Aug 24, 2008 - (1) University of Las Palmas de Gran Canaria, Department of. Animal Pathology, Parasitology ...... OF IRAN ; (2) Department of Radiology, School of Medicine, ...... (Keiser et al., J. Antimicrobial Chemother. (2006) 57, 1139-.
Xth European Multicolloquium Of Parasitology Paris - France August 24th-28th, 2008
PROGRAM & ABSTRACT BOOK
IMPORTANT NOTICE: The abstracts included in this book are the proceedings of the "Xth European Multicolloquium Of Parasitology", as provided by the authors, without modification or copy-editing. The organizers of the congress are, therefore, in any way responsible for abstract presentation or scientific content".
Editorial Dear Colleagues, Welcome to Paris! Please find here the program and the abstracts of communications to be presented to the Xth European Multicolloquium of Parasitology (EMOP-10, Paris, August 24-28, 2008). The keynote lectures are published in a special issue of “Parasite”. This issue, associated with the present Abstract book, displays a large view of the present state of parasitological research. Indeed, EMOP-10 provided an unparalleled opportunity to get a comprehensive view of diverse research trends in the field. The contributions summarized the latest advances on the knowledge of cell biology of eukaryotic pathogens, host parasite interactions, genome, genetic diversity, population genetics, and typing strategies to approach taxonomy, phylogeny, and parasite-host coevolution. Contributions dealing with innovative strategies of control and prevention as well as topics focusing on bioethics, epistemology and history of parasitology will be presented in the meeting. The EMOP-10 Scientific Committee has proposed a first list of symposia, each one including keynote papers and communications in order to ensure that topics of “general” or basic parasitology (including, for instance, parasites of invertebrates and plants) be adequately represented. Parasitology emerges as a dynamic transdisciplinary field of research that extends beyond medical or veterinary perspectives. Actually, beyond health or economic impact of parasite organisms, research focusing mainly on cell or molecular biology, parasitism ecology, hostparasite relationships, molecular taxonomy, phylogeny and phylogeography was surely a significant trend of contributions submitted to the EMOP-10 meeting. Parasitology, as a biological discipline, materializes at best the idea that medical or veterinary research should not be opposed to basic biology approaches. The ability of parasites to succeed as pathogenic agents seems to depend to a large extent upon their life history traits, ecology, genetic background and ultimately upon their evolution. Therefore, without accurate identification of hosts, reservoir, critical ecological issues, and factors involved in parasite circulation in the environment, it is usually impossible to understand parasitic diseases comprehensively. The broad scope of contributions (both keynote lectures and communications) welcomed at the EMOP10 meeting is clearly attesting of the current developments of parasitological research. Above all, the multidisciplinary character of EMOP-10 provides an unparalleled opportunity of common analysis and debate (unfortunately unusual in conventional scientific meetings) among specialists, including scientists, bioethicists, philosophers and historians of sciences. We would like to wish you a fruitful congress. Eduardo Dei-Cas (coordinator of the scientific committee)
Jean Dupouy-Camet (coordinator of the organising committee)
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Committees EMOP10 is organised under the patronage of the European Federation of Parasitology.
Executive board President: Santiago Mas-Coma (Spain) Vice-Presidents: Fabrizio Bruschi (Italy) & Yves Carlier (Belgium) Treasurer: Hans Sluiters (The Netherlands) Secretary: Jean Mariaux (Switzerland) Members at large Tor A. Bakke (Norway), Libuse Kolarova (Czech Republic), Kirill Galaktionov (Russia), Ewert Linder (Sweden), Ermanno Candolfi (France) EMOP10 is organised with the moral support of the representatives of the following societies of Parasitology: Bretislav Koudela (Czech Republic), Branislav Pet’ko (Slovakia), Ziya Alkan (Turkey), Lisa Ranford-Cartwright Charlotte Maddox-Hyttel Carmen M. Cretu (Romania), (England), (Scandinavia), Róbert Farkas (Hungary), Henk Schallig (The Netherlands), Maria-Teresa Manfredi (Italy), Vasil Golemansky (Bulgary), Thomas Schnieder (Germany), Bozena Moskwa (Poland), Stylianos Haralabidis (Greece), Peter Van den Bossche Kosta Y. Mumcuoglu (Israel), Andrew Hemphill (Switzerland), (Belgium). Lali Murvanidze (Georgia), Celia Holland (Ireland), EMOP10 Organising Committees Organising committee Jean Dupouy-Camet (coordinator), Pascal Boireau, Daniel Camus, Gérard Duvallet, Philippe Dorchies, Marie-Laure Dardé, Gilles Dreyfuss, Michel Franc,
Scientific Committee Eduardo Dei-Cas (coordinator), Joseph Schrevel, Jean-François Dubremetz, Simone Caccio, Jean-Antoine Rioux, Gérard Cuny, Robert Killick-Kendrick, Gérard Duvallet, Philippe Dorchies, Arezki Izri, Loïc Favennec, Jean Dupouy-Camet, Odile Bain, Renaud Piarroux, Raymond Pierce, Eric Viscogliosi, Jean Mariaux,
Francis Derouin, Ron Miller, Hélène Yera, Marie-France Cesbron-Delauw, Melanie Cushion, Stéphane Picot, Jamal Khalife, Ermanno Candolfi, K. Trouillas, Simonetta Mattiucci, Bahram Sayyaf Dezfuli, Andrew Shinn, Thierry de Meeûs, Michel Tibayrenc, Patrick Giraudoux, Stéphane De La Roque, Thaddeus Graczyk,
The Honorary president of EMOP10 is Jean-Antoine Rioux.
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René Houin, Jacques Le Bras, Philippe Loiseau, Gérard Luffau. Dominique Mazier, Laurence Delhaes, Isabelle Villena, Pascal Boireau, Fabrizio Bruschi, Jacques Le Bras, Jacques Cabaret, Hervé Pelloux, Michel Franc, Hubert Ferté, André Paugam, Pierre Buffet, Martin Danis, Santi Mas-Coma, David Rollinson.
Acknowledgements The congress is kindly supported by the following companies:
Institutional Sponsorship Région Île de France Université Paris Descartes INRA Université Paris-Sud Agence Française de Sécurité Sanitaire des Aliments CIRAD (EDEN project) Direction Générale de la Santé
Private Sponsorship Merial Marque Verte Bio-Rad Laboratories Sanofi Aventis Cambridge University Press Bordier Gilbert Laboratoires Laboratoire AIM LDBIO Diagnostics Janssen Animal Health ID VET Fulltec AG Merck Medication Familiale Omegapharma Biorecept Biotrin
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Summary
Editorial ......................................................................................................................................... 2 Committees .................................................................................................................................... 3 Acknowledgements ......................................................................................................................... 4 Map of the Cite Internationale Universitaire de Paris .......................................................................... 6 Program at a glance........................................................................................................................ 7 Detailed program Sunday 24th August ................................................................................................................ 9 Monday 25th August................................................................................................................ 9 Tuesday 26th August ............................................................................................................. 12 Wednesday 27th August ........................................................................................................ 18 Thursday 28th August............................................................................................................ 24 Concert in Saint-Jacques-du-Haut-Pas Church ................................................................................. 28 Oral communications..................................................................................................................... 29 Posters classification ....................................................................................................................116 Posters (Session 1) ......................................................................................................................117 Posters (Session 2) ......................................................................................................................168 Index of authors ..........................................................................................................................218 Notes..........................................................................................................................................223
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Program at a glance Insertion du prog. synoptique
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suite programme
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Centre International de Courts Séjours (CICS) : 11 Résidence André HONNORAT 38 Résidence Robert GARRIC 12 Fondation Victor LYON 13 Fondation Danoise 14 Fondation DEUTSCH DE LA MEURTHE 15 Collège d’Espagne 16 Fondation des États-Unis 17 Collège Franco-Britannique 18 Résidence André DE GOUVEIA 19 Maison Heinrich HEINE
20 Fondation Hellénique 21 Maison de l’Inde 22 Maison des Industries Agricoles et Alimentaires 23 Maison de l’ Institut National Agronomique 24 Maison de l’ Italie 25 Maison du Japon 26 Maison du Liban 27 Résidence Lucien PAYE 28 Maison du Maroc 29 Maison du Mexique 30 Fondation de Monaco
SERVICES
Collège Néerlandais Maison de Norvège Maison des Provinces de France Maison des Étudiants Suédois Fondation Suisse Maison de Tunisie
31 Centre de ressources sur le patrimoine de la Cité 37 Espace Sud / citésport
31 32 33 34 35 36
Accueil des chercheurs et des étudiants étrangers Aircup Alliance internationale Bibliothèque / Esp@ce Langues Citéculture Orchestre Théâtre 39 Pavillon administratif Délégation générale 40 Poste de contr ôle sécurité 11 Relais Social International
38 Maison Internationale
BEDROOMS: 17: Collège Franco-Britannique / 9: Maison du Cambodge / 14: Fondation Deutsch de la Meurthe / 33: Maison des Provinces de France
FONDATION VICTOR LYON: Meeting room “Victor Lyon”
MAISON DU CAMBODGE: Meeting room “Cambodge”
Map of the Cité Internationale Universitaire de Paris
MAISON INTERNATIONALE: - Registration desk - Meeting rooms: Adenauer, David-Weill, Gulbenkian, Honnorat, Nathan - Lunch/coffee breaks - Poster area PARIS - Exhibition area UNIVERSITE CLUB- Preview room (PUC / CIUP)
1 Fondation ABREU DE GRANCHER 2 Fondation Argentine 3 Maison des Étudiants Arméniens 4 Maisons des Élèves Ingénieurs des Arts et Métiers 5 Maison des Étudiants de l’Asie du Sud-Est 6 Fondation AVICENNE 7 Fondation BIERMANS-LAPÔTRE 8 Maison du Brésil 9 Maison du Cambodge 10 Maison des Étudiants Canadiens
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Detailed Program Sunday 24th August, 2008 14:00-17:30
REGISTRATION
Monday 25th August, 2008 09:00-10:30
REGISTRATION
09:00-10:15
YSA Committee Organization
Room: DAVID-WEILL
10:30-12:30
OPENING CEREMONY
Room: ADENAUER & HONNORAT
12:30-14:00
LUNCH
14:00-15:30
SY 01 Apicomplexa / 01- Cellular & molecular interactions with the host cells: Session 1 Chair: J.F. DUBREMETZ, V. HEUSSLER Room: DAVID-WEILL SY01/01-01: MOLECULAR DISSECTION OF HOST CELL INVASION BY THE APICOMPLEXANS: THE GLIDEOSOME - D. Soldati-Favre SY01/01-02: INTERACTION BETWEEN MURINE DENDRITIC CELLS AND NEOSPORA CANINUM - S. Dion SY01/01-03: COMPARATIVE DEVELOPMENT OF EIMERIA NINAKOHLYAKIMOVAE IN DIFFERENT IN VITRO CELL CULTURE SYSTEMS - A. Ruiz Discussion
14:00-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30
14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30
SY 02 Kinetoplastida / 01- Genetic & molecular biology Chair: F. BOLAS-FERNANDEZ, J. STEVENS Room: GULBENKIAN SY02/01-01: KINETOPLASTID: PHYLOGENETICS AND EVOLUTION - J. Stevens SY02/01-02: MOLECULAR CHARACTERIZATION OF TWO PROTEIN-TYROSINE PHOSPHATASES FROM LEISHMANIA INFANTUM - F. Bolas-Fernandez SY02/01-03: STRUCTURE AND FUNCTION OF PROTEINS AS PROPHYLACTIC OR THERAPEUTIC TOOLS: A CLOSER LOOK TO LEISHMANIA BRAZILIENSIS HISTONE H1 - E. Carmelo SY02/01-04: LEISHMANIA AMAZONENSIS LEISHPORIN IS A CHOLESTEROL-INDEPENDENT PORE-FORMING PROTEIN AND REQUIRES ONLY LIPIDS TO BIND TO AND LYSE NATURAL AND ARTIFICIAL MEMBRANES - T. Castro-Gomes SY02/01-05: CHROMOSOME COHESION AND SEGREGATION IN TRYPANOSOMA BRUCEI - M. Bessat SY 03 Ectoparasites, Vectors & Control Strategies / 01- Bioecology of mosquitoes, flies & fleas Chair: D. FONTENILLE, G. HAMILTON Room: ADENAUER SY03/01-01: SANDFLY PHEROMONES: THEIR BIOLOGY AND POTENTIAL FOR USE IN CONTROL PROGRAMS G. Hamilton SY03/01-02: STUDIES ON THE BIOLOGY OF MALARIA VECTOR (ANOPHELES LABRANCHIAE) IN THE TONGA LAKE - Z. Harrat SY03/01-03: BIOLOGY OF ANOPHELES MULTICOLOR (CAMBOULIU, 1902) IN OUARGLA (SOUTH OF ALGERIA) - S. Boubidi SY03/01-04: PHLEBOTOMINE SANDFLIES FROM MADAGASCAR (DIPTERA: PSYCHODIDAE) - TAXONOMY, DISTRIBUTION AND PHYLOGENY - J. Depaquit SY03/01-05: EFFICACY OF CAT TREATMENT WITH A COMBINATION OF FIPRONIL AND (S)-METHOPRENE UPON CAT FLEA (CTENOCEPHALIDES FELIS FELIS) EGG DEVELOPMENT IN EXPERIMENTAL BEDDING AREAS - M. Franc SY 04 Intestinal, urinary & liver parasites / 01- Cryptosporidium (except human infection) Chair: G. GARGALA, N. KAPEL Room: HONNORAT SY04/01-01: DRUG TREATMENT & NOVEL DRUG TARGET AGAINST CRYPTOSPORIDIUM - G. Gargala SY04/01-02: ISOLATE-DEPENDENT SHORT AND LONG TERM PATHOGENICITY OF CRYPTOSPORIDIUM PARVUM OOCYSTS IN AN IMMUNOCOMPETENT SUCKLING RAT MODEL - S. Khaldi SY04/01-03: AN ORIGINAL REGULATION OF PEPT1 INTESTINAL EXPRESSION DURING CRYPTOSPORIDIUM PARVUM IN SUCKLING RATS - P. Marquet SY04/01-04: EFFECTS OF NITAZOXANIDE THERAPY ON PATHOGENIC CONSEQUENCES OF ACUTE CRYPTOSPORIDIUM PARVUM INFECTION IN IMMUNOCOMPETENT SUCKLING RAT MODEL - L. Le Goff SY04/01-05: EXPERIMENTAL CRYPTOSPORIDIUM-INDUCED DIGESTIVE NEOPLASIA - G. Certad SY 05 Blood & Tissue Metazoan Infections / 01- Parasite migration-invasion in the early events of infection Chair: R. CURTIS, W. HAAS Room: VICTOR LYON
