, extension service and provides advices for environmental, plant, animal, and human problems. The 6th International Conference of Virology is organized by ESV in collaboration with FAO to shed light on current state of knowledge and research needs in the field of diagnosis, epidemiology and control of emerging viral diseases, new trends in vaccines and vaccinology, virus evolution and diversities, novel diagnostic methods, biotechnology, and control of viral diseases with special emphasis on the roles of international organizations.

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Welcome message to conference guests Dear Colleagues, On behalf of the Directory Board of the Egyptian Society of virology (ESV), we have the honor to welcome you to the 6th International Conference of Virology, to be held in Serenity Beach Makadi Hotel, Hurghada during the period from November 29 - December 2nd, 2016. It is such a blissful occasion to have participants from Arabian and other International Institutions and Organizations representing different disciplines including human, animal, and plant virology, a matter that will be reflected on widening and enriching the scope of sharing knowledge and scientific views. The fast overflow of information on scientific discoveries and the development of technologies, including Bio- and Nano-technology in the field of virology have made it multidisciplinary in term of close linkage with other frontier life sciences such as molecular biology, biochemistry, immunology, epidemiology and other basic and applied biological sciences. Recently, a number of potentially hazardous microbial pathogens have emerged, a matter that represent a real challenge and severe impact on the health patterns of humans, animals and plants with special reference to zoonotic viruses that could be transmitted from animal to man like avian Zika virus, influenza, Foot and mouth disease, Rabies, Ebola, Alkhurma, Rift valley fever and Noroviruses. A large variety of scientific articles that will be presented in this conference will cover the different approaches and applications of bio-Nanotechnology and molecular-based approaches in favor of diagnosis of human, animal and plant viral diseases as well as in vaccine production. ESV directory board members have the feeling that virologists and other biomedical researchers participating in this conference will draw a roadmap that could help in setting up a better strategy for rapid, prompt diagnosis as well as control and eradication of such diseases. ESV wishes you all the best and have fruitful discussions and knowledge sharing, thus giving us the opportunity for formulating applicable recommendations that we hope to be undertaken and considered during setting up the strategic plans for the control of viral diseases in our countries .We would like to thank the scientific and organizing committee, the invited speakers, all participants and sponsors without their help and support this conference would not have been possible. Prof. Dr.Mohamed A. Shalaby President of the Egyptian Society of Virology Professor of virology, Faculty of veterinary medicine, Cairo University Member of the National Comettee of Biological sciences NationalAcademy of Science and Technology, Egypt Email: [email protected] /[email protected]

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Egyptian Society of Virology

THE SIXTH INTERNATIONAL CONFERENCE OF VIROLOGY VIRAL RISKS AND ENVIRONMENTAL CHANGES

The Role of National & International Organization

(November 29 – December 2, 2016) Venue: Hurghada, Desert Rose Resort.

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Schedule-at-a-Glance Tuesday November 29, 2016

Wednesday, November 30, 2016

17:00-20:00

Registeration

09:00-13:00

Opening cermoney, Honorary Speech, Keynote Lecture Scientific Session I: Zoonosis, Human Infections, and Animal Models

15.00-19.15

Scientific Session II (a&b): Vaccines and immunity

19.15 -20.15

Poster Session

16.00-20:00 09:00-11.30 Thursday, December 1, 2016

11.30-13.00 16.00-20:30

Friday, December 2, 2016

10:00-11:00

Training course on the rapid point of need molecular system for the detection of emerging and veterinary pathogens (selected candidates) Scientific Session III: Antiviral Therapy and Potential Antiviral Targets Scientific Session IV: Diagnosis and Diagnostic Tools Scientific Session V: Epidemiology, Evolution and emerging viruses Closing Ceremony & Recommendations

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Keynote lectures 9.30 – 10.00

10.15 – 11.00

Embracing One Health approach: the role of International Organisations “One Health” - A solution to fight emerging viral threats in the future?

11.00– 11.30

Prof. Christopher C. L. Chase South Dakota State University, Brookings, SD USA

Bovine Viral Diarrhea Virus – Is it A Real World Model for Zika Virus Congenital Syndrome?

15.00 – 15.30

Prof. Falko Steinbach Animal and Plant Health Agency Weybridge, Addlestone Surrey KT15 3NB, UK & University of Surrey, Guildford, GU2 7AL, UK.

Challenges for Veterinary Vaccines

15.30 – 16.00

Prof. Mohamed Faisal Michigan State University, East Lansing, Michigan, USA

Wednesday, November 30

9.00 – 9.30

9.30 – 10.00

10.00 - 10.30

Thursday, December 1

Dr. Markos Tibbo Food and Agriculture Organization of the United Nations, Cairo, Egypt Prof. Mathias Neidrig Robert Koch Institut, Berlin, Germany

11.30 – 12.00

16-16.30

16.30-17

17-17.30

Prof. Claus-Peter Czerny Department of Animal Sciences, Georg-August-University Göttingen, Germany. Prof. Kathleen Laura Hefferon Food Sciences Dep. Cornell University Prof. Husein H Aly National Institute of Infectious Diseases, Tokyo, Japan Dr. Ahmed Abd El Wahed Division of Microbiology and Animal Hygiene, Georg-AugustUniversity Goettingen, Germany Prof. Youssry A. Radwan Former professor, FVMCU & WAHO associate member Prof. Alaa Eldin Eissa Dept of fish diseases, Faculty of Veterinary Medicine, Cairo University. Dr. Ahmed M. El-Adly Department of Botany & Microbiology, Faculty of Science, Al-Azhar University, 71524 Assiut, Egypt.

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Viral Hemorrhagic Septicemia Virus (Rhabdoviridae) DNA Vaccination Reveals the Presence of Herd Immunity in the Aquatic Environment Epitope Mapping and Engineering of Site-Directed Protective Recombinant Antibodies New Applications of Plant Virus Nanoparticles Precore Stop Codon Regulates the Stability of HBV X-mRNA The Mobile Suitcase Laboratory: A Tool for the Rapid Detection of Emerging and Endemic Infectious Diseases Equine Herpes Viruses (Risks For Arabian Horses) Emerging Viral Diseases of Aquatic Animals: Regional Perspective Distribution of Hepatitis E Virus Genotypes from Human and Animal reservoirs

Dr. Markos Tibbo, is regional Livestock Officer at the United Nations Organization for Food and Agriculture (FAO) based in the Regional Office for the Near East and North Africa. He advises 20 countries of the region in sustainable improvement of production and productivity, safe and hygienic production of animal source foods, dairy value chain, safe livestock trade and ensuring food security of farmers and pastoralists, rural livelihoods and employment (with focus in rural women, youth and the vulnerable). He earned a PhD in Animal Science (especially Genetics and Breeding) from the Swedish University of Agricultural Sciences in Uppsala, Sweden and a DVM from Addis Ababa University (Addis Ababa, Ethiopia). Dr Tibbo has over 23 years of experience in research (at CGIAR centres) and development (at FAO and NGOs) in Sub-Saharan Africa, West Asia, South Asia and North Africa. He has authored or co-authored more than 120 publications. His areas of competence include animal health and production, adaptation to climate change, smallholder dairy, animal genetic resources; sheep and goats, animal feeds, infectious and trans-boundary animal diseases and zoonoses. He received awards for leading outstanding research team, contributions to science and professional societies. Prof Niedrig graduated in 1985 from the Universities of Oldenburg, Göttingen, West Berlin (Germany), Staff position for setting up a One Health agenda for RKI, Attain his Ph.D., which performed at Robert Koch-Institute, Berlin 1988, till 1993 attain position as Laboratory chief at Behringwerke AG, Marburg (Germany), working on characterisation of a HIVVaccine safety of blood products, development of in-process controls for vaccine production (Polio, TBE, Rabies), project manager for TBE-Vaccine (Encepur) attain position as Laboratory group leader at Robert Koch-Institute, Berlin, responsible for Yellow fever virus vaccine production, research on diagnostic of imported viruses, Establish a European Network for Diagnostics of Imported viral Diseases (ENIVD, project coordinator,And responsible for Tick borne encephalitis (TBE) reference laboratory of Germany at 2008 and he is Experienced in Development of various diagnostic tests, and Broad experience in analysis of immune response after vaccination. Performance of external quality assurance studies on PCR serology diagnostic for rare and imported viral diseases. Prof. Chris Chase is a professor of veterinary science at South Dakota State University in Brookings, SD. He was involved in clinical practice and consulting for almost 30 years. His research and diagnostic interests are in bovine and porcine immunology, respiratory disease pathogenesis, emerging bovine diseases with wildlife interface and vaccine development. Dr. Chase is the President of the American College of the Veterinary Microbiology. Dr. Chase cofounded RTI (Research, Technology Innovation) LLC. An animal health contract research organization founded in 1994 and has served as President since 1998.

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Prof Steinbach graduated as a veterinarian in 1990 from the Faculty of Veterinary Medicine in Berlin, Germany, and subsequently obtained his post-graduate degrees there (1994, 2004). After a time as post-doctoral researcher at the Institute of Virology at the Free University of Berlin, he moved to the Leibniz Institute for Zoo and Wildlife Research, also in Berlin, to lead the immunology team. In 2005, he moved to what is now known as the Animal and Plant Health Agency (APHA), Weybridge. In 2001, he was a founding member of the European Veterinary Immunology Group (EVIG), which he chaired until 2012, and has organized national and international meetings for veterinary immunologists. He is a member of several national and international scientific societies and a member of the editorial board for two journals (TBED, JMMCR). In July 2014, he was appointed Professor for Veterinary Immunology at the School of Veterinary Medicine in the Faculty of Health and Medical Sciences at the University of Surrey. Prof. Mohamed Faisal is a professor of aquatic animal medicine at the Department of Pathobiology and Diagnostic Investigation of the Michigan State University (MSU)College of Veterinary Medicine and the Department of Fisheries and Wildlife of MSU-College of Agriculture and Natural Resources. Dr. Faisal joined MSU in March of 2001 and prior to that he was a professor at the School of Marine Science, The College of William and Mary, Virginia, USA. He received his doctoral degrees from the University of Ludwig-Maximillian, Munich, Germany. As a veterinarian, he devoted his career to the study of diseases affecting aquatic animals (both freshwater and Marine). In specific, he is interested in deciphering the mechanisms of the pathogen-host interactions; how the pathogens invade its host and overcoming its immune system and how the host reacts and combat the intruding microbes. Over a career that extended 37 years, Professor Faisal has made several milestone discoveries that advanced his field of specialization. During his relatively short tenure at MSU, he was successful in establishing the Aquatic Animal Medicine Program that integrated MSU, Michigan Department of Natural Resources, Michigan Department of Agriculture, and the Great Lakes Fishery Commission. Dr. Faisal’s research enabled him to develop several important tools that facilitated the diagnosis of emerging infections and finding promising targets for the biological control of invasive species. He is instrumental in providing guidance to the Great Lakes states on how to manage their fishery resources in the presence of emerging pathogens. At the international level, Dr. Faisal is the Lead Scientist and CoFounder of the Living Oceans Foundation, which is undertaking important research to mitigate the effects of diseases on coral reef biodiversity. He is also on the International Pollution Responses in Marine Organisms Association and is Consultant to the Secretary General of the UN on pollution issues. He established a central laboratory for fish disease diagnosis in Senegal 2008. Dr. Faisal has been recognized by a number of national and international awards and honorary degrees. For his outstanding research serving the community of Aquatic Animal health for several decades, Dr. Faisal has been selected as SF Snieszko Distinguished Fellow and Professor. Most recently, Dr. Mohamed Faisal has received the 2015 Jack Christie/Ken Loftus Award for his contribution toward understanding healthy great lakes ecosystems. 10

Prof. Dr. Czerny is the managing director of the Institute of Veterinary Medicine, University of Goettingen, Germany. He is also the chair of the Division of Microbiology and Animal Hygiene with a research focus on poxviruses, paratuberculosis and immunogenetics. He has published 65 international publications in peer-reviewed journals

Dr.Kathleen Hefferon received her PhD from the Department of Medical Biophysics, University of Toronto and continued her post-doctoral studies at Cornell University. Dr. Hefferon has worked on faculty at the Division of Nutritional Sciences at Cornell and has written two books on biopharmaceuticals in plants. She teaches and conducts research at both the University of Toronto and in the Department of Food Sciences, Cornell University. Kathleen has 4 patents, has edited 6 books, and has multiple research publications. Kathleen currently lives with her family near Ithaca NY.

Prof Hussein graduated as a veterinarian in 1995 from the Faculty of Veterinary Medicine, Cairo University, Egypt, and subsequently obtained his post-graduate degrees master (1999), Ph.D. (2007) in Human Tumor Viruses Laboratory, the institute for virus research, Kyoto University, Kyoto, Japan. Senior Research Scientist Virology II department, National Institute of Infectious Diseases, National Institute of Health, Toyama, Shinjuku, Tokyo, Japan, Interested in viral hepatitis, Molecular biology, and Basic Science, Viral Immunology, Cancer Biology, He is a member of several national and international scientific societies, The Japanese society for Virology, Molecular Biology society of Japan. Dr. Abd El Wahed studied veterinary medicine at Mansoura University, Egypt. He received his PhD in biology from Goettingen University, Germany on the use of peptide microarrays for mapping viral B cell epitopes. Currently, he is the head of Virology laboratory at the Division of Microbiology and Animal Hygiene, Goettingen, Germany. He has participated in the development of 30 point-of-care assays for the detection of infectious agents. In 2013, he was awarded the young investigator award from the ASTMH on the establishment of a mobile laboratory for rapid detection of hemorrhagic fever viruses at low resource settings. Recently, he established a mobile suitcase laboratory for rapid detection of viruses and parasites. The mobile setup was in field trials in Guinea, Senegal, Bangladesh and Brazil. 11

Prof. Youssry Chairman of Association for Development of Arabian Horses and Keeping of Their Genetics,Giza, Egypt, graduated as a veterinarian in 1972 from the Faculty of Veterinary Medicine in cairo Uni, Egypt, and subsequently obtained his post-graduate degrees there (1976, 1979), Professor at 1982, and Head of Department of Veterinary Medicine & Surgery, Faculty of Veterinary Medicine, Tripoli Libya, Assistant Professor, and Full Professor, Department of Medicine and Infectious Diseases Faculty of Veterinary, Cairo University, Egypt, Professor of Equine Medicine and Therapeutic, Clinical Studies Department, College of Veterinary Medicine and Animal Resources, King Faisal University, Saudi Arabia, Animal Health Consultant Expert, Ministry of Environment, Doha, Qatar. Alaa Eldin Eissa has earned a Ph.D. in Aquatic Animal Health from Michigan State University, USA in 2005 and a Masters in Vet Sciences “Fish Diseases” from Cairo University in 1997and a Bachelor of Veterinary Sciences from Cairo University in 1993. He is currently working as Professor of Fish Diseases and Management at Faculty of Veterinary Medicine, Cairo University. Dr. Eissa has over 20 years of experience in fish/ shellfish diseases diagnosis, aquaculture health management, aquatic environment risk assessment and rehabilitation of degraded aquatic wildlife habitats. With this unique mix of expertise, Alaa has excelled in delivering leading-edge aquaculture project development and management know-how to clients in USA, Canada, Egypt and Libya.Dr. Eissa was also certified as Molecular Laboratory Diagnostician (graduate diploma in Molecular Pathology from Michigan State University, USA). For his profound renowned expertise in the field of Aquatic Animal Health , Aquatic diversity conservation and Environmental rehabilitation Dr. Eissa has been selected as a council member of the Strategic and planning committee at The Egyptian Center for the Advancement of Science, Technology and Innovation (ECASTI). For distinction & innovation in aquaculture health management Dr. Eissa was awarded the “National Encouragement Award in Agricultural Sciences” from the Supreme Council of the Egyptian Academy of Science and Technology for the year 2008 and the “Scientific Distinction Award in Interdisciplinary and Multi-specialty Sciences” from the Supreme Council of Scientific Research of Cairo University for the year 2010. Dr. Ahmed Mohammed El-Adly studied at the Botany and Microbiology Dept. Faculty of Science, Al-Azhar University, Assiut branch, Egypt. He obtained PhD in Virology at the Microbiology and Virology dept., Faculty of medicine, Russian Peoples' Friendship University, Moscow. He has a research focus on HEV.

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Scientific Program All talks will be held in Hotel Conference Auditorium, Desert Rose Resort Wednesday, November 30, 2016 9.00 - 9.10: Opening Session- Chairman Welcome Professor M.A. Shalaby "Conference Chairman" 9.10 - 9.20: Welcome Speach Professor Khaled Farouk El-Amry "Head of Egyptian Veterinarians Syndicate" 9.20 - 9.30: Honorary Speech, Dr. Hussein Gadain, FAO representative in Egypt. 9.30 - 10.00: Opening Keynote Lecture, Dr. Markos Tibbo “Embracing One Health approach: the role of International Organisations” (Abst. No. 1) Scientific Session I: Zoonosis, Human Infections, and Animal Models Chairman Prof. Matthias Niedrig Co-Chairmen Prof. Christopher C. L. Chase and Prof. Ali Zaki Keynote Lecture: “One Health” - A solution to fight emerging viral threats in the future? 10.15 – 11.00 Mathias Neidrig Robert Koch Institut, Berlin, Germany (Abst. No. 2) 11.00 – 11.15 Break Keynote Lecture: Bovine Viral Diarrhea Virus – Is it A Real World Model for Zika Virus Congenital Syndrome? 11.15– 11.45 Christopher C. L. Chase South Dakota State University, Brookings, SD USA (Abst. No. 3) Seroprevalence of Falviviruses, Chikungunya virus and Rift Valley fever virus, Saudi Arabia 11.45 – 12.00 Ali M. Zaki (Abst. No. 4) 3-D structure prediction and analysis of the p7-transactivated protein1 of hepatitis C virus 12.00 – 12.15 Mahmoud M. El Hefnawi1, Mohamed E. Hasan, Amal Mahmoud, Yehia A. Khidr, El-Sayed A. El-Absawy, Alaa A. Hemeida (Abst. No. 5) Detection and comparative analysis for the tandem repeats motifs in the completed genomes of human coronavirus (hku1) by using regular expression finder tool 12.15 – 12.30 Mohamed A. Ezz, Amal M. Hussein, Fawzy A. Elfeki, and Alaa A. Hemeida (Abst. No. 6) 12.45 – 13.00 Lunch break 13.00 - 15.00

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Scientific Session IIa: Vaccines and immunity Chairman Prof. Falko Steinbach, Prof. Mohamed Faisal CoChairmen Prof.Hamed Mazyad, Prof. Seham El-Zeidy Keynote Lecture: 15.00 – 15.30

Challenges for Veterinary Vaccines Falko Steinbach Animal and Plant Health Agency Weybridge, Addlestone Surrey KT15 3NB, UK & University of Surrey, Guildford, GU2 7AL, UK (Abst. No. 7) Keynote Lecture:

15.30 – 16.00

16.00 – 16.15

16.15 – 16.30

16.30 – 16.45

16.45– 17.00 17.00 – 17.15 17.15 – 17.30

17.30 – 17.45

17.45 – 18.00

Viral Hemorrhagic Septicemia Virus (Rhabdoviridae) DNA Vaccination Reveals the Presence of Herd Immunity in the Aquatic Environment. Mohamed Faisal Michigan State University, East Lansing, Michigan, USA (Abst. No. 8) Recombinant TMV vector for development of highly immunogenic HSV- 2 antigens used as a plant-based vaccine candidate Ahmad K. El-Attar, Josefine Persson, Aboul-Ata E. Aboul-Ata, Hamed H. Mazyad, Ali M. Harandi, and Olof Olsson (Abst. No. 9) Generation of novel fusion gene-dependent DNA vaccine candidates for protection against human respiratory syncytial viruses: the role of codon optimization, truncation and immunodominant epitopes. Mohamed A. Farrag, Haitham M. Amer, Peter Öhlschläger, Fahad N. Almajhdi (Abst. No. 10) Generation of a reassortant avian influenza virus H5N2 vaccine strain capable of protecting chickens against infection with Egyptian H5N1 and H9N2 viruses. Kandeil A, Moatasim Y, Gomaa MR, Shehata MM, El-Shesheny R, Mostafa A, Barakat A, Webby RJ, Ali MA, Kayali G (Abst. No. 11) Influenza Virus Polymerase Basic 1 (PB1) Origin is A Key Determinant for High-Yield Seed Vaccine Production Ahmed Mostafa, Pumaree Kanrai, John Ziebuhr, Stephan Pleschka (Abst. No.12) Break The use of different types of Montanide adjuvant in preparation of inactivated rabbit haemorrhagic vaccine Owais, G. A. Salman and Samah, E. Abodalal (Abst. No.13) Novel aluminum hydroxide nanorods elicit enhanced immune responses to inactivated genotype VIId NewCastle disease virus vaccine formulations Walaa A. Mohammad, Ausama A. Yousif, Ahemd A. Farghali, Sama I. EL Dek, Abo Zeid A. Abo Zeid, Mohamed A. Shalaby and Ahmed A. El-Sanousi. (Abst. No. 14) Preparation and Immunological Evaluation of Inactivated Avian Influenza Virus Vaccine Encapsulated in Chitosan Nanoparticles in Chickens Samah H. Mohamed, A. Arafa, W. H. Mady, Rania Morsy, Hanan Aly, Lamiaa Omar (Abst.14No. 15)

