Prof. Dr. Abdullah INCI, Erciyes University (Turkey) Prof. Dr. Aydın S. TUNCBILEK, Erciyes University (Turkey)

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5 Honorary President Prof. Dr. H. Fahrettin KELESTEMUR Rector of Erciyes University President Prof. Dr. Abdullah INCI, Erciyes University (Turkey) Organisation Committee Prof. Dr. Aydın S. TUNCBILEK, Erciyes University (Turkey) Prof. Dr. Suleyman YAZAR, Erciyes University (Turkey) Prof. Dr. (Turkey) Aykut OZDARENDELI, Erciyes University Prof. Dr. Ibrahim NARIN, Erciyes University (Turkey) Assoc. Prof. Dr. Alparslan YILDIRIM, Erciyes University (Turkey) Assoc. Prof. Dr. Ramazan CANHILAL, Erciyes University (Turkey) Assist. Prof. Dr. (Turkey) Onder DUZLU, Erciyes University Turkish Society of Parasitology

6 Scientific Committee Prof. Dr. Peter ADLER, Clemson University (SC, USA) Prof. Dr. (Israel) Kosta Y. MUMCUOĞLU, Hebrew University Prof. Dr. Kevin Y. NJABO, California University (CA, USA) Prof. Dr. Yasuko RIKIHISA, Ohio State University (OH,USA) Dr. Elmer W. Gray, University of Georgia (GA, USA) Prof. Dr. Mehmet Ali OZCEL, Ege University (Turkey) Prof. Dr. Mehmet DOGANAY, Erciyes University (Turkey) Prof. Dr. Filiz ERTUNC, Ankara University (Turkey) Prof. Dr. Yusuf OZBEL, Ege University (Turkey) Prof. Dr. Irfan EROL, Republic of Turkey Ministry of Agriculture and Rural Affairs General Directorate of Protection and Control Prof. Dr. K. Zafer KARAER, Ankara University (Turkey) Assoc. Prof. Dr. Masum BURAK, Republic of Turkey Ministry of Food, Agricultural and Livestock General Directorate of Agricultural Research and Policy

7 MS. Bio. Abdullah YILMAZ, (Turkey) Ministry of Agriculture Secretaria of the Symposium Assoc. Prof. Dr. Alparslan YILDIRIM, Erciyes University (Turkey) Assoc. Prof. Dr. Ramazan CANHILAL, Erciyes University (Turkey) Assist. Prof. Dr. Onder DUZLU, Erciyes University (Turkey) Lecturer MS Zuhal BISKIN, Erciyes University (Turkey) Assistant Arif CILOGLU, Erciyes University (Turkey) Mol. Bio. Gozde Ozge KORKMAZ, (Turkey) Erciyes University Social Committee Vet. Med. Hakan YESILOZ, Ministry of Agriculture (Turkey) Vet. Med. Ahmet DEMIRCIOGLU, Ministry of Agriculture (Turkey) Vet. Med. Seval KORKMAZ, Erciyes University (Turkey) Vet. Med. Tugba MEYILLI, Erciyes University (Turkey)

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11 CONTENTS Rector s Message... 2 Preface 4 Message of the President of Turkish Society for Parasitology 7 Contributors 12 Symposium Program 22 Lectures of the Invited Speakers 28 Oral Presentations 85 Poster Presentations 107 Index

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13 Rector s Message It is a great pleasure for me to speak to this eminent audience and I want to say welcome to all of you as the Rector of Erciyes University and honorary president of this symposium. First of all, I would like to give very brief information about our university. Erciyes University was established in 1978 in Kayseri province which is the world's first trade capital and has taken its name from Mount Erciyes. Today, there are 16 faculties, 9 vocational colleges, 4 colleges, 6 institutes and 24 centers in Erciyes University. One of these centers is the Vectors and Vector-Borne Diseases Implementation and Research Center, which was established in This is the first and unique center in Turkey in this field. Similar centers were presented in the United States, Europe and some countries in the world. Dear participants, today we are all together here for the first scientific activity of our center titled, The 1 st National Symposium on Vectors and Vector Borne 2

14 Diseases with International Participation. Several presentations and discussions on the topics such as important vectors, biology of vectors, vector borne diseases, host-parasite evolution, epidemiology, diagnostics, insecticides-acaricides and resistance, control strategies, environmental impact, economic impact, and geographical information systems will be organized in the symposium. I strongly believe that the results generated from the presentations and the following discussions during the symposium will be useful for the benefit of humankind. Taking this belief into consideration, I would like to thank all of you for sharing your expertise with us and I wish you to have a good time in Cappadocia, which has a unique history and natural beauty. Yours sincerely. Prof. Dr. H. Fahrettin KELESTEMUR Rector of Erciyes University 3

15 Preface VBDS 12SYMPOSIUM Some of the world s most destructive diseases are vector-borne which are transmitted to humans, animals and plants by vectors such as ticks, flies, fleas or other arthropods. Emerging vector-borne infectious diseases are of increasing concern worldwide. The Vectors and Vector-Borne Diseases Implementation and Research Center is the first center established in the Erciyes University in Turkey for researching, surveillance, preventing and controlling of vectors and vector-borne diseases. The 1 st National Symposium on Vectors and Vector Borne Diseases with International participation was organized by the Vectors and Vector Borne Diseases Implementation and Research Center of the Erciyes University. The symposium was held in Avrasya Hotel, Avanos, Nevşehir, Cappadocia between 9 and 10 September In this meeting, the goal is to gather all 4

16 the national and international experts and researchers to promote the discussions on vectors and vector-borne diseases of humans, animals and plants, and to take the issue one step further. Furthermore, the symposium brought scientists together to share and discuss research results, problems encountered, technological developments, solutions, as well as theoretical and practical knowledge in the scientific environment. In this symposium, scientists working at different institutes had the opportunity to initiate future cooperation on vectors and vector borne diseases. I would like to thank The Honorable President of the symposium and the rector of the Erciyes University Prof. Dr. H. Fahrettin KELESTEMUR and Erciyes University Scientific Research Fund for all their support. In addition, I appreciate the contributions of the Ministry of Health,Ministry of Food, Agriculture and Livestock of Turkish Republic, Kayseri and Nevsehir governorships and Kayseri, Nevsehir and Avanos Municipalities. 5