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14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30
14:00-15:30 14:00-14:30 14:30-15:00 15:00-15:15 15:15-15:30 14:00-15:30
SY05/01-01: PLANT-NEMATODE INTERACTIONS: ENVIRONMENTAL SIGNALS DETECTED BY THE NEMATODE’S CHEMOSENSORY ORGANS CONTROL CHANGES IN THE SURFACE CUTICLE AND BEHAVIOUR R. Curtis SY05/01-02: PARASITISM GENES FROM THE PLANT PARASITIC ROOT-KNOT NEMATODE - M.N. Rosso SY05/01-03: ORIENTATION WITHIN HOST TISSUES: MECHANISMS OF PARASITE NAVIGATION - W. Haas SY05/01-04: INTERACTIONS BETWEEN THE LARVAL ENDOPARASITOID, EXORISTA LARVARUM (DIPTERA) AND ITS FACTITIOUS HOST GALLERIA MELLONELLA (LEPIDOPTERA) - V. Michalkova SY05/01-05: THE ABSENCE OF D6 CHEMOKINE RECEPTOR IMPROVES LITOMOSOIDES SIGMODONTIS FILARIAL INFECTION AND IS ASSOCIATED WITH MAST-CELL ACTIVATION, GRANZYME B RELEASE AND OVEREXPRESSION OF VEGF - C. Martin SY 06 Systematics, Evolution & Phylogeny of parasites / 01- Protozoa & fungi Chair: C.M. ALIOUAT, J. GUARRO, C. NOEL Room: CAMBODGE SY06/01-01: CRYPTIC DIVERSITY IN INTESTINAL PROTISTS - G. Clark SY06/01-02: MALASSEZIA YEASTS, A JUMP FROM THE NON-GENOMIC INTO THE GENOMIC ERA - T. Boekhout SY06/01-03: THE EXPERIMENTAL APPROACH FOR DIRECT INVESTIGATION OF REPRODUCTIVE ISOLATION OF PARASITES: HYBRIDIZATION OF HAEMOPROTEUS SPP. IN VITRO - G. Valkiunas SY06/01-04: PNEUMOCYSTIS OCCURRENCE IN WILD RODENTS FROM SOUTH EAST ASIA - M. Chabé
15:15-15:30
SY 11 Population Genetic Structure & Molecular Epidemiology / 01- Population genetics of parasitic & fungal diseases Chair: T. DE MEEUS, K. McCOY Room: NATHAN SY11/01-01: POPULATION GENETICS OF FUNGAL DISEASES OF PLANTS - T. Giraud SY11/01-02: POPULATION STRUCTURE OF TOXOPLASMA GONDII IN GABON - A. Mercier SY11/01-03: GENETIC DIVERSITY OF PLASMODIUM FALCIPARUM POPULATIONS FROM SITES ACROSS AFRICA INFERRED FROM SNP UNDER SELECTION VS. NEUTRAL SNP - O. Maiga SY11/01-04: MOLECULAR EPIDEMIOLOGY OF ASCARIASIS ON ZANZIBAR: DNA BARCODING OF ASCARIS LUMBRICOIDES REVEALS A HETEROGENEOUS POPULATION STRUCTURE - R. Stothard Discussion
15:30-16:00
COFFEE BREAK
16:00-17:30
SY 02 Kinetoplastida / 02- Immunology & vaccines Chair: R. KILLICK-KENDRICK, P. MARTY Room: GULBENKIAN SY02/02-01: MOLECULAR CROSSTALKS IN LEISHMANIA-SAND FLY-HOST RELATIONSHIPS - P. Volf SY02/02-02: IMMUNISATION WITH NON-PERSISTENT LEISHMANIA MAJOR PHOSPHOMANNOMUTASE DEFICIENT PARASITES PROTECTS MICE FROM VIRULENT CHALLENGE VIA INCREASED MAGNITUDE OF T CELL RESPONSES - L. Kedzierski SY02/02-03: LUMINESCENT LEISHMANIA INFANTUM PARASITES AS A TOOL FOR MONITORING PARASITIC PROCESS IN THE BALB/C MOUSE - G. Michel SY02/02-04: UNDERSTANDING T CELL-MEDIATED PROTECTION AGAINST CUTANEOUS LEISHMANIASIS USING A NOVEL REVERSE GENETICS STRATEGY - K. Kedzierska SY02/02-05: PROTECTION EFFECT OF INTRANASAL VACCINATION WITH L. INFANTUM PROMASTIGOTE ANTIGENS AND CYTOTOXIC NECROTIZING FACTOR (CNF-1) AS ADJUVANT - G. Michel
14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15
16:00-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30 16:00-17:30 16:00-16:30 16:30-17:00 17:00-17:15 17:15-17:30
16:00-17:30 16:00-16:15 16:15-16:30 16:30-16:45 16:45-17:00 17:00-17:15
SY 07 Emergent & Nosocomial Parasitic or Fungal Infections / 01- New aspects of molecular epidemiology of protozoan infections & Nosocomial risks for parasite and fungal infections Chair: F. DEROUIN, G. NEVEZ Room: CAMBODGE SY07/01-01: NOSOCOMIAL PNEUMOCYSTIS JIROVECII INFECTIONS - G. Nevez SY07/01-02: TOXOPLASMA GONDII, “NEW” GENOTYPES AND VIRULENCE - M.L. Darde SY07/01-03: PCR FOR DIAGNOSIS OF PNEUMOCYSTIS JIROVECII FROM ITALIAN PATIENTS - F. Berrilli SY07/01-04: A NEW SAMPLING METHOD FOR PNEUMOCYSTIS JIROVECII DETECTION AND QUANTIFICATION IN THE AIR SURROUNDING PATIENTS DEVELOPING PNEUMOCYSTIS PNEUMONIA - F. Choukri SY 08 Cellular or molecular biology & molecular genetics / 01- Morphogenesis, development, differentiation Chair: S. PICOT, Z. SWIDERSKI Room: ADENAUER SY08/01-01: CRYPTOSPORIDIUM AZAMI N. SP. (APICOMLEXA: CRYPTOSPORIDIIDAE) FROM HOMO SAPIENS - M. Azami SY08/01-02: IDENTIFICATION AND CHARACTERISATION OF COWP-LIKE PROTEINS IN EIMERIA SPECIES R. Walker SY08/01-03: CHARACTERISATION OF EIMERIA TENELLA PROTEASES INVOLVED IN OOCYST WALL FORMATION - M. Katrib SY08/01-04: ULTRASTRUCTURAL CHARACTERS OF SPERMIOGENESIS AND THE SPERMATOZOON IN CARYOPHYLLIDEAN AND TRYPANORHYNCH CESTODES: PHYLOGENETICAL IMPLICATIONS - Z. Swiderski SY08/01-05: COMPARATIVE TEM STUDIES ON THE INFECTIVE ONCOSPHERES OF SEVEN SELECTED SPECIES OF CYCLOPHYLLDEAN CESTODES OF MEDICAL AND VETERINARY IMPORTANCE - Z. Swiderski
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17:15-17:30
SY08/01-06: PRIMARY CELL CULTURE CHARACTERIZATION FROM ECHINOCOCCUS GRANULOSUS LARVAL STAGE - G. Denegri
16:00-17:30
SY 09 Parasite Immunology / 01- Innate immunity Chair: A. GRUDEN-MOVSESIJAN, J. KHALIFE Room: DAVID-WEILL SY09/01-01: ROLE OF NATURAL KILLER T LYMPHOCYTES DURING HELMINTHIC INFECTION - F. Trottein SY09/01-02: NEW GENES OF HARD TICK IXODES RICINUS, INVOLVED IN TICK INNATE IMMUNITY: MLDOMAIN CONTAINING PROTEIN, DER-P2-LIKE ALLERGEN AND TICK RECEPTOR FOR OSPA (TROSPA) - J. Horackova SY09/01-03: TICK(S) DEFENCE PROTEINS: SIMILARITY, SITE OF EXPRESSION AND STAGE-SPECIFIC EXPRESSION - T. Matejovska SY09/01-04: THE ROLE OF DENDRITIC CELLS IN PRESENTATION OF DIFFERENT TRICHINELLA SPIRALIS ANTIGENS AND SUBSEQUENT T CELL POLARIZATION - A. Gruden-Movsesijan SY09/01-05: DIFFERENTIAL EFFECT OF DENDRITIC CELL MATURATION BY EXCRETORY/SECRETORY ANTIGENS DERIVED FROM DIFFERENT HELMINTH SPECIES - M. Langelaar
16:00-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30 16:00-17:30 16:00-16:20 16:20-16:34 16:34-16:48 16:48-17:02 17:02-17:16 17:16-17:30 16:00-17:30 16:00-16:30 16:30-16:45 16:45-17:15 17:15-17:30 16:00-17:30 16:00-16:15 16:15-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30
SY 10 Parasites of fishes, other ectothermic vertebrates & marine mammals / 01- Systematics: from genes to faunistic Chair: T. LITTLEWOOD, S. MATTIUCCI Room: HONNORAT SY10/01-01: USE OF NUCLEAR AND MITOCHONDRIAL DNA PCR AND SEQUENCING FOR MOLECULAR IDENTIFICATION OF DIPHYLLOBOTHRIUM ISOLATES POTENTIALLY INFECTIVE FOR HUMANS - H. Yera SY10/01-02: GENETIC RELATIONSHIPS AMONG ANISAKID SPECIES OF CONTRACAECUM FROM PINNIPEDS INFERRED FROM MITOCHONDRIAL COX2 SEQUENCE: CONGRUENCE WITH ALLOZYME DATA, AND HOSTPARASITE CO-EVOLUTIONARY PROCESSES - S. Mattiucci SY10/01-03: INTRA-INDIVIDUAL VARIATION IN THE NUCLEAR RDNA IN SIBLING SPECIES OF THE ANISAKIS SIMPLEX COMPLEX (NEMATODA: ANISAKIDAE) - S. D'Amelio SY10/01-05: THREE-DIMENSIONAL VISUALISATION OF THE DEVELOPMENTAL STAGES OF AN APICOMPLEXAN FISH BLOOD PARASITE IN ITS INVERTEBRATE HOST - P. Hayes SY10/01-06: BIODIVERSITY OF PARASITES OF CORAL REEF FISH IN THE LAGOON OF NEW CALEDONIA (SOUTH PACIFIC): 10,000 SPECIES, OR MORE? - J.L. Justine Discussion SY 11 Population Genetic Structure & Molecular Epidemiology / 02- Population genetics of vectors Chair: T. DE MEEUS, P. SOLANO Room: NATHAN SY11/02-01: THE POPULATION GENETIC STRUCTURE OF VECTORS AND OUR UNDERSTANDING OF DISEASE EPIDEMIOLOGY - K. McCoy SY11/02-02: USE OF SATELLITE AND MICROSATELLITES TO DESIGN AREA-WIDE TSETSE CONTROL CAMPAIGNS: THE EXAMPLE OF SENEGAL - J. Bouyer SY11/02-03: POPULATION GENETIC STRUCTURE OF G. PALPALIS S.L. IN WEST AFRICA: CONSEQUENCES FOR TSETSE CONTROL - P. Solano Round table: Future directions for the population genetics of vectors SY 12 Innovative Epidemiology / 01- Biodiversity & parasite ecology in a changing world Chair: S. DE LA ROCQUE, S. MORAND Room: VICTOR LYON SY12/01-01: DIVERGENCE BETWEEN EAST ASIAN AND EUROPEAN POPULATIONS OF THE SWIMBLADDERNEMATODE ANGUILLICOLA CRASSUS - E. Heitlinger SY12/01-02: ECHINOCOCCOSIS: SEROLOGICAL DETECTION OF PATIENTS AND MOLECULAR IDENTIFICATION OF PARASITES - A. Ito SY12/01-03: PARASITE COMMUNITY ANALYSIS, A TOOL FOR ASSESSMENT STUDIES - R. Guerrero SY12/01-04: IS THE SPREADING OF CANINE LEISHMANIASIS IN MOROCCO DUE TO THE ECONOMIC, DEMOGRAPHIC AND ENVIRONMENTAL CHANGES? - A. Dakkak SY12/01-05: HOST AND HABITAT SPECIFICITIES OF HEMOSPORIDIAN PARASITES IN TWO AFRICAN BIRD SPECIES - C. Loiseau SY12/01-06: INSTANCES OF PARASITE TRANSMISSION FROM INTRODUCED RED-EARED SLIDER TURTLE (TRACHEMYS SCRIPTA ELEGANS) TO NATIVE EUROPEAN POND TURTLE (EMYS ORBICULARIS) IN SOUTH OF FRANCE - O. Verneau
17:30-19:00
POSTER SESSION #1
Room: ADENAUER
17:30-19:30
EFP Board Meeting
Room: DAVID-WEILL
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Tuesday 26th August, 2008 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-10:00
SY 01 Apicomplexa / 02- Cellular & molecular interactions with the host cells: Session 2 Chair: P. GRELLIER, D. MORRISON Room: DAVID-WEILL SY01/02-01: CELLULAR AND MOLECULAR INTERACTIONS BETWEEN THE APICOMPLEXAN PARASITES PLASMODIUM AND THEILERIA AND THEIR HOST CELLS - V. Heussler SY01/02-02: LCCL DOMAIN PROTEINS: HOMOLOGS OF PLASMODIUM SEXUAL STAGES GENES IDENTIFIED IN BABESIA DIVERGENS - C. Becker SY01/02-03: ROLE OF APOPTOSIS IN THE TRANSMISSION OF PLASMODIUM FALCIPARUM - A.H. Beavogui SY01/02-04: MALARIA AND OBESITY IN MICE - D. Depoix SY01/02-05: NET FORMATION AS PART OF INNATE IMMUNE REACTIONS AGAINST EIMERIA BOVIS - A. Ruiz SY 02 Kinetoplastida / 03- Molecular epidemiology Chair: G. CUNY, W. GIBSON Room: GULBENKIAN SY02/03-01: MOLECULAR EPIDEMIOLOGY OF AFRICAN TRYPANOSOMIASIS: THE CONTRIBUTIONS OF DAVID GEORGE GODFREY OBE TO THE BIOCHEMICAL CHARACTERIZATION OF TRYPANOSOMES - W. Gibson SY02/03-02: IDENTIFICATION OF THE IRON SUPEROXIDE DISMUTASE GENE REPERTOIRE AND ITS USE AS DIAGNOSTIC TOOL IN TRYPANOSOMA BRUCEI GAMBIENSE - S. Herder SY02/03-03: SENSITIVE DETECTION AND EASY TYPING OF LEISHMANIA BY REAL-TIME PCR - J. Roelfsema SY02/03-04: MICROSATELLITES CHARACTERIZATION OF LEISHMANIA ISOLATED STRAINS - S. Reale SY02/03-05: BIOCHEMICAL CHARACTERIZATION OF CAUSATIVE AGENTS OF CUTANEOUS LEISHMANIASIS (CL) IN LAST DECADE AND CURRENT STATUS OF CL IN IRAN - G.R. Hatam SY 03 Ectoparasites, Vectors & Control Strategies / 02- Ticks & tick-borne diseases Chair: N. BOULANGER, L. GERN Room: ADENAUER SY03/02-01: BORRELIA BURGDORFERI SENSU LATO, THE AGENT OF LYME BORRELIOSIS: LIFE IN THE WILDS - L. Gern SY03/02-02: INHIBITORY EFFECT OF TICK SALIVA ON THE INFLAMMATION OF RESIDENT SKIN CELLS INDUCED BY BORRELIA BURGDORFERI, THE CAUSATIVE AGENT OF LYME DISEASE - N. Boulanger SY03/02-03: MULTIPLICATION OF THE TICK ENDOSYMBIONT MIDICHLORIA MITOCHONDRII DURING THE DEVELOPMENT OF THE HOST - C. Genchi SY03/02-04: THE EPIDEMIOLOGY OF TICKS AND TICK BORNE DISEASE IN THE NETHERLANDS - J. Van Der Giessen SY03/02-05: SURVEY OF BABESIA MICROTI INFECTION IN FIELD RODENTS IN CROATIA - R. Beck SY 04 Intestinal, urinary & liver parasites / 02- Amoeba & amoebiasis Chair: J. DUPOUY-CAMET, D. MIRELMAN Room: HONNORAT SY04/02-01: TROPHOZOITES OF ENTAMOEBA HISTOLYTICA EPIGENETICALLY SILENCED IN SEVERAL GENES ARE VIRULENCE ATTENUATED - D. Mirelman SY04/02-02: IDENTIFICATION OF HB-BINDING PROTEINS IN E. HISTOLYTICA GENOME BY IN SILICO ANALYSIS - J.D.J. Olivares Trejo SY04/02-03: THE UNIQUE EPIGENETIC MACHINERY OF ENTAMOEBA HISTOLYTICA AS A TARGET FOR ANTIAMEBIC DRUGS - S. Ankri SY04/02-04: MOLECULAR IDENTIFICATION OF ENTAMOEBA (E.) HISTOLYTICA, E. DISPAR, AND E. MOSHKOVSKII IN STOOLS FROM TUNISIAN INDIVIDUALS WITH AMOEBA INFECTION - A. Bouratbine SY04/02-05: IN VITRO ENCYSTMENT OF ENTAMOEBA HISTOLYTICA BY OXIDATIVE STRESS AND OLIGOELEMENTS - J.C. Carrero SY 05 Blood & Tissue Metazoan Infections / 02- Molecular bases of host-helminth interaction Chair: C. GREVELDING, G. MITTA Room: VICTOR LYON SY05/02-01: THE COMPATIBILITY POLYMORPHISM IN INVERTEBRATE HOST/TREMATODES INTERACTIONS: RESEARCH OF MOLECULAR DETERMINANTS - G. Mitta SY05/02-02: CELLULAR AND MOLECULAR FACTORS OF HEPATIC FIBROSIS DURING METACESTODE INFECTION CAUSED BY MESOCESTOIDES VOGAE (SYN. M. CORTI) - G. Hrckova SY05/02-03: THE OUTER-SURFACE OF ADULT SCHISTOSOMA MANSONI AND TRANSFER OF ANTIGENS TO HOST LIPOPROTEINS - J. Van Hellemond SY05/02-04: SCHISTOSOMA HAEMATOBIUM TOTAL ANTIGEN IS ABLE TO INACTIVATE ESTROGEN RECEPTOR TRANSCRIPTION THROUGH THE USE OF A PLASMID CONTAINING AN ESTROGEN RESPONSIVE ELEMENT IN MAMMALIAN CELLS - M. Botelho Posters' presentation SY 06 Systematics, Evolution & Phylogeny of parasites / 02- Lophotrochozoa: Session 1 Chair: C. BLEIDORN, D. FERRIER Room: CAMBODGE SY06/02-01: ANNELIDS IN EVOLUTIONARY DEVELOPMENTAL BIOLOGY AND COMPARATIVE GENOMICS - D. Ferrier SY06/02-02: LOPHOTROCHOZOAN RELATIONSHIPS AND PARASITES - A SNAP-SHOT - C. Bleidorn
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10:00-10:10 10:10-10:20 10:20-10:30
SY06/02-03: SURPRISES AMONG THE AVIAN SCHISTOSOMES: UNKNOWN LINEAGES AND NEW CHALLENGES FOR UNDERSTANDING THEIR DIVERSIFICATION - E.S. Loker SY06/02-04: BIRD SCHISTOSOMES - NEW FINDINGS FROM NORTHERN AND CENTRAL EUROPE - J. Rudolfova SY06/02-05: UNLOCKING THE SECRETS OF THE SCHISTOSOME SEX CHROMOSOMES : MINING THE GENOME FOR MARKERS TO BE USED IN EVOLUTIONARY AND ECOLOGICAL STUDIES - S.P. Lawton
09:00-10:30
Satellite symposium: VIRTUAL MICROSCOPY FOR PARASITOLOGY Organizer: E. LINDER Room: NATHAN - From satellites to web-microscopes - J. Lundin - Parasites in stools: Problems with quality assessment? - J. Utzinger - Parasites under the Web-Microscope - the WMP - E. Linder - Diagnosis of schistosomiasis in adventure tourists - R. Bergquist, C. Thors - Computer vision for parasitemia assessment - J. Konsti - The future of the WMP - E. Linder, J. Lundin
10:30-11:00
COFFEE BREAK
11:00-12:30
SY 02 Kinetoplastida / 04- Epidemiology-Host parasite interaction Chair: A. BOURATBINE, P. VOLF Room: GULBENKIAN SY02/04-01: PHLEBOTOMINE SANDFLIES AND LEISHMANIA: COEVOLUTION AND CAPTURES - J. Depaquit SY02/04-02: CHARACTERISATION OF TRYPANOSOMES FROM AUSTRALIAN MARSUPIALS - S. Averis SY02/04-03: EPIDEMIOLOGICAL STUDIES ON LEISHMANIOSIS IN THE PROVINCE OF LLEIDA (CATALONIA, NE OF SPAIN) - M. Gallego Culleré SY02/04-04: THE IMPACT OF AGRICULTURE DEVELOPMENT IN THE SPREAD OF VISCERAL LEISHMANIASIS IN TUNISIA - K. Aoun SY02/04-05: THE DETERMINATION OF SPECIES AND GENOTYPES OF LEISHMANIA SPP. USING PCR-RFLP ASSAYS IN CLINICAL SAMPLES OF PATIENTS&RESERVOIRS IN TURKEY - S. Ozensoy Toz