Scientific Session IIb: Vaccines and immunity Chairman Prof. Falko Steinbach CoProf. Mansour Hashim, Prof. Fahd El-Majhdi Chairmen Naked DNA Vaccination with Full-Length Attachment Gene of Human Respiratory Syncytial Virus Saudi Strain Induces Protective 18.15 – Immune Response with No Enhancement of Pulmonary Inflammation 18.30 Maaweya E. Hamad, Haitham M. Amer, Mohamed A. Farra, Fahad N. Almajhdi (Abst. No.16) Field application of trivalent foot and mouth disease vaccine 18.30 – adjuvanted with Zeolite. Hiam M Fakhry; Assem A Mohamed; Sonia A Risk; Akram Z 18.45 Hegazy and Abobakr M Agor. (Abst. No. 17) The immunogenicity and protective efficacy of DNA vaccine coding for full length H5 gene of avian influenza H5N1 subtype 18.30 – Yousef A. Soliman; Eman, M.S. El- Naga; Maha A. Gamal; Salah 18.45 A. Selim; Mohamed M. El-Hady and Fekria A. El-Bordeny(Abst. No. 18) Evaluation of vaccination with local and imported vaccine against foot and mouth disease virus in Kalubeya governorate 18.45 – 19.00 Lamya, A.F.Ateya, Ahmed, S. A, Khames, A.S, H.A. Abdel-Hady1 (Abst. No. 19) Vaccination of potato plants against Alfalfa mosaic virus (AMV) by using mild strain vaccine of Potato virus Y in Upper Egypt 19.00 -19.15 Safynaz A. Mohamed , and F.G. Fahmay (Abst. No. 20) 19.15 – Poster Session 20.15

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Poster Session 19.15-20.15 Molecular, clinico-pathological and sero-diagnosis of LSDV in cattle in Sharkia and Fayoum Governorates Nashwa M. Helmy, Ahmed, S. A. and Zeinab, Y. Mohamed (Abst. No. 21) Lyophilized EHV-1 vaccine inactivated and adjuvanted by Quillaja Saponaria Molina extract Nehal S. Saleh, Eman M. Ebeid, Fatma F. Warda, Eldakhly A.T., Nashwa K Madkour, Elkabany M.A. and Soliman I.M.A. (Abst. No. 22) Some molecular assays used for identity of PPR vaccines Manar F. Seioudy, Magda M. Sayed, Ahmed A. El-Sanousi and Mohammed A. Shalaby (Abst. No. 23) Isolation and identification of lumpy skin disease virus in cattle in Kalubeya Governorate Lamya, A. F. Ateya1, Ahmed, S. A., Mansour, A, H Khames, A. S. H. A. Abdel Hady (Abst. No. 24) Molecular Characterization and Confirmation of Tomato Spotted Wilt Virus Associated with an Asymptomatic Infected Chrysanthemum Plant in Egypt Ahmed K. El-Attar, Manal A. EL-Shazly and Samah A. Mokbel (Abst. No. 25) Induction of resistance in pepper plants against Potato Virus Y (PVY) by two medicinal and aromatic plant essential oils and its major oil components. Radwa M. Shafie, Ahmed A. Kheder and Amal A. Farghaly (Abst. No. 26) Studding the physical, morphological and inclusion bodies properties of Bean yellow mosaic virus A. Wardany A., Usama M. A., Sabry Y. M. Mahmoud, Shaddad, M. A. K, Raafat F. A., (Abst. No. 27) Title: Development of multi-resistant tomato hybrids for heat and Tomato Yellow Leaf Curl Virus (TYLCV) Amro A. Farrag; Essam M. E. A. Khalil; Ahmed K. EL-Attar; A. A. Kheder and H. M. Mazyad (Abst. No. 28) Impact of Viruicide Carrageenan in Suppression of Potato Virus Y in Potato Plant Hanaa H. Gomaa (Abst. No. 29) The plant growth promoting fungus Penicillium sp. GP16-2 enhances the growth and confers protection against Cucumber mosaic virus in tobacco Mohsen M. Elsharkawy, Jehan M. Abass, Said M. Kamel and Mitsuro Hyakumachi (Abst. No. 30) Elimination of Alfalfa mosaic virus (AMV) from infected leaf potato (Solanum tuberosum) cv. Ditta by embryonic calli Maha. A. El-Abhar, Mostafa. A.S. El-Kady; Khaled. M. Ghanem and Hussieny A. Bosila (Abst. No. 31) Investigation of virus/meristem interaction by using Cre-virus vectors Mayada A.M., Badawi A.A. Othman, Taha R.M., Ali R.M and Schiemann J (Abst. No. 32) 16

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Isolation, pathotyping and genotyping of Newcastle disease virus from broiler chickens in Egypt. Ayman, S. El-Habbaa, Gabr, F. El-Bagoury, Samar, F. El-Adaway, and Susan, S. El-Mahdy (Abst. No. 33) A genotyping of a new avian infectious bronchitis virus isolated from chickens proventriculus in Egypt E.M. El-Nahas, H.S. El-Sayed, S.S. El-Basuni, G.F. El-Bagoury (Abst. No. 34) Avian influenza virus vaccine match against circulating H5N1 avian influenza viruses in Suez Canal region, Egypt Amira Mohamed Helal, Abdel Satar Arafa, Hanan Fathy Abdein, Dalia Mansour Hamed and Mohsen El Dimerdash (Abst. No. 35) Variable Pathogenicity of Avian influenza viruses H5N1 isolated from Egypt Ola. A. Khader., Arafa. A, Hassan, M. K., A.A. ElSanousi, M. A. Shalaby (Abst. No. 36) Trial for using emergency vaccination for controlling duck virus hepatitis (DVH) outbreaks in ducks Sabry, E.O., Hemat, S. Elsayed, Naglaa Hagag, Wesam Mady (Abst. No. 37) Evaluation of avian influenza H5N1 plasmid-based inactivated Egyptian vaccine and treatment with antiviral drugs Omnia M. Hassan, Ahmed Mostafa, Ahmed Kandeil, Rabeh El shesheny, Ali M. Ahmdy, Mohamed A. Ramadan, Mohamed A. Kutkat, Mohamed A. Ali (Abst. No. 38) In silico design and vitro validation of efficient si-RNAs as universal silencers of the polymerase genes of influenza A virus Sara H. Mahmoud, Rabeh El Shesheny, Mahmoud El Hefnawi, Naglaa Abdallah, Abdelhadi Abdallah, Ahmed Mostafa, Mohamed Ali (Abst. No. 39) Reassortment between avian highly pathogenic H5N1 and low pathogenic H9N2 influenza viruses and its impact on pathogenicity and infectivity in chickens Yassmin Moatasim, Ahmed Kandeil, Ahmed Mostafa, Sary Khaleel Abd elghaffar, Rabeh El Shesheny, Ghazi Kayali, Ahmed H. M. Elwahy, Mohamed Ahmed Ali (Abst. No. 40)

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Thrusday, December 1, 2016 Scientific Session III: Antiviral Therapy and Potential Antiviral Targets Chairman Prof. Claus-Peter Czerny CoDr. Kathleen Laura Hefferon, Prof. Aboulata E. Aboulata Chairmen Keynote Lecture: 9.00 – 9.30

Epitope mapping and engineering of site-directed protective recombinant antibodies Claus-Peter Czerny Department of Animal Sciences, Georg-August-University Göttingen, Germany. (Abst. No. 41) Keynote Lecture:

9.30 – 10.00

New Applications of Plant Virus Nanoparticles Kathleen Laura Hefferon Food Sciences Dep. Cornell University (Abst. No. 42) Keynote Lecture:

10.00 - 10.30

10.30 – 10.45

10.45 – 11.00

11.00- 11.15 11.15 – 11.30

Precore stop codon regulates the stability of HBV X-mRNA Husein H Aly National Institute of Infectious Diseases, Tokyo, Japan. (Abst. No. 43) Interleukin-27 Is Differentially Associated with CD4+ T Cell Counts in HIV Saudi patients under Highly Active Antiretroviral Therapy Nagwa M. Aref, Haifa M. Al-Nafea and Nuha M. Hamdy (Abst. No. 44) Dispersed gold nanoparticles potentially ruin Gold Barley Yellow Dwarf Virus and eliminate virus infectivity hazards Nagwa M. Aref and Noorah A.Alkubaisi (Abst. No. 45) In vitro Studies on antiviral activity of silver nanoparticles against Foot and Mouth Disease virus. Sonia A. Rizk; Safy E. Mahdy; Amr I. Hasanin; Hiam M Fakhry and Ahmed F. Ramadan. (Abst. No. 46) Break

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Scientific Session IV: Diagnosis and Diagnostic Tools Chairman Prof. Mohamed A. Shalaby CoProf. Khaled A. Dougdoug, Chairmen Dr. Ahmed Abd El Wahed Keynote Lecture: 11.30 – 12.00

12.00 – 12.15

12.15 – 12.35

12.35 – 12.50

12.50 – 13.00

The mobile suitcase laboratory: A tool for the rapid detection of emerging and endemic infectious disease Ahmed Abd El Wahed Division of Microbiology and Animal Hygiene, Georg-AugustUniversity Goettingen, Germany (Abst. No. 47) No additional value for the detection of Zika virus from urine of Brazilian patients in the first five days of infection Rodrigo Pessôa, João Veras Patriota, Maria de Lourdes de Souza, Ahmed Abd El Wahed and Sabri S. Sanabani (Abst. No. 48) Rapid and Quantitative Detection of some food born Viruses by RealTime Reverse Transcription-PCR in Egypt Randa M. farag, S. M. E. A. Amer, K. A. El Dougdoug (Abst. No. 49) Analyzing infection of chicken oviduct explant cultures of differentiated oviduct epithelial cells by infectious bronchitis virus in vitro Sahar Abd El Rahman, Ali El Kenawy, Christine Winter, Ulrich Neumann, and Georg Herrler (Abst. No. 50) General discussion

Lunch break 13.00- 16.00

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Scientific Session Va: Epidemiology, Evolution and emerging viruses Chairman Prof. Ahmed El- Kafrawy CoProf. Sayed Salem Chairmen Prof. Yosry A Radwan Keynote Lecture: 16.00 – 16.30

Equine Herpes Viruses (Risks for Arabian Horses) Youssry A. Radwan Former professor, FVMCU & WAHO associate member (Abst. No. 51) Keynote Lecture:

16.30 – 17.00

Emerging Viral Diseases of Aquatic Animals: Regional perspective Alaa Eldin Eissa Dept of fish diseases, Faculty of Veterinary Medicine, Cairo University. (Abst. No. 52) Keynote Lecture:

17.00 – 17.30

17.30 – 17.45

17.45 – 18.00

18.00 – 18.15

Distribution of Hepatitis E Virus Genotypes from human and animals reservoir Ahmed M. El-Adly Department of Botany & Microbiology, Faculty of Science, Al-Azhar University, Assiut, Egypt. (Abst. No. 53) Predominance and Geo-Mapping of avian influenza H5N1 in poultry sectors in Egypt Abdelsatar Arafa, Ihab El-Masry, Shereen Kholosy, Mohammed K. Hassan, Mousa Soliman, F. O. Fasina, Gwenaelle Dauphin, Juan Lubroth, Yilma Makonnen (Abst. No. 54) Surveillance for Middle East Respiratory Syndrome Coronavirus in camels from 2014 to 2016 in Egypt Mahmoud M. Shehata, Ahmed N. El-Taweel, Mokhtar R. Gomaa, Ahmed Kandeil, Rabeh El-Shesheny, Ahmed S. Kayed, Basma Elsokary, Nagla Hassen, Ola Bagato, Yasmin Moatasem, Sara Husein,Omnia Kutkat, Asmaa M. Maatouq, Pamela P. McKenzie, Ahmed Othman, Richard J. Webby, and Ghazi Kayali, Mohamed A. Ali (Abst. No. 55) Break

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Scientific Session Vb: Epidemiology, Evolution and emerging viruses Chairman Prof. Aly M. Abdel-Salam. CoProf. Hussein Ali Hussein Prof. Haitham M. Amer Chairmen

18.15 – 18.30

18.30 – 18.45

18.45 – 19.00

19.00 – 19.15

19.15 – 19.30

19.30 - 19.45

19.45 – 20.00 20.00 – 20.15

20.15 – 20.30

Molecular characterization of a Sweet Potato Leaf Curl Virus isolate from Egypt and its phylogenetic relationship with other members of the Begomovirus Aly M. Abdel-Salam, Malik Mujaddad-ur-Rehman, Salama M. ElSaghir, and Manal A. El-Shazly (Abst. No. 56) Molecular description and topotyping of the isolated FMDV in Egypt during 2015. Saudy A. F. and Safy Mahdy (Abst. No. 57) Genetic variability of human metapneumovirus among hospitalized children with acute respiratory infections in Saudi Arabia Haitham M. Amer, Mohamed A. Farrag, Maaweya E. Hamad, Fahad N. Almajhdi (Abst. No. 58) Isolation of H5 Highly Pathogenic Avian Influenza from Cattle Egret (Bubulcus Ibis) in the Vicinity of Affected Broiler Chicken Flocks in Egypt Moustafa M. El-Shazly, Basem M. Ahmed, Ahmed A. El-Sanousi, and Youssef I. Youssef (Abst. No. 59) Lemon grass Cymbopogon citratus as a host plant for Cucumber mosaic virus (CMV) in Upper Egypt Safynaz A.E. Mohamed, and F.G. Fahmy (Abst. No. 60) Mutation signature in Neuraminidase gene of avian InfluenzaAIVH9N2/G1 in Egypt Zienab Mosaad1; Abdelsatar Arafa1 Hussein A. Hussein, Mohamed A. Shalaby (Abst. No. 61) A new, widespread Emaravirus discovered in blackberry Hassan, M., Di Bello P.L., Keller, K.E., Martin R.R., Sabanadzovic S. and Tzanetakis, I. E. (Abst. No. 62) The co- infection of cucurbits with criniviruses and ipomoviruses: possible adverse effect on virus diagnosis and breeding for resistance. Aly M Abdel-Salam (Abst. No. 63) Molecular Characterization of the Newcastle Disease Virus detected in some provinces of The West Delta in Egypt Elham S.M. Ragab, Mohammed A. AboElkhair, Zakaria R. Elanawaty, Mahmoud G. Ibrahim, Hesham A. Sultan (Abst. No. 64)

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Friday, December 2, 2016 Closing Ceremony & Recommendations Prof. Dr. MA. Shalaby, Prof. Dr. Ahmed El kafrawy, Prof. Dr. Ali Zaki, Prof. Dr. Aboul-Ata El-Nady, Prof. Dr. Hussein. A. Hussein, Prof. Dr. Ausama A. Yousif, Prof. Dr. Sayed Salem, Prof. Dr. K. El-Dougdoug, Prof. Dr. Badawy Othman, Prof. Dr. Seham El Zeidy, Prof. Dr. Mansour Hashim. 10.00 – 10.30

Closing Ceremony.

10.30 – 11.00

Recommendations.

11.00 – 12.00

Check out & departure of delegates

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SESSIONS AND ABSTRACTS

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1. Embracing One Health approach: the role of International Organisations Markos Tibbo1* and Juan Lubroth2 1 Food and Agriculture Organization of the United Nations, Cairo, Egypt 2 Food and Agriculture Organization of the United Nations, Rome, Italy Over 60 percent of human infections originate in animals and 75 percent of emerging diseases are zoonotic – a disease that can be transmitted from animals to people or, more specifically, a disease that normally exists in animals but that can infect humans. In 2010, concerned from the increasing number of emerging infectious viral diseases and pandemic threats at the animal-human interface, international organisations (FAO, OIE and WHO) confirmed their commitment to a One Health approach to address the multiple factors influencing the emergence of infectious diseases. The approach is a collaborative, international, cross-sectoral, multidisciplinary mechanism to address threats and reduce risks of detrimental infectious diseases at the animal–human–ecosystem interface. A number of diseases emerged recently: Nipah virus, Severe Acute Respiratory Syndrome (SARS), Highly Pathogenic Avian Influenza virus (H5N1, H9N7), Influenza H1N1, Middle East Respiratory Syndrome Coronavirus (MERS-CoV) whereas others re-emerged (e.g. Ebola virus and Zika virus) heightening worldwide public awareness of the multidimensional linkages between wild animals, livestock production and global public health. Main global drivers that are amplifying these events include high human population growth, mobility of people, climate change, evolution and dynamics of food systems and agriculture, and encroachment of forest and game reserves. Today, pathogens can travel the world in less than a day, emerging diseases of humans, livestock or wildlife can have severe implications for public health, livelihoods, food security, international trade and tourism. Joint efforts of international organisations have been developing partnerships with implementing entities and advocating for the One Health approach in all measures that pertains to infectious diseases at the human-animal-ecosystem interface and other important public health concerns such as antimicrobial resistance and food safety. Following the Tripartite Agreement, systems such as GLEWS – the Joint FAO-OIE-WHO Global Early Warning System for health threats and emerging risks at the human– animal–ecosystems interface and OFFLU – the OIE-FAO global network of expertise on animal influenza for effective collaboration among animal health experts and the human health sector, have been established. The implementation of the approach at national level, however, has been sluggish. In 2014, the country-led Global Health Security Agenda was launched, which pursues multilateral and multi-sectoral approach to strengthen both the global capacity and nations’ capacity to prevent, detect, and respond to human and zoonotic infectious diseases threats whether naturally occurring or accidentally or deliberately spread. Among few success examples of the use of One Health approach are on Rabies control in Sierra Leone and Togo, and RVF control in Madagascar, Sudan and Tanzania. For RVF, for example, the success factors have been the establishment of a coordinated interministerial management taskforce; communication, social mobilization and at-risk population awareness; strengthening of case reporting and surveillance systems; human case management; and livestock management, i.e. animal movements, slaughtering and vaccination. Although the One Health approach – built with understanding that disease risks resulting from interactions between animals, humans and the environment need to be addressed through greater inter-sectoral collaboration, coordination among international organisations and concerned ministries of member countries – its realisation has been slow compared to the magnitude and levels of emerging pandemic threats.

* Correspondence: Food and Agriculture Organization of the United Nations, Regional Office for the Near East and North Africa, 11 Al-Eslah El-Zeraie Street, 12311 Dokki, PO Box 2223, Cairo, Egypt; [email protected]

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Scientific Session I: Zoonosis, Human Infections, and Animal Models Chairman Prof. Matthias Niedrig Co-Chairmen Prof. Christopher C. L. Chase and Prof. Ali Zaki Keynote Lecture: “One Health” - A solution to fight emerging viral threats in the future? 10.15 – 11.00 Mathias Neidrig Robert Koch Institut, Berlin, Germany (Abst. No. 2) 11.00 – 11.15 Break Keynote Lecture: Bovine Viral Diarrhea Virus – Is it A Real World Model for Zika Virus Congenital Syndrome? 11.15– 11.45 Christopher C. L. Chase South Dakota State University, Brookings, SD USA (Abst. No. 3) Seroprevalence of Falviviruses, Chikungunya virus and Rift Valley fever virus, Saudi Arabia 11.45 – 12.00 Ali M. Zaki (Abst. No. 4) 3-D structure prediction and analysis of the p7-transactivated protein1 of hepatitis C virus 12.00 – 12.15 Mahmoud M. El Hefnawi1, Mohamed E. Hasan, Amal Mahmoud, Yehia A. Khidr, El-Sayed A. El-Absawy, Alaa A. Hemeida (Abst. No. 5) Detection and comparative analysis for the tandem repeats motifs in the completed genomes of human coronavirus (hku1) by using regular expression finder tool 12.15 – 12.30 Mohamed A. Ezz, Amal M. Hussein, Fawzy A. Elfeki, and Alaa A. Hemeida (Abst. No. 6) 12.45 – 13.00 Discussion

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2. “One Health” - A solution to fight emerging viral threats in the future? Matthias Niedrig, Robert Koch Institut, Berlin, Germany The majority of recent emerging virus infections like SARS, influenza, Ebola, Hanta, MERSCov, or Rift Valley Fever either have a zoonotic background or are transmitted by arthropods as mosquitoes, ticks or sandflies transfer viruses like West Nile, Crimean Congo Hemorrhagic Fever, Dengue, Sandfly Fever, Chikungunya, Zika, and Yellow Fever. The outbreaks of these “new” emergencies cause tremendous problems because of missing diagnostic assays, unclear pathogenesis, and unclear routes of infection. The coincidence of these problems with a rudimentary, badly prepared or missing health care system can elicit a health catastrophe of major concern as we have seen for the Ebola or Yellow Fever outbreak. The bigger outbreaks provoke spreading of the disease to neighboring countries sometimes creating similar outbreaks if the health care system is badly prepared. Although outbreaks are somehow difficult to predict strengthening the health care system is a prerequisite for immediate and reliable implementation of answers and health measures. Could a widespread One Health view help to improve the health situation? The analysis of animal’s health and trading routs might provide some insights into transmissible diseases with potential risk for human infections. Surveillance studies in domestic and wild animals could provide insides into distribution of animal and human pathogens. Ideas for targeted sampling and strategies for diagnostic will be discussed by some examples.