17 Finally, I also want to thank the members of the organizing committee for their effort to make this meeting a success, especially the members of the scientific committee for their helpful guidance, and all the invited speakers, their families and attendee for their valuable contributions to the meeting. Prof. Dr. Abdullah INCI President of VBDS 12 Director of the Vectors and Vector-Borne Diseases Implementation and Research Center 6

18 Message of the President of TurkishSociety for Parasitology Distinguished Quests, Dear Colleagues and Friends, It is my great pleasure to extend my warmest welcome to all participants of the First National Vectors and Vector-Borne Disease Symposium with international participation. On behalf of Turkish Society for Parasitology it is also great pleasure for us to have you all in Avrasya Hotel Cappadocia, Avanos, Nevsehir,Turkey. My special thanks to Prof. Dr. Fahrettin Kelestemur Rector of Erciyes University and the honorary president of the symposium for his excellent support of the symposium. I also would like to thank to Prof. Dr. Abdullah Inci president of the symposium and the members of organizing committee for their excellent work on scientific social program of this symposium. 7

19 Now, you are in the center of Cappadocia which is unique in the world, it is miraculous nature wonder and covers the provinces of Nevsehir, Aksaray, Nigde and Kayseri in the Central Anatolia Region. The Cappadocia Region has been created as results of the volcanic eruptions which accrued in Erciyes, Hasandag and Gulludag mountains. This region was formed as a large tableland from the volcanic tufts together with eruption of Kizilirmak River and the wind ten thousands of years which resulted in the appearance of chimney rocks which are considered wonders of the nature. Cappadocia was one of the most important places in the first period of the Christian Religion. The first Christians trying to escape from Roman soldiers who wanted to avoid the spreading of this religion. They settled in the region of Cappadocia which was so suitable for hiding in cave houses and underground cities. These Christians were able to continue their normal life all spread their 8

20 religion. More information will be given to you during the excursion to Cappadocia. Let me talk about the vectors and vector-borne diseases. As the president of Turkish Society for Parasitology, first we try to have members medical and veterinary Parasitologist. Now, we have 421 members. We have been organizing National Parasitology Congresses every two year; so far we have organized 17 National Parasitology Congress in different cities, with different universities in Turkey. We also organized EMOP IV in 1984, ICOPA VIII in 1994 in Izmir, Turkey. We have been organizing four World Leish Meetings, Istanbul, Greek (Greece), Sicily (Italia), and Luck now (India). That means our society is quite active for organizing meetings, but not so active to encourage to our members to work on vectors. Personally I believe that if you want to control of parasitic diseases, first we have the control vectors, without controlling vectors there is no way to control 9

21 not only parasitic diseases but all vector-borne infection diseases. Now, I would like to say welcome to the world of vectors. This symposium will bring us a large window you can get in and trying to understand what is going on in vectors of the diseases. If you want to know much about these diseases you have to have vectors in your laboratory. When you start to have more information everyday that means you start to understand life cycle of the parasites and also morphology, biology and the psychology of the vectors. This symposium will give us a chance to enter to world of vectors. This is going to be the beginning, let s get in to the world of black flies, mosquitoes, human body louse, ticks, sand flies and other vectors. That is why I want to thanks a lot to the President and organizing committee of this symposium, my sincere thanks to the scientists came from USA, Israel and different universities from Turkey. 10

22 It is true that success of this symposium depend on a great deal of the work and the cooperation of the scientists who devoted their time to prepare their conferences. Let s get together to know and understand more about vectors and the vector- borne diseases. I am sure your participation will highlight this symposium and advocate our mission of improving animal and human health. Thank you. Prof. Dr. M. Ali OZCEL The President of Turkish Society for Parasitology and Former President of World Federation of Parasitology 11

23 Contributors Prof. Dr. H. Fahrettin KELESTEMUR, Rector of Erciyes University, Kayseri,TURKEY Prof. Dr. Abdullah INCI, President of VBDS 12 and Director of Vectors and Vector-Borne Diseases Implementetion and Research Center, Erciyes University, Kayseri, TURKEY Prof. Dr. Halit CANATAN, Dean of Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Prof.Dr. Huseyin ALTINDIS, Dean of Faculty of Science, Erciyes University, Kayseri, TURKEY Prof. Dr. Muberra KOSAR, Dean offaculty of Pharmacy, Erciyes University, Kayseri, TURKEY Prof. Dr. Muhammet GUVEN, Dean offaculty of Medicine,Erciyes University, Kayseri, TURKEY Prof. Dr. Ali Irfan ILBAS, Dean of Faculty of Agriculture, Erciyes University, Kayseri,TURKEY 12

24 Prof. Dr. Peter ADLER,Entomology Program, Clemson University, SC, USA Prof. Dr. Kosta Y. MUMCUOGLU,Department of Microbiology and Molecular Genetics, The Kuvin Center for the Study of Infectious and Tropical Diseases, The Hebrew University-Hadassah Medical School, Jerusalem, ISRAEL Prof. Dr. Kevin Y. NJABO, Africa Director of Center for Tropical Research Institute of the Environment and Sustainability, University of California, Los Angeles, USA Prof. Dr. Yasuko RIKIHISA, National Academy of Sciences, The Ohio State University, Columbus, Ohio, USA Dr. Elmer W. GRAY, The University of Georgia Entomology Department, Athens, GA, USA Prof. Dr. Mehmet Ali OZCEL, The President of Parasitology Association of Turkey and Former President of World Federation of Parasitology, Izmir, TURKEY 13