11:00-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:30 11:30-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:25
SY 07 Emergent & Nosocomial Parasitic or Fungal Infections / 02- Cyclospora, Cryptosporidium & Sarcocystis infections in humans Chair: S. CACCIO, R. CHALMERS Room: CAMBODGE SY07/02-01: CRYPTOSPORIDIUM: FROM LABORATORY DIAGNOSIS TO SURVEILLANCE AND OUTBREAKS - R. Chalmers SY07/02-02: GENOTYPING OF CRYPTOSPORIDIUM FROM HIV PATIENTS AND FROM CATTLE IN THAILAND C. Nuchjangreed SY07/02-03: THE EPIDEMIOLOGY OF CRYPTOSPORIDIUM SPECIES IN CHILDREN IN NIGERIAN VILLAGES: TEMPORAL VARIABILITY, MOLECULAR SPECIES IDENTIFICATION AND THE IDENTIFICATION OF RISK FACTORS FOR INFECTION - S.F. Molloy SY07/02-04: CRYPTOSPORIDIOSIS AND GIARDIASIS IN FRANCE. FIRST REPORT FROM THE ANOFEL CRYPTOSPORIDIUM FRENCH NETWORK - F. Derouin SY07/02-05: A NEW COMPREHENSIVE TESTING APPROACH FOR FOOD-BORNE PROTOZOAN OOCYSTS OF PUBLIC HEALTH IMPORTANCE - A. Gajadhar SY 08 Cellular or molecular biology & molecular genetics / 02- Apoptosis Chair: H. HURD, S. PICOT Room: ADENAUER SY08/02-01: INDUCTION AND EXECUTION OF APOPTOSIS-LIKE CELL DEATH IN PLASMODIUM BERGHEI OOKINETES - H. Hurd SY08/02-02: HOST-CELL APOPTOSIS IN TAENIA SOLIUM INDUCED BRAIN GRANULOMAS IN NATURALLY INFECTED PIGS - C.S. Sikasunge SY08/02-03: FEATURES OF PLASMODIUM FALCIPARUM APOPTOSIS AND ROLE OF THE PFMCA1 METACASPASE PROTEIN - B. Meslin SY08/02-04: APOPTOSIS AND MALARIA: FROM BENCH TO BED - S. Picot Discussion SY 09 Parasite Immunology / 02- Immune responses against protozoan & metazoan infections Chair: F. BRUSCHI, J. KHALIFE Room: DAVID-WEILL SY09/02-01: MUCOSAL IMMUNITY IN TOXOPLASMA GONDII INFECTION - D. Buzoni-Gatel SY09/02-02: PROTECTIVE IMMUNITY AGAINST MALARIA LIVER STAGE AFTER VACCINATION WITH LIVE PARASITES - L. Renia SY09/02-03: IN VIVO PROTECTIVE ROLE OF IMMUNOGLOBULIN E AND EOSINOPHILS IN MURINE TRICHINELLOSIS AS SHOWN BY DOWN REGULATING TH2 RESPONSES - F. Bruschi SY09/02-04: NATURALLY ACQUIRED ANTIBODIES TO THE VARO VARIANT FROM P. FALCIPARUM PFEMP1 PROTEIN IN BENINESE CHILDREN - A.G. Lokossou SY 10 Parasites of fishes, other ectothermic vertebrates & marine mammals / 02- Fish parasites in aquaculture and mariculture Chair: A. SHINN, C. SOMMERVILLE Room: HONNORAT SY10/02-01: FISH IMMUNE RESPONSE TO MYXOZOAN PARASITES: MINI-REVIEW - A. Sitja-Bobadilla
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11:25-11:38 11:38-11:51 11:51-12:04 12:04-12:17 12:17-12:30
11:00-12:30 11:00-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:30 11:30-11:42 11:42-11:54 11:54-12:06 12:06-12:18 12:18-12:30
SY10/02-02: MYXOZOAN PARASITES IN TWO IMPORTANT CANDIDATE SPECIES IN MEDITERRANEAN AQUACULTURE: STRIPED SEA BREAM, LITHOGNATHUS MORMYRUS AND SHARPSNOUT SEA BREAM, DIPLODUS PUNTAZZO - G. Alama-Bermejo SY10/02-03: CHARACTERISATION, LOCALISATION AND INFECTION DYNAMICS OF THE RENAL MYXOZOANS GADIMYXA ATLANTICA AND ZSCHOKKELLA HILDAE IN CULTURED ATLANTIC COD GADUS MORHUA L. USING MOLECULAR APPROACHES - A.S. Holzer SY10/02-04: THE IMPACT OF THE COPEPOD PARASITE, LERNAEOCERA BRANCHIALIS, ON CULTURED JUVENILE COD, GADUS MORHUA - C. Sommerville SY10/02-05: OCCURANCE AND INTENSITY OF NITZSCHIA STURIONIS (MONOGENEAN: CAPSALIDAE) IN THE STURGEONS (CHONDROSTEI: ACIPENSERIDAE) OF SOUTH OF CASPIAN SEA - A. Halajian SY10/02-06: ULTRASTRUCTURE OF THE DIGESTIVE SYSTEM OF MACROGYRODACTYLUS CONGOLENSIS PRUDHOE, 1957, A MONGENEAN SKIN PARASITE OF THE NILE CATFISH CLARIAS GARIEPINUS - M. ElNaggar SY 11 Population Genetic Structure & Molecular Epidemiology / 03- Co-structures of hosts, parasites & vectors Chair: T. DE MEEUS, F. PRUGNOLLE Room: NATHAN SY11/03-01: PARASITE CO-STRUCTURE: BROAD AND LOCAL SCALE APPROACHES - C. Criscione SY11/03-02: POPULATION STRUCTURE AND GEOGRAPHICAL SUBDIVISION OF THE LEISHMANIA MAJOR VECTOR PHLEBOTOMUS PAPATASI (DIPTERA: PSYCHODIDAE) AS REVEALED BY MICROSATELLITE VARIATION - O. Hamarsheh SY11/03-03: STRIKING HUMAN INFLUENCE ON THE DISTRIBUTION AND DIVERSITY OF TRICHINELLA SPIRALIS - B. Rosenthal SY11/03-04: WITHIN POPULATION STRUCTURE OF IXODES RICINUS, THE EUROPEAN VECTOR OF LYME BORRELIOSIS - F. Kempf SY11/03-05: POPULATION GENETIC STRUCTURE OF OSTERTAGIINAE OF CERVIDAE AND HISTORICAL HOST DISPLACEMENTS - E. Lienard SY 12 Innovative Epidemiology / 02- Multidisciplinary approach of transmission studies & epidemiology Chair: P. GIRAUDOUX, M. WOOLHOUSE Room: VICTOR LYON SY12/02-01: MULTIDISCIPLINARY STUDIES, SYSTEMS APPROACHES AND ECO-EPIDEMIOLOGY: SOMETHING OLD, SOMETHING NEW - P. Giraudoux SY12/02-02: SENSORY SYSTEM OF REDIAE AND CERCARIAE OF PARAGONIMUS OHIRAI, DIGENEA, PARAGONIMIDAE, INTEREST IN EPIDEMIOLOGY - C. Bayssade-Dufour SY12/02-03: INFRAPOPULATION DYNAMICS OF GYRODACTYLUS SALARIS INFECTING ARCTIC CHARR FRY A.C. Winger SY12/02-04: ANTI-MOSQUITO SALIVA ANTIBODY RESPONSE OF HORSES TO ANOPHELES BITES IN SENEGAL - D. Boulanger SY12/02-05: INFERRING TRANSMISSION PATTERNS OF SCHISTOSOMA MANSONI USING MOLECULAR MARKERS AND PARENTAGE ANALYSIS - M. Steinauer SY12/02-06: MULTIPLE EFFECTS OF A NATIONWIDE LONG-LASTING INSECTICIDE-TREATED NETS (LLINS) DISTRIBUTION IN NIGER - C. Czeher
12:30-14:00
LUNCH
14:00-15:30
SY 01 Apicomplexa / 03- Gene expression & regulation Chair: J.F. DUBREMETZ, D. SOLDATI-FAVRE Room: DAVID-WEILL SY01/03-01: TOXOPLASMA GONDII GENE EXPRESSION IS UNDER THE CONTROL OF REGULATORY PATHWAYS ACTING THROUGH CHROMATIN STRUCTURE - M.A. Hakimi SY01/03-02: CAN VAR GENE MODULATION SERVE AS A NOVEL TARGET FOR MALARIA THERAPY? - E. Rosenberg SY01/03-03: IDIOSYNCRASY OF MITOCHONDRIAL TRANSLATION IN APICOMPLEXANS - P. Pino SY01/03-04: DRUG-INHIBITION OF HDAC ACTIVITY: EPIGENETIC CONTROL OF CELL CYCLE AND DIFFERENTIATION IN APICOMPLEXAN PARASITES - A. Bougdour SY01/03-05: SEQUENCING AND ANNOTATION OF THE GENOME OF NEOSPORA CANINUM - J.M. Wastling
14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30
SY 03 Ectoparasites, Vectors & Control Strategies / 03- Vector control & live/scabies Chair: A. IZRI, K.Y. MUMCUOGLU Room: VICTOR LYON SY03/03-01: TREATMENT OF SCABIES INFESTATIONS - K.Y. Mumcuoglu SY03/03-02: BIONOMICS OF MALARIA VECTORS IN A MALARIOUS AREA, IRAN - H. Vatandoost SY03/03-03: HABITAT, DISTRIBUTION AND STRUCTURE OF RIVERINE TSETSE POPULATIONS OF THE MOUHOUN RIVER BASIN IN BURKINA FASO: IMPLICATIONS FOR CONTROL CAMPAIGNS - J. Bouyer SY03/03-04: MICROORGANISMS AND MOSQUITO VECTORS: PERSPECTIVES FOR THE CONTROL OF MOSQUITO BORNE DISEASES - G. Favia SY03/03-05: ACARICIDAL EFFECT OF PELARGONIUM ROSEUM AND EUCALYPTUS GLOBULUS ESSENTIAL OILS AGAINST ADULT STAGE OF RHIPICEPHALUS (BOOPHILUS) ANNULATUS - K. Pirali Kheirabadi
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14:00-15:30 14:00-14:15 14:15-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30
14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30
SY 04 Intestinal, urinary & liver parasites / 03- Epidemiology of giardasis Chair: A.G. BURET, L. FAVENNEC Room: CAMBODGE SY04/03-01: EVIDENCE FOR GENETIC EXCHANGES AMONG HUMAN ISOLATES OF GIARDIA DUODENALIS S. Caccio SY04/03-02: GENOTYPIC CHARACTERISATION OF GIARDIA DUODENALIS ISOLATES FRON FRENCH DIARRHEIC AND/OR DYSPEPTIC PATIENTS - J. Bonhomme SY04/03-03: MOLECULAR AND EPIDEMIOLOGICAL STUDY ON HUMAN HEALTH IMPACTS FROM GIARDIA DUODENALIS IN DRINKING WATER IN HUNGARY - J. Plutzer SY04/03-04: GIARDIA ZOONOTIC ASSEMBLAGE CIRCULATING AMONG CHILDREN AND DOGS IN A SOCIALLY DISADVANTAGED SMALL COMMUNITY - F. Berrilli SY04/03-05: GIARDIA DUODENALIS GENETIC ASSEMBLAGES DIFFER IN DIFFERENT DOG POPULATIONS IN BELGIUM - E. Claerebout SY04/03-06: SWEDISH CATS SHARE GIARDIA SUB-GENOTYPES WITH RUMINANTS BUT NOT WITH HUMANS - M. Lebbad SY 10 Parasites of fishes, other ectothermic vertebrates & marine mammals / 03- Ecology I: parasite life-cycles and host-specificity Chair: A. FALTYNKOVA, M. KALBE Room: HONNORAT SY10/03-01: ALTERNATIVE LIFE CYCLE STRATEGIES OF MEGALODISCUS TEMPERATUS IN TADPOLES AND METAMORPHOSED ANURANS - M. Bolek SY10/03-02: TRANSMISSION ECOLOGY OF POLYSTOMA GALLIENI (PLATHYHELMINTHES, MONOGENEA), AN OPPORTUNIST PARASITE OF HYLA MERIDIONALIS (ANURA, HYLIDAE) - M. Badets SY10/03-03: LINK BETWEEN INTRASPECIFIC VARIABILITY AND HOST SPECIFICITY IN LAMELLODISCUS MONOGENEANS - Y. Desdevises SY10/03-04: LAMELLODISCUS (MONOGENEA, DIPLECTANIDAE) PARASITES OF SEVERAL SPARID FISH HOSTS: TRUE GENERALISTS OR ONGOING SPECIATIONS? - T. Poisot Discussion SY 13 Innovative Strategies in Diagnosis / 01- Advances in malaria diagnosis & molecular biology Chair: D. MAZIER, I. VILLENA Room: NATHAN SY13/01-01: PCR-BASED METHODS TO THE DIAGNOSIS OF IMPORTED MALARIA - A. Berry SY13/01-02: MOBILE BIOLOGY LABORATORY FOR ANTIMALARIAL DRUG SUSCEPTIBILITY EVALUATION IN THE FIELD - V. Sinou SY13/01-03: OOCYST DETECTION OF TOXOPLASMA IN WATER: AN EXAMPLE OF DETECTION FROM WATER IN CHAMPAGNE-ARDENNE, FRANCE - I. Villena SY13/01-04: CLINICAL VALUE OF CURRENT PCR FOR DETECTION OF PNEUMOCYSTIS JIROVECII IN INVASIVE VERSUS NON-INVASIVE CLINICAL SAMPLES - I. Durand-Joly SY13/01-05: HUMAN-VECTOR IMMUNE INTERACTIONS DURING MALARIA: IDENTIFICATION OF AN IMMUNOLOGICAL MARKER OF EXPOSURE TO ANOPHELES GAMBIAE BITES BASED ON A PEPTIDE DESIGN OF THE GSG6 SALIVARY PROTEIN - A. Poinsignon SY 15 Innovative Prevention & Therapeutic Strategies / 01- Immunotherapy Chair: J. GOLENSER, N. TUIKUE NDAM Room: GULBENKIAN SY15/01-01: TOWARDS A VACCINE AGAINST PREGNANCY-ASSOCIATED MALARIA - N. Tuikue Ndam SY15/01-02: FOLLOW UP OF PLASMODIUM FALCIPARUM GENOTYPE AND ANTI-VAR2CSA IMMUNE RESPONSE DURING PREGNANCY IN SENEGAL - J. Guitard SY15/01-03: IMMUNE INTERVENTION FOR TREATMENT OF CEREBRAL MALARIA - J. Golenser SY15/01-04: IL-13 INDUCES EXPRESSION OF CD36 RECEPTOR IN HUMAN MONOCYTES THROUGH PPAR ACTIVATION AND CONTRIBUTES TO THE ENHANCEMENT OF PLASMODIUM-PARASITIZED ERYTHROCYTES PHAGOCYTOSIS - D. Olagnier SY15/01-05: PROGRESS WITH MALARIA ELIMINATION IN THE WHO EUROPEAN REGION - R. KurdovaMintcheva
14:00-15:30
Satellite symposium: GEOSPATIAL HEALTH IN PARASITOLOGY AND VECTOR BORNE DISEASES (1) sponsored by the Global Network on Geospatial Health & the IRD Unité Espace Organizers: G. CRINGOLI, C. ROMANA Room: ADENAUER - Geospatial Health: a Timely Approach in a Changing Environment - R. Bergquist - Health Sentinel Data: Progress in Medical Applications of Earth Observing Satellites - J. Malone - CNES Strategy towards Operationnal Tools for Vector Borne Diseases – A. Guelle
15:30-16:00
COFFEE BREAK
16:00-17:30
SY 01 Apicomplexa / 04- Diversity & evolution from ancestral to highly specialized Apicomplexa Chair: S. CACCIO, J. SCHREVEL Room: DAVID-WEILL SY01/04-01: PROSPECTS FOR ELUCIDATING THE PHYLOGENY OF THE APICOMPLEXA - D. Morrison SY01/04-02: EUGREGARINES AND CRYPTOSPORIDIA: PECULIAR PARASITES WITH EPICELLULAR DEVELOPMENT - A. Valigurová
16:00-16:30 16:30-16:45
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16:45-17:00 17:00-17:15 17:15-17:30 16:00-17:30 16:00-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30 16:00-17:30 16:00-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30
16:00-17:30 16:00-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30 16:00-17:30 16:00-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30 16:00-17:30
SY01/04-03: BLASTOGREGARINES (ALVEOLATA: APICOMPLEXA: SPOROZOA) ARE A PLESIOMORPHIC GROUP OF SPOROZOANS AS INFERRED FROM ULTRASTRUCTURAL AND MOLECULAR DATA - T. Simdyanov SY01/04-04: PHYLOGENY OF HAEMOSPORIDIA WITH BAT PARASITES FROM MADAGASCAR AND CAMBODIA - F. Ariey SY01/04-05: THE METHYLCITRATE CYCLE, AN UNEXPECTED METABOLIC PATHWAY IN THE MITOCHONDRION OF COCCIDIAN PARASITES - T. Fleige SY 03 Ectoparasites, Vectors & Control Strategies / 04- Other ecto- & meso-parasites Chair: G. DUVALLET, D. OTRANTO Room: VICTOR LYON SY03/04-01: BIODIVERSITY AND EXTINCTION VS. CONTROL OF OESTRID CAUSING MYIASIS IN MEDITERRANEAN AREA - D. Otranto SY03/04-02: EKBOM SYNDROM OR ECTOPARASITOSIS? A CASE REPORT - A.P. Bellanger SY03/04-03: BACTERIAL TAXA ASSOCIATED WITH THE HEMATOPHAGOUS MITE, DERMANYSSUS GALLINAE BY 16S RDNA PCR AMPLIFICATION AND TTGE FINGERPRINT - L. Zenner SY03/04-04: IDENTIFICATION OF FORENSICALLY IMPORTANT BEETLES ON EXPOSED HUMAN LEFT OVER PARTS IN ASSIUT, EGYPT - L. Galal SY03/04-05: SCLERODERMA DOMESTICUM - AN UNKNOWN INDOOR BITING INSECT - P. Delaunay SY 04 Intestinal, urinary & liver parasites / 04- Giardia: physiopathology & treatment Chair: A.G. BURET, M.G. ORTEGA-PIERRES Room: CAMBODGE SY04/04-01: PATHOPHYSIOLOGY OF ENTERIC INFECTIONS WITH GIARDIA DUODENALIS - A.G. Buret SY04/04-02: GIARDIA DUODENALIS: ADHESION DEFICIENT CLONES HAVE REDUCED ABILITY TO ESTABLISH INFECTION IN MONGOLIAN GERBILS - M.G. Ortega-Pierres SY04/04-03: INTESTINAL ALKALINE PHOSPHATASE ISOENZYME AUGMENTATION FOLLOW GIARDIA INFECTION - E. Razmjou SY04/04-04: IDENTIFICATION OF DIFFERENTIALLY EXPRESSED GENES IN A GIARDIA LAMBLIA STRAIN RESISTANT TO NITAZOXANIDE AND METRONIDAZOLE - N. Mueller SY04/04-05: DIFFERENTIAL DRUG METABOLIZATION AND EXPRESSION OF AN ANTIOXIDANT PEROXIREDOXIN IN ALBENDAZOLE-SENSITIVE AND RESISTANT GIARDIA DUODENALIS CLONES - R. Arguello-Garcia SY 12 Innovative Epidemiology / 03- Molecular markers & sentinel organisms for environmental monitoring Chair: D.B. CONN, T. GRACZYK Room: HONNORAT SY12/03-01: MOLECULAR MARKERS AND SENTINEL ORGANISMS FOR ENVIRONMENTAL MONITORING - T. Graczyk SY12/03-02: ANTIBODY RESPONSES TO SALIVA OF TRIATOMA INFESTANS: THEIR POTENTIAL AS EPIDEMIOLOGICAL TOOL FOR CHAGAS DISEASE SURVEILLANCE - A. Schwarz SY12/03-03: INCIDENCE RATE AND ENVIRONMENTAL DETERMINANTS OF CANINE VISCERAL LEISHMANIASIS IN ALPES-MARITIMES, FRANCE - R. Pin-Diop SY12/03-04: INFECTION WITH HEPATOZOON IN THE SMALL CROATIAN RODENTS APODEMUS SYLVATICUS, APODEMUS FLAVICOLIS, APODEMUS AGRARIUS AND CLETHRIONOMYS GLAREOLUS - L. Vojta SY12/03-05: FIRST MOLECULAR CHARACTERIZATION OF CANINE HEPATOZOONOSIS IN CROATIA USING A SCREENING PCR AND SEQUENCING METHODOLOGIES - L. Vojta SY 15 Innovative Prevention & Therapeutic Strategies / 02- Anthelminthic drug strategies Chair: J. CABARET, P. JACQUIET Room: GULBENKIAN SY15/02-01: PRO AND CONS OF TARGETED SELECTIVE TREATMENT AGAINST DIGESTIVE-TRACT STRONGYLES OF RUMINANTS - J. Cabaret SY15/02-02: NEMATODE TARGETED TREATMENTS IN GOATS UNDER TEMPERATE CLIMATE USING BODY CONDITION SCORING - C. Paraud SY15/02-03: LEVAMISOLE-SENSITIVE NICOTINIC ACETYLCHOLINE RECEPTOR SUBUNIT GENE INVESTIGATION IN THE TRICHOSTRONGYLINA NEMATODES HAEMONCHUS CONTORTUS, TELADORSAGIA CIRCUMCINCTA AND TRICHOSTRONGYLUS COLUBRIFORMIS - C. Neveu SY15/02-04: RECONSTITUTING THE LEVAMISOLE-SENSITIVE ACHR OF NEMATODES IN XENOPUS OOCYTES - T. Boulin SY15/02-05: EOSINOPHILS IN HAEMONCHUS CONTORTUS-INFECTED RESISTANT AND SUSCEPTIBLE BREEDS OF SHEEP: ABOMASAL TISSUE RECRUITMENT AND IN VITRO FUNCTIONAL STATE - P. Jacquiet Satellite symposium: GEOSPATIAL HEALTH IN PARASITOLOGY AND VECTOR BORNE DISEASES (2) sponsored by the Global Network on Geospatial Health & the IRD Unité Espace Organizers: G. CRINGOLI, C. ROMANA Room: ADENAUER - Geospatial Tools in Veterinary Parasitology - L. Rinaldi - Landscape and Modelling Spatial Ecosystemic Approach - C. Romana - Climate and Ecological Niches of Ticks and Tick-borne Diseases - A. Estrada-Pena