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3. Bovine Viral Diarrhea Virus – Is it A Real World Model for Zika Virus Congenital Syndrome? Christopher CL Chase Department of Veterinary and Biomedical Sciences, South Dakota State University, Brookings, SD USA BVDV and Zika are both Flaviviruses with many similar characteristics. Often, infection with either Zika or BVDV, in immunologically competent adults, is subclinical. However, infection during pregnancy, while inapparent or mild in the mother, can have profound negative impact on the fetus. Both Zika and BVDV readily cross the placenta and cause lesions of the brain and CNS in the developing fetus. Outbreaks are frequently not noted until the birth of offspring with congenital defects. With over 50 years of information on the use of vaccination to prevent BVDV fetal infections, the level of immune protection that is efficacious for the protection against acute infections with other pathogens is not adequate for protection against BVDV fetal infections. Lessons we have learned with BVDV suggests that sterilizing immunity is required for fetal protection. Recent publications (Dowd et al., 2016, Cell Reports 16, 1485-1491) suggest an over estimation of protection by current Zika vaccines based on experience with BVDV vaccines. In general, vaccines work because the infecting virus is recognized after initial replication by the immune system and eliminated. BVDV infection of the fetus is the result of transport of the virus through maternal tissues to the amniotic fluid to the fetus. The transport of the virus is accomplished by the infection of immune cells that traffic throughout the body. Thus BVDV hijacks transport in the cells that are the primary defense against viral infection. When a dam is infected with BVDV during the second trimester of pregnancy the virus infects the cells of the placenta and crosses into fetal circulation. This unique route of entry may be in part responsible for the development of persistent infection. The BVDV antigen was exclusively found in cells that were Kupffer cells at day 89 (14 days following infection). Kupffer cells were not uniformly infected with BVDV at this stage of infection; as uninfected Kupffer cells were also observed in all tissues tested. Other hepatic cell populations such as hepatocytes, sinusoidal endothelial cells, hematopoetic precursors and lymphocytes were not positive for BVDV antigen. Kupffer cell cytokine response to infection was decreased. Kupffer cells could be responsible for presenting antigen to lymphocytes, and in the context of the specific microenvironment of the liver, the presentation may result in systemic tolerance rather than immune activation. Infection of Kupffer cells could be opportunistic for the BVDV and may be important for the development of persistent infection. There appears to be a window of time in which infection with BVDV results in brain and CNS lesions. However, epidemiological data indicates that individuals infected with BVDV in utero outside this window while appearing normal have compromised health and reproduction later in life. No fetal infection is without its price. Based on the BVDV epidemiology data it is likely the impact of Zika infection is being underestimated as detection has focused on congenital defects but the long-term implications of fetal infections has yet to be determined. Both the brain and the thymus in the early stage fetus are uniquely vulnerable to viral infections. Neither “tissue” can employ typical immune responses, which are based on the destruction of infected cells, to defend itself. Both rely on natural barriers that prevents most pathogens from entering protected tissues. In the case of BVDV, that barrier is broken resulting in brain lesions and the destruction of thymic tissue. Further understanding of BVDV in utero infections will be a valuable model for understanding Zika virus congenital infections.

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4. Seroprevalence of Falviviruses, Chikungunya virus and Rift Valley fever virus, Saudi Arabia Ali M. Zaki Professor of Medical Microbiology and Immunology Ain Shams University Cairo Egypt Abstract Background: Arboviruses are emerging zoonotic viral infections that infect their host by arthropod vector. Saudi Arabia have been hit by four emerging viral infections during the last two decades, namely dengue fever virus, Alkhurma virus, Rift valley fever virus, and Chikungunya virus. Objective: The aim of this work was to determine the prevalence of antibodies to these viruses among resident in Saudi Arabia. Samples and methods: Left over sera from patients using the laboratory of Dr. Soliman Fakeeh hospital were randomly selected and coded and stored at -20 until tested. The total number of sera tested was 1505 over three years’ period 2009-2011 Sera were tested for Dengue fever virus, Yellow fever virus, Alkhurma virus, chikungunya virus and Rift valley fever virus using IgG ELISA and or Immunoflourescent assay. Results: Total samples tested were 1505 from 855 males and 650 females. Antibodies for dengue fever were detected in 20% of cases followed by yellow fever 12. 6%, Alkhurma virus 9.6 %, Chikungunya virus 1.8%, and lastly Rift valley fever virus 0.9% Conclusion: Flaviviruses represent the most common arboviruses infecting peoples in Saudi Arabia with dengue the most active. Yellow fever virus antibodies may indicate silent unnoticed yellow fever virus circulation and should be further investigated. Key words: Flaviviruses, Chikungunya, Alkhurma, Rift Valley Fever, Saudi Arabia

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5. 3-D structure prediction and analysis of the p7-transactivated protein1 of hepatitis C virus Mahmoud M. El Hefnawi1, Mohamed E. Hasan2*, Amal Mahmoud2, Yehia A. Khidr3, El-Sayed A. El-Absawy2, Alaa A. Hemeida2 1

Informatics and Systems Department, Division of Engineering Research Sciences, the National Research Centre, Egypt 2 Bioinformatics Department, Genetic Engineering and Biotechnology Research Institute, Sadat City University, Sadat city, Egypt 3 Plant Biotechnology Department, Genetic Engineering and Biotechnology Research Institute, Sadat City University, Sadat city, Egypt Abstract: Background: The p7-transactivated protein of Hepatitis C virus is a small integral membrane protein of 127 amino acids, which is crucial for assembly and release of infectious virions. Ab initio and comparative modelling, is an essential tool to solve the problem of protein structure prediction and to understand the physicochemical principle of how proteins fold in nature. Results: Only one domain (1-127) of p7 had been predicted using the systematic in silico approach, ThreaDom. I-TASSER was ranked as the best server for full-length 3-D protein structural predictions of p7 where the benchmarked scoring system such as C-score, TM-score, RMSD and Z-score are used to obtain quantitative assessments of the I-TASSER models. Scanning protein motif databases, along with secondary and surface accessibility predictions integrated with post translational modification sites (PTMs) prediction revealed functional and protein binding motifs. Three protein binding motifs (two Asp/Glutamnse, CTNNB1-bd_N) with high sequence conservation and two PTMs prediction: Camp_phospho_site and Myristyl site were predicted using BLOCKS and PROSITE scan. These motifs and PTMs were related to the function of p7 protein in inducing ion channel/pore and release of infectious virions. Using SCOP, only one hit matched protein sequence at 71-120 and was classified as small proteins and FYVE/PHD zinc finger superfamily. Conclusion: Integrating this information about the p7 protein with SCOP and CATH annotations of the templates facilitate the assignment of structure–function/ evolution relationships to known and newly determined protein structures. Keywords: Hepatitis C virus, p7 protein, Ab initio, protein structure prediction, motifs and PTMs.

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6. Detection and comparative analysis for the tandem repeats motifs in the completed genomes of human coronavirus (hku1) by using regular expression finder tool Mohamed A. Ezz 1*, Amal M. Hussein1, Fawzy A. Elfeki2 and Alaa A. Hemeida1 1.

Bioinformatics Department, Genetic Engineering and Biotechnology Research Institute, Sadat City University, Egypt 2. Biotechnology Department, Faculty of Agriculture, Al-Azhar University, Egypt

Abstract Background: Relationship between the level of repetitive sequences in genomic sequence and genome size has been studied and investigated including complete prokaryotic and eukaryotic genomes, but relevant studies are fewer made in virus genomes. Objective: To design a specific tool to find different motif types of tandem repeats in the completed genomes of human coronavirus hku1. To search for a relationship between the virus repeats and the genome size. To find a specific pattern in coronavirus hku1. Methods: A total of 29 records of different genotypes of human coronavirus hku1 strain completed genomes were retrieved from the Genbank then downloaded in fasta format file. (Repeater Finder Regular Expression (RFRE) was used to find different extracted types of tandem repeats motifs in the completed genomes of human corona virus hku1. The designed tool gave the user a chance to write different specific regular expression patterns and combined patterns. To ensure the sensitivity of the RFRE tool in finding perfect repeats a YASS tool (http://bioinfo.lifl.fr/yass) was used for this purpose. To detect recombination among genomes the RAT tool was used to check for the recombination among the genomes of corona viruses. PRATT tool (http://www.ebi.ac.uk/Tools/pfa/pratt/) was used to expect a specific motif in human corona virus. Results: Different patterns of tandem repeats were founded in different lengths and different positions where the output of the tool gave a different tandem repeats length (240bp, 300bp and 480bp). A picture view for the direct repeats of the accession number (KF430201.1) of Human corona virus was produced by the dot plot and presented in diagonal lines paralleled to the main diagonal line. The RAT tool was used to check for the recombination among the genomes of corona viruses and identified a recombination site 3012 which was identified at the same time as a starting position of the retrieved repeats. A specific motif (VVTGDNDDEDVVTGDNDDE) was predicted from the translated extracted repeats of Human Corona Virus. There was a correlation between the total length of repeats and genome size of Human Corona Virus (R2=0.451). Conclusion: In conclusion, this study focused on the importance of identifying the tandem repeats in of human coronavirus hku1 and make a relation between the genome sizes of hKu1 types and repeats length. Recombination between the repeated regions in the completed genomes of different CoV HKU1 strains was observed in the repeated position (3012) among the human corona virus and expected to be a key role of virus evolution. Key words: HKU1; Recombination; RFRE; SSRs.

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Scientific Session IIa: Vaccines and immunity Chairman Prof. Falko Steinbach CoProf. Mohamed Faisal Chairmen Prof. Seham El-Zeidy Keynote Lecture: 15.00 – 15.30

Challenges for Veterinary Vaccines Falko Steinbach Animal and Plant Health Agency Weybridge, Addlestone Surrey KT15 3NB, UK & University of Surrey, Guildford, GU2 7AL, UK (Abst. No. 7) Keynote Lecture:

15.30 – 16.00

16.00 – 16.15

16.15 – 16.30

16.30 – 16.45

16.45– 17.00 17.00 – 17.15 17.15 – 17.30

17.30 – 17.45

17.45 – 18.00

Viral Hemorrhagic Septicemia Virus (Rhabdoviridae) DNA Vaccination Reveals the Presence of Herd Immunity in the Aquatic Environment. Mohamed Faisal Michigan State University, East Lansing, Michigan, USA (Abst. No. 8) Recombinant TMV vector for development of highly immunogenic HSV2 antigens used as a plant-based vaccine candidate Ahmad K. El-Attar, Josefine Persson, Aboul-Ata E. Aboul-Ata, Hamed H. Mazyad, Ali M. Harandi, and Olof Olsson (Abst. No. 9) Generation of novel fusion gene-dependent DNA vaccine candidates for protection against human respiratory syncytial viruses: the role of codon optimization, truncation and immunodominant epitopes. Mohamed A. Farrag, Haitham M. Amer, Peter Öhlschläger, Fahad N. Almajhdi (Abst. No. 10) Generation of a reassortant avian influenza virus H5N2 vaccine strain capable of protecting chickens against infection with Egyptian H5N1 and H9N2 viruses. Kandeil A, Moatasim Y, Gomaa MR, Shehata MM, El-Shesheny R, Mostafa A, Barakat A, Webby RJ, Ali MA, Kayali G (Abst. No. 11) Influenza Virus Polymerase Basic 1 (PB1) Origin is A Key Determinant for High-Yield Seed Vaccine Production Ahmed Mostafa, Pumaree Kanrai, John Ziebuhr, Stephan Pleschka (Abst. No.12) Break The use of different types of Montanide adjuvant in preparation of inactivated rabbit haemorrhagic vaccine Owais, G. A. Salman and Samah, E. Abodalal (Abst. No.13) Novel aluminum hydroxide nanorods elicit enhanced immune responses to inactivated genotype VIId NewCastle disease virus vaccine formulations Walaa A. Mohammad, Ausama A. Yousif, Ahemd A. Farghali, Sama I. EL Dek, Abo Zeid A. Abo Zeid, Mohamed A. Shalaby and Ahmed A. ElSanousi. (Abst. No. 14) Preparation and Immunological Evaluation of Inactivated Avian Influenza Virus Vaccine Encapsulated in Chitosan Nanoparticles in Chickens Samah H. Mohamed, A. Arafa, W. H. Mady, Rania Morsy, Hanan Aly, Lamiaa Omar (Abst. No. 15)

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7. CHALLENGES FOR VETERINARY VACCINES Falko Steinbach Virology Department, Science Division, Animal and Plant Health Agency Weybridge, Addlestone Surrey KT15 3NB, UK & School of Veterinary Medicine, University of Surrey, Guildford, GU2 7AL, UK Vaccines represent some of the greatest success stories of immunology as evident through the eradication of Smallpox and Rinderpest. However, a closer look also reveals some limitations i.e. we still lack ideal vaccines. More so, for some diseases we do not have effective and/or safe vaccines. Taking the history of some existing vaccines into account I will review some stories of success and failure that could lead the way towards a rational and safe design of veterinary vaccines for mammals in the future. Important considerations include the selection of pathogens, but equally the question what purpose the vaccine shall serve. In this context, I will address some requirements for quality control as well as looking towards alternatives for animal experiments for the in the development of vaccines.

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8. Viral Hemorrhagic Septicemia Virus (Rhabdoviridae) DNA Vaccination Reveals the Presence of Herd Immunity in the Aquatic Environment. Mohamed Faisal and Isaac F. Standish

Professor, Department of Pathobiology and Diagnostic Investigation. College of Veterinary Medicine. Department of Fisheries and Wildlife. College of Agriculture and Natural Resource College of Veterinary Medicine, Michigan State University, East Lansing, Michigan 48824, USA Viral hemorrhagic septicemia virus genotype IVb (VHSV-IVb) is presently found throughout the Laurentian Great Lakes region of North America. We recently developed a DNA vaccine preparation containing the VHSV-IVb glycoprotein (G) gene with a cytomegalovirus (CMV) promoter that proved highly efficacious in protecting muskellunge (Esox masquinongy) and three salmonid species. This study was conducted to determine whether cohabitation of VHSV-IVb immunized fishes could confer protection to non-vaccinated (i.e., naïve) fishes upon challenge. The experimental layout consisted of multiple flow-through tanks where viral exposure was achieved via shedding from VHSV-IVb experimentally infected muskellunge housed in a tank supplying water to other tanks. The mean cumulative mortality of naïve muskellunge averaged across 8 trials (i.e., replicates) was significantly lower when co-occurring with immunized muskellunge than when naïve muskellunge were housed alone (36.5% when co-occurring with vaccinated muskellunge versus 80.2% when housed alone), indicating a possible protective effect based on cohabitation with vaccinated individuals. Additionally, vaccinated muskellunge when co-occurring with naïve muskellunge had significantly greater anti-VHSV antibody levels compared to vaccinated muskellunge housed alone suggesting that heightened anti-VHSV antibodies are a result of cohabitation with susceptible individuals. This finding could contribute to the considerably lower viable VHSV-IVb concentrations we detected in surviving naive muskellunge when housed with vaccinated muskellunge. Our research provides initial evidence of the occurrence of herd immunity against fish pathogens.

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9. Recombinant TMV vector for development of highly immunogenic HSV- 2 antigens used as a plant-based vaccine candidate Ahmad K. El-Attara,d, Josefine Perssonb, Aboul-Ata E. Aboul-Ataa, Hamed H. Mazyada, Ali M. Harandib, and Olof Olssonc,d

a

Department of Plant Virus and Phytoplasma Research Section, Plant Pathology Research Institute, ARC, P.O. Box 12619, Giza Egypt. b Department of Microbiology and Immunology, Institute of Biomedicine, University of Gothenburg, Medicinaregatan 7A, SE40530, Gothenburg, Sweden. c Department of Pure and Applied Biochemistry, Lund University, Getingevägen 60, Box 124, SE22100 Lund, Sweden. d Department of Plant and Environmental Sciences, Carl Skottsbergsgata 22B, Box461, University of Gothenburg, SE40530 Gothenburg, Sweden. Abstract Background: Genital herpes, caused mainly by herpes simplex virus type 2 (HSV-2), is a sexually transmitted disease with a high prevalence in both developed and developing countries. No vaccine is currently available against genital herpes. Objectives: In this study we modify the tobacco mosaic virus to be used for expression of HSV-2 gD protein in plants. Methods: we introduced and expressed two genes encoding HSV-2 glycoprotein D (gD) and VP16 protein and a GFP marker gene (control) in tobacco seedlings. Positive plant infection was assessed through characteristic tobacco mosaic virus disease symptoms on leaves and by monitoring the fluorescence emitted by the expressed GFP protein. Expression of the HSV-gD2 and VP16 antigens was verified by RT-PCR, ELISA and Western blot. As a proof of concept, the immunogenicity and the protection ability of the plant produced gD antigen was tested in a mouse model of genital herpes and compared to gD antigen produced in a mammalian expression system. Results: This research showed that the plant-gD preparation, when used in combination with a CpG oligodeoxynucleotide as adjuvant, was highly immunogenic and capable of inducing complete protective immunity to an otherwise lethal vaginal HSV-2 challenge in mice. Conclusion: The data presented here may have implications for the development of a production system for highly immunogenic plant-based HSV-2 vaccine antigens. Keywords: TMV, plant vaccine, genital herpes, HSV-2, gD, VP16, GFP, mouse model genital herpes.

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10. Generation of novel fusion gene-dependent DNA vaccine candidates for protection against human respiratory syncytial viruses: the role of codon optimization, truncation and immunodominant epitopes. Mohamed A. Farrag1*, Haitham M. Amer1,2, Peter Öhlschläger3, Fahad N. Almajhdi1 1

Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia 2 Virology Department, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt 3 Institute of Nano- and Bio-technology, Department of Chemistry and Biotechnology, Aachen University of Applied Sciences, Juelich, Germany Abstract:

Background: Human respiratory syncytial virus (HRSV) infections continue to threat human beings where the virus causes deadly infections in infants less than one year of age. Although extensive endeavor to generate safe and effective HRSV vaccines, licensed vaccines do not exist till now. Objectives: In this study, the safety, immunogenicity and protection efficiency of three DNA vaccine candidates that encode different forms of codon-optimized fusion glycoprotein (F) of HRSV were assessed. Methods: Three different forms of codon-optimized HRSV F gene were cloned individually into the pPOE-CpG vector; full length F gene (pPOE-CpG-Fopt), truncated F gene that lacks the transmembrane domain (pPOE-CpG-TFopt), and full length F gene with the nucleotide sequence that encodes the immunodominant epitope of the second matrix protein (pPOE-CpG-FM2opt). In vitro expression of the different F gene forms was verified by ELISA and immunofluorescent assays. Seven groups of BALB/c mice were injected twice (days 0 and 10) via the intramuscular route with the different pPOE-CpG constructs and relevant controls. Humoral and cellular immune responses were evaluated using ELISA, ELISPOT and chromium release assays. At day 24, mice were challenged intranasally with the wild-type HRSV and lungs were harvested 2, 4, 6, and 8 days later. The viral load was tracked in the lung tissue by end-point titration assay using immunofluorescence. Lung sections were also stained with Hematoxylin and Eosin to record signs of pulmonary abnormalities. The extent of protection efficiency was evaluated by measuring lung cytokine profile using RT² Profiler PCR Array. Results: A distinctive expression of the F protein was identified in HEp-2 cells transfected with the different pPOE-CpG constructs. All constructs induced high level of antibody (specifically neutralizing antibodies) and CD+8 T cell responses. In particular, pPOE-CpG-TFopt showed higher level of gene expression in cell culture and higher induction of humoral and cellular immune responses. Following mice challenge, the different pPOE-CpG constructs protected immunized mice from disease enhancement with no obvious clinical signs and no mortalities as compared to negative controls. They also showed lower lung virus titer, restrictive pulmonary histopathology and lower level of inflammatory cytokines in lungs. As expected, pPOE-CpG-TFopt was superior in mice protection than the others did. Conclusions: pPOE-CpG constructs are promising HRSV DNA vaccine candidates as inferred from preclinical evaluation in terms of safety, immunogenicity and protection efficiency with no pulmonary inflammation. Keywords: DNA vaccine; fusion protein; human respiratory syncytial virus; pPOE-CpG; truncated form

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11. Generation of a reassortant avian influenza virus H5N2 vaccine strain capable of protecting chickens against infection with Egyptian H5N1 and H9N2 viruses. Kandeil A1, Moatasim Y1, Gomaa MR1, Shehata MM1, El-Shesheny R1, Mostafa A1, Barakat A2, Webby RJ3, Ali MA4, Kayali G5 1

Center of Scientific Excellence for Influenza Viruses, National Research Centre, Giza, Egypt. 2 Microbiology Department, Ain Shams University, Egypt. 3 Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, TN, United States. 4 Center of Scientific Excellence for Influenza Viruses, National Research Centre, Giza, Egypt. 5 Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, TN, United States. Abstract Background: Avian influenza H5N1 viruses have been enzootic in Egyptian poultry since 2006. Avian influenza H9N2 viruses which have been circulating in Egyptian poultry since 2011 showed high replication rates in embryonated chicken eggs and mammalian cells. Methods: To investigate which gene segment was responsible for increasing replication, we constructed reassortant influenza viruses using the low pathogenic H1N1 PR8 virus as backbone and included individual genes from A/chicken/Egypt/S4456B/2011(H9N2) virus. Then, we invested this finding to improve a PR8-derived H5N1 influenza vaccine strain by incorporation of the NA segment of H9N2 virus instead of the NA of H5N1. The growth properties of this virus and several other forms of reassortant H5 viruses were compared. Finally, we tested the efficacy of this reassortant vaccine strain in chickens. Results: We observed an increase in replication for a reassortant virus expressing the neuraminidase gene (N2) of H9N2 virus relative to that of either parental viruses or reassortant PR8 viruses expressing other genes. Then, we generated an H5N2 vaccine strain based on the H5 from an Egyptian H5N1 virus and the N2 from an Egyptian H9N2 virus on a PR8 backbone. This strain had better replication rates than an H5N2 reassortant strain on an H9N2 backbone and an H5N1 reassortant on a PR8 backbone. This virus was then used to develop a killed, oil-emulsion vaccine and tested for efficacy against H5N1 and H9N2 viruses in chickens. Results showed that this vaccine was immunogenic and reduced mortality and shedding. Discussion: Our findings suggest that an inactivated PR8-derived H5N2 influenza vaccine is efficacious in poultry against H5N1 and H9N2 viruses and the vaccine seed replicates at a high rate thus improving vaccine production.

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12. Influenza Virus Polymerase Basic 1 (PB1) Origin is A Key Determinant for High-Yield Seed Vaccine Production Ahmed Mostafa1, Pumaree Kanrai2, John Ziebuhr2, Stephan Pleschka2 1 2

Center of Scientific Excellence for Influenza Viruses, National Research Centre, Giza, Egypt.