25 Prof. Dr. Mehmet DOGANAY, Department of Infectious Diseases, Faculty of Medicine, Erciyes University, Kayseri, TURKEY Prof. Dr. Filiz ERTUNC, Department of Plant Protection, Faculty of Agriculture, Ankara University, Ankara, TURKEY Prof. Dr. Yusuf OZBEL, Department of Parasitology, Faculty of Medicine, Ege University, Bornova, Izmir, TURKEY Prof. Dr. Irfan EROL, Republic of Turkey Ministry of Agriculture and Rural Affairs General Directorate of Protection and Control, Ankara, TURKEY Prof. Dr. K. Zafer KARAER, Department of Parasitology, Faculty of Veterinary Medicine, Ankara University, Ankara, TURKEY Prof. Dr. Levent AYDIN, Department of Parasitology,Faculty of Veterinary Medicine, Uludag University, Bursa, TURKEY 14

26 Prof. Dr. Munir AKTAS, Department of Parasitology, Faculty of Veterinary Medicine, Firat University, Elazıg, TURKEY Prof. Dr. Abdurrahman GUL, Faculty of Science and Literature, Bingol University, Bingol, TURKEY Prof. Dr. Osman TIRYAKI, Department of Plant Protection, Faculty of Agriculture, Kayseri, TURKEY Erciyes University, Prof. Dr. Aydin S. TUNCBILEK, Department of Biology, Faculty of Science, Erciyes University, Kayseri, TURKEY Prof. Dr. Suleyman YAZAR, Department of Parasitology, Faculty of Medicine, Erciyes University, Kayseri, TURKEY Prof. Dr. Izzet SAHIN, Department of Parasitology, Faculty of Medicine, Erciyes University, Kayseri, TURKEY Prof. Dr. Aykut OZDARENDELI, Department of Microbiology, Faculty of Medicine, Erciyes University, Kayseri, TURKEY 15

27 Prof. Dr. Erkut TUZER, Department of Parasitology, Faculty of Veterinary Medicine, Istanbul University, Istanbul, TURKEY Prof. Dr. Ibrahim NARIN, Department of Analytical Chemistry, Faculty of Pharmacy, Erciyes University, Kayseri, TURKEY Prof. Dr. Ahmet AKSOY, Department of Biology, Faculty of Science, Erciyes University, Kayseri, TURKEY Prof. Dr. Nusret AYYILDIZ, Department of Biology, Faculty of Science, Erciyes University, Kayseri, TURKEY Assoc. Prof. Dr. Alparslan YILDIRIM, Department of Parasitology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Assoc. Prof. Dr. Salih KUK, Department of Parasitology, Faculty of Medicine, Erciyes University, Kayseri, TURKEY Assoc. Prof. Dr. Kursat ALTAY, Department of Parasitology, Faculty of Veterinary Medicine, Cumhuriyet University, Sivas, TURKEY 16

28 Assoc. Prof. Dr. Murat KANBUR,Department of Pharmacology-Toxicology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Assoc. Prof. Dr. Semih GUMUSSOY,Department of Microbiology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Assoc. Prof. Dr. Ayse GENCAY,Department of Virology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Assoc. Prof. Dr. Savas SARIOZKAN,Department of Animal Health Economics and Management, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Assoc. Prof. Dr. Aysegul TAYLAN OZKAN, National Reference Laboratory for Parasitic Diseases, Microbiology Reference Laboratories Department, Ministry of Health, Public Health Intitution of Turkey, Ankara, TURKEY 17

29 Assoc. Prof. Dr. Yavuz UYAR, Public Health Intitution of Turkey, Ankara, TURKEY Assoc. Prof. Dr. Masum BURAK, Republic of Turkey Ministry of Food, Agricultural and Livestock General Directorate of Agricultural Research and Policy, Ankara, TURKEY Assoc. Prof. Dr. Ramazan CANHILAL, Department of Plant Protection,Faculty of Agriculture, Erciyes University, Kayseri, TURKEY Assoc. Prof. Dr. Yeliz YILDIRIM, Department of Food Hygiene and Technology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Assist. Prof. Dr. Onder DUZLU, Department of Parasitology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Assist. Prof. Dr. Nuran AYSUL, Department of Parasitology, Faculty of Veterinary Medicine, Adnan Menderes University, Aydin, TURKEY 18

30 Assist. Prof. Dr. Mustafa Necati MUZ, Department of Parasitology, Faculty of Veterinary Medicine, Mustafa Kemal University, Hatay, TURKEY Assist. Prof. Dr. Aysel KEKILLIOGLU, Department of Biology, Faculty of Science and Literature, Nevsehir University, Nevsehir, TURKEY Dr. Aytac AKCAY, Department of Biometrics, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Dr. Bekir CELEBI, Public Health Institute, Ankara, TURKEY Dr. Mehmet ARSLAN, Department of Field Crops, Faculty of Agriculture, Mustafa Kemal University, Hatay, TURKEY Dr. Kerem OTER, Department of Parasitology, Faculty of Veterinary Medicine, TURKEY Istanbul University, Istanbul, MS. Bio. Abdullah YILMAZ, Ministry of Agriculture, Ankara, TURKEY 19

31 Vet. Med. Akif BASOL, Directorate of Kayseri Veterinarian Association, Kayseri, TURKEY Lecturer MS Zuhal BISKIN, Department of Parasitology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Assistant Arif CILOGLU, Department of Parasitology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY Assistant Muhammet KARAKAVUK, Department of Parasitology, Faculty of Veterinary Medicine, Mustafa Kemal University, Hatay, TURKEY Mol. Bio. Gozde Ozge KORKMAZ, Department of Stem Cell Science, Graduate School of Health Sciences,Erciyes University, TURKEY PhD Student Gozde COSKUNSERCE, Department of Parasitology, Faculty of Veterinary Medicine, Uludag University, Bursa, TURKEY 20

32 PhD Student Mehmet KARAKUS, Graduate School of Natural and Applied Science, Ege University, Izmir, TURKEY Vet. Med. Hakan YESILOZ, Agriculture Department of Nevşehir, Ministry of Agriculture, Nevsehir, TURKEY Vet. Med. Ahmet DEMIRCIOGLU, Avanos Municipality, Nevsehir, TURKEY Vet. Med. Tugba MEYILLI, Department of Parasitology, Faculty of Veterinary Medicine, Erciyes University, Kayseri, TURKEY 21