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16:00-17:30
Satellite symposium: FILARIASIS: NEW QUESTIONS AND INTEREST FOR OTHER PARASITIC DISEASES Organizers: O. BAIN, P. BOIREAU, R. MAIZELS Room: NATHAN - Update on anti-wolbachial chemotherapy of human filariasis - where do we stand? - A. Hoerauf - Preliminary results of our experience with the community directed treatment with doxycycline in the rain forest villages of Cameroon - S. Wanji - Vector transmission and low microfilarial density - M.G. Basanez, O. Bain - Current Controversies in Antibiotic Chemotherapy - B. Makepeace, S. Trees - Immunoregulation with regards to malaria infection as well as asthma - S. Specht - Co-infection filariae and oxyurids - C. Martin - Unexpected consequences of vaccine-immunity to filariasis - S. Babayan - Analysis of filarial secreted proteins - R. Maizels
17:30-19:00
WFP Meeting
Room: GULBENKIAN
17:30-19:00
POSTER SESSION #1
Room: ADENAUER
17:30-19:30
Presentation of YSA candidates
Room: DAVID-WEILL
17
Wednesday 27th August, 2008 09:00-10:30 09:00-09:15 09:15-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30
SY 04 Intestinal, urinary & liver parasites / 05- Waterborne protozoa & helminths Chair: L. LE GOFF, M.G. ORTEGA-PIERRES Room: HONNORAT SY04/05-01: DETECTION AND GENOTYPING OF GIARDIA DUODENALIS IN ORETO RIVER (SICILY, SOUTH ITALY) - F. Di Piazza SY04/05-02: IDENTIFICATION OF PARAMETERS INFLUENCING CRYPTOSPORIDIUM SPP. OOCYST RESTITUTION FRON KARSTIC CHALK AQUIFERS IN HAUTE-NORMANDIE - L. Le Goff SY04/05-03: OCCURRENCE OF A NEW PARASITE AMOEBOZOAN IN GAMMARID AMPHIPODA FROM BALTIC SEA - D. Codreanu Balcescu SY04/05-04: THE SCHOOL HEALTH PROGRAMME IN EGYPT: THE IMPACT OF A TWELVE-YEAR INTEGRATED HELMINTH CONTROL PROGRAMME IN THE NILE DELTA (1996-2007) - F. Curtale SY04/05-05: CONTROL OF PARASITIC DISEASES THROUGH SCHOOL-BASED HEALTH EDUCATION AND DEWORMING: A FIVE YEARS ACTIVITIES IN ADA-FOAH, GHANA - K.M. Bosompem SY04/05-06: BLASTOCYSTIS: A NEGLECTED PATHOGEN? - C.R. Stensvold SY 05 Blood & Tissue Metazoan Infections / 03- Current trends in tissular helminth propagation. Zoonoses & environmental changes Chair: C. GENCHI, P. HORAK Room: VICTOR LYON SY05/03-01: CURRENT TRENDS IN TISSUE-AFFECTING HELMINTHS - B. Gottstein, R. Piarroux SY05/03-02: SONOLOGIC AND SEROLOGICAL SURVEY OF HYDATID DISEASE IN RURAL REGIONS OF SHAHDAD AND CHATROUD, KERMAN PROVINCE - M. Fasihi Harandi SY05/03-03: PENETRATION OF TRICHOBILHARZIA CERCARIAE INTO MAMMALS: DANGEROUS OR NEGLIGIBLE EVENT? - P. Horak SY05/03-04: RARE ARTHROPOD-TRANSMITTED HELMINTHES WITH REGARDS TO ECO-BIOLOGICAL FACTORS IN IRAN - G. Mowlavi SY 06 Systematics, Evolution & Phylogeny of parasites / 03- Lophotrochozoa: Session 2 Chair: J. MARIAUX, O. VERNEAU Room: DAVID-WEILL SY06/03-01: PLATYHELMINTH SYSTEMATICS AND THE EMERGENCE OF NEW CHARACTERS - T. Littlewood SY06/03-02: GYRODACTYLID (MONOGENEA) RECORDS ON NORWEGIAN ARCTIC CHARR (SALVELINUS ALPINUS) - T.A. Bakke SY06/03-03: CALIBRATING A MOLECULAR CLOCK IN PLATYHELMINTH PARASITES (MONOGENEA, POLYSTOMATIDAE) USING THE VERTEBRATE HOSTS - O. Verneau SY06/03-04: GENETIC CHARACTERIZATION OF DIPLOZOID SPECIES FROM BRITISH FRESHWATER FISH - D. Thenet SY06/03-05: EGG-SHAPE DIVERSITY IN CESTODES: ADAPTATION TO TRANSMISSION? - F. Lefebvre SY 07 Emergent & Nosocomial Parasitic or Fungal Infections / 03- Health and economic impact of human microsporidian infections & Epidemiology and diagnosis of free living amoebae Chair: T. VAN GOOL, H. YEAR Room: NATHAN SY07/03-01: ENTEROCYTOZOON BIENEUSI IN HUMAN AND ANIMALS, FOCUS ON LABORATORY IDENTIFICATION AND MOLECULAR EPIDEMIOLOGY - M. Thellier SY07/03-02: IDENTIFICATION OF ACANTHAMOEBA T4 AND T15 GENOTYPES IN KERATITIS PATIENTS IN ITALY - D. Di Cave SY07/03-03: STRESS 'TOXIC' REACTION: A NEWLY-IDENTIFIED PROTECTION OF ACANTHAMOEBAE AGAINST ORGANIC SOLVENTS. - J. Kliescikova SY07/03-04: ASYMPTOMATIC CARRIAGE OF CRYPTOSPORIDIUM SPP. IN PRE-SCHOOL AGE CHILDREN IN THE UK – R. Chalmers Discussion SY 08 Cellular or molecular biology & molecular genetics / 03- Host-parasite molecular interaction Chair: P. CAVAILLES, E.S. LOKER Room: CAMBODGE SY08/03-01: HOW THE TOXO1 LOCUS DIRECTS TOXOPLASMOSIS OUTCOME - P. Cavailles SY08/03-02: IDENTIFICATION OF ANOPHELES GAMBIAE GENES THAT AFFECT PLASMODIUM INFECTION G. Jaramillo Gutierrez SY08/03-03: FREP 4: A MODEL OF A TREMATODE-RESPONSIVE POLYPEPTIDE IN BIOMPHALARIA GLABRATA HEMOLYMPH - E.S. Loker SY08/03-04: PROTEOMIC ANALYSIS OF LOW ABUNDANCE PROTEINS INVOLVED IN HOST-PARASITIC INTERACTION IN SHEEP SELECTED FOR RESISTANCE AND/OR SUSCEPTIBILITY TO TRICHOSTRONGYLUS COLUBRIFORMIS - S. Binos SY08/03-05: MOLECULAR CHARACTERISATION OF TOTAL AND DIFFERENTIALLY EXPRESSED SECRETOME OF TRYPANOSOMA CONGOLENSE: IDENTIFICATION OF NOVEL VIRULENCE AND/OR PATHOGENIC FACTORS - P. Holzmuller
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09:00-10:30 09:00-09:15 09:15-09:30 09:30-09:45 09:45-10:00 10:00-10:15
SY 09 Parasite Immunology / 03- Vaccines Chair: D. BUZONI-GATEL, L. RENIA Room: GULBENKIAN SY09/03-01: FIRST FIELD TRIAL OF A RECOMBINANT VACCINE AGAINST ONCHOCERCIASIS: EVALUATION IN CATTLE EXPOSED TO ONCHOCERCA OCHENGI - B. Makepeace SY09/03-02: EVALUATION OF PROTECTIVE EFFECT OF RECOMBINANT PROTEIN OF EIMERIA USED AS AN ADJUVANT AGAINST CHRONICAL TOXOPLASMOSIS IN MICE - D. Hedhli SY09/03-03: CD8+ BUT NOT CD4+ T CELLS MEDIATE ANTI-TOXOPLASMA GONDII IMMUNITY IN A CHRONIC MURINE MODEL BY IMMUNIZATION WITH PULSED DENDRITIC CELLS - R. Guiton SY09/03-04: THE CELLULAR RESPONSE OF SHEEP VACCINATED WITH A RECOMBINANT FASCIOLA HEPATICA PHOSPHOGLYCERATE KINASE (PGK) AND INFECTED WITH FLUKE METACERCARIAE - M. Kozak SY09/03-05: OVERCOMING ANTIGENIC DIVERSITY OF THE MALARIA VACCINE CANDIDATE APICAL MEMBRANE ANTIGEN-1 - S. Dutta
09:00-10:30
Satellite symposium: CHANGES AND SPATIAL SHIFTS IN ARTHROPOD BORNE DISEASES IN EU? FOCUS ON TICKS sponsored by MERIAL Organizer: F. BEUGNET Room: ADENAUER - Reasons for modification of tick Ecology in EU - F. Beugnet - Situation of 3 unrecognised zoonotic diseases in EU: granulocytic ehrlichiosis, tick borne encephalitis and flea borne rickettsiosis - J.L. Marie - Wind as an important factor to explain the spreading of some vector borne diseases - G. Hendrickx
10:30-11:00
COFFEE BREAK
11:00-12:30
SY 06 Systematics, Evolution & Phylogeny of parasites / 04- Nematoda & acanthocephala Chair: O. BAIN, M. BLAXTER Room: DAVID-WEILL SY06/04-01: THE NEMATODA FILARIOIDEA: CRITICAL ANALYSIS LINKING MOLECULAR AND TRADITIONAL APPROACHES - O. Bain SY06/04-02: MOLECULAR INSIGHT INTO HOST SPECIFICITY, LIFE CYCLES AND GEOGRAPHICAL DISTRIBUTION OF THE RHABDIASIDAE (NEMATODA) - Y. Kuzmin SY06/04-03: OXYURID NEMATODES FROM TAPOLES: GYRINICOLA SPECIES AND ADAPTATIONS OF THEIR BIOLOGY TO THE HOST LIFE-CYCLE - B. Planade SY06/04-04: A COMPUTER-ASSISTED IDENTIFICATION KEY FOR THE HELIGMONELLINAE (NEMATODA, TRICHOSTRONGYLINA) - R. Vignes-Lebbe SY06/04-05: AN UPDATE ON RHABDIAS NEMATODES FROM LUNGS OF CHAMAELEONIDAE: DIVERSITY AND BIOLOGY - N. Lhermitte-Vallarino
11:00-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:15 11:15-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:20 11:20-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30
SY 08 Cellular or molecular biology & molecular genetics / 04- Gene regulation & polymorphisms Chair: C. GRUNAU, M.A. HAKIMI Room: CAMBODGE SY08/04-01: MOLECULAR DETERMINATION OF GAMETOCYTOGENESIS IN PLASMODIUM FALCIPARUM - B. Witkowski SY08/04-02: THE ROLE OF EPIGENETICS IN THE GENERATION OF PHENOTYPIC VARIABILITY IN HOSTPARASITE INTERACTIONS: THE BIOMPHALARIA GLABRATA / SCHISTOSOMA MANSONI MODEL - C. Cosseau SY08/04-03: AN EXPERIMENTAL APPROACH FOR THE SCREENING OF TRYPANOSOMATID CONSERVED SECRETED PROTEINS - R.M. Corrales SY08/04-04: HIGHLY POLYMORPHIC MICROSATELLITES IN NEOSPORA CANINUM - C. Bazzocchi SY08/04-05: GENOTYPING OF ECHINOCOCCUS GRANULOSUS SENSU LATO: OVERVIEW OF THE MITOCHONDRIAL COX1 AND NAD1 POLYMORPHISM AND EVALUATION OF THE MULTILOCUS MICROSATELLITE EMSB - S. Maillard SY08/04-06: MULTIPLE ORIGIN OF PLASMODIUM VIVAX DHFR AND DHPS MUTANT ISOLATES FROM MADAGASCAR - C. Barnadas SY 10 Parasites of fishes, other ectothermic vertebrates & marine mammals / 04- Ecology II: host-parasite interaction & parasite communities Chair: M. BOLEK, Y. DESDEVISES Room: HONNORAT SY10/04-01: PARASITES AS SELECTIVE FORCES IN ECOLOGICAL DIVERSIFICATION - EXAMPLES FROM STICKLEBACKS - M. Kalbe SY10/04-02: WHAT CAN WE LEARN FROM PARASITES ABOUT SWORDFISH (X. GLADIUS) STOCK ASSESSMENT AND HEALTH STATE? - S. Damiano SY10/04-03: TREMATODE COMMUNITY STRUCTURE IN VALVATA MACROSTOMA WITH A SPECIAL REFERENCE TO TRANSMISSION PATTERNS OF ICHTHYOCOTYLURUS PILEATUS - A. Faltynkova SY10/04-04: PARASITES OF APOLLONIA MELANOSTOMA (NEOGOBIUS MELANOSTOMUS) (PALLAS) AND NEOGOBIUS KESSLERI (GÜNTHER, 1861) (GOBIIDAE: OSTEICHTHYES) FROM THE DANUBE RIVER IN AUSTRIA - J.M. Muhlegger SY10/04-05: HOST INTRODUCTION AND PARASITES: A CASE STUDY ON THE PARASITE COMMUNITY OF THREE FISH SPECIES IN THE HAWAIIAN ISLANDS - M. Vignon SY10/04-06: MONITORING OF TREMATODE COMMUNITIES (CERCARIAE) IN LYMNAEA STAGNALIS (GASTROPODA, PULMONATA) BY MARK-RECAPTURE METHOD - M. Soldanova
19
11:00-12:30 11:00-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:30 11:30-11:42 11:42-11:54 11:54-12:06 12:06-12:18 12:18-12:30 11:00-12:30 11:00-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30 11:00-12:30
SY 11 Population Genetic Structure & Molecular Epidemiology / 04- Impact of clonality on population genetics Chair: T. DE MEEUS, T. GIRAUD Room: NATHAN SY11/04-01: THE IMPACT OF CLONALITY ON PARASITE POPULATION GENETIC STRUCTURE - F. Prugnolle SY11/04-02: INBREEDING IN PROTOZOAN PARASITES OF THE GENUS LEISHMANIA - V. Rougeron SY11/04-03: COMPARATIVE POPULATION GENETICS OF TOXOPLASMA GONDII AND RELATED PARASITES B. Rosenthal SY11/04-04: VARIATION IN GIARDIA: TOWARDS A TAXONOMIC REVISION OF THE GENUS - A. Thompson Round table: Future direction in the population genetics of parasitic clones SY 12 Innovative Epidemiology / 04- Spatial & temporal modelling for transmission studies and risk assessment Chair: F.M. DANSON, P. GIRAUDOUX Room: VICTOR LYON SY12/04-01: SPATIAL AND TEMPORAL MODELLING FOR TRANSMISSION STUDIES AND RISK ASSESSMENT F.M. Danson SY12/04-02: PREDICTIVE MAPPING OF HOST ASSEMBLAGES: AN EXAMPLE WITH SMALL MAMMALS IN WESTERN CHINA - A. Vaniscotte SY12/04-03: SPATIO-TEMPORAL VARIATION IN ECHINOCOCCUS MULTILOCULARIS INFECTIONS IN TIBETAN DOGS - D. Pleydell SY12/04-04: ECHINOCOCCUS MULTILOCULARIS, AN EMERGING PARASITE IN THE MOST WESTERN EUROPEAN BORDER LINE - J. Van Der Giessen SY12/04-05: IMPROVING A SPATIAL MODEL FOR ECHINOCOCCUS MULTILOCULARIS TRANSMISSION STUDY - T. Quintaine SY12/04-06: REMOTE SENSING, LANDSCAPE EPIDEMIOLOGY AND RISK ASSESSMENT IN THE ONCHOCERCIASIS AMAZONIAN FOCUS OF SOUTHERN VENEZUELA - C. Botto Abella SY 17 Parasites, pregnancy & congenital infections / 01- Congenital toxoplasmosis: models & epidemiology Chair: P. FLORI, H. PELLOUX Room: GULBENKIAN SY17/01-01: KINETICS OF TISSUE PARASITE LOAD AND SWITCH AFTER ORAL INFECTION OF MICE WITH TOXOPLASMA GONDII - G. Chene SY17/01-02: CONSEQUENCES OF TOXOPLASMA GONDII INFECTION ON REPRODUCTIVE PARAMETERS OF MALE RATS EXPERIMENTALLY INFECTED BY THE PARASITE - K. Terpsides SY17/01-03: REAL-TIME QUANTITATIVE PCR STUDY OF TOXOPLASMA GONDII BURDEN IN DIFFERENT TISSUES IN A MURINE MODEL OF CHRONIC TOXOPLASMOSIS - L. Halos SY17/01-04: LARGE SCALE STUDY OF THE SEROPREVALENCE OF TOXOPLASMA GONDII IN OVINE MEAT FOR HUMAN CONSUMPTION IN FRANCE - L. Halos Discussion Satellite symposium: EMERGING DISEASES IN A CHANGING EUROPEAN ENVIRONMENT (1) Organizer: R. LANCELOT Room: ADENAUER
The EDEN project – Emerging diseases in a changing European environment – is an integrated project funded under FP6 for a period of five years (2004-2009). This large research project aims at understanding and modelling the risk of occurrence, establishment and spread of human, vector-borne diseases in European ecosystems, in a context of climate and other global and local changes. The research consortium includes more than 80 scientific teams from 49 consortium partners, and from 24 countries in Europe, the Middle East and Africa. The purpose of this symposium is to present and discuss some of the methods developed and results obtained by EDEN teams during the past three years. In addition, a group of 24 EDEN and associated partners have undertaken recently an expertise on the assessment of magnitude and importance of vector-borne diseases in Europe, for the European Centre for Disease Prevention and Control (ECDC, Stockholm). An original risk assessment method was developed for this purpose. This approach will also be presented and illustrated by some key results.