Institute of Medical Virology, Justus Liebig University Giessen, Schubertstrasse 81, 35392 Giessen, Germany. Abstract

Influenza vaccine strains (IVSs) contain the haemagglutinin (HA) and neuraminidase (NA) genome segments of relevant circulating strains in the genetic background of influenza A/PR/8/1934 virus (PR8). Previous work has shown that the nature of the PB1 segment may be a limiting factor for the efficient production of IVSs. Here, we showed that the PB1 segment (PB1 Gi) from the 2009 pandemic influenza A virus (IAV) A/Giessen/06/2009 (Gi wt, H1N1pdm) may help to resolve (some of) these limitations. We produced a set of recombinant PR8-derived viruses that contained (i) the HA and NA segments from representative IAV strains (H3N2, H5N1, H7N9, H9N2); (ii) the PB1 segment from PR8 or Gi wt, respectively; and (iii) the remaining five genome segments from PR8. Viruses containing the PB1Gi segment, together with the heterologous HA/NA segments and five PR8 segments (5+2+1), replicated to higher titres compared with their 6+2 counterparts containing six PR8 segments and the equivalent heterologous HA/NA segments. Compared with PB1PR8-containing IVSs, viruses with the PB1Gi segment replicated to higher or similar titres in both cell culture and embryonated eggs, most profoundly IVSs of the H5N1 and H7N9 subtype, which are known to grow poorly in these systems. IVSs containing either the PB1Gi or the cognate PB1 segment of the respective specific HA/NA donor strain showed enhanced or similar virus replication levels. This study suggests that substitution of PB1PR8 with the PB1Gi segment may greatly improve the large-scale production of PR8-derived IVSs, especially of those known to replicate poorly in vitro.

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13. The use of different types of Montanide adjuvant in preparation of inactivated rabbit haemorrhagic vaccine Owais, G. A. Salman and Samah, E. Abodalal Newcastle disease department, Veterinary Serum and Vaccine Research Institute, Abbasia, Cairo, Egypt, P.O.B.131 Abstract Background: Rabbit viral haemorrhagic disease (RVHD) is characterized by high mortality in adult rabbits causing severe economic losses. Vaccination against rabbit haemorrhagic disease virus (RHDV) is the main preventive method. Objective: The current study was conducted to investigate the immuno-enhancing effects of some Montanide adjuvants incorporated in inactivated RHDV vaccine on the vaccinated rabbits in comparing to the currently produced aluminum hydroxide (Al OH) gel adjuvanted vaccine. Methods: Four experimental batches of inactivated RHDV oil emulsified vaccines were prepared as water-in-oil emulsion using 4 different types of Montanide adjuvants (SEPPIC, France) (ISA 70 VG, ISA 71 VG, ISA 206 VG and ISA 760 VG) in addition to another batch adjuvanted with Al OH gel. Aqueous to oil ratio differed according to Montanide type. The efficacy of the prepared vaccines was studied in five groups of vaccinated and one group of unvaccinated rabbits. The efficacy was based on antibody response that measured by HI test and ELISA in addition to the resistance to challenge with virulent RHDV. Results: All of the prepared vaccines induced specific RHDV-antibody titers, with variable values, detected from the 1st week post vaccination (WPV) in the vaccinated rabbits in the different groups. The induced RHDV-antibody titers increased reached the highest level in 3 rd month post vaccination (MPV) for all groups except group 5 in which the maximum level was attained at 5 th MPV. The vaccinated rabbits could resist the challenge against virulent RHDV as early as at the 3 rd WPV and at 6th MPV with 100% protection in contrary to unvaccinated group in which all rabbits get died (0% protection). Conclusion: According to the results of our study it could be concluded that all Montanide ISA adjuvants used in this study could be used to produce inactivated oil emulsion RHDV vaccine locally and it found to be more preferable than Al OH gel for long term immunity especially Montanide ISA70. Keywords: Inactivated vaccine; Montanide ISA VG Adjuvants; Rabbit Haemorrhagic Disease Virus

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14. Novel aluminum hydroxide nanorods elicit enhanced immune responses to inactivated genotype VIId NewCastle disease virus vaccine formulations Walaa A. Mohammad1, Ausama A. Yousif2, Ahemd A. Farghali3, Sama I .EL Dek3, Abo Zeid A. Abo Zeid1, Mohamed A.Shalaby2 and Ahmed A.El-Sanousi2 1Avian Viral Vaccines Department, Veterinary Serum and Vaccine Research Institute, Abbasia, Cairo, Egypt 2Department of Virology, Faculty of Veterinary Medicine, Cairo University, Giza, Egype. 3College of Graduate Studies, Beni Suef University, Beni Suef, Egypt. Abstract Background Aluminum salts are still extensively used as vaccine adjuvant due to their good track rec ord of safety, low cost, and proper adjuvanticity with a variety of antigens. Adsorption of antigens ont o aluminum compounds depends heavily on electrostatic forces and ligand exchange between adjuvant and antigen. Unloaded traditional aluminum hydroxide gel form particulates of 1–20 µm. There is evid ence that antigen-carrier nanoparticles around or less than 200 nm have a more potent adjuvant activit y than large microparticles. Objective This study was designed to test the adjuvant properties of locally synthesized aluminum hy droxide nanoparticles. Inactivated NDV was used as model to test the adjuvant properties of the synthe sized materials because the virus causes serious problems to the poultry industry, has a clearly defined challenge model, and has commercial vaccines that can be used for comparison. Methods: Aluminum hydroxide nanoparticles with a mean hydrodynamic diameter of 191.9 nm were synthesized. Newcastle genotype VIId and LaSota were characterized and propagated. Viruses were in activated using binary ethylenimine (BEI) and a cross linker. The adjuvant properties of traditional alu minum hydroxide(1.8-2.2mg/dose), aluminum hydroxide nanoparticles(1mg/dose) alone or with Immu nopotentiator Factor X (final concentration 1%) and Montanide ISA71 70%in a vaccine formulation was evaluated in 6 weeks old seronegative chickens of spf origin The formulations were administered sub cut. Control birds groups received saline, antigen alone, Montanide ISA71 or aluminum hydroxide nanoparticles. Birds were challenge 15 days post vaccination by intranasal administration of NDV gen otype VIId. Antibody titers were monitored at 0, 7 and 15 days post vaccination. Protective percentage s 15 days after challenge were determined. Results: NDV inactivation was confirmed by the absence of HA activity after 3 blind passages in emb ryonated SPF eggs. Hydrodynamic diameter scanning of NDV antigen-loaded aluminum hydroxide pr eparations showed the presence of two dispersion peaks, 123 nm and 1470 nm. Birds receiving antigen alone, or adjuvant alone failed to produce NDV-specific antibodies, and were not protected following NDV challenge. Mean protective antibody titer which obtained with aluminu m hydroxide nanoparticles formulation were significantly higher than traditional aluminum hydroxide gel and nearly equal to montanide ISA71. Aluminum hydroxide nanoparticles with Immunopotentiator the protective mean of antibody titer of aluminum hydroxide nanoparticles with Immunopotentiator w as approximately 2 folds higher than all adjuvants. Protective percentage in aluminum hydroxide nano particles alone and Montanide ISA71 was 80% but in traditional aluminum hydroxide gel was71.42%a nd in aluminum hydroxide nanoparticle with Immunopotentiator was86.68%. Conclusion: aluminum hydroxide nanoparticles with Immunopotentiator showed more adjuvanticity t han aluminum hydroxide gel, Montanide ISA71, and aluminum hydroxide nanoparticles alone in inact ivated Newcastle vaccine. Keywords: Inactivated; Formulations; Nanorods; NewCastle disease virus; Protection; Vaccine.

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15. Preparation and Immunological Evaluation of Inactivated Avian Influenza Virus Vaccine Encapsulated in Chitosan Nanoparticles in Chickens Samah H. Mohamed1, A. Arafa2, W. H. Mady2, Rania Morsy3, Hanan Aly4, Lamiaa Omar5 1

2

Immunology Department, Animal Health Research Institute, Dokki, Giza, Egypt. Reference Laboratory for Veterinary Quality Control on Poultry Production (RLQP), Animal Health Research Institute, Giza, Egypt. 3 Petrolium Research Institute, Cairo, Egypt. 4 Biotechnology Department, Animal Health Research Institute, Dokki, Giza, Egypt. 5 Central Laboratory for Evaluation of Veterinary Biologics (CLEVB), Abbassia, Cairo, Egypt. Abstract

The aim of this study is to maximize the efficacy of the inactivated avian influenza virus vaccine by using safe and novel adjuvants such as chitosan nano and microparticles. The results showed that the degree of deacetylation and the molecular weight of the extracted chitosan were 85% and 2122 kDa respectively. The average size of the chitosan nanoparticles was 150 nm with spherical and good dispersed nature as revealed by TEM and DLS and its Zeta potential was 11.5 mV whereas after encapsulation with AIV vaccine, the average size was 397 nm and its Zeta potential was 4.29 mV. The encapsulation efficiency of AIV vaccine on chitosan nanoparticles was 100 %. Sterility and safe property of the prepared vaccines were tested. The highest HI antibody titers results were shown in group 2 (G2) which was vaccinated with inactivated AIV chitosan micoparticles followed by group 1 (G1) vaccinated with inactivated AIV chitosan nanoparticles then group 3 (G3) vaccinated with oil inactivated AIV vaccine, on using S1 chicken antigen at 2 weeks post second vaccination, but on using S2 duck antigen, the highest HI antibody titers was shown in G3 followed by G1 then G2. The results of phagocytic % activity and phagocytic index of both G1 and G2 at 3 days’ post first vaccination were increased significantly in comparison with other groups, whereas at 14 days post first vaccination G1 only showed significant increase in phagocytic % activity and phagocytic index compared with other groups. The stimulation indices of lymphocytic proliferation test indicated that chickens in G1 induced the best T cell immune response results among all vaccinated groups. Finally, this study indicates that chitosan nano and microparticles are promising adjuvants for AIV vaccine in chickens by enhancing the humoral and cell mediated immunity. However, further investigations are required to explore the mechanism of action and to evaluate the protection trials for using of chitosan nano and microparticles as adjuvants. Keywords: Avian influenza virus, vaccine, adjuvants, chitosan, nanoparticles, humoral immunity, cell mediated immunity.

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Scientific Session IIb: Vaccines and immunity Chairman Prof. Falko Steinbach CoProf. Mansour Hashim, prof. Fahd El-Majhdi Chairmen Naked DNA Vaccination with Full-Length Attachment Gene of Human Respiratory Syncytial Virus Saudi Strain Induces Protective Immune Response with No Enhancement of Pulmonary Inflammation 18.15 – 18.30

18.30 – 18.45

18.30 – 18.45

18.45 – 19.00

19.00 -19.15 19.15 – 20.15

Maaweya E. Hamad, Haitham M. Amer, Mohamed A. Farra, Fahad N. Almajhdi (Abst. No.16) Field application of trivalent foot and mouth disease vaccine adjuvanted with Zeolite. Hiam M Fakhry; Assem A Mohamed; Sonia A Risk; Akram Z Hegazy and Abobakr M Agor. (Abst. No. 17) The immunogenicity and protective efficacy of DNA vaccine coding for full length H5 gene of avian influenza H5N1 subtype Yousef A. Soliman; Eman, M.S. El- Naga; Maha A. Gamal; Salah A. Selim; Mohamed M. El-Hady and Fekria A. El-Bordeny(Abst. No. 18) Evaluation of vaccination with local and imported vaccine against foot and mouth disease virus in Kalubeya governorate Lamya, A.F.Ateya, Ahmed, S. A, Khames, A.S, H.A. Abdel-Hady1 (Abst. No. 19) Vaccination of potato plants against Alfalfa mosaic virus (AMV) by using mild strain vaccine of Potato virus Y in Upper Egypt Safynaz A. Mohamed , and F.G. Fahmay (Abst. No. 20) Poster Session

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16. Naked DNA Vaccination with Full-Length Attachment Gene of Human Respiratory Syncytial Virus Saudi Strain Induces Protective Immune Response with No Enhancement of Pulmonary Inflammation Maaweya E. Hamad1*, Haitham M. Amer1,2, Mohamed A. Farrag1, Fahad N. Almajhdi1 1

Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia 2 Virology Department, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt Abstract

Background: Human respiratory syncytial virus (HRSV) is a common cause of acute lower respiratory tract infection in infants and young children worldwide. The development of HRSV vaccine that confers better protection than natural infection remains a global challenge. Vaccination with naked DNA is currently considered as a promising approach for the prevention and control of many viral diseases owing to its considerable safety, stability, ease of construction and targeted immunity. Objectives: In this study, the potential of DNA vaccination using full-length HRSV attachment (G) gene was evaluated in mouse model. Methods: The complete G gene sequence of HRSV type A strain Riyadh 38/2008 was amplified in a High fidelity RT-PCR and agarose gel purified. G gene was cloned in pcDNA3.1+ vector via 5`HindIII and 3`EcoRI (pcDNA/GA). The expression potential of pcDNA/GA was confirmed in HEp-2 cells on both RNA and protein levels using RT-PCR and immunofluorescence, respectively. DNA immunization of Balb/C mice with pcDNA/GA was followed by evaluation of both antibody and cytotoxic T cell responses using ELISA, micro-neutralization and ELISPOT assays. Mice were further challenged by wild-type virus and physiological parameters, clinical signs and mortalities were daily recorded. Seven days postchallange, lungs were harvested for evaluation of the pulmonary pathogenicity using histopathological examination, virus titration and cytokine profiling. Results: pcDNA/GA DNA construct was expressed to high level in HEp-2 cells as indicated by the distinct RT-PCR product and intra-cytoplasmic fluorescence granules in comparison to control cells. Immunized mice with pcDNA/GA exhibited high antibody titers in ELISA with superior neutralization activity. ELISOPT assay revealed the ability of pcDNA/GA to induce HRSVspecific CD8+ T cells in immunized mice. Following challenge, immunized mice showed no virus propagation and no distinct histopathology compared to control mice (formalin-inactivated RSV vaccine and pcDNA3.1+ immunized mice). The pulmonary cytokine profile in immunized mice after challenge displayed notable upregulation of Th1-associated cytokines such as GM-CSF, IL2, TLR6, while that of control mice exhibited high levels of Th2-associated cytokines like IL4, IL5, IL6, Il10, IL13, IL25, and, IL27. Conclusions: The generated DNA vaccine candidate (pcDNA/GA) has elicited potent antibody and CTL responses and confirmed pulmonary Th1 biased immune response postchallange in mice. These preliminary data suggested the compatibility of the DNA vaccine candidate for further testing in clinical trials. Keywords: attachment glycoprotein; challenge; DNA vaccination; human respiratory syncytial virus; mice immunization, T-helper biased response.

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17. Field application of trivalent foot and mouth disease vaccine adjuvanted with Zeolite. Hiam M Fakhry; Assem A Mohamed; Sonia A Rizk; Akram Z Hegazy and Abobakr M Agor Veterinary Serum and Vaccine Research Institute, FMD Department, Abbasia, Egypt. Abstract Background: Zeolite is a mineral micro particle that in earlier studies has shown adjuvant activity against different antigens. Clinoptilolite zeolite is safe, economic and effective. Objective: Comprehensive field sero-immunological study was conducted to reveal the adjuvant's effect of zeolite on the immune response of oil adjuvanted trivalent Foot and mouth disease (FMD) vaccine in cattle. Methods: This study was conducted on 53 cattle in Kaliobia governrate - Egypt. Animals was vaccinated intramuscularly (I/M) with trivalent FMD zeolite (1 μg/dose) vaccine, serum samples were collected from vaccinated animals for 40 weeks. The humeral immune responses were monitored by Serum Neutralization Test (SNT) and ELIZA technique. The results revealed that vaccinated cattle reach the protective level at 2nd to 3rd week post vaccination (WPV) and continued up to 35 week. Results: Our results show that the incorporation of zeolite into FMDV vaccine induces an early & long period of high specific protective immune response in cattle. Conclusion: Finally we recommended to use a ground zeolite alone as a potential and highly economic adjuvant in FMD vaccine in cattle.

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18. The immunogenicity and protective efficacy of DNA vaccine coding for full length H5 gene of avian influenza H5N1 subtype Yousef, A. Soliman1; Eman, M.S.El- Naga*2; Maha, A. Gamal1; Salah, A. Selim3; Mohamed, M. El-Hady3 and Fekria, A. El-Bordeny2 1 Department of biotechnology, Central Lab. for Evaluation of Vet . Biologics, Abassia, Cairo. Egypt 2 New casle disease department. Vet. Serum and vaccine research institute, Abassia, Cairo. Egypt 3 Department. Of microbiology, Vet. Collage, Cairo University. Cairo. Egypt Abstract Control of avian influenza epidemics can be achieved through both bio-security measures and massive vaccination. DNA vaccines represent a new approach for delivery of the most immunogenic compartments of the virus without virus handling required by traditional inactivated vaccine production. Heamagglutinin gene represent the main protein molecule required by the virus for cell entry and initiation of infection. Here, a DNA vaccine coding for the full H5 gene was produced against HPAI H5N1 subtype and adjuvanated with the genetic immune-modulating plasmid pcDNA3.1/esat6 that target the T-cell to enhance the cellular immunity. Full length H5 gene was amplified, sequenced, cloned in pENTER/D/SD/Topo entry clone and homologues recombination with pDEST40 mammalian expression destination vector was done to produce the DNA vaccine pDEST40/H5. SPF chickens were vaccinated either pDEST40/H5 alone or co-administrated with the immune-modulator pcDNA3.1/esat6 the third group received the commercially available reassorted inactivated vaccine and the forth group remained unvaccinated negative control. the survival rate were 84% and 88% for the groups received pDEST40/H5 alone and pDEST40/H5 plus pcDNA3.1/esat6 respectively while the survival rate for the group vaccinated with the inactivated vaccine was 80% only. shedding was 2.84 and 0.78 Log 10 as measured by EID50 for the cloacal swaps for group pDEST40/H5 and pDEST40/H5 plus pcDNA3.1/esat6 respectively while the group received the inactivated vaccine the shedding was 3.52 Log 10 indicating the high protective efficacy in protecting chickens against viral infection and the efficacy of pcDNA3.1/esat6 to reduce the shedding of the virus and reducing environmental load of the virus. Key words: Avian influenza; H5N1; DNA vaccine; esat 6; pENTER/D/SD/Topo and pDEST40.

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19. Evaluation of vaccination with local and imported vaccine against foot and mouth disease virus in kalubeya governorate *Lamya, A. F. Ateya, **Ahmed, S. A, *** Khames, A.S.****H.A. Abdel.Hady *Animal Health Research Institute – Dept.of virology, Benha branch **Elisa Unit and virus strain bank *** Shebeen El-Kome branch **** Alexandria branch Summary FMD virus poses a serious threat to the inter-national animal trade. The preventive vaccinations followed by sero-survey has been implemented for the control of FMD. 3ABC ELISA is highly effective for early identification of infected and vaccinated animals. Our study was designed for assessment of vaccination by local and imported FMD vaccine in cattle sera in Kalubeya Governorate. 220 serum samples were tested for FMD NSP by 3ABC Trapping ELISA.20 serum samples were used as control (no vaccination),100 serum samples collected from animals received local vaccine and 100 serum samples from animals received imported vaccine. 34/220(15.5%) were +ve and 186/220(84.5%) were –ve for NSP test preinoculation with vaccines. Serotyping of FMD antibodies were adopted on examined sera for imported and local vaccine using solid phase competitive ELISA 2,4,8, and 10 weeks post vaccination(wpv). Results were 45(24.2%), 65(34.9%) 2WPVand 55(29.5%) ,68(36.5%) 4WPV- 58(31.2%),71 (38.2%) 8WPV and 58(31.2%), 69(37.1%) 10 WPV for serotype A. For serotype O: 55(29.5%) ,58(31.2%)2WPVand60(32.3%), 62(33.3%) 4WPV and 62(33.3.5%), 62(33.3%) 8 WPV 60(32.3%),63(33.9%) 10 WPV. For serotype SAT2:30(.16.1%), 22(11.8%)2WPV and 39(20.9%),41(22%) 4WPV and 37(19.9%),41(22%) 8WPVand 27(14.5%), 35(18.8%) 10 weeks post vaccination. In conclusion: vaccines are a fundamental strategies aimed at global control and eradication of FMD. Control of FMD requires vaccines that will provide differentiating infected from vaccinated animals (DIVA). Key words: FMD; ELISA; NSP; (DIVA); Week Post vaccination; Cattle.

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20. Vaccination of potato plants against Alfalfa mosaic virus (AMV) by using mild strain vaccine of Potato virus Y in Upper Egypt Safynaz A. Mohamed *, and F.G. Fahmay Department of Plant Pathology, Faculty of Agriculture, Assiut University, Assiut, Egypt Abstract Background: Cross-protection is considered One of the first descriptions of cross- protection was by Mc Kinney (1929) who showed that severe yellow symptom of one strain of tobacco mosaic virus (TMV) did not appear in inoculated plants pre-infected with a more normal strain that produce a green mosaic. Many researchers had been studied the Mechanisms of cross-protection such as (Cassells, and Herriek, 1977) who has been demonstrated that challenge virus (eg. TMV) can actually accumulate in the infected plant without expressing the symptoms that would normally appear with a similar level of expression in non-protected plant). Objective: The goal of following studies is to reveal the effect of vaccination or cross-protection of potato against Alfalfa mosaic virus under greenhouse and field conditions and also elucidate this effect within the plant tissues in vaccinated and non-vaccinated plants with mild strain of Potato Y virus, which is considered non-related to the severe tested virus. Methods: Different methods such as, indicator plants, serological techniques and electron microscope were used for identification of the mild strain and the causal virus. Data revealed that inoculation of potato plants cultivar "Burna" with mild strain of Potato virus Y produced vaccination on potato plants which gave no visible syndrome of yellowing mosaic and leaf necrosis on potato leaves. Results: Vaccination of potato plants had been done after 0, 1, 2 and 3 days with mild strain. Data also revealed that the vaccination of potato plants was induced after inoculation it firstly with mild strain and followed with the challenging severe strain of Alfalfa mosaic virus after three days. Results of transverse ultra-thin section of electron microscope of vaccinated potato Burna variety with mild strain of Potato virus Y (PVY) against severe strain of Alfalfa mosaic virus was demonstrated that some of cell organelles like mitochondria, vacuole, and endoplasmic reticulum were noticed intact and it liked healthy ones. Conclusions: Possibility using of mild isolate of Potatovirus Y which is capable to protect potato plants against severe non-related virus of Alfalfa mosaic virus in the greenhouse and field experiments. Keywords: Vaccination, mild strain, AMV, vaccine, cross-protection.