33 Symposium Program Program: Registration Get together party Dinner Program: Registration I. SESSION Chairmen: Prof. Dr. Kevin Y. NJABO, Prof.Dr. Mehmet Ali OZCEL Opening Speech by the Erciyes University Rector Prof. Dr. Fahrettin H. KELESTEMUR, President of the Symposium Prof. Dr. Abdullah INCI, President of the Turkish Society of Parasitology Prof. Dr. Mehmet Ali OZCEL 22

34 World perspective on black flies as pests and vectors, Prof. Dr. Peter ADLER, Department of Entomology, Soils, and Plant Sciences, Clemson University, Clemson, South Carolina, USA Coffee Break II. SESSION Chairmen: Prof. Dr. Peter H. ADLER, Prof. Dr. Filiz ERTUNC Vectors of avian malaria in Africa and their blood feeding habits, Prof. Dr. Kevin Y. NJABO, Center for Tropical Research Institute of the Environment and Sustainability, University of California, Los Angeles, CA, USA The vectorial capacity of the human body louse, Pediculus humanus humanus, Prof. Dr. Kosta Y. MUMCUOGLU, Department of Microbiology and 23

35 Molecular Genetics, Faculty of Medicine, Hebrew University, Jerusalem, Israel Coffee Break III. SESSION Chairmen: Prof. Dr. Kosta MUMCUOGLU, Prof. Dr. Mehmet DOGANAY Ehrlichiae of public health and veterinary importance: How can they infect phagocytes?, Prof. Dr. Yasuko RIKIHISA, Department of Veterinary Biosciences The Ohio State University, Columbus, Ohio, USA Standardized methodsfor black fly rearing and product development for the operational control of black flies, Dr. Elmer GRAY, Department of Entomology Georgia University, Athens, GA, USA Lunch Program: (Continued) 24

36 IV. SESSION Chairmen: Prof. Dr. Yasuko RIKIHISA, Prof. Dr. Levent AYDIN Ecology of Hyalomma marginatum and Crimean-Congo hemorrhagic fever, Prof. Dr. Zati VATANSEVER, Department of Parasitology Faculty of Veterinary Medicine, Kafkas University, Kars, TURKEY Development of an inactivated vaccine against Crimean-Congo hemorrhagic fever virus, Prof. Dr. Aykut OZDARENDELI, Department of Microbiology Faculty of Medicine, Erciyes University, Kayseri, TURKEY Coffee Break V. SESSION Chairmen: Dr. Elmer W. GRAY, Prof. Dr. Yusuf OZBEL Reemerging diseases: Tularemia, Prof. Dr. Mehmet DOGANAY, Clinical Infectious Diseases and 25

37 Microbiology Faculty of Medicine, Erciyes University, Kayseri, TURKEY Control programs of the Simulium species in the Center Kızılırmak Basin of Turkey, MS. Bio. Abdullah YILMAZ, Ministry of Agriculture, TURKEY Coffee Break VI. SESSION Chairmen: Prof.Dr. Zati VATANSEVER, Prof. Dr. Aydin S. TUNCBILEK The infections transmitted by sand flies in Turkey, Prof. Dr. Yusuf OZBEL, Department of Parasitology Faculty of Medicine, Ege University, Izmir, TURKEY A new threat for Turkish horticulture: Phytoplasma diseases and their vectors, Prof. Dr. Filiz ERTUNC, Department of Plant Protection Faculty of Agriculture, Ankara University, Ankara, TURKEY 26

38 Coffee Break VII. SESSION Chairmen: Prof.Dr. Suleyman YAZAR, Prof. Dr. Aykut OZDARENDELI Oral Presentations Session Poster Session Welcome Party Dinner Program: Assesment of VBDS 12 and Closing Excursion to Cappadocia Hotel check out Turkish Folklore and Dinner 27

39 Lectures of the Invited Speakers

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41 World Perspective on Black Flies as Pests and Vectors Peter H. Adler Entomology Program, Clemson University Clemson, SC , USA, Black flies (Simuliidae) generally rank third in medical and veterinary importance among the arthropods. A total of 2,127 living, formally described species of black flies are currently recognized as valid, with new species continuing to be discovered at a rapid rate. About 98% of all species are capable of acquiring blood from birds and mammals, and most of these species are probably vectors of vertebrate parasites. About 30 species of vertebrate parasites have been formally described from black flies, although many more species have been documented by molecular techniques. These parasites are responsible for 9 simuliid-borne diseases, but only 1.5% of all black fly species are vectors of parasites that cause the two human diseases, mansonellosis and 29

42 onchocerciasis. Onchocerciasis is the premiere simuliidborne disease of humans, affecting people in Africa, Latin America, and Yemen, whereas mansonellosis is a mild disease in the Neotropics. An understanding of the evolutionary factors that have promoted pest and vector status is important for the control of vector-borne simuliid diseases. Black flies probably have a Pangean origin, although the oldest fossils are from the late Jurassic Period. Anautogeny and multiple gonotrophic cycles, which are prerequisites for vector efficiency, evolved early in the Simuliidae. Consequently, pests and vectors are found throughout the family. The evolution of multivoltinism was associated with movement of black flies into warmer climatic areas, where human populations ultimately would be centered, increasing the probability of repeated exposure to infection by simuliid-borne parasites. The ability to colonize large rivers, which appears multiple times in the family Simuliidae, contributed to the ability to produce large populations of flies, exacerbating the potential for pest 30

43 and vector problems. Development of anthropophily further increased the risk of vector-borne diseases by simuliids. Anthropophilic species are found throughout the family Simuliidae, although no species of black fly feeds exclusively on humans. 31