- EDEN: European ecosystems and emergence of vector-borne diseases: general presentation, integrated approach and selected results - R. Lancelot - Environmental changes and emerging, vector-borne diseases in Europe: methodological advances; from statistical to biological models - D.J. Rogers - Modelling the risk of installation of vector-borne diseases in Europe: methodological advances to account for changes in environment in risk models - N. Hartemink
12:30-14:00
LUNCH
14:00-15:30
SY 01 Apicomplexa / 05- New therapeutic targets Chair: M.A. HAKIMI, J. SCHREVEL Room: CAMBODGE SY01/05-01: DISCOVERY OF NEW TARGETS FOR ANTIMALARIAL CHEMOTHERAPY - P. Grellier SY01/05-02: NEOSPORA CANINUM AND NEOSPOROSIS: NOVEL TARGETS FOR PREVENTION OF INFECTION AND TREATMENT OF DISEASE THROUGH MOLECULAR STUDIES ON HOST-PARASITE INTERACTIONS - A. Hemphill SY01/05-03: IN VITRO GROWTH INHIBITION OF PLASMODIUM FALCIPARUM BY CATESTATIN, AN ANTIBACTERIAL CHROMOGRANIN A-DERIVED PEPTIDE - A. Akaddar
14:00-14:30 14:30-14:45 14:45-15:00
20
15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:15 14:15-14:30 14:30-14:45 14:45-15:00 15:00-15:30 14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:15 14:15-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30
SY01/05-04: MASLINIC ACID: SERIN PROTEASE INHIBITOR THAT BLOCKS SERIN PROTEASE ACTIVITY, GLIDING MOTILITY, CELL INVASION, AND SECRETORY PATHWAY OF TACHYZOITES OF TOXOPLASMA GONDII - L.M. De Pablos Torró Discussion SY 15 Innovative Prevention & Therapeutic Strategies / 03- Antiprotozoan drug strategies Chair: S. CROFT, P. LOISEAU Room: VICTOR LYON SY15/03-01: KINETOPLASTIDA: NEW THERAPEUTIC STRATEGIES - S. Croft SY15/03-02: MECHANISM OF ACTION OF MILTEFOSINE IN LEISHMANIA DONOVANI: MEMBRANE AFFINITY AND EFFECT ON PHOSPHOLIPID METABOLISM - P. Loiseau SY15/03-03: GLIBENCLAMIDE-RESPONSIVE ABC TRANSPORTERS IN LEISHMANIA: THEIR ROLE IN GLUCANTIME DISPOSITION AND ACTIVITY - A. Ponte-Sucre SY15/03-04: CHEMOSENSITIVITY TO ANTIMONY OF LEISHMANIA INFANTUM ISOLATED FROM DOGS AND SANDFLIES IN THE REGION OF MONTPELLIER (SOUTH OF FRANCE) - B. Oury SY15/03-05: BACTERIA VERSUS PARASITES: LACTOBACILLUS RHAMNOSUS CELL-FREE SUPERNATANT INHIBIT THE SPORULATION OF EIMERIA OOCYSTS IN VITRO - A.L. Molan SY 16 Tropical clinical parasitology Chair: P. BUFFET, J.P. DEDET Room: DAVID-WEILL SY16/01-01: THE RELATIONSHIP BETWEEN ASCARIS LUMBRICOIDES AND MALARIA IN CHILDREN AGED 1-4 YEARS - P. Kirwan SY16/01-02: ONCHOCERCIASIS AS A SIGNIFICANT CAUSE OF EPILEPSY: EVIDENCE FROM MODELLING THE RELATIONSHIP BETWEEN ONCHOCERCIASIS AND EPILEPSY IN 7 AFRICAN COUNTRIES - S. Pion SY16/01-03: ELEPHANTIASIS OF NON-LYMPHATIC FILARIASIS ORIGIN (PODOCONIOSIS) IN THE HIGHLAND AREA OF NORTH WEST CAMEROON - S. Wanji SY16/01-04: EPIDEMIOLOGICAL FEATURES OF CUTANEOUS LEISHMANIASIS FOCI OF AFRICA, NEAR AND MIDDLE EAST, BASED ON THE ISOENZYMATIC ANALYSIS OF 1,048 STRAINS - J.P. Dedet Discussion SY 17 Parasites, pregnancy & congenital infections / 02- Human congenital toxoplasmosis Chair: C. COSTACHE, H. PELLOUX Room: NATHAN SY17/02-01: CONTRIBUTION OF IMMUNOBLOTTING, REAL TIME PCR AND GOLDMANN-WITMER COEFFICIENT TO THE DIAGNOSIS OF ATYPICAL TOXOPLASMIC RETINOCHOROIDITIS - H. Talabani SY17/02-02: THE IMPORTANCE OF ANTITOXOPLASMIC IGA DETECTION IN NEWBORNS - C. Costache SY17/02-03: SEROLOGICAL STUDY FOR TOXOPLASMA GONDII INFECTION IN WOMEN WITH SPONTANEOUS ABORTION IN SOUTHWEST OF IRAN - K. Manouchehri Naeini SY17/02-04: DETECTION OF TOXOPLASMA GONDII ANTIGEN IN CEREBROSPINAL FLUID OF HYDROCEPHALIC CHILDREN BY WESTERN BLOT - L. Galal Discussion SY 18 Parasites of veterinary importance / 01- Trematoda Chair: M. FRANC, B. LOSSON Room: HONNORAT SY18/01-01: THE EFFECT OF TRICLABENDAZOLE SULPHOXIDE ON EGG PRODUCTION AND HATCH RATE IN THE LIVER FLUKE, FASCIOLA HEPATICA - L. Shaw SY18/01-02: ADULT TRICLABENDAZOLE-RESISTANT FASCIOLA HEPATICA: MORPHOLOGICAL RESPONSES TO IN VIVO TREATMENT WITH ARTEMETHER IN THE RAT MODEL - J. O'Neill SY18/01-03: TRANS-TEGUMENTAL OR ORAL UPTAKE OF TRICLABENDAZOLE: WHICH ROUTE PREDOMINATES IN THE LIVER FLUKE? - E. Toner SY18/01-04: SEM OBSERVATIONS ON THE IMPACT OF METABOLIC INHIBITORS ON THE DRUG SENSITIVITY OF A TRICLABENDAZOLE (TCBZ)-RESISTANT ISOLATE OF FASCIOLA HEPATICA - C. Devine SY18/01-05: TEM OBSERVATIONS ON THE IMPACT OF METABOLIC INHIBITORS ON THE DRUG SENSITIVITY OF A TRICLABENDAZOLE (TCBZ)-RESISTANT ISOLATE OF FASCIOLA HEPATICA - G. Brennan SY18/01-06: HELMINTH FAUNA OF BLACK COOT FULICA ATRA IN PAKISTAN - N.A. Birmani SY 21 3rd KE Mott symposium on Flukes & Schistosomes / 01- Fasciolasis: Epidemiology, treatment & control Chair: I. FAIRWEATHER, S. MAS-COMA Room: GULBENKIAN SY21/01-01: THE WORLDWIDE INITIATIVE AGAINST HUMAN FASCIOLIASIS - S. Mas-Coma SY21/01-02: RELATIVE ACTIVITY OF TRICLABENDAZOLE METABOLITES AGAINST THE LIVER FLUKE, FASCIOLA HEPATICA - L. Halferty SY21/01-03: THE ROLE OF DRUG EFFLUX PUMPS IN THE DEVELOPMENT OF RESISTANCE TO TRICLABENDAZOLE IN FASCIOLA HEPATICA - I. Fairweather SY21/01-04: THE USE OF COMMUNITY WASHING UNITS TO REDUCE TRANSMISSION OF LIVER FLUKES AND OTHER WATER BORNE PARASITES IN EGYPT - F. Curtale Discussion
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14:00-15:30
Satellite symposium: EMERGING DISEASES IN A CHANGING EUROPEAN ENVIRONMENT (2) Organizer: R. LANCELOT Room: ADENAUER - Assessment of magnitude and importance of vector-borne diseases in Europe: general presentation, method and main results of the risk assessment - G. Hendrickx, L. Vial
15:30-16:00
COFFEE BREAK
16:00-17:30
SY 04 Intestinal, urinary & liver parasites / 06- Liver parasites Chair: G. DREYFUSS, L. FAVENNEC Room: CAMBODGE SY04/06-01: BIODIVERSITY OF FLUKES - G. Dreyfuss SY04/06-02: SPATIAL DISTRIBUTION OF CYSTIC ECHINOCOCCOSIS OF HUMAN AND ANIMALS IN IRAN: A SYSTEMATIC REVIEW - M. Fasihi Harandi SY04/06-03: SITUATION OF HYDATID CYST INFECTION DURING LAST TWO DECADES (1985-2005) IN IRAN (REVIEW OF ARTICLES) - H. Yousofidarani SY04/06-04: HEPATIC AND PULMONARY CYSTIC ECHINOCOCCOSIS (CE) WITH GENOTYPING CHARACTERIZATION IN A PATIENT FROM CENTRAL AFRICAN REPUBLIC - A. Angoulvant SY04/06-05: TRICLABENDAZOLE-RESISTANCE IN THE LIVER FLUKE, FASCIOLA HEPATICA: SCANNING CANDIDATE GENES FOR MUTATIONS AND MRNA-EXPRESSION LEVELS - M. Fuchs
16:00-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30 16:00-17:30 16:00-16:15 16:15-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30 16:00-17:30 16:00-16:20 16:20-16:40 16:40-16:53 16:53-17:06 17:06-17:19 17:19-17:32 16:00-17:30 16:00-16:30 16:30-17:00 17:00-17:30 16:00-17:30 16:00-16:15 16:15-16:30 16:30-16:45 16:45-17:00 17:00-17:15 17:15-17:30
SY 06 Systematics, Evolution & Phylogeny of parasites / 05- Lophotrochozoa: Session 3 Chair: T. LITTLEWOOD, J. MARIAUX Room: DAVID-WEILL SY06/05-01: FASCIOLA SYSTEMATICS REVISITED: IS THE HEPATICA/GIGANTICA DIVIDE TOO CONSERVATIVE! - S. Walker SY06/05-02: EUPARAGONIMUS GONDWANENSIS N.SP. AND PARAGONIMUS KERBERTI N. SP. DIGENEA, PARAGONIMIDAE, IN CAMEROON - C. Bayssade-Dufour SY06/05-03: EVOLUTIONARY RESPONSES THAT ENHANCE DIGENEAN TRANSMISSION IN A VARIETY OF SPECIFIC ECOSYSTEMS - K. Galaktionov SY06/05-04: EVIDENCE OF INCOMPLETE EVOLUTIONARY DIVERGENCY OF ECHINOCOCCUS GRANULOSUS AND E. ORTLEPPI - D.A.S. Graichen SY06/05-05: RAILLIETINA (FUHRMANN, 1920) BY MOVSESSIAN, 2003: CENTRES OF ORIGIN AND CHARACTERISTICS OF HOST DISTRIBUTION - S. Movsessian Discussion SY 10 Parasites of fishes, other ectothermic vertebrates & marine mammals / 05- Interactions between parasites & pollutants in the aquatic environment Chair: R.A. KHAN, B. SAYYAF DEZFULI Room: ADENAUER SY10/05-01: ENVIRONMENTAL PARASITOLOGY: INTERACTIONS BETWEEN PARASITES AND POLLUTANTS IN THE AQUATIC ENVIRONMENT - B. Sures SY10/05-02: THE INFLAMMATORY RESPONSE OF FISH TO HELMINTH PARASITES - B. Sayyaf Dezfuli SY10/05-03: CHANGES IN GENE EXPRESSION IN EUROPEAN EELS (ANGUILLA ANGUILLA) INDUCED BY INFECTION WITH SWIMBLADDER NEMATODES (ANGUILLICOLA CRASSUS) - G. Fazio SY10/05-04: HEAVY METALS IN THE TAPEWORM CARYOPHYLLAEUS LATICEPS (PALLAS 1781) AND ITS HOST THE NASE CHONDROSTOMA NASUS (L. 1758) FROM TWO AUSTRIAN RIVERS: BIOINDICATIVE ASPECTS - F. Jirsa SY10/05-05: PARASITES, HEAVY METALS AND SANITARY RISK BOUND TO THE CONSUMPTION OF A PRIMITIVE FISH (PROTOPTERUS ANNECTENS) IN IVORY COAST - M. Cisse SY10/05-06: INFLUENCE OF SEDIMENT CONTAMINATED WITH UNTREATED PULP AND PAPER EFFLUENT ON WINTER FOUNDER, PLEURONECTES AMERICANUS: DOSE RESPONSE AND GRADIENT STUDIES - R.A. Khan SY 14 Ethical & epistemological issues in Parasitology / 01- Epistemological issues along of Parasitology History Chair: F. DELAPORTE, V. NUROCK Room: VICTOR LYON SY14/01-01: MANSON’S TRIPLE ERROR - F. Delaporte SY14/01-02: THE SCIENTIFIC RESEARCH PROGRAMMES OF LAKATOS AND APPLICATIONS IN PARASITOLOGY - G. Denegri, J. Cabaret SY14/01-03: ETHICS AND RESEARCH METHODS - S. Gazzola SY 18 Parasites of veterinary importance / 02- Cestoda - Nematoda of companion animal Chair: F. BEUGNET, A. DAKKAK Room: HONNORAT SY18/02-01: STUDY OF BACTERIAL INFECTION OF HYDATID CYSTS IN SLAUGHTERED ANIMALS OF WEST OF IRAN AND, BACTERIAL EXOTOXIN'S EFFECT ON CYST STERILIZATION - M. Fallah SY18/02-02: MOLECULAR CHARACTERIZATION OF ECHINOCOCCUS GRANULOSUS IN TUNISIA AND MAURITANIA BY MITOCHONDRIAL RRNS GENE SEQUENCING - M. Busi SY18/02-03: UPDATING ON CYSTIC ECHINOCOCCOSIS IN NORTHERN ITALY - G. Poglayen SY18/02-04: CANINE HEARTWORM DISEASE: A RETROSPECTIVE STUDY OF 98 CASES - C.M. Cretu SY18/02-05: THE DAILY EGG PRODUCTION OF ANCYLOSTOMA CANINUM AND THE DISTRIBUTION OF THE WORM ALONG THE DIGESTIVE TRACT OF THE DOG - O. Sowemimo SY18/02-06: THE PREVALENCE AND INTENSITY OF GASTROINTESTINAL PARASITES OF DOG AND THEIR POTENTIAL PUBLIC HEALTH SIGNIFICANCE IN ILE-IFE, NIGERIA - O. Sowemimo
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16:00-17:30
Satellite symposium: IMMUNOMODULATION BY HELMINTHS Organizer: W. HARNETT Room: GULBENKIAN - Macrophage activation by helminth parasites - J. Allen - Secreted products from H. polygyrus which induce regulatory T cells - R. Maizels - Heligmosomoides polygyrus antigens inhibit lymphocyte apoptosis provoked by dexamethasone - K. Donskow-Schmelter - The relationship between Ascaris lumbricoides and malaria in children aged 1-4 years - P. Kirwan - Chronic parasite infection has beneficial effects on the course of experimental autoimmune encephalomyelitis - A. Gruden-Movsesijan - Anti-inflammatory activity of the filarial nematode immunomodulator, ES-62 - W. Harnett
16:00-17:30
Satellite symposium: ROUND TABLE GIARDIA Organizer: M.G. ORTEGA-PIERRES Room: NATHAN - Impact of variation in Giardia on molecular typing, taxonomy and molecular epidemiology - A. Thompson, S. Caccio - Genes and novel proteins related to drug resistance in Giardia - N. Mueller, M.G. Ortega-Pierres
17:30-19:00
WFP Meeting
Room: GULBENKIAN
17:30-19:00
POSTER SESSION #2
Room: ADENAUER
17:30-19:30
YSA Selection
Room: DAVID-WEILL
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Thursday 28th August, 2008 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:42 09:42-09:54 09:54-10:06 10:06-10:18 10:18-10:30 09:00-10:30 09:00-09:15 09:15-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:15 09:15-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30 09:00-10:30 09:00-09:30 09:30-09:45 09:45-10:00 10:00-10:15 10:15-10:30
SY 04 Intestinal, urinary & liver parasites / 07- Intestinal parasites & soil transmitted helminths Chair: K. BREHM, G. GARGALA Room: DAVID-WEILL SY04/07-01: THE INFLUENCE OF HOST HORMONES AND CYTOKINES ON ECHINOCOCCUS MULTILOCULARIS SIGNALLING AND DEVELOPMENT - K. Brehm SY04/07-02: SENSITIVITY OF HOST RESILIENCE AND RESISTANCE TO HELIGMOSOMOIDES BAKERI INFECTION PRESSURE AND PROTEIN NUTRITION - J. Coltherd SY04/07-03: EXPLOITING THE ASCARIS SUUM MOUSE MODEL TO IDENTIFY THE MECHANISTIC BASIS OF RESISTANCE/SUSCEPTIBILITY - C. Dold SY04/07-04: GLUCOSAMINE: FRUCTOSE-6-PHOSPHATE AMINOTRANSFERASE (ETGFAT) IDENTIFIED AS A FEMALE-SPECIFIC ENZYME IN EIMERIA TENELLA: IMPLICATIONS IN OOCYST WALL FORMATION - R. Walker SY04/07-05: WHY ARE SCHISTOSOMES MONOGAMOUS? - S. Beltran SY 12 Innovative Epidemiology / 05- Evaluating host and parasite densities & population parameters in transmission systems Chair: M. GILBERT, D. HEINZMANN Room: VICTOR LYON SY12/05-01: EVALUATING PARASITE DENSITIES AND ESTIMATION OF PARAMETERS IN TRANSMISSION SYSTEMS – D. Heinzmann SY12/05-02: PREY DENSITY, PREDATOR-PREY INTERACTION AND CESTODE TRANSMISSION: ECHINOCOCCUS MULTILOCULARIS AS A STUDY CASE - F. Raoul SY12/05-03: A QUANTITATIVE EVALUATION OF ECHINOPARYPHIUM ACONIATUM TREMATODA PARASITE POPULATION IN CHANY LAKE ECOSYSTEM, WESTERN SIBERIA, RUSSIA - N. Yurlova SY12/05-04: HUMAN AND ANIMAL SEROPREVALENCE OF TOXOPLASMA GONDII IN GABON: COMPARISON BETWEEN A RURAL AND A URBAN AREA - A. Mercier SY12/05-05: A VEGETATION INDEX QUALIFYING PASTURE EDGES IS RELATED TO TICK DENSITY AND REFLECTS THE RISK OF HIGH BABESIA DIVERGENS SEROPREVALENCE IN DAIRY CATTLE HERDS - A. Agoulon SY12/05-06: FLEATICKRISK: A CLIMATIC MODEL DEVELOPED TO MONITOR AND PREDICT THE ACTIVITY AND THE DENSITY OF 3 TICKS SPECIES AND THE CAT FLEA IN FRANCE - F. Beugnet SY 18 Parasites of veterinary importance / 03- Nematoda Chair: V. COZMA, M. FRANC Room: HONNORAT SY18/03-01: STUDY (PREVALENCE, PATHOGENICITY, FAUNA) OF INTESTINAL HELMINTHES OF CATTLES IN NORTH OF IRAN COAST OF CASPIAN SEA - A. Halajian SY18/03-02: PARASITE DISEASE RESISTANCE: CAN WE EXPLOIT OVINE BREEDS FOR PARASITE PROTECTIVE GENES? - D. Piedrafita SY18/03-03: SURVEY ON THE CLINICAOPATHOLOGICAL CHANGES IN EXPERIMENTAL INFESTATION TO HAEMONCUS CONTORTUS IN SHEEP - M. Zaeemi SY18/03-04: POPULATION DYNAMICS OF TRICHURIS SUIS IN TRICKLE INFECTED PIGS - P. Nejsum SY18/03-05: THE DYNAMICS OF GENETICALLY MARKED ROUNDWORM INFECTIONS IN PIGS - P. Nejsum SY18/03-06: MACRO- AND MICRO-PARASITES OF THE RUMINANT ABOMASUM - I. Scott SY 20 Non tropical clinical parasitology / 01- Trichinellosis Chair: F. BOLAS-FERNANDEZ, I. VALLEE Room: CAMBODGE SY20/01-01: TRICHINELLA OF WILD ANIMALS IN UKRAINE - J. Didyk SY20/01-02: FIRST CASE OF TRICHINELLA PSEUDOSPIRALIS IN A WILD BOAR IN BULGARIA - I. Bankov SY20/01-03: GEOGRAPHICAL VARIABILITY WITHIN TRICHINELLA GENUS: STUDY CASE OF A HIGH ENDEMIC AREA - R. Blaga SY20/01-04: THE USEFULNESS OF AN ELISA TEST FOR THE SEROLOGICAL DIAGNOSTICS OF TRICHINELLA INFECTION IN DOMESTIC PIGS AND WILD BOARS - J. Bien SY20/01-05: EARLY TRICHINELLA ANTIGENS FOR THE DESIGN OF AN INDIRECT ELISA TEST - S. Lacour SY20/01-06: IMMUNOPROTEOMIC ANALYSIS OF SERA FROM PATIENTS SUFFERING TRICHINELLOSIS AND TREATED WITH ALBENDAZOLE - F. Bolas-Fernandez SY 21 3rd KE Mott symposium on Flukes & Schistosomes / 02- Schistosomes: Epidemiology & control Chair: D. ROLLINSON, A. THERON Room: ADENAUER SY21/02-01: PIGA VITA KICHOCHO: CONTROL OF SCHISTOSOMIASIS AND SOIL-TRANSMITTED HELMINTHIASIS ON UNGUJA ISLAND, (ZANZIBAR) SINCE 2003 - R. Stothard SY21/02-02: EXTRAVASATION OF SCHISTOSOME EGGS: AN OBSERVATION AN INTERPRETATION - E. Linder SY21/02-03: SCHISTOSOMA MANSONI AND S. HAEMATOBIUM CO-INFECTION IN NDER, SENEGAL; TRANSMISSION DYNAMICS AND GENETIC EPIDEMIOLOGY - B. Webster SY21/02-04: HOST CHOICE AND PENETRATION OF SCHISTOSOMA HAEMATOBIUM MIRACIDIA - F. Allan SY21/02-05: COMMERCIAL WESTERN BLOT KIT FOR THE DETECTION OF ANTI-SCHISTOSOMA ANTIBODIES: ADVANTAGES AND DISADVANTAGES - C. Thors
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09:00-10:30
Satellite symposium: REPRODUCTIVE AND DEVELOPMENTAL BIOLOGY OF PARASITES Organizer: Z. SWIDERSKI Room: GULBENKIAN - Importance of TEM data on cestode eggs for diagnostics and disease dissemination: Comparative TEM studies - D.B. Conn - Comparative TEM studies on the infective oncospheres of seven selected species of cyclophyllidean cestodes of medical and vetereinary importance - Z. Swiderski - Ultrastructural characters of spermiogenesis and the spermatozoon in caryophyllidean and trypanorhynch cestodes: phylogenrtical implications - Z. Swiderski - Morphological "junction" between Trichinella spiralis larva and mouse skeletal muscle cell - J. Dabrowska - Ultrastructural characteristics of particular developmental stages of the bird schistosome Trichobilharzia regent - J. Bulantova
09:00-10:30
Satellite symposium: TEACHING PARASITOLOGY Organizer: F. BRUSCHI
Room: NATHAN
During this round table, the results of a questionnaire distributed at the beginning of the conference will be presented and discussed, furthermore proposals will be presented to increase the appeal of Parasitology not only for students, but also, even more important, for colleagues.