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Poster Session 19.15-20.15 Molecular, clinico-pathological and sero-diagnosis of LSDV in cattle in Sharkia and Fayoum Governorates (Poster) Nashwa M. Helmy, Ahmed, S. A. and Zeinab, Y. Mohamed (Abst. No. 21) Lyophilized EHV-1 vaccine inactivated and adjuvanted by Quillaja Saponaria Molina extract (Poster) Nehal S. Saleh, Eman M. Ebeid, Fatma F. Warda, Eldakhly A.T., Nashwa K Madkour, Elkabany M.A. and Soliman I.M.A. (Abst. No. 22) Some molecular assays used for identity of PPR vaccines (Poster) Manar F. Seioudy, Magda M. Sayed, Ahmed A. El-Sanousi and Mohammed A. Shalaby (Abst. No. 23) Isolation and identification of lumpy skin disease virus in cattle in Kalubeya Governorate (Poster) Lamya, A. F. Ateya1, Ahmed, S. A., Mansour, A, H Khames, A. S. H. A. Abdel Hady (Abst. No. 24) Molecular Characterization and Confirmation of Tomato Spotted Wilt Virus Associated with an Asymptomatic Infected Chrysanthemum Plant in Egypt (Poster) Ahmed K. El-Attar, Manal A. EL-Shazly and Samah A. Mokbel (Abst. No. 25) Induction of resistance in pepper plants against Potato Virus Y (PVY) by two medicinal and aromatic plant essential oils and its major oil (Poster) components. Radwa M. Shafie, Ahmed A. Kheder and Amal A. Farghaly (Abst. No. 26) (Poster)

(Poster)

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(Poster)

Studding the physical, morphological and inclusion bodies properties of Bean yellow mosaic virus A. Wardany A., Usama M. A., Sabry Y. M. Mahmoud, Shaddad, M. A. K, Raafat F. A., (Abst. No. 27) Title: Development of multi-resistant tomato hybrids for heat and Tomato Yellow Leaf Curl Virus (TYLCV) Amro A. Farrag; Essam M. E. A. Khalil; Ahmed K. EL-Attar; A. A. Kheder and H. M. Mazyad (Abst. No. 28) Impact of Viruicide Carrageenan in Suppression of Potato Virus Y in Potato Plant Hanaa H. Gomaa (Abst. No. 29) The plant growth promoting fungus Penicillium sp. GP16-2 enhances the growth and confers protection against Cucumber mosaic virus in tobacco Mohsen M. Elsharkawy, Jehan M. Abass, Said M. Kamel and Mitsuro Hyakumachi (Abst. No. 30) Elimination of Alfalfa mosaic virus (AMV) from infected leaf potato (Solanum tuberosum) cv. Ditta by embryonic calli Maha. A. El-Abhar, Mostafa. A.S. El-Kady; Khaled. M. Ghanem and Hussieny A. Bosila (Abst. No. 31) Investigation of virus/meristem interaction by using Cre-virus vectors Mayada A.M., Badawi A.A. Othman, Taha R.M., Ali R.M and Schiemann J (Abst. No. 32) Isolation, pathotyping and genotyping of Newcastle disease virus from broiler chickens in Egypt. Ayman, S. El-Habbaa, Gabr, F. El-Bagoury, Samar, F. El-Adaway, and Susan, S. El-Mahdy (Abst. No. 33) A genotyping of a new avian infectious bronchitis virus isolated from chickens proventriculus in Egypt E.M. El-Nahas, H.S. El-Sayed, S.S. El-Basuni, G.F. El-Bagoury (Abst. No. 34)

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(Poster)

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(Poster)

(Poster)

Avian influenza virus vaccine match against circulating H5N1 avian influenza viruses in Suez Canal region, Egypt Amira Mohamed Helal, Abdel Satar Arafa, Hanan Fathy Abdein, Dalia Mansour Hamed and Mohsen El Dimerdash (Abst. No. 35) Variable Pathogenicity of Avian influenza viruses H5N1 isolated from Egypt Ola. A. Khader., Arafa. A, Hassan, M. K., A.A. El-Sanousi, M. A. Shalaby (Abst. No. 36) Trial for using emergency vaccination for controlling duck virus hepatitis (DVH) outbreaks in ducks Sabry, E.O., Hemat, S. Elsayed, Naglaa Hagag, Wesam Mady (Abst. No. 37) Evaluation of avian influenza H5N1 plasmid-based inactivated Egyptian vaccine and treatment with antiviral drugs Omnia M. Hassan, Ahmed Mostafa, Ahmed Kandeil, Rabeh El shesheny, Ali M. Ahmdy, Mohamed A. Ramadan, Mohamed A. Kutkat, Mohamed A. Ali (Abst. No. 38) In silico design and vitro validation of efficient si-RNAs as universal silencers of the polymerase genes of influenza A virus Sara H. Mahmoud, Rabeh El Shesheny, Mahmoud El Hefnawi, Naglaa Abdallah, Abdelhadi Abdallah, Ahmed Mostafa, Mohamed Ali (Abst. No. 39) Reassortment between avian highly pathogenic H5N1 and low pathogenic H9N2 influenza viruses and its impact on pathogenicity and infectivity in chickens Yassmin Moatasim, Ahmed Kandeil, Ahmed Mostafa, Sary Khaleel Abd elghaffar, Rabeh El Shesheny, Ghazi Kayali, Ahmed H. M. Elwahy, Mohamed Ahmed Ali (Abst. No. 40)

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21. Molecular, clinico-pathological and sero-diagnosis of LSDV in cattle in Sharkia and Fayoum Governorates 1

Nashwa M. Helmy, 2Ahmed, S. A. and3Zeinab, Y. Mohamed 1

Biotechnology Dept., 2ELISA Unit and 3 Pathology Dept. Animal Health Research Institute, Dokki, Giza, Egypt Abstract:

A total of 30 biopsy samples from cutaneous nodules were obtained from infected animals at different stages during the course of the Lumpy skin disease (LSD), and (100) Peripheral blood samples without anticoagulant were drawn from apparently healthy non vaccinated cattle against LSDV and (100) serum samples were drawn from cattle 4 weeks post vaccination with local attenuated sheep pox virus vaccine located in Sharkia and Fayoum governorates. Lumpy skin disease virus (LSDV) was isolated from skin biopsies collected from clinically infected cattle. The virus was isolated on MDBK cell line and identified by agar gel precipitation test (AGPT) and indirect fluorescent antibody technique (IFAT) using specific hyper immune serum against LSDV. Further identifications were carried out by polymerase chain reaction (PCR) and clinco-pathological investigation. The results showed that 11/30 biopsies were positive by AGPT, 19/30 by IFAT and 30/30 by PCR. While results of sero-diagnosis showed that 45/100 from apparently healthy non vaccinated cattle and 68/100 from vaccinated cattle were positive by SNT respectively and in general 90/200 of tested cattle give protective antibody titer, while 23/200 gave non protective titer and 87/200 have no antibody againest LSDV. The results of clinco-pathological revealed highly significant increase in ALT, AST, ALP, GGT, urea and uric acid, while the level of total protein, albumin and calcium showed significant decrease and non-significant reaction in creatinine and non-organic phosphorus in infected cattle. The results of antioxidant both malondialdehyde (MDA) and catalase enzyme (CAT) showed significant increase while level of gloutathion (GSH), total antioxidant capacity (TAC) and gloutathion peroxidas (GPX) showed significant decrease in infected cattle. Sero-survey, conventional techniques and PCR assay should be applied besides clinco-pathological for any cases with skin lesions as early as possible to diagnosis and apply adequate control measures. The results encountered in the present study revealed that cattle infected with LSD exposed to strong oxidative stress so recommended to use antioxidants in infected animals during treatment. Key words: IFAT; LSDV; PCR.

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22. Lyophilized EHV-1 vaccine inactivated and adjuvanted by Quillaja Saponaria Molina extract Nehal S. Saleh*, Eman M. Ebeid*, Fatma F. Warda, Eldakhly A.T*., Nashwa K Madkour*, Elkabany M.A*. and Soliman I.M.A*. *Veterinary Serum and Vaccine Research Institute, Abbasia, Cairo, Egypt Abstract Background: Equine Herpes virus-1 (EHV-1) is one of the most common respiratory pathogens of the horses. EHV-1 induces several clinical signs of disease ranging in severity, from mild respiratory distress to abortion in pregnant mares, neonatal foal death and neuropathogenic disorders. Whole inactivated EHV-1 vaccines, which provide variable levels of protection against the disease through the induction of antibodies, have been the main type of vaccine commercially available. Objectives: The current report designed to evaluate the production of EHV-1 inactivated vaccine in lyophilized form which enhance the stability and thus the marketability of a product Methods: Three different lyophilized vaccines formula, inactivated and/or adjuvanted with saponin were prepared. Formula 1 contain Lyophilized EHV-1 reconstituted at time of inoculation in saponin as inactivator and adjuvant, Formula 2 contain Lyophilized EHV-1 with saponin as inactivator and adjuvant then reconstituted at time of inoculation in saline, Formula 3 contain Lyophilized EHV-1 inactivated with Binary Ethyleneimine (BEI) and reconstituted at time of inoculation in saponin as adjuvant. Different saponin concentration were used with each formula (1, 1.5, 2 mg/dose). and comparing between immune responses stimulate by such vaccines in mice and horses using ELISA and CFT. Also shelf life of the prepared vaccines was examined. Results: That the highest ELISA antibody titer in mice was (2100, 2250, and 3000) which induced by the prepared formulae 1, 2 and 3 respectively were obtained by 1mg Saponin /dose. Immunization of horses with two dose (one-month interval) from the prepared three vaccines formula contain 1mg/ dose saponin, revealed that the mean CF antibody titers against EHV-1 were increased 4- fold within two weeks, from 8, 8, 10 at time of booster dose (4week post inoculation -WPI) to 32, 32, 48 at 6WPI in sera of horses inoculated with formula 1, 2 and 3 respectively. Also the immune response of the inoculated horses was monitored up to seven months’ post inoculation (MPI) by ELISA. There was significant increasing in EHV-1 antibody titers at 21 days’ post booster (DPB) dose than the prebooster (from 300, 330, and 570 at 4WPI to 730, 880, and 1170 at 21DPB) for the three vaccines formula respectively which indicate good immunogenicity of the vaccines. The antibody titer reached its peak at 2MPI then began to decline gradually till 6MPI for the three formulas. The three formulas of lyophilized vaccine kept its antigenic potency when stored at 4˚C until one yare. Conclusions: The three vaccine formulas were good immunogenic. but formula 2 is more safe as it contains less chemicals and can be produced by lower costs than formula 3 which given higher antibody titers but contain BEI and saponin. Keywords. EHV-1; Lyophilized vaccine; Saponin; BEI; Mice model; Vaccine preparation

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23. Some molecular assays used for identity of PPR vaccines Manar F. Seioudy1*, Magda M. Sayed1, Ahmed A. El-Sanousi2 and Mohammed A. Shalaby2 1 2

Central Laboratory for Evaluation of Veterinary Biologics (CLEVB), Cairo, Egypt Department of Virology, Faculty of Veterinary Medicine, Cairo University, Egypt

Abstract Peste des petits ruminants (PPR) is a highly contagious, economically important viral disease of sheep and goats with high morbidity and mortality rates. A live attenuated PPR vaccine has and still been utilized to control PPR disease in endemic areas of the Arabian Gulf. In this study, the identity of four batches of PPR vaccines was tested as per the Office International des Epizooties (OIE) guidelines using RT-PCR technique that based on the amplification of fragments of F-protein and fragments of N-protein. All four batches are also tested for identity using Taq Manbased, one-step, real time RT-PCR. Results in this study showed that molecular techniques could be used for rapid evaluation of PPR vaccine including RT-PCR and real time RT-PCR for identity testing. However, the one-step real-time RT-PCR is proved to be the most rapid, sensitive and specific assay for identification of PPRV in PPR vaccines. In the meanwhile, the conventional RT-PCR using primer set that targets N gene is more sensitive in identification of PPRV than RT-PCR using primer set that targets F gene which could give false negative results. Key words: PPR vaccine; Identity test; RT-PCR; real time RT-PCR

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24. Isolation and identification of lumpy skin disease virus in cattle in Kalubeya Governorate *Lamya,A.F.Ateya1, **Ahmed, S. A1, ***Mansour, A, H1 **** Khames,A.S1.*****H.A.Abdel.Hady1 *Animal Health Research Institute – Dept.of virology, Benha branch **Elisa Unit ***Dept. of Biotechnology Doky, Giza, **** shebeen El-Kome branch***** Alexandria branch Abstract Lumpy skin disease virus (LSDV) is an infectious viral disease which is an important serious skin disease of cattle. LSDV was isolated from skin biopsies collected from cattle which are clinically infected at Kaluobia governorate. A total 23 skin biopsies were used for virus isolatation on chorioallantoic membrane (CAM) of embryonated chicken eggs SPF at 9-11 day which showed pock lesions and CPE on MDBK cell culture. Isolated virus was identified by indirect fluorescent antibody technique (IFAT) using specific hyper immune serum against Lumpy Skin Disease Virus. Further identifications were carried out by polymerase chain reaction (PCR).15 samples were +ve for LSDV by conventional PCR.The results showed that 11/23 biopsies were positive by IFAT. Molecular identification of LSD virus by using RT-PCR, revealed positive for amplification of bands at the predicted molecular size (1926bp). Neutralizing antibodies against LSDV were (55) out of total 100 serum samples by serum neutralization test. Selection and processing of clinical specimens, viral isolation and PCR assay applied, for LSDV are much sensitive and rapid diagnostic tool of LSD reflecting their importance in controlling the rapid spread of disease in Egypt. Key words: LSDV; CAM; MDBK; IFAT; PCR.

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25. Molecular Characterization and Confirmation of Tomato Spotted Wilt Virus Associated with an Asymptomatic Infected Chrysanthemum Plant in Egypt Ahmed K. El-Attar, Manal A. EL-Shazly and Samah A. Mokbel Virus and Phytoplasma Research Department, Plant Pathology Research Institute, Agricultural Research Center (ARC), Giza, Egypt. Abstract The detection of Tomato spotted wilt virus (TSWV) in locally traded asymptomatic chrysanthemums plants constitutes a severe threat to the Egyptian farming. The detection and identification of TSWV in asymptomatic chrysanthemums plants were confirmed by double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) using a specific polyclonal antiserum and virus-specific primers, respectively. These results demonstrated that TSWV might exist in asymptomatic plants in different locations in the Giza governorate, and remain undetected or unidentified. RT-PCR accurately identified 26% (28 vs. 15, out of 50) more symptomless samples than that the ELISA did. The molecular characterization of the Egyptian isolate of TSWV has been performed successfully. Total RNA isolated from asymptomatic infected chrysanthemum plants was subjected to RT-PCR using primers specific to the full coat protein (CP) gene of TSWV and the amplified product was cloned and sequenced. Nucleotide sequence of the TSWV-clone was deposited to the NCBI GenBank. Phylogenetic analysis based on cp gene sequences of the Egyptian TSWV-isolate revealed different identity levels when compared with other isolates on GenBank from different countries. The pathogenicity of TSWV isolate was confirmed by inducing characteristic local lesions and systemic symptoms in a set of different vegetable crops and some ornamental or herbaceous plants. This is a first experimental demonstration of TSWV associated with circular whitish spots and necrotic regions on onion and sweet potato plants. The information generated in this study will be useful in formulating effective control measures for this economically important virus. Keywords: Chrysanthemums; Tomato spotted wilt virus; DAS-ELISA; RT-PCR; Cloning and Phylogenetic analysis.

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26. Induction of resistance in pepper plants against Potato Virus Y (PVY) by two medicinal and aromatic plant essential oils and its major oil components. Radwa M. Shafie, Ahmed A. Kheder and Amal A. Farghaly Virus and Phytoplasma Research Department, Plant Pathology Research Institute, Agriculture Research Center (ARC), Giza, Egypt. Abstract Background: Potato Virus Y (PVY) causes enormous economic loss wherever pepper is grown. The antiviral activity of essential oils extracted from clove and fennel seeds oils and its major components as foliar treatment were screened for their inhibitory effect against Potato Virus Y (PVY). Objective: Systemic acquired resistance could be induced in pepper plants using different concentration of essential oils extracted from clove and fennel seeds oils and its major components against Potato Virus Y (PVY). Methods: In vitro in both systemic and local lesion hosts were treated at different concentrations from antiviral clove and fennel seeds oils. Transmission of Potato Virus Y (PVY) to pepper by the green peach aphid, Myzus persicae (sulzer), was evaluated by means of DAS-ELISA as well as the severity of symptom were assessed by visual inspection. Also, Polyacrylamide gel electrophoresis (PAGE) was used to determine the qualitative changes in the soluble proteins of pepper plants (healthy or infected with PVY) related to inducers. Vegetative growth characters (leaves area. Plant-1) of the pepper plants were taken to determine the effect of PVY on plants depending on date of infection and compared with control (healthy) pepper plants. Results: At conc. 15%, clove seeds oil gave the highest inhibitory effect against PVY infection than fennel oil, the percentages of inhibition were (85%) and (80%) respectively on inoculated pepper and number of local lesions were reduced up to (90%) and (87%) respectively on Chenopodium amaranticolor when applied 48 hrs. before virus inoculation. When the major components of essential oils applied individually, eugenol gave the highest inhibitory effect (90%) on PVY- systemically infected pepper plants and (91%) on Ch. amaranticolor respectively when applied 48 hrs. before virus inoculation followed by anethol. While, limonene was less effective in reducing PVY infection, moreover SDS-PAGE inducing new proteins band 20 KDa was found only in pepper plants sprayed with clove oil at conc. of 15%. Also a new protein 20 KDa and 25 KDa were found only in pepper plants sprayed with eugenol. It has been suggested that, the induced proteins may help to limit virus spread or multiplication. Conclusion: Essential oil of clove and its major components was more effective in reducing the local lesions produced by PVY on Ch. amaranticolor than essential oil of fennel and its major components. Key words: Clove seed oil; Fennel seed oil; Induce systemic resistance; Major oil components; Potato Virus Y (PVY) and SDS-PAGE.

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27. Studying the physical, morphological and inclusion bodies properties of Bean yellow mosaic virus A. Wardany A1., Usama M. A1., Sabry Y. M. Mahmoud 2, Shaddad, M. A. K3, Raafat F. A.1 1Botany and Microbiology department, Faculty of Science, Al-Azhar University, Assiut branch, Egypt, 2Agricultural Microbiology department, Faculty of Agriculture, Sohag University, Sohag 82786, Egypt, 3Botany and Microbiology department, Faculty of Science, AL-Azhar University, Assiut, Egypt. Abstract Bean yellow mosaic virus (BYMV) has been recognized as one of the major economic disease and reduced a considerable destructive in seed yield, particularly, when plants were infected at the beginning stage of Vicia faba growing. The study of physical properties, types of forming inclusion bodies and structure of virus was the most important methods used in the definition of the virus. In this study, thermal inactivation point, dilution end point and longevity in vitro that virus becomes inactivated at which were experimented as physical properties of BYMV affecting V. faba (Vicia faba cv. Sakha 1). On the other hand, under BYMV stress, viral morphology and characters of inclusion bodies forming inside infected cells of V. faba were examined by electron microscope. Practical experiments of studding physical characteristics of BYMV indicate that the thermal inactivation point, dilution end point and longevity in vitro were at 60 º C, 10 -4 of dilution point and three days, respectively, in which no virus infection occurs. Electron microscopy technique provides strong evidence about morphology of Bean yellow mosaic virus. Particles of BYMV are flexuous rods with a modal length of 700 -750 nm and about 12 nm in diameter. Transmission electron microscopic observations on leaves of V. faba plants (Vicia faba cv. Sakha 1) infected with BYMV revealed a variety of potyvirus-like inclusions. Potyvirus specific inclusion bodies were detected in the cytosol of the infected cells. Laminated aggregates (La), bundle (Bu) and dense bodies (D) loosely distributed in the cytoplasm.

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28. Development of multi-resistant tomato hybrids for heat and Tomato Yellow Leaf Curl Virus (TYLCV) Amro A. Farrag1*; Essam M.E.A. Khalil2; Ahmed K. EL-Attar1; A. A. Kheder1 and H. M. Mazyad1 1

Virus and Phytoplasma Research Department, Plant Pathology Institute Agriculture Research Center 2 Breeding Vegetable Crops & Medicinal and Aromatic Plants Research Department, Horticulture Research Institute, Agriculture Research Center, Giza, Egypt.