44 Vectors of Avian Malaria in Africa and Their Blood Feeding Habits Kevin Y. Njabo, Thomas B. Smith University of California, Los Angeles, USA Malaria parasites use vertebrate hosts for asexual multiplication and Culicidae mosquitoes for sexual and asexual development, yet the literature on avian malaria remains biased towards examining the asexual stages of the life cycle in birds. To fully understand parasite evolution and mechanism of malaria transmission, knowledge of all three components of the vector-hostparasite system is essential. Little is known about avian parasite-vector associations in African rainforests where numerous species of birds are infected with avian haemosporidians of the genera Plasmodium and Haemoproteus. Secondly, mosquito blood feeding behavior is a very significant component of pathogen transmission and determinant of disease epidemiology. Yet, knowledge of foraging ecology of mosquitoes often 32

45 depends on the presence of undigested blood in the mosquito midgut. Approximately 36 hours after feeding, the blood meal is sufficiently digested to make identification by molecular techniques difficult leaving a very narrow window in which these methods can be used. In this study, we apply high resolution melt qpcrbased techniques and nested PCR to examine the occurrence and diversity of mitochondrial cytochrome b gene sequences of haemosporidian parasites in wildcaught mosquitoes sampled across 12 sites in Cameroon. We also investigate the feeding habits of these mosquitoes from four genera based on the isotopic ratios of nitrogen (δ15n), carbon (δ13c) and sulphur (δ34s). In all, we screened 3134 mosquitoes representing 27 species. Four genera were infected with twenty-two parasite lineages (18 Plasmodium spp. and 4 Haemoproteus spp.). Presence of Plasmodium sporozoites in salivary glands of Coquillettidia aurites further established these mosquitoes as likely vectors. Unfed mosquitoes were also found to have a lower 33

46 δ13c, δ15n and middle δ34s values than fed mosquitoes with. δ13c appearing to be the best element to differentiate between mosquito species that fed on different host species. Isotopic analyses show that the different mosquito genera may be separated based on their diets, suggesting that linking stable isotope-based assays and DNA analysis may be a powerful new tool to investigate mosquito feeding ecology and the dynamics of vector-borne pathogens. 34

47 The Vectorial Capacity of the Human Body Louse, Pediculus humanus humanus Kosta Y. Mumcuoglu Department of Microbiology and Molecular Genetics, The Kuvin Center for the Study of Infectious and Tropical Diseases, The Hebrew University - Hadassah Medical School, Jerusalem, ISRAEL, kostam@cc.huji.ac.il The head louse, Pediculus humanus capitis, the body louse, Pediculus humanus humanus and the pubic louse, Pthirus pubis, are specific parasites of humans. Three pathogenic bacteria are transmitted by the body louse: a) Rickettsia prowazekii, the agent of epidemic typhus; b) Borrelia recurrentis, the agent of relapsing fever; and c) Bartonella quintana, the agent of trench fever. Historic reports of outbreaks of epidemic typhus date back to the Peloponnesian War, but more accurate descriptions begin in 1489 with the Spanish siege of Granada and Napoleon s invasion to Russia. The last important outbreak of epidemic typhus was observed 35

48 during the civil war in Burundi ( ), when approximately 100,000 died because of it. Today, this pathogen is endemic in Sudan, Burundi and Ethiopia. Brill Zinsser disease is a delayed relapse of epidemic typhus. In patients with epidemic typhus, the rickettsia can remain latent and reactivate months or years later, with symptoms similar to the original attack of typhus, including a maculopapular rash. This reactivation event can be transmitted by the body louse, and form the focus for a new epidemic of typhus. The number of cases with endemic typhus dropped dramatically in the last years, nevertheless, WHO still classify it as a "disease under surveillance". In the last century, successive epidemics of louse-borne relapsing fever spread from Asiatic Russia into Europe. Now, this disease has all but vanished in all areas except the Andean foothills, the Ethiopian highlands, and nearby Rwanda. In the last decades, B. quintana infections have emerged in diverse regions of the world such as Russia, France and USA, predominantly involving socially disadvantaged persons. 36

49 B. quintana is now recognized as an agent of bacillary angiomatosis bacteremia, trench fever, endocarditis, and chronic lymphadenopathy among homeless people. Recently, Acinetobacter baumannii was isolated from body lice collected worldwide and it was shown that they are able to transmit this pathogen. Under experimental conditions with laboratory animals it was also shown that body lice could transmit Yersinia pestis, the causative agent of plague, Rickettsia typhi (endemic or murine typhus) and Rickettsia conorii (Mediterranean spotted fever). Head lice might have a vectorial role during outbreaks of epidemic typhus. Although B. quintana has been also found in head lice, its vectorial capacity is still unclear. Pubic lice are not knot to be vectors of any human pathogenic microorganisms. 37

50 Ehrlichiae of Public Health and Veterinary Importance: How Can They Infect Phagocytes? Yasuko Rikihisa Member, National Academy of Sciences, The Ohio State University, Columbus, Ohio, USA Ehrlichia and Anaplasma spp. (collectively called ehrlichiae) are obligatory intracellular bacteria that persist in nature by cycling between varieties of mammals and ticks. Human infection with Ehrlichia chaffeensis and Anaplasma phagocytophilum via the bite of infected ticks could lead to fatal emerging zoonoses called human monocytic ehrlichiosis and human granulocytic anaplasmosis, respectively. Ehrlichiae also cause important diseases in domestic animals such as canine ehrlichiosis, bovine anaplasmosis, and heartwater. By using blood cells as the primary target cell of infection, ehrlichiae increase the chance of bloodborne transmission through ticks. However, monocytes (host cells of E. chaffeensis) and neutrophils (host cell of 38

51 A. phagocytophilum) also are a primary host defense cells equipped with powerful innate immune mechanisms. How can ehrlichiae thrive inside phagocytes? Studies revealed ehrlichiae have a remarkable ability to subvert innate immune mechanisms and host cell homeostasis to facilitate infection. First of all these bacteria got rid of genes encoding lipopolysaccharides and peptidoglycans which activate phagocytes. Second, ehrlichiae have evolved a type IV secretion system to deliver bacterial molecules to the host cell cytoplasm in order to manipulate host cell signaling. Several of such molecules have been identified thus far and shown to have critical roles in subversion of intracellular host cell defenses. Ehrlichiae also alter host cellular homeostasis and vesicular traffic so that molecules essential for their growth and survival are enriched in the infected cells and delivered to intracellular bacteria. For example, cholesterol, an essential component of ehrlichiae, is enriched in infected cells and transported to bacterial inclusions 39