10:30-11:00
COFFEE BREAK
11:00-12:30
SY 14 Ethical & epistemological issues in Parasitology / 02- Ethical issues in Parasitology, Teaching parasitology Chair: J. CABARET, L. DELHAES Room: NATHAN SY14/02-01: CURRENT BIOETHICAL ISSUES IN PARASITOLOGY - D. Boury SY14/02-02: RESEARCH ETHICS AND DECISION-MAKING PROCESS - L. Leveque SY14/02-03: HOW PARASITOLOGY IS TAUGHT IN MEDICAL FACULTIES IN EUROPE? PARASITOLOGY LOST? - F. Bruschi
11:00-11:30 11:30-12:00 12:00-12:30 11:00-12:30 11:00-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:15 11:15-11:30 11:30-12:00 12:00-12:15 12:15-12:30 11:00-12:30 11:00-11:15 11:15-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30
SY 15 Innovative Prevention & Therapeutic Strategies / 04- Models & drug design Chair: M. BOUSSINESQ, J. LE BRAS Room: VICTOR LYON SY15/04-01: ONCHOCERCIASIS CONTROL: BIOLOGICAL RESEARCH IS STILL NEEDED - M. Boussinesq SY15/04-02: DYNAMICS OF ONCHOCERCA VOLVULUS MICROFILARIAL LOADS AFTER IVERMECTIN TREATMENT OF PATIENTS WITH EITHER HIGH OR LOW PREVIOUS DRUG PRESSURE - M. Boussinesq SY15/04-03: ANTIPLASMODIAL ACTIVITY ON BOTH SEXUAL AND ASEXUAL PLASMODIUM STAGES OF TRIOXAQUINES, NEW ANTIMALARIAL DRUGS - F. Benoit-Vical SY15/04-04: ANTIPLASMODIAL ACTIVITY OF GIROLLINE, A NEW LEAD STRUCTURE FOUND IN NATURAL PRODUCTS - P. Njomnang Soh SY15/04-05: ANALYSIS OF 34 SNPS ASSOCIATED TO P.FALCIPARUM RESISTANCE BY DNA-CHIP IN NIGER I. Maman Laminou SY 19 Wild life & parasitic fauna: epidemiological aspects / 01- Birds schistosomes Chair: H. FERTE, K. SKIRNISSON Room: GULBENKIAN SY19/01-01: IN VITRO TRANSFORMATION AND DEVELOPMENT OF BIRD SCHISTOSOMES - M. Chanova SY19/01-02: ULTRASTRUCTURAL CHARACTERISTICS OF PARTICULAR DEVELOPMENTAL STAGES OF THE BIRD SCHISTOSOME TRICHOBILHARZIA REGENTI - J. Bulantova SY19/01-03: CERCARIAL DERMATITIS AND BIRD SCHISTOSOMES IN EUROPE: THE PRESENT STATE - L. Kolarova SY19/01-04: SWIMMER'S ITCH AND THE OCCURRENCE OF BIRD SCHISTOSOMES IN ICELAND - K. Skirnisson SY19/01-05: AVIAN SCHISTOSOMES IN FRENCH AQUATIC BIRDS: MOLECULAR APPROACH - D. Jouet SY 20 Non tropical clinical parasitology / 02- Echinococcosis Chair: A. HEMPHILL, A. PAUGAM Room: CAMBODGE SY20/02-01: MOLECULAR CHARACTERIZATION OF ECHINOCOCCUS GRANULOSUS STRAINS IN PORTUGAL S. Beato SY20/02-02: CYSTIC ECHINOCOCCOSIS IN ROMANIA: AN ANALYSIS OF 13 YEARS OF EXPERIENCE - C.M. Cretu SY20/02-03: HUMAN HYDATIDOSIS: GENOTYPING, SEROLOGY AND MORPHOLOGICAL DATA ON FIFTEEN CYSTS - S. Gabrielli SY20/02-04: CONSEQUENCES OF LONG CHEMOPREVENTION WITH ALBENDAZOLE IN EXPERIMENTAL HYDATID INFECTION IN SHEEP - I.L. Mitrea SY20/02-05: IN VITRO AND IN VIVO TREATMENT OF ECHINOCOCCUS METACESTODES WITH ARTEMISININ DERIVATIVES ARTESUNATE AND DIHYDRO-ARTEMISININ - A. Hemphill SY20/02-06: CHEMOTHERAPY OF HYDATIDOSIS WITH ALBENDAZOLE, COMBINED WITH AN IMMUNOSTIMULATOR - ISOPRINOSIN - D. Vutchev
25
11:00-12:30 11:00-11:15 11:15-11:30 11:30-11:45 11:45-12:00 12:00-12:15 12:15-12:30
SY 21 3rd KE Mott symposium on Flukes & Schistosomes / 03- Immunity, diagnosis & hostparasite interactions Chair: F. MUTAPI, D. ROLLINSON Room: ADENAUER SY21/03-01: CELL SIGNALLING IN SNAIL-SCHISTOSOME INTERACTIONS: RECIPROCAL INTERPLAY REVEALED THROUGH ANALYSIS OF KINASE PATHWAYS - A. Walker SY21/03-02: EXPLORING THE EFFECTS OF SCHISTOSOMA MANSONI EXCRETORY-SECRETORY PRODUCTS ON GENE EXPRESSION IN BIOMPHALARIA GLABRATA HAEMOCYTES, USING A CDNA MICROARRAY - Z. Zahoor SY21/03-03: WHY DOES IMMUNITY TO SCHISTOSOMES TAKE SO LONG TO DEVELOP: THE THRESHOLD HYPOTHESIS - F. Mutapi SY21/03-04: INVESTIGATING THE CAUSES OF THE SLOW DEVELOPMENT OF HUMAN ANTI-SCHISTOSOME IMMUNITY - K. Mitchell SY21/03-05: GLYCAN-INDUCED IMMUNOPATHOLOGY IN SCHISTOSOMIASIS - M. Meevissen SY21/03-06: ROLE OF PLASMIN IN SCHISTOSOME PERIOVAL EXTRACELLULAR MATRIX DEGRADATION AND EGG EXCRETION? - E. Linder
11:00-12:30
Satellite symposium: CANINE LEISHMANIASIS Organizer: A. DAKKAK Room: DAVID-WEILL - General introduction - A. Dakkak - Canine, feline and human leishmaniasis relationships : Situation in Iran - G.R. Hatam - Canine and human leishmaniasis relationships: Situation in Tunisia - K. Aoun - Risk factors of (re)emergenge - A. Dakkak - In vitro diagnosis with kits - C. Bordier - Difficulties in the management & treatment - A. Dakkak - Waiting for the vaccine: canine leishmaniasis and Scalibor - R. Killick-Kendrick - Vaccines - A. Dakkak
11:00-12:30
Satellite symposium: EUROPEAN SUPPORT TO RESEARCH IN PARASITOLOGY Organizer: P. LOISEAU Room: HONNORAT - General EC funding mechanisms - O. Olesen - Future activities of EC in Parasitology - O. Olesen - Med Vet Net a European Network of Excellence on Zoonoses Research - A. Jestin - From an EU research project toward an Integrated Project: example of the Trichinella network - P. Boireau - European Collaborative projects, the vision of a consultancy company specialised in projects setting-up and management - C. Germain
12:30-14:00
LUNCH
14:00-15:30
SY 18 Parasites of veterinary importance / 04- Protozoa Chair: B. GOTTSTEIN, S. SOTIRAKI Room: HONNORAT SY18/04-01: SEROPREVALENCE OF TOXOPLASMA GONDII INFECTION IN SHEEP AND GOATS FROM TRANSYLVANIA (ROMANIA) - A. Titilincu SY18/04-02: EPIDEMIOLOGICAL STUDIES OF AN EMERGING BESNOITIOSIS EPIDEMIC IN FRENCH ALPS - L. Zenner SY18/04-03: NCGRA2 AS A MOLECULAR TARGET TO ASSESS THE PARASITICIDAL ACTIVITY OF TOLTRAZURIL AGAINST NEOSPORA CANINUM - B. Gottstein SY18/04-04: STUDY ON THE EFFECT OF TOLTRAZURIL-BASED PREVENTION ON PIGLETS COCCIDIOSIS - S. Sotiraki SY18/04-05: PREVALENCE AND ECONOMIC IMPACT OF TROPICAL THEILERIOSIS ON HEALTH AND THE BOVINE PRODUCTION - H. Ziam SY18/04-06: DETECTION OF TRITRICHOMONAS FOETUS INFECTIONS IN SWISS CATS BY CULTURE SYSTEM AND A NOVEL PCR - B. Gottstein SY18/04-07: FACTORS INFLUENCING CLINICAL EXPRESSION OF HISTOMONOSIS IN TURKEY - M.P. CallaitCardinal Discussion
14:00-14:12 14:12-14:24 14:24-14:36 14:36-14:48 14:48-15:00 15:00-15:12 15:12-15:24 15:24-15:30 14:00-15:30 14:00-14:15 14:15-14:30 14:30-14:45 14:45-15:00 15:00-15:15
SY 19 Wild life & parasitic fauna: epidemiological aspects / 02- Other parasites of wildlife Chair: H. FERTE, K. SKIRNISSON Room: DAVID-WEILL SY19/02-01: INFECTION KINETICS AND PARASITE STRATEGY INFLUENCED BY HIBERNATION IN ALPINE MARMOTS - M.P. Callait-Cardinal SY19/02-02: BABESIA SPECIES INFECTING ROE DEER (CAPREOLUS CAPREOLUS). SEROPREVALENCE AND PCR DETECTION - S. Bastian SY19/02-03: A NOVEL TRYPANOSOMA SP AND ITS ROLE IN THE DECLINE OF A THREATENED SPECIES OF AUSTRALIAN MARSUPIAL, THE BRUSH-TAILED BETTONG (BETTONGIA PENICILLATA) - A. Thompson SY19/02-04: THE FIRST EVIDENCE OF NEOSPORA CANINUM INFECTION IN FARMED AND FREE LIVING DEER IN POLAND - K. Gozdzik SY19/02-05: THE DEVELOPMENT OF PLASMODIUM RELICTUM (LINEAGE P-SGS1) IN EXPERIMENTALLY INFECTED PASSERINE BIRDS - V. Palinauskas
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15:15-15:30
SY19/02-06: IDENTIFICATION OF THE TREMATODE ALARIA ALATA IN WILD BOAR IN EAST OF FRANCE - I. Vallee
14:00-15:30
SY 20 Non tropical clinical parasitology / 03- Toxocarosis, Taenia & Cystecercosis and others Chair: V. COZMA, C.P. RACCURT Room: CAMBODGE SY20/03-01: TOXOCARIASIS-RELATED SKIN DISORDERS: A CASE CONTROL STUDY USING IMMUNOBLOT A.P. Bellanger SY20/03-02: OCULAR TOXOCARIASIS IN PERU: A REPORT OF 16 CASES - L. Getaz SY20/03-03: SEROPREVALENCE OF HUMAN TAENIA SOLIUM CYSTICERCOSIS IN HAITI - C.P. Raccurt SY20/03-04: FREQUENCY OF TAENIASIS IN HOSPITALIZED PATIENTS FROM WESTERN ROMANIA - A.M. Neghina SY20/03-05: PARASITIC DISEASES IN FEDERATION OF BOSNIA AND HERZEGOVINA - Z. Obradovic SY20/03-06: HUMAN DIROFILARIA REPENS INFECTION IN SERBIA - A. Dzamic
14:00-14:15 14:15-14:30 14:30-14:45 14:45-15:00 15:00-15:15 15:15-15:30 14:00-15:30 14:00-14:30 14:30-14:45 14:45-15:00 15:00-15:30 14:00-15:30
SY 21 3rd KE Mott symposium on Flukes & Schistosomes / 04- Genetics & biology of flukes and lymnaeid snails Chair: M.D. BARGUES, R. HOUIN Room: ADENAUER SY21/04-01: GENETICS AND PHYLOGEOGRAPHY OF FASCIOLA HEPATICA CAUSING HUMAN FASCIOLIASIS ENDEMIC AREAS IN THE AMERICAS - M.D. Bargues SY21/04-02: FASCIOLOSIS IN THE NETHERLANDS - A PHYLOGENETIC APPROACH - S. Walker SY21/04-03: DIVERSITY OF DNA SEQUENCES AND SPECIES IDENTIFICATION OF CERCARIAE OF BIRD SCHISTOSOMES OBTAINED FROM RUSSIAN AND BELARUSIAN WATER PONDS - S. Semyenova Discussion Satellite symposium: EUROPEAN CLUB OF ECTOPARASITOLOGISTS Organizer: A. IZRI Room: VICTOR LYON
During this symposium, Areski Izri will show how lice, scabies and all these small animals that poison people's life are problems at the European level. He will propose to organise a club and/or a network with all interested European ectoparasitologists with the aim to reinforce collaborative studies and informal exchanges.
14:00-15:30
Satellite symposium: DNA BARCODING: AN INTEGRATED TOOL IN PARASITE IDENTIFICATION Organizer: O. BAIN, M. CASIRAGHI, S. SAMEDI Room: NATHAN - DNA barcoding: a molecular tool for organisms identification - S. Samedi - Molecular tools in phylogenetic inference. DNA barcoding is something different - S. Williams - Identification of Parasites, a necessary but difficult task - O. Bain - DNA barcoding as a clinical approach in veterinarian context - C. Genchi - DNA barcoding: applications for the parasites identification - M. Casiraghi
14:00-15:00
Satellite symposium: LAST MINUTE SATELLITE
15:30-16:00
COFFEE BREAK
15:30-17:30
EFP Meeting of Society Delegates
Room: GULBENKIAN
16:15-17:30
“From Parasites to Cognac”
Room: HONNORAT
A place for last minute breaking news!!! Please contact Pr Jean Dupouy-Camet.
Room: GULBENKIAN
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You are invited to join us at this Special satellite symposium which will be held on Thursday 28 at 4.15 pm in room Honnorat
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After a short scientific introduction, producers will present us different Cognacs and explain us how to taste them…
Please register before 6 pm on Wednesday 27 at the Congress desk. A symbolic fee of 2 will be asked.
16:00-17:30
POSTER SESSION #2
Room: ADENAUER
17:30-19:00
CLOSING CEREMONY
Room: ADENAUER & HONNORAT
27
Concert in Saint-Jacques-du-Haut-Pas Church 252 rue St Jacques, 75005 Paris Tuesday 26th August, 2008 at 8:30pm
How to get there: From RER B station Cité Universitaire, take the train towards Gare du Nord. Exit at Luxembourg stop. Then follow rue Gay Lussac and turn right in rue St Jacques. Historical features: The ground on which the church was built belonged since 1180 to hospitallers coming from Alto Pascio close to Lucca (Italy). When Catherine de Médicis, wife of king Henri IV, decided in 1572 to install the Benedictines of Saint-Magloire, there were around only fields and meadows. In 1620, the seminar of Oratorians replaced the Benedictines. The increasing surrounding population practising their faith in the small convent chapel became rapidly an inconvenience for the Oratorians. The bishop gave them the authorisation to build a small church joined with the convent chapel. But, this small church became also quickly too small, and in 1630, Gaston d'Orleans, brother of the king Louis XIII, decided to carry out important works. At the same period, Angélique Arnaud, abbess of Port-Royal, hearth of the Jansenist movement, set up nearby an appendix of the monastery of Port-Royal des Champs. Very close links were tied with the parish of Saint-Jacques-du Haut-Pas which will play an important role in the diffusion of jansenism. The duchess of Longueville (1619-1679), sister of the Grand Condé and protective of Port-Royal offered substantial gifts for the construction of the building. Thanks to them, t work could thus begin again in 1675. The architect, Daniel Gittard, was selected to finish the building. This architect had already built the choir of Saint-Sulpice church, latter very well known thanks to the Da Vinci Code novel. On May 6, 1685, the principal work were completed. In the church nice paintings from the XVIIth to the XIXth century can be seen and particularly a masterpiece “L’Annonciation” painted by the famous Le Nain brothers. In the choir is buried Du Vergier de Haurane (1581-1643), known as the abbot of Saint-Cyran founder of the jansenism with Jansen. Jansenism, a branch of Catholic Gallican thought, emphasised original sin, human depravity, the necessity of divine grace, and predestination. The philosopher Blaise Pascal and tragedy writer Jean Racine were linked to this movement. The Concert: The concert will include works from the two last centuries : Bellon (3 quintets), Berio (Call from the St. Louis Fanfare), Boukhitine (Total Recall, tribute to Berio), Arnold (Brass Quintet N°.1); Bernstein (Dance Suite) Turbulence Quintet: In 1998, five students of the Higher National Academy of Lyon met in order to prepare the international contest of Narbonne. Finalist of the competition in 2000, the quintet, then anonymous, finds his identity in an press article which qualifies them the “turbulent ones”. They become then quite naturally: “Turbulences” and gain in 2001 the 1st Price of the contest Philip Jones. David GUERRIER: international concert trumpetist, solo French horn with the Orchestre National de France Guillaume JEHL: solo trumpet with the Orchestre Symphonique de Bâle (Switzerland) Antoine DREYFUSS: solo French horn with the Orchestre Philharmonique de Radio-France Antoine GANAYE: solo trombone with the Orchestre Philharmonique de Radio-France Arnaud BOUKHITINE: solo tuba with the Ensemble InterContemporain, composer Tickets are still available at the Welcome desk. 28
Xth European Multicolloquium Of Parasitology Paris - France August 24th-28th, 2008
Oral presentations
29
development of schizonts and the production of merozoites. SY01/01-02
SY01/02-02
INTERACTION BETWEEN MURINE DENDRITIC CELLS AND NEOSPORA CANINUM
LCCL DOMAIN PROTEINS : HOMOLOGS OF PLASMODIUM SEXUAL STAGES GENES IDENTIFIED IN BABESIA DIVERGENS
DION S., GUITON R., DIMIER-POISSON I. UMR 0483 Université-INRA, TOURS, FRANCE
BECKER C.(1), BISCHOFF E.(2), MALANDRIN L.(1), CHAUVIN A.(3), DAVID P.(2), BONNET S.(4) (1) INRA, NANTES, FRANCE ; (2) Institut Pasteur, PARIS, FRANCE ; (3) ENVN, NANTES, FRANCE ; (4) INRA, MAISONS ALFORT, FRANCE
The apicomplexa Neospora caninum has emerged as a major cause of reproductive failure in livestock worldwide since its discovery and represents a major economic problem. Few are known on the immune response induced by this parasite. Therefore, a better knowledge of the immunological mechanisms involved after a N. caninum infection would help to finalize an efficient vaccine strategy. Our work was to determine the ability of N. caninum to activate dendritic cells (DC), the major population cells implicated in immune response of Toxoplasma gondii, a close parasite. We use a dendritic cell line named SRDC established and characterized in our laboratory from murine splenic DCs with similar morphology, phenotype and activity to CD4-CD8+CD205+CD11b- DCs purified ex vivo. We test the ability of live N. caninum to infect SRDC, to follow up the phenotype changes and the IL-12 production when they interacted with live, killed or a total antigen of N. caninum. Results show that N. caninum infect DC but not altered the phenotype of them. We determine that only N. caninum antigen activates DC with an upregulation of the CD40 and CMHII surface molecules associated with a production of IL-12. This study demonstrates the potential role of DC in the host immune response after infection by N. caninum and will be purchased by evaluating the ability of DC to activate the lymphocyte population.
Babesia divergens, a protozoan parasite of bovine red blood cells, is the main agent of bovine babesiosis in Europe. Babesia development includes asexual and sexual multiplications in the vertebrate and arthropod hosts, respectively. Very few data are available on this Apicomplexa, especially concerning its life cycle inside its tick vector, Ixodes ricinus. To follow and study the parasite development inside the tick, our aim was to find markers of B. divergens vector stages. We used an in silico post-genomic approach, analysing genome, transcriptome and proteome of a close apicomplexan genus, Plasmodium, to select proteins that are specifically expressed during sexual stages inside the vector, and conserved within the phylum. Thanks to degenerated primers designed on sequence identity analysis including other Babesia spp. sequences, we amplified by PCR and RT-PCR, fragments of three B. divergens homologs. We finally identified and characterized three novel genes of B. divergens, encoding members of a highly conserved apicomplexan protein family, defined by the presence of LCCL domains. The role of these proteins, potentially implicated in adhesive functions, has now to be investigated in parasite-tick interactions during vector transmission.
SY01/01-03
COMPARATIVE DEVELOPMENT OF EIMERIA NINAKOHLYAKIMOVAE IN DIFFERENT IN VITRO CELL CULTURE SYSTEMS
SY01/02-03
ROLE OF APOPTOSIS IN THE TRANSMISSION OF PLASMODIUM FALCIPARUM.