Abstract Background: Tomato (Solanum lycopersicon) is affected with many biotic and abiotic stresses those may become major limiting factors for growth, yield quality and quantity. High temperature, as an abiotic stress, affecting stigma tube elongation, pollen germination, pollen tube growth and carbohydrate level which cause significant decrease in fruit set and fruit yield. TYLCV, as biotic stress, infects tomato and dramatically affect production causing losses may reach 80% at early infection. Molecular marker assisted breeding was used to develop tomato hybrids with combined resistance for both heat and viral infection Objective: Development of multi-resistant tomato hybrids for heat tolerant by conventional breeding and molecular marker-assisted selection for Tomato Yellow Leaf Curl Virus (TYLCV). Methods: Twenty tomato F1 hybrids were produced by crossing ten pure lines as source of Tomato Yellow Leaf Curl Virus (TYLCV) resistance with two heat resistant lines LA 2662 and LA 3120 in line x tester mating fashion. Produced hybrids and parents were evaluated to heat tolerant and TYLCV resistance during the late summer season year 2015 and 2016 then, the yield component and Ty marker were recorded. Results: Recorded data showed a significant General combining ability (GCA) and Specific combining ability (SCA) of 4 different hybrids those showed a good yield with a good firmness and medium fruit weight. Data for TYLCV resistance evaluation showed that all hybrids with LA 3120 showed mild symptoms from which five selected hybrids showed no symptoms with a relatively high resistance for TYLCV infection. Different hybrids derived from LA2662 showed moderate TYLCV symptoms in addition to one hybrid that showed mild symptoms and four selected hybrids showed no symptoms with a significant resistance level. Screening for Ty marker confirmed the resistance of sex hybrids due to the existence of two different markers (Ty1 and Ty3) and the resistance of one more hybrid due to the presence only one marker (Ty2). Conclusion: Results showed that there are four promising hybrids contain multi-resistance for both heat and TYLCV those resulted in a high yield with good firmness and medium fruit size .Plants contain both Ty1 and Ty3 in a dominant heterozygous or homozygous did not show any symptoms while plants contain only one TY marker showed mild or moderate symptoms. Keywords: Tomato, Solanum lycopersicon, General combining ability (GCA), Specific combining ability (SCA), Line × Testers, Yield, TY marker.

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29. Impact of Viruicide; Carrageenan in Suppression of Potato Virus Y in Potato Plant Hanaa H. Gomaa Botany Department, Faculty of Science, Suez Canal University, Ismailia, Egypt Abstract Potential effect of viruicide (carrageenan) which was extracted from red algae (Acanthophora specifira) in induction of systemic acquired immunity in potato against Potato Virus Y was detected via virus infectivity, potato growth and biochemical indicators. Potato virus Y belongs to group of potyvirus that causes serious diseases to potato plants. Reduction in the disease severity was recorded and inhibition PVY infectivity as result to periodically foliar potato plants with viruicide. An improvement by foliar periodically with viruicide in morphological characters. PVY infection resulted significant decreased in shoot length, leave area; fresh and dry weight of shoots. Chlorophyll and carotinoids contents were increased in PVY infected potato plants foliar treatment with viruicide. Biochemical markers as indicators for systemic acquire resistance were detected via significant increased in total proteins, phenol compounds, free proline (PO), polyphenol oxidase (PPO) and superoxide dismatase (SOD) enzyme activities. As well as expressed proteins of induced potato plants were determined using SDS-PAGE. The results indicated that new expressed proteins were produced and different in number and density of bands in foliar treatment plants with viruicide compared with healthy ones. It has been suggested that, the induction of systemic acquired resistance (SAR) was successfully achieved could also protected potato plants against PVY infection. Key words: Carrageenan, Polyphenol oxidase (PPO), Potato virus Y (PVY), potato, Proline (PO), Systemic acquired resistance (SAR), SDS- PAGE, Superoxide dismutase (SOD).

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30. The plant growth promoting fungus Penicillium sp. GP16-2 enhances the growth and confers protection against Cucumber mosaic virus in tobacco Mohsen M. Elsharkawy1*, Jehan M. Abass2, Said M. Kamel3 and Mitsuro Hyakumachi4 1 Department of Agricultural Botany, Faculty of Agriculture, Kafrelsheikh University, Kafr El-sheikh 33516, Egypt. 2 Department of Virus and Phytoplasma Research, Plant Pathology Research Institute, Agricultural Research Center, (ARC), Giza, Egypt 3 Plant Pathology institute, Agriculture Research Center, Giza, Egypt 4 Laboratory of Plant Pathology, Faculty of Applied Biological Sciences, Gifu University, 1-1 Yanagido, Gifu City 501-1193, Japan. Tel: +81-58-2993-2847; Fax: +81-58-293-2847b Abstract Background: Plant growth promoting fungi (PGPF) have received considerable attention in the recent years due to not only promote plant growth but also potentiate systemic resistance against different plant pathogens. Objective: The objective of this study is to evaluate the ability of Penicillium sp. GP16-2 to stimulate growth and Cucumber mosaic virus (CMV-Y) resistance in tobacco plants. Methods: Tobacco plants were pre-inoculated with GP16-2 and the leaves were inoculated with CMV. An indirect enzyme-linked immunosorbent assay (ELISA) was used to measure CMV accumulation in tobacco leaves and the expression levels of defence-related genes were tested using semi-quantitative RT-PCR analysis. Results: The use of the barley grain inoculum GP16-2 significantly enhanced fresh weight and dry weight of tobacco plants 6 weeks after planting. Our results demonstrated that tobacco plants treated with barley grain inoculum (BGI) of GP16-2 or its culture filtrate (CF) showed significant reduction in disease severity and CMV titre as compared with control plants. RT-PCR results showed increased expression levels of SA-inducible genes, but not JA -inducible gene, in leaves of BGI treated plants. Moreover, expression levels of SA- and JA inducible genes were increased in leaves of CF treated plants. Conclusion: In conclusion, BGI treatment induced systemic resistance against CMV through SA signaling cascade in tobacco plants. While, treatment with CF of GP16-2 mediated the expression of a majority of the various pathogen related genes, which led to the increased defense mechanism against CMV infection. Key words: Cucumber mosaic virus; plant growth promoting fungi; induced systemic resistance; Penicillium sp.; tobacco.

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31. Elimination of Alfalfa mosaic virus (AMV) from infected leaf potato (Solanum tuberosum) cv. Ditta by embryonic calli Maha. A. El-Abhar1*; Mostafa. A. S. El-Kady 1; Khaled. M. Ghanem2 and Hussieny A. Bosila3. 1

Virus and phytoplasma Res. Dep., Plant Path. Res. Inst., ARC, Giza, Egypt 2 Environment and Bio-Agriculture Dep., Agri. Fac., Azhar Univ., Egypt. 3 Horticultural Dep., Agri. Fac., Azhar Univ., Egypt

Background: Potato (Solanum tuberosum L.) is considered as one of most important and economic vegetable crops. In 2104, four million and eight thousand 4.8 million tones of tubers were harvested from 4.4 thousand Fadden of potato grown in Egypt. Alfalfa mosaic virus (AMV) has been isolated from a number of crop plants including potato in Egypt. Callus cultures may prove useful in raising plants free of virus. the callus derived from infected tissues does not carry the pathogen uniformly in all cells. Callus culture to eliminate virus was occurred beging from 60s. Embryonic callus and/or somatic embryogenesis pathway have practical applications, including crop improvement such as virus elimination from potato Objective: Elimination of Alfalfa Mosaic Virus (AMV) in potato Solanum tuberosum CV. Ditta via Embryonic calli. Method: AMV was isolated from potato and characterized using Indirect-ELISA .Vrus-infected plants grown under greenhouse conditions were used as a source for virus elimination. leaves excised from arial parts of infected potato to produce pro-embryogenic masses (PEMs) were controlled by MS basal medium supplemented with specific growth regulators 2,4-D concentrations at 0,1,2,3,4,5 and combination of 5mgl-1 2,4-D + 0.5 of Kin, BAP, TDZ or Zeiten mgl-1). Pro-embryo development stages for embryonic calli were subjected to histological via light microscopy and serological tests by ELISA technique. Result: After incubation period 30 – 35 days, 100 % of creamy-green granular callus with 81.17% pro-embryonic mass were obtained with 5 mgl-1 2, 4-D + 0.5 mg-l Kin. Vice versa, the combination of 5 mgl-1 2,4-D + 0.5 mgl-1 TDZ had the lowest effect 11.11 % of brown rigid callus formed. Proembryo development stages for embryonic calli were showed 100% virus-free at all treatmens. Conculosion: AMV was eliminated by embryonic calli and pro-embryo development from potato (Solanum tuberosum L.) CV. Ditta which characterized features by microscopic or histological means. Outcome of this study demonstrated the effectiveness of embryonic calli at various stages of the proembryo development as procedure to eliminate AMV from infected leaves as a source material. Key words: Potato (Solanum tuberosum), embryonic callus, growth regulators, Alfalfa mosaic virus (AMV)

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32. Investigation of virus/meristem interaction by using Cre-virus vectors Mayada A.M1., Badawi A.A. Othman2, Taha R.M3., Ali R.M4 and Schiemann J5. 1,3and4 Faculty of Science, Botany department, Fayoum University 2 Faculty of Agriculture, Microbiology department, Ain Shams University 5 Plant Biotechnology, Institute for Biosafety, Julius Kuhn-Institut, and Federal Research Center for cultivated Plants, Quedlinburg, Germany Abstract Developing convenient defense approaches against invading plant pathogens is gaining more interest in recent years to palliate their impact on many important crops. To develop such approaches, investigation and understanding of underlying mechanisms for plant-virus interaction is indispensable to achieve such a goal. Potato virus X (PVX) like most plant viruses has been thought to be excluding from plant growing points containing the shoot apical meristem. In the current work, a new experimental system consisting of PVX and recombinant PVX-Cre (P1 recombinase) has developed, to study the effect of Cre recombinase on virus entry into growing points. PVX-Cre infected plants were more severly diseased, heavily stunted as compared to PVX infected Nicotiana benthamiana plants and has showed a "recovery" phenotype. Florescence microscpic investigations could show PVX in meristematic tissue of host plant in presence of Cre. Although, it has not any RNA silencing suppression activity, RT-qPCR investigation showed that Cre protein might has an effect on the endogenous polymerase RDR1 activity. The here reported results give more insights for understanding the mechanism of plant infection process. Keywords: Cre-virus vectors, PVX, RT-qPCR, RNA silencing endogenous polymerase RDR1 and Nicotiana benthamiana.

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33. Isolation, pathotyping and genotyping of Newcastle disease virus from broiler chickens in Egypt. Ayman, S. El-Habbaa 1, Gabr, F. El-Bagoury1, Samar, F. El-Adaway2, and Susan, S. El-Mahdy2 1 2

Department of Virology, Faculty of Veterinary Medicine, Benha University. Central Laboratory for Evaluation of Veterinary Biologics, Abbassia, Cairo. Abstract

In Egypt, outbreaks of Newcastle disease (ND) have been occurring in vaccinated chickens with great economic losses. This work aimed for diagnosis and characterization of Newcastle Disease Virus (NDV) from infected broiler chickens in Egypt. Suspected virus isolates on embryonated chicken eggs was detected by Hemagglutination (HA) test and identified using hemagglutination inhibition (HI) test. F protein encoding gene of NDV was amplified using Reverse Transcription-Polymerase Chain Reaction (RT-PCR) then subjected for nucleotide and amino acid sequence detection. NDV Giza 2014 isolate and NDV Qualubiya 2014 were characterized as velogenic and lentogenic strains, respectively using Mean Death Time (MDT), Intracerebral Pathogenicity Index (ICPI) and studying amino acid motif of the F protein cleavage site. Phylogeny put NDV Giza 2014 in a separate branch independent from other Egyptian isolates, however it is more related to Lasota (genotype II) and Clone 30 vaccinal strains, while NDV Qualubiya 2014 was grouped more related to Ulster strain (genotype I) and Australian isolates originating from the same ancestral node however it is distantly related to other Egyptian strains 2005 and 2006 grouped together with a common ancestral node but on a separate branch. These results proved diversity between the isolated velogenic NDV Giza 2014 strain and other vaccinal and circulating NDV strains. It is concluded that sequencing of the complete genome of NDV is required to study genetic relatedness among NDV strains circulating in Egypt as well as further studies on the antigenic characters are required. Keywords: HI; ICPI; Lentogenic; MDT; NDV; PCR, Velogenic; phylogeny.

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34. A genotyping of a new avian infectious bronchitis virus isolated from chickens proventriculus in Egypt E.M. El-Nahas 1*, H.S. El-Sayed 2, S.S. El-Basuni 3, G.F. El-Bagoury 1 1

Department of Virology, Faculty of Veterinary Medicine, Benha University, 13736 Moshtoher , Benha, Egypt 2 National Laboratory for Veterinary Quality Control on Poultry Production, Animal Health Research Institute, Dokki, Giza. 3 Department of Poultry disease, Faculty of Veterinary Medicine, Benha University, 13736 Moshtoher , Benha, Egypt Abstract A novel avian infectious bronchitis virus (IBV), Egypt/Qal/014p was isolated from 14-day-old broiler chickens with proventriculitis in Qaluobia province of Egypt in 2014. The isolate was identified by a real time RT-PCR targeting the nucleocapsid (N) gene. Sequence analysis of partial S1 gene of Egypt/Qal/014p had close relation with that of the Massachusetts prototypes. Comparisons nucleotide and amino acid sequence of partial S1 gene showed that the recent isolate had 98.2% and 95.2% nucleotide similarities and 96.8% and 91.4% amino acid similarities with the commonly used IBV vaccine Massachusetts strains H120 and M41 respectively. Egypt/Qal/014p had a unique amino acid substitution at residues 39 (Serine), 40 (Tyrosine), 41 (Lysine), 64 (Glutamate) and 69 (Valine) of the S1 proteins from H120 and M41. The cross-neutralization test revealed antigenic relatedness of Egypt/Qal/014p with Massachusetts serotypes. The commercial H120 vaccine conferred partial protection against proventriculitis induced by Egypt/Qal/014p. In conclusion, IBV strains exhibiting proventriculitis were found co-circulating in broiler chickens in Egypt and optimum control of IBV in Egypt require preparation of vaccine from indigenous isolates beside periodic evaluation of crossprotective capabilities of such vaccine. Keywords: Egypt/Qal/014p; Partial S1 gene analysis; Antigenic relatedness; Protection test

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35. Avian influenza virus vaccine match against circulating H5N1 avian influenza viruses in Suez Canal region, Egypt Amira Mohamed Helal1, Abdel Satar Arafa2, Hanan Fathy Abdein3 Dalia Mansour Hamed3 and Mohsen El Dimerdash3 1

National laboratory for veterinary quality control on poultry production, Animal Health Research Institute, Ismailia, Egypt 2 National laboratory for veterinary quality control on poultry production, Animal Health Research Institute, Dokki, Giza, Egypt 3 Avian and Rabbit Medicine, Suez Canal University, Ismailia, Egypt Abstract Different types of inactivated imported and locally produced AI vaccines have been used in Egypt in order to control the AIV infection in poultry and regardless the countrywide control plan and vaccination policy of poultry to stop the widespread of HP H5N1 AIVs, continuous circulation and evolution of the viruses in all poultry sectors is still recorded every season. This reflects the need to review efficacy of vaccination strategy and commercially available vaccines used in the field. Therefore, in this study, samples were collected from different poultry production sectors in Suez Canal region during 2014-2015 then viral RNA was extracted and subjected to real time reverse transcription polymerase chain reaction (RT-qPCR) then positive samples were further subtyped for detection of the H5 gene then sequence analysis for HA gene and sequence similarity to match the recently circulating Egyptian viruses against the currently used imported and local vaccines seed strains. Our results revealed that the amino acid identity percent of vaccine seed strains of American and European origin showed the lowest similarity (76-77%) while the Chinese strains showed higher similarity (91-92%). However, the highest amino acid identity percent was observed with Egyptian seed strains (93-99%). Consequently, our study verified that there are wide range of genetic similarity of the Egyptian vaccine seed strains against the currently circulating field viruses in Egypt, this reflect the importance of continuous evaluation and updating of the vaccine seed strains to determine its efficacy against the currently circulating field virus.

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36. Variable Pathogenicity of Avian influenza viruses H5N1 isolated from Egypt Ola.A.Khader 1, Arafa. A.1, Hassan, M.K1, A.A. El-Sanousi 2, M.A. Shalaby 2 1

Reference Laboratory for Veterinary Quality Control on poultry production, P.O. Box264, Dokki, Giza 2 Department of Virology, Faculty of Veterinary Medicine, Cairo University, 12211.Giza, Egypt Abstract The highly pathogenic avian influenza virus H5N1 (HPAIV- H5N1) represents an important

poultry pathogen and constitutes a threat to mammals including humans. Egypt has been affected by HPAI-H5N1 since 2006 and the virus still circulates in poultry population until now. The virulence determinants for highly pathogenic avian influenza viruses (AIVs) are considered multigenic, although the best characterized virulence factor is the hemagglutinin (HA) gene. In the present study, pathogenicity of ten isolates (9 classic strains of 2.2.1 clade and one variant of 2.2.1.1) was examined in specific pathogen-free chickens (SPF). As a part of this investigation, sequence analysis of 2 genes HA, Nucleoprotein (NP) related to the virulence was carried out. The findings revealed that the 9 classical viruses contained multiple basic amino acids structure (PQGEKRRKKRGLF) in addition to the presence of one deletion of amino acid serine at position 129 (∆129). The variant virus contained cleavage site structure (PQGEGRRKKRGLF). The NP gene has one substitution in position 109 (I109V) in classic viruses instead of Isoluecine in variant virus. In conclusion, this study indicates the importance to trace virus pathogenicity markers in relation to other genes such as NP with the main pathogenicity determinants in HA gene in different clades of HPAI-H5N1 viruses.

Key words: Highly pathogenic avian influenza (HPAI), Hemagglutinin (HA), Nucleoprotein (NP), pathogenic signature.

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37. Trial for using emergency vaccination for controlling duck virus hepatitis (DVH) outbreaks in ducks Sabry, E.O.1, Hemat, S. Elsayed1, Naglaa Hagag2, Wesam Mady2 1

Department of Poultry Diseases, Animal Health Research Institute, Benha Branch, Benha, Egypt 2 National Laboratory for Veterinary Quality Control on Poultry Production, Animal Health Research Institute, Ministry of Agriculture, Dokki, Giza, Egypt

Abstract This study was conducted to evaluate the use of emergency vaccination to control DVH in ducks. Five liver samples were collected from Pekin ducks showing clinical symptoms of DVH from Kalubia province in summer 2016. The virus was isolated in specific pathogenic free (SPF) embryonated chicken eggs (ECES). The clinical samples were examined by generic RT-PCR followed by partial sequencing of the 3D gene; the results revealed that the isolate was characterized as Duck hepatitis A virus resembling the recent Egyptian virus (DHV/Duck/Egypt/F139/2016). Sequence and phylogenetic analysis indicated that the DHV/Duck/Egypt/F139/2016 virus is clustered in serotype-1 with highest percent identity of (97.9%) with DHV/Duck/Egypt/AL-Gharbia/2014 and with lowest identity (79.4%) with Duck hepatitis–A virus vaccine. The results obtained from emergency vaccination experiment indicated that emergency vaccination against DVH in young duckling in the experimentally infected and emergently vaccinated group gave good protection against infection with no deaths after emergency vaccination till the end of the experiment (40 days). In contrast, the second group (experimentally infected and not emergently vaccinated) showed deaths reached up to 40% of the ducklings and the lesions in dead and survived ducklings were severe. Therefore, emergency vaccination can be used effectively to face DVH outbreaks as a tool to control the disease. Keywords: DVH, SPF, RT-PCR and emergency vaccination.

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38. Evaluation of avian influenza H5N1 plasmid-based inactivated Egyptian vaccine and treatment with antiviral drugs Omnia M. Hassan1, Ahmed Mostafa1, Ahmed Kandeil1, Rabeh El shesheny1, Ali M. Ahmdy2, Mohamed A. Ramadan2, Mohamed A. Kutkat3, Mohamed A. Ali1 1

Center of Scientific Excellence for Influenza Viruses, National Research Centre, Giza, Egypt. Microbiology and Immunology Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt. 3 Veterinary Research Division, National Research Centre, Giza, Egypt.