52 through membrane vesicles containing Niemann-Pick type C1 proteins. Although the detailed mechanisms responsible for these effects at the pathogen and host interface remain to be elucidated, studies along these lines are expected to provide new insights into intracellular parasitism and targets for prophylactic and therapeutic intervention. 40

53 Standardized Methods for Black Fly Rearing and Product Development for the Operational Control of Black Flies Elmer W. Gray, Ray Noblet, Joseph P. Iburg, Roger D. Wyatt The University of Georgia Entomology Department, Athens, GA 30602, USA Black flies (Diptera: Simuliidae) are serious pests of man and animals worldwide. In addition, black flies serve as the vector for the causative agent of human onchocerciasis, the filarial nematode Onchocerca volvulus. Due to their public health significance and worldwide pest status, substantial efforts have been made to develop efficient methods to rear black flies in the laboratory environment. The University of Georgia Black Fly Rearing and Bioassay Laboratory currently operates such a colony. The original field-collected material for this colony, Simulium vittatum cytospecies IS-7, was collected as eggs from trailing grasses in 41

54 Flaxmill Creek, Cambridge, NY, USA in The colony currently produces 200, ,000 larvae per week. The primary components of the colony system are nine aquatic rearing units. Each unit forms a closedcirculation trough system, to create an ideal larval environment. Water is pumped from a lower reservoir to upper chambers where it flows over a runway where larval development occurs. Associated systems have been developed to inject food material into the unit, capture emerging adults, induce mating and provide suitable substrate for oviposition. The primary purpose of the colony is to produce uniform larvae for a variety of research activities associated with the insecticidal proteins of Bacillus thuringiensis subsp. israelensis (Bti) contained in the Vectobac 12 AS product. Such activities include product development, quality control, and other research associated with the feeding activity of larval black flies. As part of this work, a controlled current toxicity test (CCTT) has been developed that uses a gyrotory shaker and 250 ml flat-bottomed 42

55 extraction flasks to create multiple microcosms that simulate the natural Simuliidae larval habitat. A standardized technique has also been developed, and used extensively, to conduct field evaluations of Bti based larvicides and to evaluate operational applications. 43

56 Ecology of Hyalomma marginatum and Crimean-Congo hemorrhagic fever Zati Vatansever Kafkas University, Faculty of Veterinary Medicine, Department of Parasitology, 36100, Kars, TURKEY, e- mail: CCHF is the most widespread tick-borne viral disease of humans. It has been recorded from more than 30 countries (Hoogstraal 1979,Ergonul 2006,Whitehouse 2004). The causative agent is single stranded RNA virus, which belongs to the Nairovirus genus in the Bunyaviridae family (Whitehouse 2004). The virus circulates in nature by tick-animal-tick enzootic cyle. Although it causes viraemia without any clinical signs in its animal hosts, it leads to severe even lethal disease in humans (Hoogstraal 1979). Humans acquire the infection via tick bites, crushing the infected ticks or contact with viraemic animals blood or tissues. Nosocomial infections due to contact with infected 44

57 patients are also common (Hoogstraal 1979,Whitehouse 2004). Numerous studies have shown presence of anti- CCHFv antibodies in many wild and domestic vertebrates such as cattle, sheep, goats, horses, camels, ostriches, hedgehogs, hares and some rodents and carnivores (Hoogstraal 1979,Nalca and Whitehouse 2007). However the natural infection (viraemia or antigen in tissues) was demonstrated in limited number of animals such as hares, hedgehogs (Hoogstraal 1979,Vatansever, et al. 2007), wild boars (Vatansever, et al. 2007) and rooks (Vodyanitskaya, et al. 2008). It has been accepted for long time that the birds are refractive to the virus and their role in CCHF epidemiology is restricted to support and spread of tick populations (Hoogstraal 1979). But recent findings stress out the possible reservoir role of ground feeding birds in CCHF epidemiology (Vodyanitskaya, et al. 2008,Zeller, et al. 1994,Zeller, et al. 1994) Although viraemia can be demonstrated in some animals and hares are long time accepted as reservoir for 45

58 infection, the role of ticks in maintenance and survival of the virus from one year to other crucial. While vertebrate animals can be viraemic for up to 2 weeks, ticks can harbour the virus lifelong (1 year) and can also maintain it by transstadial/transovarial passages (Hoogstraal 1979,Zeller, et al. 1994,Turell 2007). Although CCHFv has been isolated from about 30 tick species, the vector competence has been demonstrated only for limited number of tick species (Amblyomma variegatum, Hyalomma marginatum, H. rufipes, H. anatolicum, H. asiaticum, H. truncatum, H. impeltatum, Dermacentor marginatus, Rhipicephalus evertsi, Rh. rossicus), among which Hyalomma species are strictly associated with the global distribution of the disease (Hoogstraal 1979,Turell 2007,Camicas, et al. 1994,Dohm, et al. 1996,Logan, et al. 1989,Korsunova and Petrova- Pointkovskaya 1949). The main tick species associated with the CCHF outbreaks in Turkey, Balkans, Crimea and Southern Federal Districts of the Russian Federation (SFDRF) is H. marginatum. It is important to mention 46