RUIZ A.(1), BEHRENDT J.H.(2), ZAHNER H.(2), TAUBERT A.(2), HERMOSILLA C.(2), GUEDES A.(1), MUNOZ M.C.(1), MOLINA J.M.(1) (1) University of Las Palmas de Gran Canaria, Department of Animal Pathology, Parasitology Unit, LAS PALMAS, SPAIN ; (2) Justus Liebig University Giessen, Institute of Parasitology, GIESSEN, GERMANY
BEAVOGUI A.H., MESLIN B., PICOT S. Université Claude Bernard, LYON, FRANCE
Apoptosis is known to play a critical role in development of multicellular organisms and to be involved in the biology of several protozoan parasites including Plasmodium. Plasmodium falciparum is responsible for malaria severe cases in human. It has previously been demonstrated that its asexual stages exhibit features of apoptosis in vitro under drug pressure. Chloroquine could trigger parasite apoptosis with a significant difference observed between sensitive and resistant strains. Gametocytes, the parasites sexual forms, are the key stage for an effective transmission from human to mosquitoes. Only few antimalarial drugs (primaquine and artemisinin derivatives) are actives against these stages, but mechanisms are still unclear. Here, we report the first study of apoptosis induction by antimalarial drugs on gametocytes from reference strains of Plasmodium falciparum. Morphological features of apoptosis were observed in gametocytes from in vitro culture using fluorescence and microscopy: DNA fragmentation (TUNEL labelling), mitochondrial depolarization (JC1 labelling) and phosphatidylserines exposure (annexin V labelling). Understanding gametocyte apoptosis will help developing new strategies in the fight against malaria.
The infections induced by different species of the protozoa Eimeria, reported as coccidiosis in veterinary medicine, are considered among the parasitoses the most prevalent and spread within the goat production systems. These parasites, which, regardless of the management system, may affect up to 100% of goat kids between 4-10 weeks of age, represent one of the main reasons of economic losses for the goat industry all around the world. During the endogenous development, the coccidian parasites are subjected to a strong immune control which leads to a protective immunity after primary infection. This immunoprotection does not last for the whole life of the animal and is only restricted to the species responsible of this immunity. E. ninakohlyakimovae, one of the most pathogenic species, develops the first schizogony in endothelial cells of the intestinal lymph capillaries, which are considered as highly immunoreactive cells, able to produce a broad range of adhesion molecules, cytokines and proinflammatory chemokines upon activation. In the present study we have evaluated the development of E. ninakohlyakimovae in different in vitro cultures for further identification of the pathogenic mechanisms and the immune interaction between the parasite and the host cell. Comparative analyses have been performed by microscopic examination of the infection rate, the 30
been shown yet, these data support the idea that NET formation is involved in the innate immune response against coccidiosis in cattle. This is the first report indicating parasite induced NET formation.
SY01/02-04
MALARIA AND OBESITY IN MICE
DEPOIX D.(1), BOURGOUIN C.(2), LOMBARD M.N.(3), BACCAM D.(1), THOUVENOT C.(2), GRELLIER P.(1), ROBERT V.(4) (1) MNHN USM 504, PARIS, FRANCE ; (2) CEPIA, Institut Pasteur, PARIS, FRANCE ; (3) MNHN, PARIS, FRANCE ; (4) IRD UR 77 et UR 16; MNHN USM 504, PARIS, FRANCE
SY01/03-02
CAN VAR GENE MODULATION SERVE AS A NOVEL TARGET FOR MALARIA THERAPY?
Very little is known about a possible relation between obesity and responses to malarial infection. We have compared responses of the mutated obese ob/ob mouse (leptin deficient) and the lean ob/+ heterozygous control to a single iv. injection of Plasmodium berghei ANKA. Control mice developed the expected rapid neurological syndromes leading to death after 6 days in absence of high parasitaemia ; on the contrary, obese mice did not develop cerebral malaria and responded by an increasing parasitaemia (up to 40%) ending up with severe anaemia leading to death 18-25 days post injection (PI). Parasitaemia was higher in obese mice versus controls from day 3 to day 8 PI, with significant differences at day 3 (0.039% vs 0.017% in controls). Mature hepatic schizonts in obese mice appeared to be smaller in size (average : 1917 μm2 vs 3526 μm2 in controls) contrarily to measures previously reported on obese fa/fa rats injected with Plasmodium yoelii yoelii. Furthermore, in ob/ob mice, the first blood stages appeared delayed (53 h PI vs 49 h in controls). We therefore conclude that the ob/ob obese mouse : - is a permissive model to malarial infection - can open the way for new investigations on cerebral malaria - offers an in vivo model to study hepatic schizont development in lipid-enriched environment. A possible link between obesity and responses to malarial infection in humans is suggested.
POLLACK Y., ROSENBERG E., BEN-SHMUEL A., SHALEV O., SINAY R. Ben-Gurion University of the Negev, BEER-SHEVA, ISRAEL The Plasmodium falciparum heterogeneous var gene family encodes the PfEMP1 protein which is exported to the surface of the infected erythrocyte (IE) mediating the adherence of IEs to the endothelium of the host microvasculature. PfEMP1 also mediate antigenic variation through the constant change of the displayed PfEMP1 from the repertoire of var genes, effectively avoiding host immune system clearance. var gene regulation has been extensively studied although no cues have been found which influence var gene transcription. Here we demonstrate that biological factors act as an additional layer of var gene regulation, possibly affecting the widely proposed allelic exclusion mechanism. We show the existence of a differential expression pattern of these genes upon exposure to stress (oxidative stress and glucose deprivation) in relation to their positional placement on the chromosome. These forms of stress, which the parasite encounters throughout its complex intra-erythrocytic life-cycle, induce expression of centrally located var genes while repressing the sub-telomeric copies of the family, suggesting a stress-induced controlled modulation of var gene transcription orchestrated by the previously described histone deacetylase pfsir2 which is crucial for this phenomenon. Moreover, pfsir2 activation with resveratrol was shown to dramatically decrease var gene expression, thereby it may serve as a potential adjuvant therapy.
SY01/02-05
NET FORMATION AS IMMUNE REACTIONS BOVIS
PART OF INNATE AGAINST EIMERIA
SY01/03-03
IDIOSYNCRASY OF MITOCHONDRIAL TRANSLATION IN APICOMPLEXANS
BEHRENDT J.H.(1), RUIZ A.(2), ZAHNER H.(1), TAUBERT A.(1), HERMOSILLA C.(1) (1) Justus Liebig University Giessen, Institute of Parasitology, GIESSEN, GERMANY ; (2) University of Las Palmas de Gran Canaria, Department of Animal Pathology, Parasitology Unit, LAS PALMAS, SPAIN
PINO P.(1), SHEINER L.(1), CHARRIERE F.(2), SCHNEIDER A.(2), SOLDATI D.(1) (1) University of Geneva, GENEVA, SWITZERLAND ; (2) University of Berne, BERNE, SWITZERLAND In most Apicomplexans, protein synthesis occurs in three compartments, the cytosol, the mitochondrion, and the apicoplast. The extremely reduced mitochondrial genome of Apicomplexans codes for 3 ORFs but not for any components of the translation machinery. The complete absence of tRNA genes is compensated for by the import of the nuclear encoded cytosolic tRNAs. Moreover, these parasites possess an insufficient number of genes coding for the aminoacyl-tRNA synthetases (aaRSs) that are responsible for charging their cognate tRNAs with the appropriate amino acid. In Toxoplasma gondii 13 aaRSs are encoded by two genes, whereas the rest of them exist as single copy genes. Subcellular localization studies by epitope-tagging revealed that some aaRSs are uniquely targeted to the apicoplast or to the cytosol while few of them are likely shared between these two compartments by dual targeting. The absence of aaRSs in the mitochondrion suggests that charged aminoacylated tRNAs are imported to the organelle and used only once. To prevent accumulation of uncharged tRNAs in mitochondrion uncharged tRNAs are either degraded or exported to be re-charged in the cytosol.
Neutrophil Extracellular Traps (NETs) have recently been demonstrated to act as a killing mechanism of neutrophils against several pathogens, i.e. bacteria and fungi. Up to date, there has been no indication of NETs being involved in virus or parasite killing. In the present study, the interaction between bovine neutrophils and sporozoites of Eimeria bovis was investigated in vitro. SEM micrographs revealed thin fibres to appear when bovine neutrophils were co-cultivated with E. bovis sporozoites. Fluorescence microscopy, using the Sytox Orange nucleic acid stain, proved the emerging fibres to consist of DNA. Consequently, DNAse treatment abolished these structures indicating it being NET. Quantification of the Sytox Orange signal was used to estimate the extent of NET formation. DPI, an inhibitor of NADPH oxidase, previously described to inhibit NET formation, prevented the reaction as expected. As reported in other cases, serum did also interfere with NET formation. Extracellular fibres in close contact with sporozoites strongly suggest that NETs may immobilize E. bovis sporozoites and thereby prevent them from infecting host cells. Even if direct killing of E. bovis sporozoites by NETs has not 31
obtained from proteomic analysis of Neospora tachyzoites were used to refine the annotation. We also report on preliminary comparative genomic analysis with T. gondii, suggesting high synteny between the two genomes.
SY01/03-04
DRUG-INHIBITION OF HDAC ACTIVITY: EPIGENETIC CONTROL OF CELL CYCLE AND DIFFERENTIATION IN APICOMPLEXAN PARASITES
SY01/04-02
BOUGDOUR A.(1), MAUBON D.(2), BALDACCI P.(3), BARALE J.C.(4), PELLOUX H.(2), MÉNARD R.(3), HAKIMI M.A.(1) (1) UMR5163 - CNRS/UJF - Laboratoire Adaptation et Pathogénie des Microorganismes, GRENOBLE, FRANCE ; (2) Parasitologie-mycologie, département des agents infectieux, centre hospitalier universitaire, GRENOBLE, FRANCE ; (3) Unité de Biologie et Génétique du Paludisme, Institut Pasteur, PARIS, FRANCE ; (4) Department of Immunology, Institut Pasteur, CNRS, URA1961, PARIS, FRANCE
EUGREGARINES AND CRYPTOSPORIDIA: PECULIAR PARASITES WITH EPICELLULAR DEVELOPMENT
VALIGUROVÁ A.(1), KOUDELA B.(2) (1) Department of Botany and Zoology, Faculty of Science, Masaryk University, BRNO, CZECH REPUBLIC ; (2) Department of Parasitology, University of Veterinary and Pharmaceutical Sciences, BRNO, CZECH REPUBLIC
Toxoplasma gondii is an obligate intracellular Apicomplexan parasite responsible for encephalitis in immunocompromised individuals and birth defects when the parasite infects the fetus. T. gondii can differentiate from the replicative form, the tachyzoite, to the cystic form, the bradyzoite. The differentiation process is believed to be at the basis for regulation of virulence. Although different set of proteins are specifically expressed in each stage, little is known about regulation of gene expression in Apicomplexa. A recent study from our lab showed that acetylation of histones is regulated during differentiation suggesting a role for chromatin structure in the control of gene transcription. Acetylation of histones is controlled by histone acetyltransferases and histone deacetylases (HDAC). We found that HDAC inhibitors (HDACi) interfere with parasite growth. Treatment of parasite with HDACi induces the expression of bradyzoite related proteins suggesting that HDACs are involved in the control of differentiation. ChIP-Chip experiments showed that HDACi induces modifications of the pattern of histone acetylation at level of multiple bradyzoites related genes. A genetic approach showed that mutations in a HDAC were sufficient to render T. gondii resistant to HDACi indicating that inhibition of a specific HDAC is responsible for growth arrest. Overall, our data indicate that HDAC is a key regulator of gene expression during parasite conversion.
Gregarines and cryptosporidia are often considered to be peculiar apicomplexans, as they exhibit unique features in their attachment strategy, and some phylogenetic analyses have pointed out their affinity. This study focuses on the ultrastructural analysis of their early development and hostparasite interactions, and searches for explicit characteristics common to both apicomplexans. The formation of attachment site was studied in two eugregarines, Gregarina polymorpha and G. steini from laboratory reared insects, and obtained data were directly compared with comparable features observed during attachment process of Cryptosporidium muris in the stomach of experimentally infected rodents. In both apicomplexans, the apical complexes disappear early during cell invasion and parasites are attached to the host cell by their apical processes, i.e. epimerite in eugregarines and the feeder organelle in cryptosporidia. Data presented herein support the term epicellular to reflect cryptosporidian localization, rather than the term intracellular-extracytoplasmic traditionally used throughout the literature. The cryptosporidian attachment strategy is very similar to that of the eugregarines, which are also epicellularly located on the microvillous surface of host epithelial cells, although not enveloped by host cell membrane folds. Despite a few differences found among the studied eugregarines and cryptosporidia, our data support their close relationship.
SY01/03-05
SY01/04-03
SEQUENCING AND ANNOTATION GENOME OF NEOSPORA CANINUM
OF
BLASTOGREGARINES (ALVEOLATA: APICOMPLEXA: SPOROZOA) ARE A PLESIOMORPHIC GROUP OF SPOROZOANS AS INFERRED FROM ULTRASTRUCTURAL AND MOLECULAR DATA
THE
WASTLING J.M.(1), TREES A.J.(1), PAIN A.(2), SOHAL A.(2), PRIETO H.(1), LATHAM S.M.(1) (1) Faculty of Veterinary Science, University, LIVERPOOL, UNITED KINGDOM ; (2) Wellcome Trust Sanger Institute, HINXTON, UNITED KINGDOM
SIMDYANOV T.G.(1), PASKEROVA G.G.(2), MIKHAILOV K.V.(1), ALEOSHIN V.V.(1) (1) Moscow State University, MOSCOW, RUSSIAN FEDERATION ; (2) St. Petersburg State University, ST. PETERSBURG, RUSSIAN FEDERATION
We present the first sequence and annotation of the genome of Neospora caninum. Over 900,000 reads were obtained from the NC-Liverpool isolate giving an estimated 8.2x coverage of the 62MB genome. Sequence was assembled using PHARP into 1,114 contigs with a N50 of 200 kb (http://www.sanger.ac.uk/cgibin/blast/submitblast/n_caninum). A total of 14 pseudo chromosomes were predicted for Neospora by PROMER alignment with T. gondii. Gene finding tools using Jigsaw found an initial 5,591 putative protein coding genes and this first pass semi-automated annotated sequence is available in GeneDB (http://www.genedb.org/genedb/ncaninum/). The mean length of each gene was 6,182 bp, containing on average, 7.28 introns and with a mean intergenic distance of 11,036 bp. Gene prediction was supported by mapping over 21,000 ESTs back to the sequence. In addition, 9,000 peptides
Blastogregarines are a small group of extracellular parasites of the polychaete gut. This group has a single genus Siedleckia including 4 species and is traditionally grouped with gregarines based on their superficial morphological similarity. No ultrastructural and molecular data on bastogregarines were available, which allowed no reliable inference of their phylogeny. We have described the ultrastructure and analyzed the molecular data on the Siedleckia trophozoites found in the gut of polychaete Scoloplos armiger from the littoral zone of the White Sea. The ultrastructural data confirm that blastogregarines are apicomplexans really. The common features include the trimembrane pellicle and a regular pattern of subpellicular 32
microtubules radiating from the polar ring at the apical end that also includes the rhoptries and a large mucronal vacuole. However, neither the conoid nor micropores were found. The phylogeny of Siedleckia was assessed by the analysis of SSU and LSU rRNA genes. Siedleckia grouped with parasitic apicomplexans (i.e. sporozoans) and represented an independent lineage separated early in their evolution. The position of Siedleckia among sporozoans could not be accurately established due to the low support of some tree nodes and limited taxonomic sampling for LSU rRNA gene. Thus, blastogregarines represent a basal lineage of parasitic apicomplexans according to the ultrastructural and molecular data.
amino acids can be converted into acetyl-CoA and serve as carbon source for the TCA-cycle and FAS I. Consistent with this model, T. gondii and other coccidians possess the branchedchain amino acid degradation pathway leading to the production of acetyl-CoA in the mitochondrion and an ATP citrate lyase, capable of converting the citrate produced by the TCA-cycle into acetyl-CoA in the cytosol. The degradation of branched-chain amino acids leads to the production of propionyl-CoA, a toxic compound, which inhibits cell growth. Like fungi and bacteria, the coccidians have the capability to detoxify propionyl-CoA into pyruvate via the 2-methylcitrate cycle. We have localized the enzymes of the 2-MCC between the mitochondrion and cytosol and produced biochemical evidence that this pathway is active in T. gondii tachyzoites.
SY01/04-04
SY01/05-02
PHYLOGENY OF HAEMOSPORIDIA WITH BAT PARASITES FROM MADAGASCAR AND CAMBODIA
NEOSPORA CANINUM AND NEOSPOROSIS: NOVEL TARGETS FOR PREVENTION OF INFECTION AND TREATMENT OF DISEASE THROUGH MOLECULAR STUDIES ON HOSTPARASITE INTERACTIONS
DUVAL L.(1), ROBERT V.(2), ARIEY F.(3) (1) Muséum National d'Histoire Naturelle, USM 504 and UMR 5202 & Institut Pasteur du Cambodge, Unité Epidémiologie moléculaire, PARIS & PHNOM PENH, FRANCE & CAMBODIA ; (2) Muséum National d'Histoire Naturelle, USM 504 and UMR 5202 & Institut de Recherche pour le Développement, UR 77 and UR 16, PARIS, FRANCE ; (3) Institut de Recherche pour le Développement, UR 77 and UR 16, PARIS, FRANCE
HEMPHILL A., LEEPIN A., DEBACHE K., MUELLER J., MUELLER N. University of Berne, BERNE, SWITZERLAND Neospora caninum is an apicomplexan parasite that causes abortion in cattle, neuromuscular disease, and also infects a wide range of other domestic and wildlife animals. The importance of neosporosis as the most frequent infectious cause of bovine abortion has prompted studies on potential novel preventive and treatment measures. Both, vaccination and chemotherapeutical treatment have been noted as viable and cost-effective measures. On one hand, secretory and surface-associated proteins of the infective stages of N. caninum have been identified, and expressed as recombinant antigens and/or respective DNA-vaccines have been generated. These antigens are being characterized in vitro, and the protective potential of these antigens is currently being studied in mouse models that mimic acute cerebral infection and/or fetal infection upon pregnancy. On the other hand, we have characterized a number of compound classes (thiazolides, isoflavonoids, pentamidine-derivatives and others) for their activity against N. caninum and other protozoan parasites (Toxoplasma, Giardia) using cell culture-based approaches, and we have identified a number of parasite-, but also host cellassociated, drug targets by applying an affinitychromatography-based method employing epoxy-sepharosecoupled drugs and mass spectrometry.
There have been reported cases of host-switching in avian and lizard species of Plasmodium (Apicomplexa, Haemosporidia), as well as in those infecting different primate species. However, no evidence has previously been found for host-swapping between wild birds and mammals. We present the results of the sampling of blood parasites of wild-captured bats from Madagascar and Cambodia. The presence of Haemosporidia infection in these animals is confirmed and cytochrome b gene sequences were used to construct a phylogenetic analysis. The introduction of the 7 new genetic sequences from chiropteran hemoparasites reveals at least three different and independent Haemosporidia evolutionary histories in three different bat lineages from Madagascar and Cambodia. Plasmodium falciparum is confirmed to present a deep branching within the mammalian clade and to have different and independent evolutionary history vis-à-vis other human malaria parasites. In conclusion, phylogenetic analysis strongly suggests multiple host-switching of Haemosporidia parasites (Plasmodium and Hepatocystis) in bats with those from avian and primate hosts.