2

Abstract Background:Although several essential preventive and therapeutic strategies could help to combat severe IAV pandemics and epidemics, vaccination is still the primary defence line to combat IAV infection. Objective: this work aimed to study the potency of plasmid-based H5N1 inactivated vaccine in controlling H5N1virus infection; and the antigenic variation as well as the cross-reactivity of the currently applied imported avian influenza vaccines. Besides, we performed a deliberate screening for the antiviral activity of commercial FDA approved antiviral drugs (acyclovir, ribavirin, amantadine HCl and Oseltamivir) to control H5N1infection. Also, commercial disinfectants (Chlorine and chloroxylenol) anti-septic activity was investigated in controlling of IAV H5N1infection. Methods: After waning of MA, 400 broiler experimental chickens were divided into 8 groups which received 0.5 ml of CEVAC vaccine intramuscular (IM), Re-5strainAI vaccine, ME FLUVAC vaccine, AI-VAC H5 vaccine, Kemiflu vakAI KV vaccine, and Re-1 strain AI vaccine, group was left as blank control, rg (M2583D) AI vaccine, respectively. The levels of raised antibodies in the sera of the vaccinated chicken were monitored with the HI assays using rg A/Duck/Egypt/M2383A/2010 (H5N1) as antigen. Results: Vaccines preparations did not provide any detectable titers until day 35 including the commercial version of the laboratory-made rg (M2583A) except for the laboratory made rg (M2583D) vaccine (≥80 HIU). Secondly ribavirin at different concentrations, ≥ 60µg/µl, provides full inhibition on viral propagation, amantadine showed resistance even with high concentrations. Acyclovir did not show any significant activity against avian influenza. Finally, popular disinfectants Chlorine achieved99% and 100% and chloroxylenol achieved 50% and 99.6% reduction in TCID 50of H5N1. Conclusion: 1) KEMI-FLU and the inactivated RG vaccine provided high cross-reactive antibodies with classical 2.2.1 Egyptian isolate during the period of study. 2) Ribavirin is a potent anti-influenza drug in vitro. 3) Chlorine(Clorox) and chloroxylenol (Dettol) showed potent anti-viral activity against avian influenza A/H5N1 Keywords: Avian influenza; H5N1; vaccines; disinfectants; antiviral drugs; hemagglutination; plasmid-based; reverse genetics

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39. In silico design and vitro validation of efficient siRNAs as universal silencers of the polymerase genes of influenza A virus Sara H.Mahmoud1*, Rabeh El Shesheny1, Mahmoud El Hefnawi2*, Naglaa Abdallah3, Abdelhadi Abdallah3, Ahmed Mostafa2, Mohamed Ali2 1

Influenza Research Laboratory, Environmental Research Division, National Research Center (NRC), El-Bohous Street, Cairo, Egypt 2 Biomedical Informatics and chemoinformatics group, Informatics and Systems Department, Division of Engineering Research and Centre of Excellence for Advanced Sciences, National Research Centre, El-Bohous Street, 12311 Cairo, Egypt 3 Genetics Department, faculty of agriculture, Giza square, 12311 Cairo, Egypt Abstract Background: Influenza epidemics are caused mainly by influenza A viruses (IAVs) and result in 0.25-0.5 million deaths annually. The numbers of cases and deaths are multiplied into millions in influenza pandemic events. Besides, IAVs has broad spectrum host range with highly dynamic evolution rate developing new IAV variants of unknown characteristics. Objective: Therefore, there is an urgent need for new medications to combat Influenza epidemics and pandemics. RNA interference (RNAi) is a naturally occurring endogenous biological posttranscriptional cellular mechanism that regulates against foreign genetic elements. Small interfering RNA (siRNA) molecules utilize this mechanism to promote homology dependent messenger RNA (mRNA) degradation. siRNA triggers the formation of RNA induced silencing complex (RISC) in which the double stranded siRNA is incorporated and unwounded then binds to the target mRNA sequence resulting in its cleavage from the genome. Methods: By analysis of the IAV genome, available in Genbank, we designed and performed a combinatorial exhaustive systematic methodology for optimal design of universal therapeutic shortinterfering RNA (siRNA) molecules targeting all diverse IAVs. Here, we focused on the selection and validation of optimal siRNAs targeting IAV polymerase genes. Results: The selection of PB1-CR08_032 and PA-CR17_021 as candidate optimal siRNAs for IAV silencing was based on our novel in silico bioinformatics methodology for siRNA design. Briefly, siRNAs (20-40 pmol) were transfected individually into MDCK cells using Lipofectamine followed by cell-infection with H1N1 and H5N1 strains at MOI=0.01. Interestingly, a significant drop in viral load was observed in siRNAs-treated cells when compared to non-transfected/infected MDCK cells (control) even at the lowest concentration of 20 pmol at 6 hrs post-infection (h p.i.). Conclusion: Based on our results, we assume that siRNAs could serve as potential universal therapeutics for IAVs. Keywords: Influenza Viruses; siRNA; Polymerases; Silencing.

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40. Reassortment between avian highly pathogenic H5N1 and low pathogenic H9N2 influenza viruses and its impact on pathogenicity and infectivity in chickens Yassmin Moatasim1, Ahmed Kandeil1, Ahmed Mostafa1, Sary Khaleel Abd elghaffar2, Rabeh El Shesheny1,3, Ghazi Kayali4,5, Ahmed H. M. Elwahy6, Mohamed Ahmed Ali1 1

Center of Scientific Excellence for Influenza Viruses, National Research Centre (NRC), Dokki, Giza, Egypt. 2

Pathology and Clinical Pathology Department, Faculty of Veterinary Medicine, Assuit University, Assuit, Egypt. 3

4

Department of Infectious Diseases, St. Jude Children’s Research Hospital, Memphis, TN, USA

Department of Epidemiology, Human Genetics, and Environmental Sciences, University of Texas Health Sciences Center, Texas, USA 5 6

Human Link, Hazmieh, Lebanon

Chemistry Department, Faculty of Science, Cairo University, Giza, Egypt

Background: H5N1 and H9N2 influenza viruses have been extensively circulating in various avian species and frequently infecting mammals including humans. This synchronous circulation of both subtypes of influenza A viruses (IAVs) provides a scope for these viruses to reassort their genome, posing an unpredictable threat to the global public health and leading occasionally to a pandemic situation. Being a prospective epicenter for the next pandemic caused either by H5N1 and H9N2 viruses, reassortment between HPAIV H5N1 and LPAI H9N2 is likely emerging in Egypt. Objective: To assess the potential risk of the IAV reassortants generated by inclusion of different segments from a HPAIV H5N1 in the genetic background of an Egyptian LPAIV H9N2 strain. Methods: using reverse genetics technology of IAV, we generated a set of IAV reassortants carrying the genetic segments of clade 2.2.1.2 influenza A/duck/Egypt/Q4596D/2012 (H5N1, most prevalent clade in Egypt) in the genetic backbone of an influenza A/chicken/Egypt/S4456B/2011 (H9N2) strain, a representative of G1-like H9N2 lineage which is widely circulating in Egypt. Furthermore, the genetic compatibility, growth kinetics and virulence were evaluated using MDCK and A549 cell lines and SPF embryonated eggs. Results: Out of eight H9-reassortants, we could rescue only 5 reassortant viruses, either due to difficulty in cloning (H5-PB1) or genetic incompatibility (H5-NP and H5-NA). Results revealed higher replication rates of the H9N2 virus having NS segment of H9N2 virus. Survival results showed that the lowest rate belongs to H5N1 parent virus followed by H9N2 virus having HA of H5N1. Conclusion: Our findings also suggest that all forms of generated reassortant viruses are lower in their pathogenicity than H5N1 virus. Keywords: Avian influenza virus, Egypt, H5N1, H9N2, reverse genetics and Reassortment.

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Thrusday, December 1, 2016 Scientific Session III: Antiviral Therapy and Potential Antiviral Targets Chairman Prof. Claus-Peter Czerny Co-Chairmen Dr. Kathleen Laura Hefferon, Prof. Aboulata E. Aboulata Keynote Lecture: 9.00 – 9.30

Epitope mapping and engineering of site-directed protective recombinant antibodies Claus-Peter Czerny Department of Animal Sciences, Georg-August-University Göttingen, Germany. (Abst. No. 41) Keynote Lecture:

9.30 – 10.00

New Applications of Plant Virus Nanoparticles Kathleen Laura Hefferon Food Sciences Dep. Cornell University (Abst. No. 42) Keynote Lecture:

10.00 - 10.30

10.30 – 10.45

10.45 – 11.00

11.00- 11.15

11.15 – 11.30

Precore stop codon regulates the stability of HBV X-mRNA Husein H Aly National Institute of Infectious Diseases, Tokyo, Japan. (Abst. No. 43) Interleukin-27 Is Differentially Associated with CD4+ T Cell Counts in HIV Saudi patients under Highly Active Antiretroviral Therapy Nagwa M. Aref, Haifa M. Al-Nafea and Nuha M. Hamdy (Abst. No. 44) Dispersed gold nanoparticles potentially ruin Gold Barley Yellow Dwarf Virus and eliminate virus infectivity hazards Nagwa M. Aref and Noorah A.Alkubaisi (Abst. No. 45) In vitro Studies on antiviral activity of silver nanoparticles against Foot and Mouth Disease virus. Sonia A. Rizk; Safy E. Mahdy; Amr I. Hasanin; Hiam M Fakhry and Ahmed F. Ramadan. (Abst. No. 46) Break

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41. Epitope mapping and engineering of site-directed protective recombinant antibodies Claus-Peter Czerny Professor of Animal Hygiene & Managing director of the Department of Animal Sciences at the Georg-August-University in Goettingen Abstract Genomic and proteomic research are currently generating increased knowledge on antigenic and functional sites of infectious agents. Antibodies are considered to be the first choice to determine those structures as possible targets if delivered by vaccines. Antibodies are valuable compounds, as well, for passive immunization and inhibit virus replication by blocking functional important structures. However, species-specific antibodies are required for this reason to compass severe allergic side effects. The establishment of immunoglobulin libraries is a new approach to engineer target optimized and species-specific recombinant antibodies for prophylactic and therapeutic reasons. The technique is meanwhile well established for the application in human and veterinary medicine. Species-specific antibody libraries can be used to generate high affine antibodies as promising future therapeutics, where other strategies fail or do not exist. The principle will be demonstrated on the development of recombinant human antibodies against vaccinia virus (VACV) and recombinant bovine antibodies against the bovine coronavirus (BoCV). By screening of a genome expression library, functional antibody assays and epitope mapping, we identified sequential and conformation dependent antigenic sites on the vaccinia virus envelope proteins A27 and D8. A naïve human library established from lymphocytes of blood donors led to the generation of a recombinant human single chain antibody (scFv), which confirmed in vitro neutralization and partial protection in a monkeypox virus model with macaques (Macacus mulatta). For the development of further OPXV specific human scFvs, the IgG repertoire from 4 donors vaccinated intracutaneously with “Dryvax” (Wyeth Laboratories, USA) was amplified. The scFvassortment was cloned into the phagemid pCANTAB5E allowing the soluble scFv expression in E. coli. This library displayed a diversity of ≥4x108 independent colonies. Different immunoscreening protocols against VACV Elstree revealed a predominant selection of scFv-clones specifically binding to the 32 kDa D8 protein. The epitope of the scFv was mapped on the D8 protein by truncated and recombinant OPXV envelope proteins, as well as by competitive ELISAs. The scFv was also engineered into the larger human Fc- and IgG1-formats and transiently expressed in HEK293T and HEK293-6E cells. Similar binding affinities of 1.4 nM and 5.4 nM, respectively, were shown by the scFv and scFv-Fc antibody molecules, whereas the recombinant IgG1 antibody was much more efficient (60.6 pM). None of the purified recombinant antibodies was able to neutralize 100 pfu of VACV Munich 1 in vitro. However, neutralization was achieved by cross-linking of the scFv molecules with a polyclonal goat anti-E tag-antibody. A scFv concentration of 0.39 - 0.78 µg/ml was able to block plaque formation to 50% under these conditions. The larger antibody formats scFv-Fc and IgG1 neutralized VACV in vitro with similar concentrations after addition of 1% human complement. The recombinant antibodies were also tested in an in vivo NMRI-mouse-model, and gave partial protection after passive intraperitoneally immunization. We also constructed a bovine scFv phage display library using a new universal primer set. An isolated recombinant bovine scFv antibody against an envelope protein of Bovine Coronavirus showed excellent affinity and neutralization abilities, making it a promising candidate for further therapeutic applications in newborn calf diarrheas.

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42. New Applications of Plant Virus Nanoparticles Kathleen Hefferon, Department of Food Sciences, Cornell University Abstract Plants have been explored for their potential to act as production platforms for biopharmaceuticals, such as vaccines and monoclonal antibodies, for over two decades. Plant viruses have played an integral role in the emergence of plants as inexpensive and facile systems for the generation of therapeutic proteins. Plant viruses have more recently been engineered into nanoparticles which can target a variety of cancers by empowering the immune system to block tumor progression. This presentation describes the employment of plant virus expression vectors to treat some of the most challenging diseases known today, and the potential impact of this for developing countries

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43. Precore stop codon regulates the stability of HBV X-mRNA Hussein H Aly1, Junya Suzuki1, Koichi Watashi1, Kazuaki Chayama2, and Takaji Wakita1 1

2

National Institute of Infectious Diseases, Tokyo, Japan Gastroenterology and Hepatology department, Hiroshima University, Japan Abstract

Background: Hepatitis B virus (HBV) is a stealth virus, minimally inducing the Interferon system required for efficient induction of both innate and adaptive immune responses. However, 90% of acutely infected adults can clear the virus, suggesting the presence of other, interferon-independent pathways leading to viral clearance. Objective: The identification of unknown interferon-independent anti-viral pathways suppressing HBV infection. Methods Helicases are nucleic acid interacting proteins, and some of them like RIG-I and MDA5 induce the anti-viral response. Using shRNA library, we analyzed there effect on HBV life cycle. Results: We identified HBV suppression by SKIV2L RNA helicase. SKIV2L, an RNA exosome cofactor, showed a direct and preferential interaction with HBV X-mRNA. SKIV2L expression was essential for the degradation of HBV X-mRNA at the RNA exosome complex through a 3’ antisense mechanism. The HBV X-mRNA was targeted for degradation by the translation-dependent Non-Stop Decay (NSD) RNA quality control pathway. The initiation of translation from the redundant 3’ precore translation initiation site without a proper stop codon at the 3’ end of HBV X-mRNA transcript was identified by NSD, leading to the degradation of HBV-X mRNA. On the opposite hand, introduction of a stop codon (via the G1896A mutation) to the 3’ precore ORF increased the stability of this transcript. Interestingly, G1896A mutation is frequently observed in HBV-eAg negative patients, a subgroup characterized by high levels of HBV replication and more severe liver disease. HBV X protein is a transcription transactivator that is important for the transcription of HBV-mRNAs and HBV life cycle. The precore stop codon enables the virus to escape the degradation of HBV X-mRNA.. Conclusion: Due to the compact genomic structure, some viruses like HBV exhibits overlapping ORFs. This viral character is identified by NSD as an abnormal mRNA, leading to its degradation at the SKIV2L/RNA-exosome system.

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44. Interleukin-27 Is Differentially Associated with CD4+ T Cell Counts in HIV Saudi patients under Highly Active Antiretroviral Therapy Nagwa M. Aref 1* Haifa M. Al-Nafea2 and Nuha M. Hamdy 3 1

.Ain Shams University, Faculty of Agriculture, Microbiology Department, Cairo Egypt & King Saud University, College of Science, Botany and Microbiology Departement, Riyadh, 11495 SAUDI ARABIA. 2 Department of Clinical Laboratory Sciences, College of Applied Medical Sciences King Saud University, Riyadh, King of Saudi Arabia. 3King Saud Medical City, Clinical Pathology, College of Medicine, Riyadh. KSA. Abstract

Background: The IL-27 is a pleiotropic essential host factor for HIV- infection that modulates both the innate and adaptive immune responses. Objective: The potential use of plasma cytokines concentrations during acute HIV-1 infection especially IL-27 that could predict subsequent disease progression. Methods: The study design incorporated sixty-six HIV-1/AIDS Saudi patients under Highly Active Antiretroviral Therapy (HAART1 and HAART2) and other twenty healthy individuals as a control. Flow cytometer estimated lymphocyte immunophenotyping which resembles the total of absolute counts of mature human lymphocyte subsets in whole blood. While plasma IL-27and other cytokines; INF-γ, IL-2, IL-10, IL-4, IL-6, and TNF-α measured in pg/ml by Flow Immunoassay (Luminex 200). COBAS AmpliPrep assessed the association and quantitation of HIV-1 RNA viral load in plasma. Results: The results showed enormous differences between control and HAART2 groups with elements (INF-γ, IL-10, IL-2, IL-4, IL-6, IL-27, TNF-α).All of them had strong values at the level of 0.01 while IL-27 were significant at the level of 0.05. The IL-12 family included a heterodimeric cytokine of Interleukin 27 (IL27) which antigen-presenting cells produce it. It plays an important function in regulating the activity of B- and T-lymphocytes. IL-27 had the first protective effect against HIV-1 infection. Conclusion: ARV therapies induce quantitative and qualitative alterations in the cytokine profile of persons that could be warrants attention which causes reductions in AIDS deaths. It is an important Immune-biomedical study for AIDS Saudi patients under medication to be followed up by physicians for managing the AIDS morbidity and mortality in KSA. . Keywords: IL-27, HAART, HIV, CD4+ T cells, KSA.

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45. Dispersed gold nanoparticles potentially ruin Gold Barley Yellow Dwarf Virus and eliminate virus infectivity hazards Nagwa M. Aref 1* and Noorah A.Alkubaisi2 1.

Ain Shams University, Faculty of Agriculture, Microbiology Department, Cairo Egypt & King Saud University, College of Science, Botany and Microbiology Departement, Riyadh, 11495 SAUDI ARABIA.2. King Saud University, College of Science, Botany and Microbiology Departement, Riyadh, 11495 SAUDI ARABIA Abstract

Background: Gold nanoparticles (AuNPs) application melted Barley Yellow Dwarf Virus-PAV (BYDVPAV) spherical nanoparticle capsids. Objective: Synergistic therapeutic effects for plant virus resistance induced by interaction with binding units of prepared AuNPs in a water solution. Methods: AuNPs was characterized and evaluated by Zeta Sizer, Zeta Potential and Transmission Electron Microscopy (TEM). The yield of purified nanoparticles of BYDV-PAV obtained from Hordeum vulgare (Barley) cultivars; Local and Giza 121/Justo. Results: Virus yield was 0.62 mg/ml from 27.30g of infected leaves at A260/A280 ratio. Virus nanoparticle has a spherical shape with 30 nm in size by TEM. BYDV-PAV combined with AuNPs to challenge virus function in-vivo and in-vitro. Dual AuNPs existence in-vivo and in-vitro affected compacted configuration of viral capsid protein in the interior surface of capsomers, the outer surface, or between the interface of coat protein subunits for 24h and 48h incubation period in-vitro at room temperature. The size of AuNPs that had a potentially dramatic deteriorated effect possess sizes 3.151 and 31.67 nm with a different intensity of 75.3% for the former and 24.7% for the later which enhances optical sensing applications to eliminate virus infectivity. Damages of capsid protein due to AuNPs on the surface of virus subunits caused variable performance in four different types in TEM named: puffed, deteriorated and decorated, ruined and vanished. Viral yield showed remarkably highintensity degree of particle symmetry and uniformity in Local cultivar greater than in Giza 121/Justo cultivar. Conclusion: It noticed the high yield of ruined VLPs in Local cuv. than Justo cuv. AuNPs indicated complete lysed of VLPs and some deteriorated VLPs at 48h. Keywords: Dispersed Gold nanoparticles; Gold Barley Yellow Dwarf Virus; Plasmonic resonant energy; Agri-nanotechnology; Viral bio templates; Optical sensing; Optical scattering energy; AuNPs optical antenna

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46. In vitro Studies on antiviral activity of silver nanoparticles against Foot and Mouth Disease virus. Sonia A. Rizk; Safy E. Mahdy; Amr I. Hasanin; Hiam M Fakhry and Ahmed F. Ramadan.

Veterinary Serum and Vaccine Research Institute, FMD Department Abstract Background: The advance in nanotechnology has enabled us to utilize particles in the size of the nanoscale. Nanoparticles excel as therapeutic agents due to their unique physiochemical properties and a universally applicable physical mode of action. Silver nanoparticles (AgNPs) have received tremendous attention for their antimicrobial properties. The use of metal nanoparticles provides an interesting opportunity for novel antiviral therapies. Since metals may attack a broad range of targets in the virus there is a lower possibility to develop resistance as compared to conventional antivirals virus infections pose significant global health challenges as viral infections constitute one of the main health problems, especially in view of the fact that the emergence of resistant viral strains and the adverse side effects associated with prolonged use continue to slow down the application of effective antiviral therapies. Silver nanoparticles exert anti-viral activity at an early stage of viral replication, most likely as a virucidal agent or as an inhibitor of viral entry binding, fusion, and infectivity, acting as an effective virucidal agent against cell-free virus and cell-associated virus. Besides, silver nanoparticles inhibit post-entry stages of the virus life cycle. These properties make them a broadspectrum antiviral agent. Objective: Evaluation of the antiviral activity of Silver nanoparticles (AgNPs) against Foot and Mouth Disease virus in lab. Methods: Determination of the effect of different concentrations of silver nanoparticles (AgNPs) 10 nm particle size measuring with Electron microscope, against different types of FMDV on Baby Hamster Kidney cell line using different concentration (15- 420 μg / ml) of AgNPs. Results: AgNPs concentrations 30-60 μg / ml had no cytotoxicity effect on cells also revealed 100 % reduction in the virus infectivity. Conclusion: This study confirmed the antiviral activity of silver nanoparticles against Foot and Mouth Disease virus.