59 that Dermacentor marginatus may play role in enzootic cycle, in the above mentioned regions where H. marginatum is involved in human cases. Hyalomma anatolicum in Iran, Pakistan, Turkmenistan and Tadjikistan; Hyalomma asiaticum in Central Asia and China, Hyalomma rufipes in Africa are suggested as the main vectors of CCHF. It is also interesting that CCHF cases out of H. marginatum s areal seem to be more associated with animal butchering instead of the tick bites (Hoogstraal 1979,Turell 2007,Emelianova 2006,Chinikar, et al. 2010). Hyalomma ticks can transmit the virus transstadially and transovarially (Hoogstraal 1979,Zeller, et al. 1994,Turell 2007,Camicas, et al. 1994,Dohm, et al. 1996,Logan, et al. 1989,Faye, et al. 1999,Faye, et al. 1999,Pak, et al. 1974). Non-viraemic transmission among co-feeding ticks (Logan, et al. 1989,Gonzalez, et al. 1992,Gordon, et al. 1993) and venereal transmission have also been demonstrated (Gonzalez, et al. 1992). 47

60 Hyalomma marginatum is associated with wildlife and is adapted to steppe climate. It is a 2 host tick which immature (larvae and nymphs) stages feed on small mammals (hare, hedgehog) and ground frequenting birds (rooks, partridges). Fed larvae moult on the host and became nymphs which drop-off the ground when engorge. The feeding period of immatures (larvae, nymphs) takes about days. Engorged nymphs moult to adults in about 4-20 days on the ground. The time when newly formed adults appear is about late August-September. Those adults prefer to hide and overwinter in nature and become active in the spring of the following year. The adults of H. marginatum are of hunter character and instead of vertical climbing on vegetation, they actively seek/wait hosts horizontally on the ground. They mostly prefer artiodactyls (cattle, sheep/goats, horses, wild boars) but also aggressively attack humans as well. When attached to a host they feed for about 9-14 days. Engorged females drop-off on the ground and lay about 7000 or more eggs before they 48

61 die. The whole life cycle of H. marginatum takes about a year (Emelianova 2006,Pomerantsev 1950,Berezin 1971,Petrova-Piontkovskaya 1947,Ouhelli 1994). The ability of H. marginatum to overwinter as unfed adult is one of the main factors which is allowing the transmission of CCHF virus from year to year. (Hoogstraal 1979,Berezin 1971)(Fig 1). Fig 1.Hyalomma marginatum biology and CCHF virus circulation. 49

62 Adults of H. marginatum are responsible of CCHFv transmission to humans in the Balkans, Crimea, SFDRF and Turkey. Adult ticks become active in spring when average temperatures reach 10.5 C. They actively seek/wait for a host when average daily temperatures are C and humidity is %. When air temperature increases above 30 C and soil temperature above 45 C, ticks prefer to hide or even burry themselves in the soil (Emelianova 2006,Ouhelli 1994,Tokhov Iu 2009). CCHF epidemics in Balkans, Crimea, Southern Federal Districts of Russia have been always associated with ecological changes leading to increase of wild animals and H. marginatum population (Hoogstraal 1979,Berezin 1971). It was demonstrated that hares in Crimea and rooks in Rostov region of former USSR played vital role as being main hosts for immatures of H. marginatum ticks. In both regions the main hosts for 50

63 adult ticks were cattle. In Turkey first cases were diagnosed in 2002 and until the end August 2012 a total of 7157 laboratory confirmed cases were reported from 2200 rural settlements (Fig 2). Fig 2.Spatial distribution and density of CCHF cases in Turkey ( ). The overall mortality was 5 % (Fig 3) (Data provided by the Ministry of Health of Turkey). The number of cases are stabilized in 2008 and 2009 at around 1300 and decreased to 859 in 2010, which may indicate that the outbreak reached the plateau and further decrease in cases can be expected in the following years. Similar trend has been observed in SFDRF since

64 (Onishchenko 2009). In Turkey it has been shown that H. marginatum is the dominant species in the CCHF areas. (Tonbak, et al. 2006,Vatansever, et al. 2007). It has been also demonstrated that H. marginatum is transmitting the virus both transovarrialy and transstadially. (Vatansever, et al. 2007,Vatansever, et al. 2008) and 16.43% of host seeking H. marginatum ticks were CCHFv infected (Vatansever, et al. 2010). Fig 3.Recorded CCHF cases and deaths in Turkey. Tick population dynamics are influenced by biotic and abiotic factors (Randolph and Rogers 2006,Randolph 52

65 2004). The habitat suitability maps for Hyalomma marginatum were prepared and analyzed against spatial distribution of CCHF in Turkey. The disease risk was strongly associated with presence of H. marginatum and landscape fragmentation. (Vatansever, et al. 2007,Estrada-Pena, et al. 2007). The empirical observations on habitat regeneration and its influences on wildlife and tick abundance still need to be discussed. Hare (for immature ticks) and cattle (for adult ticks) seem to be main hosts involved in the H. marginatum biology in the epidemic area in Turkey. Both animals support tick population and are also involved in viral circulation (Vatansever, et al. 2008). We still need data on the role of ground feeding birds and other wild animals such as wild boars, which population is dramatically increased in the region. Similarly there is a need on data abaout the possible contribution of other tick species (e.g. D. marginatus) on the enzootic cycle of CCHF in Turkey. Also changes in socio-economics (migration from rural areas to urban areas) and animal 53

66 husbandry (e.g. decrease of sheep population) should be studied in details. Obtaining such data will help decision makers to understand the complex epidemiology of the disease and develop effective intervention stategies. References Hoogstraal H. The epidemiology of tick-borne Crimean-Congo hemorrhagic fever in Asia, Europe, and Africa. J Med Entomol 1979;15: Ergonul O. Crimean-Congo haemorrhagic fever. Lancet Infect Dis 2006;6: Whitehouse CA. Crimean-Congo hemorrhagic fever. Antiviral Res 2004;64: Nalca A, Whitehouse C. Crimean-Congo Hemorrhagic Fever Virus Infection among Animals. Crimean-Congo Hemorrhagic Fever2007: Vatansever Z, Midilli K, Ergin S, Gargılı A. Investigation of CCHFv in blood and tick samples collected from wild animals. Yaban hayvanlarından toplanan kan ve kene örneklerinin Kırım-Kongo kanamalı ateşi virüsü yönünden araştırılması. (In Turkish). 3rd National Virology Congress. Ağaoğlu My Resort, Uludağ-Bursa, Turkey2007:222. Vodyanitskaya SY, Moskvitina EA, Pichurina NL, Zabashta AV, Orekhov IV, Mishan kin BN, Vodopyanov SO, Suchkov IY. Crimean Hemorrhagic Fever: 54