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Scientific Session IV: Diagnosis and Diagnostic Tools Chairman Prof. Mohamed A. Shalaby Co-Chairmen Prof. Khaled A. Dougdoug, Prof. Hussein H. Aly 11.30 – 12.00 Keynote Lecture: The mobile suitcase laboratory: A tool for the rapid detection of emerging and endemic infectious disease Ahmed Abd El Wahed Division of Microbiology and Animal Hygiene, Georg-August-University Goettingen, Germany (Abst. No. 47) No additional value for the detection of Zika virus from urine of Brazilian 12.00 – 12.15 patients in the first five days of infection Rodrigo Pessôa, João Veras Patriota, Maria de Lourdes de Souza, Ahmed Abd El Wahed and Sabri S. Sanabani (Abst. No. 48) 12.15 – 12.35 12.35 – 12.50

12.50 – 13.00

Rapid and Quantitative Detection of some food born Viruses by Real-Time Reverse Transcription-PCR in Egypt Randa M. farag, S. M. E. A. Amer, K. A. El Dougdoug (Abst. No. 49) Analyzing infection of chicken oviduct explant cultures of differentiated oviduct epithelial cells by infectious bronchitis virus in vitro Sahar Abd El Rahman, Ali El Kenawy, Christine Winter, Ulrich Neumann, and Georg Herrler (Abst. No. 50) General discussion

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47. The mobile suitcase laboratory: A tool for the rapid detection of emerging and endemic infectious disease Ahmed Abd El Wahed1, Pranav Patel2, Oumar Faye3, Mohamed A. Shalaby4, Matthias Niedrig5, Claus-Peter Czerny1, Amadou A. Sall3, Frank T. Hufert6, Manfred Weidmann7 1Division of Microbiology and Animal Hygiene, Georg-August-University Goettingen, Germany 2TIB MOLBIOL Syntheselabor GmbH, Berlin, Germany 3Arbovirus unit, Pasteur Institute, Dakar, Senegal 4Department of Virology, Faculty of Veterinary Medicine, Cairo University, Egypt 5Robert Koch Institute, Berlin, Germany 6Brandenburg Medical School Fontane, Senftenberg, Germany 7Institute of Aquaculture, University of Stirling, Stirling, Scotland, UK The classical method for the diagnosis of infections is the isolation of their causative agents, which takes several days and is only possible in a few specialized laboratories. Detection of the infectious agent by using the rapid antigen lateral flow test is an easy to be applied at the point-of-care. Nevertheless, the results must be confirmed by additional laboratory tests. Laboratory diagnosis mainly depends on nucleic acid detection by real-time polymerase chain reaction (PCR), which is available in central laboratories and has a turnaround time of more than two hours. For decentralizing of the molecular diagnostics, there is a need for a simple molecular point-of-need test. We have developed a mobile suitcase laboratory (62x49x30 cm) containing all reagents and equipment for the detection of the nucleic acid of infectious agents using the recombinase polymerase amplification (RPA) technology. The RPA assay run at a constant temperature (42°C) for a maximum of 15 minutes. Moreover, all reagents were cold-chain independent and the mobile laboratory was operated by a solar power battery. The nucleic acid extraction was performed by a magnetic bead based method, in which a simple fast lysis protocol was applied. In case of highly infectious agents such as Ebola virus, the inactivation step was performed in a glovebox. We further have developed RPA tests for the detection of pathogens of relevant medical and veterinary importance such as Dengue, Zika, Chikungunya, Ebola, Corona-, foot and mouth disease, and Lumpy Skin Disease viruses as well as Tuberculosis and paratuberculosis. In conclusion, the mobile suitcase laboratory is ideal for rapid sensitive and specific detection of pathogens especially at low resource settings, quarantine stations or during outbreak situations.

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48. No additional value for the detection of Zika virus from urine of Brazilian patients in the first five days of infection Rodrigo Pessôa1, João Veras Patriota2, Maria de Lourdes de Souza2, Ahmed Abd El Wahed3 and Sabri S. Sanabani4 1Department of Pathology, School of Medicine, University of São Paulo, São Paulo, Brazil 2Municipal Hospital of Tuparetama, Pernambuco, Brazil 3Division of Microbiology and Animal Hygiene, Georg-August-University, Goettingen, Germany Zika virus (ZIKV) belongs to the genus flavivirus, a group of RNA viruses transmitted by arthropods. In 2007, ZIKV caused an outbreak of relatively mild disease characterized by rash, arthralgia, and conjunctivitis in Yap State, Federated States of Micronesia. The first cases in the Americas were detected in Brazil in May 2015. Considering the serological diagnosis of ZIKV is challenging due to the cross-reactivity between members of the flavivirus genus, virus isolation and/or detection of ZIKV RNA during the acute phase is favorable. Despite the availability of the molecular tests, the laboratory diagnosis is challenging because of the low-level or absence of viremia. Recently, Gourinat et al (1) observed that ZIKV RNA in six patients from the French Polynesia is detectable in urine at higher levels and for longer periods than the corresponding blood samples. These results were confirmed by another group from returning travelers to the USA (2) and opened the door for the utilization of urine for the diagnosis of ZIKV infection. To investigate the consistency of these results, we have screened plasma and corresponding urine samples from 61 patients that were collected during the large outbreak that occurred in Tuparetama, Brazil. All samples were collected during the first five days of onset of symptoms. Among the 61 patients, 46 (75.4%) were positive in plasma specimens and 37 (60.6%) in urine using real-time RT-PCR. In 28 (45.9%) cases, ZIKV RNA was detected in both samples. Surprisingly, no difference between the Ct values of ZIKV RNA in plasma and urine was recorded. In contrast to others, our results demonstrated that the majority of patients with active infection of ZIKV had a better detection rate in plasma than in urine. Moreover, our data do not support the recent finding that indicates higher level of ZIKV RNA in urine than in serum during the acute phase of the disease (2). Nevertheless, the urine sample has one advantage that the ZIKV persists for longer period (1). In conclusion, we recommend the simultaneous testing of blood and urine samples in ZIKV infected individuals with focus on plasma samples in the first five days of infection. Further investigations will be required to determine other factors influencing the ZIKV pathogenesis.

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49. Rapid and Quantitative Detection of some food born Viruses by Real-Time Reverse Transcription-PCR in Egypt Randa M. farag1; S. M. E. A. Amer 2; K. A. El Dougdoug 3 1 Health Sci., Dept., Fac. of Health Sci. and Physiotherapy, PNU, Riyadh, KSA. 2 Botany and Microbiol. Dept. Fac. of Sci. Zagazeg Univ., Egypt. 3Microbiol. Dept. Fac. of Agric. Ain Shams Univ., Egypt Introduction: In recent years it has been recognized that viruses are an important cause of food-borne disease .There are more than 12 families of viruses transmitted to humans by eating contaminated food. It is this Picornaviridae and Reoviridiae families, which include a total of Enteroviruses addition to hepatitis A virus epidemic. Viruses do not grow or multiply in or on foods, but foods may become contaminated with human viruses and transmit infection. Methods: Five samples from fresh fruit and vegetables (strawberry, Lettuce, green onion, and watercress) were collected and washed from the site of drainage water, (El menofiya; Baher El-baker and El- Manzala). The samples were extracted and concentrated for Rotavirus and Enterovirus HAV , HEV detection and assessment qualitatively and quantitatively by molecular, with RT-PCR and Real time rt-PCR ; serological with ELISA and Rapid chromatographic and biological by cultivated on Vero,Caco and HEp-2 cell line methods. Results : The food borne viruses were assayed qualitatively and quantitatively by (Real time PCR) in concentrated drainage water of Baher El-baker , El menofiya and El- Manzala but not detect in drainage water sabal , Tala and El-Rahawy ,as well as it was detected in concentrated extract of fruit and vegetables samples. On the other hand concentrated extracts of the vegetative food and drainage water samples whish gave negative results with Rapid chromatographic Immunoassay were cultivated on Vero,Caco and HEp-2 cell line for food borne viruses . ELISA kit and Rapid chromatographic Immunoassay were used for food borne viruses antigens detection in propagation samples. It was found that, the extracts of vegetative food crops detected with Elisa test gave positive result and not detected with Rapid chromatographic Immunoassay tests. The isolated food borne viruses on cell line Vero cell , Caco cell and HEp-2 were identified depended on some biological properties .It was found that, the titration of Rotavirus and Enterovirus particles was estimated by plaque assay on Caco and HEp-2 cell (9.5x106) and (1.2x104).Cytopathic effects were studied on Coca and Vero cells. The viruses caused cell death such as death and survival cell round cell and gianted cell. Haematoxylin and Eosin stain showed the characteristic changes post infection. Rotavirus and Enterovirus particles or clarified infected cell culture showed naked with Icosahedral symmetry using Phosphotungstic acid by Electron microscope. The amplification fragmented of VP6 gene of Rotavirus using conventional RTPCR was expected size 231 bp by RT-PCR using specific primer vp6-F/vp6-R, and the amplification fragmented of vp1gene of Enterovirus using conventional RT-PCR was expected size 242bp by RTPCR using specific primers vp1-F/ vp1-R5'. Conclusion: Based on the previous results it could be concluded that rt-RT-PCR and RT-PCR could be used as sensitive tools for confirmation of Rotavirus isolation from water and vegetable concentrates. ELISA and rapid chromatography immunoassay card test is somewhat poorly sensitive compared with rt-RT-PCR for detection of Rotavirus. Cell line culture useful for recovery, propagation, and titration and study the cytopathic effect of Rotaviruses and Enterovirus. Key Wards. Rotavirus, Enterovirus, Drains water, Foodborne viruses, RT-PCR, rt-RT-PCR, serology cellline, plaque assay, Electron microscope, Haematoxylin and Eosin stain.

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50. Analyzing infection of chicken oviduct explant cultures of differentiated oviduct epithelial cells by infectious bronchitis virus in vitro Sahar Abd El Rahman1, Ali El Kenawy1, Christine Winter2, Ulrich Neumann3, and Georg Herrler2 Department of Virology1, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt. Institute of Virology2, and Clinic for Poultry3, University of Veterinary Medicine Hannover, Hannover, Germany. Abstract Infectious bronchitis virus (IBV), an avian coronavirus, replicates not only in the respiratory epithelium, but also in other tissues e.g. kidney, intestine, gonads and the oviduct. Especially infection of the reproductive system has a great economic impact on the poultry industry. Here, we established a culture system for cells from the oviduct, which can be considered as a main target organ of IBV infections. Our method allows virus infection of cells within their natural tissue composition under in vitro conditions. Virus antigen was detected already at 8 hours post-infection in most parts of the oviduct. Lectin staining revealed that the sensitive epithelial cells express alpha2, 3-linked sialic acid. This is the first study, using chicken oviduct epithelial cell explants to study viral infection and sialic acid distribution. Keywords: (IBV; chicken oviduct epithelial cells; Sialic acid; QX strain)

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Scientific Session Va: Epidemiology, Evolution and emerging viruses Prof. Ahmed El- Kafrawy Chairman Prof. Sayed Salem Co-Chairmen Prof. Yosry A Radwan Keynote Lecture: 16.00 – 16.30

Equine Herpes Viruses (Risks for Arabian Horses) Youssry A. Radwan Former professor, FVMCU & WAHO associate member (Abst. No. 51) Keynote Lecture:

16.30 – 17.00

Emerging Viral Diseases of Aquatic Animals: Regional perspective Alaa Eldin Eissa Dept of fish diseases, Faculty of Veterinary Medicine, Cairo University. (Abst. No. 52) Keynote Lecture:

17.00 – 17.30

17.30 – 17.45

17.45 – 18.00

18.00 – 18.15

Distribution of Hepatitis E Virus Genotypes from human and animals reservoir Ahmed M. El-Adly Department of Botany & Microbiology, Faculty of Science, Al-Azhar University, Assiut, Egypt. (Abst. No. 53) Predominance and Geo-Mapping of avian influenza H5N1 in poultry sectors in Egypt Abdelsatar Arafa, Ihab El-Masry, Shereen Kholosy, Mohammed K. Hassan, Mousa Soliman, F. O. Fasina, Gwenaelle Dauphin, Juan Lubroth, Yilma Makonnen (Abst. No. 54) Surveillance for Middle East Respiratory Syndrome Coronavirus in camels from 2014 to 2016 in Egypt Mahmoud M. Shehata, Ahmed N. El-Taweel, Mokhtar R. Gomaa, Ahmed Kandeil, Rabeh El-Shesheny, Ahmed S. Kayed, Basma Elsokary, Nagla Hassen, Ola Bagato, Yasmin Moatasem, Sara Husein,Omnia Kutkat, Asmaa M. Maatouq, Pamela P. McKenzie, Ahmed Othman, Richard J. Webby, and Ghazi Kayali, Mohamed A. Ali (Abst. No. 55) Break

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51. RISKS OF (EHV) FOR ARABIAN HORSE Youssry A. Radwan Faculty of Veterinary Medicine, Cairo University, Egypt (Former) & World Arabian Horse Organization, IAM # 1427,UK Arabian horse is one of the oldest, beautiful and pure bred species among all other worldwide horses. In addition to being admired for its beauty – in show events - the Arabian horse is an ideal all-round other horses for endurance and stamina. Arabian horse developed special qualities which make it an extremely very highly important valuable horse. Equine Herpes Virus (EHV) is a common DNA virus that occurs in horse populations worldwide.To date, nine EHVs have been identified, three of these, EHV-1, EHV-3 and EHV-4, pose the most serious health risks for domesticated horses. EHV-1, which causes abortion, respiratory disease and neurologic disease; EHV-3, which causes a venereal disease called coital exanthema that affects the external genitalia and EHV-4, which usually causes respiratory disease only but can occasionally cause abortion and rarely neurological disease. In this context, I will address Equine herpes virus type 1 causing epidemic abortion storms and considerable economic losses of pregnancies in the very valuable Arabian mares.

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52. Emerging Viral Diseases of Aquatic Animals: Regional perspective

Alaa Eldin Eissa Department of Fish Diseases and Management, Faculty of Veterinary Medicine, Cairo University, Giza, Egypt. Abstract The expanding rise in aquaculture “blue revolution” to compensate the catastrophic overexploitation of global fisheries have unintentionally changed the dynamics of natural biotic elements (pathogenic or non pathogenic) in aquatic environment. Such change have widely involved the introduction of new aquatic species into new environments with subsequent transfer of their associated flora which imposed deep negative impact on the local aquatic species in this environment. Further, the swiftly evolving environmental pollution has potentially triggered colossal forms of climate changes such as global warming, sea level rise destruction of breeding habitats, disruption in genetic pools of vulnerable aquatic animals , mass migration of invasive aquatic species and finally emergence / resurgence of pathogens causing unexpected epidemics among aquatic creatures. Further, the outgrowing maritime transport , development of multi-species integrated aquaculture systems , domestication of marine mammals offshore, ease in illegal trafficking / smuggling of live aquatic animals and their products from country / region to another are all the main triggering factors in emergence of diseases among aquatic creatures. Aquatic animal epidemiologists have proposed the following criteria for a disease to be considered as an emerging disease: (i) new or previously unknown disease; (ii) known disease appearing for first time in a new species (expanding host range); (iii) known disease appearing for the first time in a new location (expanding geographic range); and (iv) known disease with a new presentation (different signs) or higher virulence due to changes in the causative agent). Thus, several viral diseases have been considered emerging diseases of fish, shellfish and marine mammals when the emergence criteria has been utilized. Regionally, Spring viremia of carp (SVC), Koi herpes virus (KHV) , Red seabream iridovrus (RSIV), Viral nervous necrosis (VNN) of grouper and recently tilapia, White spot syndrome virus of Shrimp / crayfish (WSSV), Taura syndrome virus of shrimp (TSV), Yellow tail disease virus (YTDV), Influenza virus marine mammals (avian, swine , human) (IVD), West Nile virus (WNV) of marine mammals and Morbilliviruses of marine mammals (MV) are all potential emerging viral diseases in the regional aquatic environments. Keywords: Emerging viruses, Fish viruses, Shellfish viruses, Aquatic animals, Marine mammals.

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53. Distribution of Hepatitis E Virus Genotypes from human and animal reservoirs Ahmed M. El-Adly 1* and Naeima M H Yousef2 1Department of Botany & Microbiology, Faculty of Science, Al-Azhar University, Assiut, Egypt. 2Department of Botany & Microbiology, Faculty of Science, Assiut University, Assiut, Egypt. Abstract Hepatitis E virus (HEV) belongs to the genus Hepevirus in the family Hepeviridae. HEV is a small, icosahedral, spherical particles of 27-34 nm, non-enveloped virus with a single-stranded, positivesense RNA genome of approximately 7.2 kb containing three open reading frames (ORFs), ORF1, ORF2 and ORF3, where ORF3 partially overlaps ORF2. HEV is distributed all around the world, and classified into four genotypes and at least two putative new genotypes belong to only one serotype that can infect mammals, named after the place of isolation of the reference strains. Two of the genotypes contain strains which infect only humans (HEV-1 "Burma", HEV-2 "Mexico"); the other two genotypes (HEV-3 "USA", HEV-4 "China") contain viruses isolated from both human and animal samples. HEV has been genetically identified from rat, wild boar, domestic swine, mongoose, rabbits, chickens, ferrets, cutthroat trout, bats, and deer. HEV is considered hyperendemic in many developing countries of prevalence high 25 %, acute hepatitis cases which transmitted by the fecal-oral route and associated with contamination of drinking water by HEV genotypes 1 and 2. While, HEV is considered endemic in industrialized countries, where there is a prevalence of HEV less than 25% acute hepatitis cases. Zoonotic transmission of HEV occurs either by consumption of contaminated meat and meat products or by contact with infected animals. There are at least three main approaches to the future control of HEV infection. These are sanitation, immunoprophylaxis, and vaccination measures. This review summarize the published data on distrubution, characterization and route of transmission of hepatitis E virus genotypes between human and animal species. Keywords: Genotype; HEV; Hyperendemic; Zoonotic transmission.

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54. Predominance and Geo-Mapping of avian influenza H5N1 in poultry sectors in Egypt Abdelsatar Arafa1,2, Ihab El-Masry1, Shereen Kholosy2, Mohammed K. Hassan2 Mousa Soliman4, F. O. Fasina5, Gwenaelle Dauphin3, Juan Lubroth3, Yilma Makonnen1 1 Food and Agriculture Organization of the United Nations (FAO) - Emergency Center of Transboundary Animal Diseases, Giza, Egypt National Laboratory for Veterinary Quality Control on Poultry Production (NLQP), Animal Health Research Institute, Dokki, Giza, Egypt 3 Food and Agriculture Organization of the United Nations (FAO), Viale delle Terme di Caracalla, Rome, Italy 4 General Organization of Veterinary Services, Dokki, Giza, Egypt. 5 Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Onderstepoort, Pretoria, South Africa

2

Abstract Highly pathogenic avian influenza (HPAI) virus of H5N1 subtype has been enzootic in the Egyptian poultry with significant human infections since 2008. Detailed epidemiological study on H5N1 is important in order to understand the transmission, persistence and evolution of the virus as well as to plan control strategies. This work describes the available epidemiological and virological information from Egypt, from February 2006 to May 2015 in spatial and temporal terms, and reports our findings. Tracheal/oropharyngeal, cloacal and tissue samples were collected and tested for avian influenza virus (AIV) H5 subtype using real time RT-PCR and specific primers for M and H5 genes on extracted RNAs. The results were matched with the epidemiological data from various spatially and temporally-dispersed surveillances implemented between 2006 and 2015. Spatio-temporal analysis was conducted on a total of 3,338 confirmed H5N1 HPAI poultry cases and outputs were described based on transmission patterns, poultry species and production types affected, trade, geographic and temporal distributions in Egypt. HPAI H5N1 viruses remain persistent in the Egyptian poultry and annual occurrences displayed a seasonal pattern with peak prevalence occurring in the early months of January to March using normalised data. No specific geographic pattern prevailed and both chickens and duck species were more affected. However, relatively higher disease incidences were recorded in the Nile Delta (Lower Egypt) where poultry and human densities are higher, and where poultry trades are more intense. The phylogenetic studies of the HA sequences of Egyptian H5N1 viruses indicated that multiple clusters circulated between 2006 and 2015 and significant deviations in the circulating viruses have been observed. Since 2007, the virus epidemiological dynamics have changed and the origins of the majority of the outbreaks have shifted to the household poultry, but outbreaks continued to be reported in the live bird markets and commercial poultry. The persistence of infections of HPAI H5N1 in poultry in Egypt with recurrent and sporadic infections in humans can influence virus evolution spatio-temporally. Household poultry played significant roles in the H5N1 virus transmission to poultry and humans, but the role of commercial poultry in Egypt needs some clarifications. While poultry trading can support the persistence and further long distant transmission within Egypt, the role of individual species may warrant further investigation. Surveillance should be focused using multi-sectoral approach. Keywords: HPAI H5N1, poultry, epidemiology, Spatial and temporal analysis, evolution.

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55. Surveillance for Middle East Respiratory Syndrome Coronavirus in camels from 2014 to 2016 in Egypt Mahmoud M. Shehata1*, Ahmed N. El-Taweel1, Mokhtar R. Gomaa1, Ahmed Kandeil1, Rabeh El-Shesheny1, Ahmed S. Kayed1, Basma Elsokary2, Nagla Hassen2, Ola Bagato1, Yasmin Moatasem1, Sara Husein1 ,Omnia Kutkat1, Asmaa M. Maatouq1, Pamela P. McKenzie3, Ahmed Othman4, Richard J. Webby3, and Ghazi Kayali3,5,6, Mohamed A. Ali1 1

Center of Scientific Excellence for Influenza Viruses, National Research Centre, Giza, Egypt. General Organization of Veterinary Services, Ministry of Agriculture and Land Reclamation, Giza, Egypt. 3 Department of Infectious Diseases, St. Jude Children’s Research Hospital, Memphis, Tennessee, USA. 4 Department of Biochemistry, Faculty of Science, Ain Shams University, Cairo, Egypt. 5 Department of Epidemiology, Human Genetics, and Environmental Sciences, University of Texas, Houston, Texas, USA 6 Human Link, Baabda, Lebanon. 2

Abstract Background: Middle East respiratory syndrome coronavirus (MERS-CoV) was detected in September 2012 in Saudi Arabia as of August, 2016, 1791 laboratory confirmed cases have been reported to the World Health Organization (WHO), including at least 640 fatality case. Objective: an active surveillance study was conducted to determine how MERS-CoV enters to and the dynamic of virus movement and spreading in Egypt. Methods: A total of 2825 nasal swabs, 114 rectal swabs, 187 milk samples, 26 urine samples, and 2541 sera were collected from camels through two years survey. Sampling locations included a government operated quarantine at the border with Sudan, 2 camel markets, and camel abattoirs where imported camels are found. Local free-roaming herds, and local farmed breeding herds including a herd of about 90 camels that is raised and bred in confinement on the Mediterranean coast were also sampled. Samples were subjected to serological and molecular investigation for the presence of MERS-CoV. Results: Through surveillance for MERS-CoV in camels in Egypt, we comparing local to imported camels, our data showed that imported camels had higher seroprevalence and higher RT-PCR detection rates. Camels sampled at quarantines and markets had the highest seroprevalence (>92%), while camels from farms had the lowest seroprevalence (