67 Ecologic and Epizootic Role of the Corvidae Family (In Russian). Problems of Particularly Dangerous Infections 2008: Zeller HG, Cornet JP, Camicas JL. Crimean-Congo haemorrhagic fever virus infection in birds: field investigations in Senegal. Res Virol 1994;145: Zeller HG, Cornet JP, Camicas JL. Experimental transmission of Crimean- Congo hemorrhagic fever virus by west African wild ground-feeding birds to Hyalomma marginatum rufipes ticks. Am J Trop Med Hyg 1994;50: Turell M. Role of Ticks in the Transmission of Crimean-Congo Hemorrhagic Fever Virus. In: Ergonul O, Whitehouse CA, eds. Crimean-Congo Hemorrhagic Fever: A Global Perspective2007: Camicas JL, Cornet JP, Gonzalez JP, Wilson ML, Adam F, Zeller HG. [Crimean- Congo hemorrhagic fever in Senegal. Latest data on the ecology of the CCHF virus]. Bull Soc Pathol Exot 1994;87:11-6. Dohm DJ, Logan TM, Linthicum KJ, Rossi CA, Turell MJ. Transmission of Crimean-Congo hemorrhagic fever virus by Hyalomma impeltatum (Acari:Ixodidae) after experimental infection. J Med Entomol 1996;33: Logan TM, Linthicum KJ, Bailey CL, Watts DM, Moulton JR. Experimental transmission of Crimean-Congo hemorrhagic fever virus by Hyalomma truncatum Koch. Am J Trop Med Hyg 1989;40:

68 Korsunova OS, Petrova-Pointkovskaya SP. A virus isolated from Hyalomma marginatmn marginatum Koch, ticks. Zool. Zh. 1949;28: (Translation 793 (T793), Medical Zoology Department, US NAMRU, Cairo, Egypt). Emelianova IN. Hyalomma Koch, 1844 (ACARI: IXODIDAE) ticks of Central Precaucasia and surrounding territories. (Distribution, ecology, role in the natural foci of Crimean-Congo heamorrhagic fever). MSc thesis in Biology. Stavropol: State University of Stavropol; 2006:146p. Chinikar S, Ghiasi SM, Moradi M, Goya MM, Shirzadi MR, Zeinali M, Meshkat M, Bouloy M. Geographical distribution and surveillance of crimean-congo hemorrhagic Fever in iran. Vector Borne Zoonotic Dis 2010;10: Faye O, Cornet JP, Camicas JL, Fontenille D, Gonzalez JP. [Experimental transmission of Crimean-Congo hemorrhagic fever virus: role of 3 vector species in the maintenance and transmission cycles in Senegal]. Parasite 1999;6: Faye O, Fontenille D, Thonnon J, Gonzalez JP, Cornet JP, Camicas JL. [Experimental transmission of Crimean-Congo hemorrhagic fever virus by Rhipicephalus evertsi evertsi (Acarina:Ixodidae)]. Bull Soc Pathol Exot 1999;92: Pak TP, Daniyarov OA, Kostyukov MA, Buluchev V, P,, Kuima AU. Biocenotic interrelationships between Crimean hemorrhagic fever, ixodid ticks, and their hosts. Report 1. Results from virological investigations of ixodid and 56

69 argasid ticks in Tadzhik SSR. Sborn. Trud. Ekol. Virus. 1974;2: (Translation 783 (T783), Medical Zoology Department, US NAMRU, Cairo, Egypt). Gonzalez JP, Camicas JL, Cornet JP, Faye O, Wilson ML. Sexual and transovarian transmission of Crimean-Congo haemorrhagic fever virus in Hyalomma truncatum ticks. Res Virol 1992;143:23-8. Gordon SW, Linthicum KJ, Moulton JR. Transmission of Crimean-Congo hemorrhagic fever virus in two species of Hyalomma ticks from infected adults to cofeeding immature forms. Am J Trop Med Hyg 1993;48: Pomerantsev BI. [Ixodid ticks (Ixodidae)]. Fauna of the USSR, Paukoobraznyye. new series 41, 4(2), 224 pp Berezin VV. Investigation of the ecology of arboviruses in river deltas of the Caspian and Azov sea basins.. (Avtoref. Diss. Soisk. Uchen. Step. Dokt. Biol. Nauk). Inst. Polio. Virusn. Entsefalitov, Akad. Med. Nauk SSSR, Moscow. 37 p. (In Russian). (In English, NAMRU3-T1160) Petrova-Piontkovskaya SP. Biological and ecological data on the tick Hyalomma marginatum Koch in the northwestern Crimean hemorrhagtc fever focus.. Nov. Med. 1947;5: (Translation 864 (T864), Medical Zoology Department, US NAMRU, Cairo, Egypt). Ouhelli H. Comparative development of Hyalomma marginatum (Koch, 1844), H. detritum (Schulze, 1919), H. anatolicum excavatum (Koch, 1844), 57

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72 Development of an Inactivated Vaccine against Crimean-Congo Hemorrhagic Fever Virus Aykut Ozdarendeli Erciyes University, Faculty of Medicine, Microbiology Department, Kayseri,TURKEY Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus of the genus Nairovirus family Bunyaviridae, which are enveloped viruses containing tripartite, negative polarity, single-stranded RNA. CCHF is characterized by high case mortality, occurring in Asia, Africa, the Middle East and Europe. Currently, there are no specific treatments or licensed vaccines available for CCHFV. To determine the efficacy of vaccine candidates it is important to conduct serological studies that can accurately measure levels of protective antibodies. In the present study, a pseudo-plaque reduction neutralization test (PPRNT) based on enzyme-catalyzed color development of infected cells probed with anti- CCHFV antibodies were developed to measure 60