SY01/04-05
THE METHYLCITRATE CYCLE, AN UNEXPECTED METABOLIC PATHWAY IN THE MITOCHONDRION OF COCCIDIAN PARASITES
SY01/05-03
IN VITRO GROWTH INHIBITION OF PLASMODIUM FALCIPARUM BY CATESTATIN, AN ANTIBACTERIAL CHROMOGRANIN ADERIVED PEPTIDE
FLEIGE T.(1), LIMENITAKIS J.(1), RIEZMAN H.(2), SOLDATI D.(1) (1) Centre Medical Universitaire, GENEVA, SWITZERLAND ; (2) Faculty of Biology, GENEVA, SWITZERLAND
AKADDAR A.(1), JEAN-FRANCOIS F.(2), DODERER C.(1), GUEROLD B.(3), MOUSLI M.(1), MAHATA S.(4), ABDELRAHMAN T.(1), AUNIS D.(3), DUFOURC E.J.(2), METZ-BOUTIGUE M.H.(3), CANDOLFI E.(1) (1) Institut de Parasitologie et de Pathologie Tropicale, EA 3950, Université Louis Pasteur, 67000 STRASBOURG, FRANCE ; (2) UMR 5248 CBMN, CNRS-Université Bordeaux1-ENITAB, IECB, 33607 PESSAC, FRANCE ; (3) INSERM U575, Physiopathologie du Système Nerveux, 67084 STRASBOURG, FRANCE ; (4) Departement of Medicine, University of California at San Diego, 92093 LA JOLLA, CALIFORNIA, UNITED STATES
Toxoplasma gondii is an obligate intracellular protozoan parasite, which actively enters the host cell and develop in a unique non-fusiogenic parasitophorous vacuole. Fast intracellular multiplication critically depends on access to energy and carbon sources, in particular for lipid biosynthesis. T. gondii harbours the two types of fatty acid synthesis (FAS) pathways. While glucose is anticipated to serve as carbon source for FAS II localised to the apicoplast, the carbon source for FAS I is not determined yet. We hypothesize that host 33
DEA-AYUELA M.A.(1), CUELLAR C.(1), JIMENEZ I.(2), BOLASFERNANDEZ F.(1) (1) Universidad Complutense (UCM), MADRID, SPAIN ; (2) Centro Nacional de Microbiología, Instituto de Salud Carlos III, MADRID, SPAIN
Bovine Catestatin and its active domain Cateslytin (bCGA344-358) display powerful antimicrobial activities. We have tested the activity of bovine and human synthetic peptides corresponding to Catestatin, Cateslytin and other related peptides on in vitro growth of Plasmodium falciparum. A growth inhibition was observed with Catestatin for the sensitive and resistant P. falciparum strains. Fluorescence microscopy showed that Catestatin co-localizes solely with the merozoites, extra-erythrocytic stage of this parasite, demonstrating the specificity of action of Catestatin for P. falciparum. Catestatin was able to inhibit partially the red blood cells invasion by merozoites. However parasites were still viable as shown by a persistent basic production of the metabolic enzyme pLDH. A relation structure-function analysis demonstrates the role of the LSFR median region for this activity. Biochemical methods indicate a specific protein-protein interaction between Catestatin and two Plasmodium proteins (27 kDa and 43 kDa). Furthermore, we characterized interactions of Cateslytin with model mimicking Plasmodium membrane showing aggregation of peptides into beta-sheets. Finally, we observe two complementary mechanisms at merozoite level: (i) a specific interaction between Plasmodium proteins and Catestatin and (ii) a disordering (reduction in thickness) of the phosphatidylcholine component of the Plasmodium membrane by the active domain Cateslytin.
Recently it has been shown that protein-tyrosine phosphatases (PTPs) play a role in virulence and pathogenesis in Leishmania and they could be exploited as suitable targets and vaccine candidates. A proteomic approach was applied towards further characterization of Leishmania (PTPs). Crude extracts from stationary promastigote stages of L. infantum were submitted to two-dimensional electrophoresis and thereafter probed in western-blot (WB) with two anti- PTP antibodies (anti-PTP-1B and anti-Syp/SH PTP2). Among proteins recognized by both PTP antibodies, enolase-phosphatase and ATPase beta subunit were identified by mass spectrometry (MALDI-TOF and MALDITOF/TOF). Genes encoding both proteins were amplified by PCR from genomic DNA prepared from L. infantum (MCAN/ES/98/LLM-877) using primers derived from sequences LinJ36.6410 (enolase-phosphatase) and LinJ25.1220 (ATPase beta subunit) available in the geneDB (http://www.sanger.ac.uk/Projects/L_infantum/). PCR products were purified and cloned into PCR®-TOPO® and later sequenced. Afterwards the ORFs were subcloned into pGEX4T1 and pQE30 for further expression of the recombinant proteins.
SY01/05-04
MASLINIC ACID: SERIN PROTEASE INHIBITOR THAT BLOCKS SERIN PROTEASE ACTIVITY, GLIDING MOTILITY, CELL INVASION, AND SECRETORY PATHWAY OF TACHYZOITES OF TOXOPLASMA GONDII
SY02/01-03
STRUCTURE AND FUNCTION OF PROTEINS AS PROPHYLACTIC OR THERAPEUTIC TOOLS: A CLOSER LOOK TO LEISHMANIA BRAZILIENSIS HISTONE H1.
DE PABLOS TORRÓ L.M. Universidad de Granada, GRANADA, SPAIN
CARMELO E., BORGES V.M., VALLADARES B. I. U. Enfermedades Tropicales y Salud Pública de Canarias, LA LAGUNA, SPAIN
Proteases and their role in proteolytic maturation, surface trimming, and surface shedding of proteins released during the invasion of Toxoplasma gondii in its host cell, are a clear target for the development of new inhibitory drugs. In this work, we describe the in vitro activity of maslinic acid (2-, 3-dihydroxiolean-12-en-28-oic acid) against the intracellar protozoan belonging to the phylum Apicomplexa, Toxoplasma gondii. We made a cytotoxicity study on purified tachyzoites and Vero cells, finding a IC50 with lower value in tachyzoites, comparable to that found for PMSF and AEBSF. After observing a clear inhibition of the protease enzymatic activity of the excretion-secretion product (ESP) for maslinic acid and comparable to that of PMSF, we performed zymograms of the ESP of the micronemal product and of the parasite-cell interaction, finding a dosage-dependent relation for the inhibition of several bands with proteolytic activity. The gliding motility of the parasite is also visibly reduced, in number and trail length in response to several concentrations of maslinic acid, finding a gliding-inhibition percentage of 100% for 50M. Finally, we found extensive morphological changes at 62.5 M such as swelling of the nuclear envelope, partial collapse of the Golgi complex, accumulation of myelinic bodies, and in some parasites, the partial rupture of the outer plasma membrane.
The term Histone H1 comprises a family of basic low-molecular weight proteins that take part in the maintenance of chromatin structure and function. H1s from early eukaryotes, such as Kinetoplastids, show a series of particular structural and functional characteristics, very divergent to human H1. These features give this protein a remarkable interest as a potential therapeutic of prophylactic tool against leishmaniosis that prompted us to open a research line towards the characterization of Leishmania braziliensis Histone H1. As a first step, the purification of non-denaturing recombinant L. braziliensis histone H1 has been achieved, in order to perform biochemical activity assays. Among these assays, the interaction of the protein with linear and supercoiled DNA in vitro has been evaluated using EMSAs. In addition, the aggregation state in solution of the native protein has been determined by crosslinking studies, as well as MALDI-TOF spectrometry. Our results show that L. braziliensis Histone H1 is basically a monomeric protein, although higher aggregation states are seen in the presence of DNA. Besides, the protein is able to interact with supercoiled DNA in vitro, showing higher affinity with that substrate than with linear DNA. This work has been funded by “Instituto del Metabolismo Celular” (IMC), La Laguna, Tenerife, Spain. SY02/01-04
LEISHMANIA AMAZONENSIS LEISHPORIN IS A CHOLESTEROL-INDEPENDENT POREFORMING PROTEIN AND REQUIRES ONLY LIPIDS TO BIND TO AND LYSE NATURAL AND
SY02/01-02
MOLECULAR CHARACTERIZATION OF TWO PROTEIN-TYROSINE PHOSPHATASES FROM LEISHMANIA INFANTUM 34
CHALLENGE VIA INCREASED MAGNITUDE OF T CELL RESPONSES
ARTIFICIAL MEMBRANES
CASTRO-GOMES T.(1), CALZAVARA-SILVA C.E.(1), ALMEIDACAMPOS F.R.(1), SILVA PEREIRA R.(1), VILELA J.M.(2), SPANGLER M.(2), FRÉZARD F.(1), HORTA M.F.(1) (1) Universidade Federal de Minas Gerais, BELO HORIZONTE, BRAZIL ; (2) Centro Tecnológico de Minas Gerais, BELO HORIZONTE, BRAZIL
KEDZIERSKI L.(1), CURTIS J.(1), DOHERTY P.(2), HANDMAN E.(1), KEDZIERSKA K.(2) (1) Walter and Eliza Hall Institute of Medical Research, PARKVILLE, AUSTRALIA ; (2) University of Melbourne, PARKVILLE, AUSTRALIA
Leishporin is a pore-forming protein from the protozoan L .amazonensis, a causative agent of leishmaniasis. Because it is optimally active at pH 5.5 and 37°C it is a good candidate to rupture the macrophages, an infection-amplifying mechanism that is poorly understood. In a previous work, we showed that leishporin binds to cell membrane before lysis. Here, we have studied the binding requirements of leishporin to lyse erythrocytes. To investigate whether leishporin requires a cell surface protein, we have treated human erythrocytes with various proteases before incubation with L. amazonensis promastigotes membrane soluble extract (ms-ext), which contains leishporin. This treatment did not reduce erythrocytes susceptibility to lysis, showing that leishporin does not bind to target cell protein. Inhibition assays showed that, similarly, carbohydrates are not required for leishporin-mediated lysis. These results were confirmed by the fact that calceincontaining dipalmitoilphosphatidilcholine (DPPC) liposomes were lysed by ms-ext in a temperature-dependent manner. Inclusion of cholesterol in the liposome did not increase lysis, indicating that leishporin is a cholesterol-independent cytolysin. Additionally, we visualized by atomic-force microscopy putative pore-formation in erythrocytes membranes and in DPPC lipidic films. Taken together, our results demonstrate that leishporin can form pores by binding directly to the lipids in target membranes.
Leishmaniasis, caused by parasites of the Leishmania species, is a global health problem. Although there are no vaccines against leishmaniasis, recent evidence suggests that immunisation with non-persistent parasites may protect against subsequent infection by targeting central memory CD4+ T cells. The present study asks whether immunisation with nonpersistent L. major PMM parasites leads to protection against infection and to the recruitment of T cells of a specific phenotype. Our study shows that short- and long-term vaccination of susceptible BALB/c mice with PMM parasites leads to protection against infection. Such protection is due to a higher number of CD44hiCD4+ and CD44hiCD8+ T cells recruited into draining lymph nodes and the ability to suppress IL-10 production following infection. Immunisation with PMM parasites does not alter expression of effector and memory markers on CD4+ and CD8+ T cells, including CD62L, IL-7R and IL-2R, when compared to unvaccinated controls. Thus, expansion of T cell numbers elicited by immunisation and their rapid recruitment to lymph nodes upon infection, and not their phenotypic characteristics play an important role in protection against Leishmania challenge. Since successful vaccines need to generate immunological memory, a better understanding of the formation and maintenance of memory T cells in both animal models and human disease is critical for their development.
SY02/01-05
SY02/02-03
CHROMOSOME COHESION AND SEGREGATION IN TRYPANOSOMA BRUCEI
LUMINESCENT LEISHMANIA INFANTUM PARASITES AS A TOOL FOR MONITORING PARASITIC PROCESS IN THE BALB/C MOUSE
BESSAT M. University of hull, HULL, UNITED KINGDOM
MICHEL G.(1), LANG T.(2), FERRUA B.(1), LEMICHEZ E.(1), MARTY P.(3) (1) INSERM, NICE, FRANCE ; (2) Institut Pasteur, PARIS, FRANCE ; (3) CHU de Nice, NICE, FRANCE
The nuclear genome of the African trypanosome, Trypanosoma brucei, is composed of megabase- (MBC), intermediate- (IC) and mini-chromosomes (MC). MCs segregate with high fidelity during mitosis, but little is known about the molecular basis of their segregation. The highly conserved cohesin complex is involved in the mitotic process in probably all eukaryotes and T. brucei has homologs of all four cohesin subunits, SMC1, SMC3, Scc1 and Scc3. Polyclonal antibodies were raised against TbSMC3 recombinant protein, and IF microscopy showed an intranuclear localisation of the protein throughout the cell cycle. When the soluble proteins were detergent-extracted prior to fixation, SMC3-staining was absent from anaphase and telophase cells, but reappeared after cytokinesis. RNA interference combined with fluorescence in situ hybridization analysis indicated that SMC3 depletion had a deleterious effect on MBC segregation but no or little effect on MC segregation. However, RNAi-based depletion of separase, a protease that cleaves the cohesin component Scc1 to enable metaphase to anaphase progression, affected the segregation of both MBC and MC populations.
To facilitate the follow-up of parasitic process in L.infantum infected BALB/c mouse, real time monitoring methods are needed. To this end we have prepared L.infantum clones stably expressing the firefly luciferase gene. Eleven clones were generated and analysed in two different systems i) in vitro bioluminescent screening on promastigotes stage and ii) the monitoring of the parasitic process in living animals. Parasite burdens were also quantified ex vivo by measuring luminescence from liver or spleen detergent extracts. Temporal analyses were performed following intraperitoneal inoculation of luminescent L.infantum promastigotes into BALB/c. The bioluminescent light transmitted through the tissue was imaged externally using a charge coupled device camera (CCD). Our results showed that these parasites produced significant bioluminescent signals for both in vitro studies and the development of an in vivo model. Bioluminescent signals, measured in the liver and spleen of parasites-harbouring mice well agreed with the classical parasite number estimates. In conclusion this study presents a new model for visualising the phases that L.infantum establish in BALB/c mice allowing the course of the parasitism to be readily monitored in real time longitudinally on each Leishmania-infected mouse. Additionally this model may facilitate vaccine trials against leishmaniases in particular by limiting the number of experimentally infected
SY02/02-02
IMMUNISATION WITH NON-PERSISTENT LEISHMANIA MAJOR PHOSPHOMANNOMUTASE DEFICIENT PARASITES PROTECTS MICE FROM VIRULENT 35
mice.
marked protection (85% and 76% in liver and spleen, respectively) was observed in mice vaccinated with low PL dose and CNF-1 WT.
SY02/02-04
UNDERSTANDING T CELL-MEDIATED PROTECTION AGAINST CUTANEOUS LEISHMANIASIS USING A NOVEL REVERSE GENETICS STRATEGY
SY02/03-02
IDENTIFICATION OF THE IRON SUPEROXIDE DISMUTASE GENE REPERTOIRE AND ITS USE AS DIAGNOSTIC TOOL IN TRYPANOSOMA BRUCEI GAMBIENSE
KEDZIERSKA K.(1), CURTIS J.(2), HATTON L.(1), VALKENBURG S.(1), HANDMAN E.(2), DOHERTY P.(1), KEDZIERSKI L.(2) (1) University of Melbourne, PARKVILLE, AUSTRALIA ; (2) The Walter and Eliza Hall Institute, PARKVILLE, AUSTRALIA
HERDER S.(1), SIMO G.(2), CUNY G.(1), ASONGANYI T.(2), NJIOKOU F.(3) (1) IRD, MONTPELLIER, FRANCE ; (2) Faculty of Medicine and Biomedical Sciences, University of Yaoundé 1, YAOUNDÉ, CAMEROON ; (3) Faculty of Science, University of Yaoundé 1, YAOUNDÉ, CAMEROON
Protection against Leishmania has been attributed to CD4+ Th1-type immune response. Thus, a successful antileishmanial vaccine needs to stimulate strong Th1-type CD4+ T cell response and long-lasting protective antileishmanial immunity. In this study, we have utilised the cutting edge reverse genetics strategy to manipulate influenza A virus in a way that it provides a novel and ideal delivery vector for Leishmania vaccine capable of stimulating highly desirable Th1-type CD4+ T cell responses. We engineered a single, immunodominant Leishmania-specific LACK CD4+ T cell epitope within the neuraminidase (NA) segment of influenza viruses. These recombinant viruses were used to vaccinate highly susceptible BALB/c mice and to investigate LACK-specific CD4+ T cell responses in correlation with protection against L. major challenge. Our immunisation strategy resulted in marked 2-4 log reduction in parasite burdens and reduced lesion scores associated with increased IFN- production by LACK-specific CD4+ T cells. This indicates a strong Th1 response following a single-dose immunisation in the context of non-replicative influenza delivery. Manipulating the nature of Leishmaniaspecific CD4+ T cells increases our understanding of CD4+ T cell effector function and long-lasting protective antileishmanial immunity that is central to both parasite clearance and the design of CD4+ T cell-based prophylactic and therapeutic vaccines.
Superoxide dismutase (SOD) forms part of the defense mechanism that helps to protect organisms from superoxide anions. This enzyme is one of the isoenzyme systems commonly used to differentiate T.b. gambiense from T.b. brucei and T.b. rhodesiense. To understand the genetic basis of the differences observed between SOD electrophoretic profiles of T. brucei sub-species, we undertook the identification and the characterization of SOD gene repertoire in T.b. gambiense. This study was performed on seven stocks (4 T.b. gambiense group 1 and 3 group 2) showing different SOD profiles. Four SOD genes (soda, sodb1, sodb2 and sodc) were identified in T.b. gambiense genome. These genes were cloned and their predicted amino acid sequences were deduced. Few differences were observed between nucleotide sequences of the four SOD genes of T.b. gambiense group 1 and 2 stocks. Even with T.b. brucei, few differences were observed. Several amino acids specific to FeSOD were found in the four SODs sequences of T.b. gambiense. Aligning the four T.b. gambiense protein sequences with those of others organisms, important differences were found with MnSOD and Cu/ZnSOD, but high similarity with FeSOD; indicating that the SODs of T.b. gambiense are FeSOD. High similarity exists between the proteins sequences of T.b. gambiense and T.b. brucei. Nevertheless, these sequences were used to design primers that are diagnostic for T. b. gambiense group1 & 2.
SY02/02-05
PROTECTION EFFECT OF INTRANASAL VACCINATION WITH L. INFANTUM PROMASTIGOTE ANTIGENS AND CYTOTOXIC NECROTIZING FACTOR (CNF-1) AS ADJUVANT.
SY02/03-03
SENSITIVE DETECTION AND EASY TYPING OF LEISHMANIA BY REAL-TIME PCR
MICHEL G.(1), FERRUA B.(1), FLATAU G.(1), LEMICHEZ E.(1), MARTY P.(2) (1) INSERM, NICE, FRANCE ; (2) CHU de Nice, NICE, FRANCE Because of the limited number of available anti-leishmania drugs and their limitations (toxicity, resistance), the leishmaniasis are considered by the WHO as an emergent and uncontrolled disease. Therefore, a vaccine against leishmaniasis, particularly against the visceral form, fatal if untreated, is highly desirable. In this context, we assessed, in the BALB/c model, a process of nasal vaccination against Leishmania infantum using leishmania antigens (crude promastigote lysate, PL) co-delivered with the toxin CNF-1 (cytotoxic necrotizing factor) from uropathogenic E. coli. Two vaccinal trials, with low (3x15g) and high antigens doses (4x60g) were conducted with CNF-1 WT (1g) as adjuvant. Biologically inactive CNF-1 was used as control. Mice were challenged with IP injection of 108 L. infantum promastigotes. Nasal vaccination with PL induced cellular and antibody systemic responses, augmented in the presence of CNF-1 WT. Vaccination with high dose of PL caused an exacerbation of the infection inhibited in the presence of CNF-1 WT. Conversely,
ROELFSEMA J., NOZARI N., KORTBEEK L., PINELLI E. National Institute of Public Health, BILTHOVEN, NETHERLANDS In the case of leishmaniasis, routine molecular diagnostics should preferably be accomplished by a single PCR performed on DNA isolated directly from patient material. This PCR should be as sensitive as possible and ideally amplify a target that is present in all species but divergent enough to distinguish these species. In our laboratory we receive a variety of patient samples. We perform a PCR on the 18S gene and in case of a positive result we digest the product with CfoI and RsaI for typing to genus level (Meredith et al. 1993). Clinicians, however, often demand more precise typing to species level. Therefore, we compared three alternative loci that could be amplified without culturing the parasites from patient samples: the intergenic region of the mini-exon gene, the repeat described by Piarroux and the ITS1 region. We found that the PCR on ITS1, as described by Schönian et al (2003), was as sensitive as the PCR on 18S. Digesting the ITS1 PCR product with HaeIII and CfoI enables us to distinguish 36
between most species. However, New World species like L.braziliensis, L.panamensis or L.guyanensis cannot be differentiated this way. When such a distinction is needed we perform the mini-exon PCR and subsequent sequencing. We have adapted the ITS1 PCR so that it works well on a Roche LightCycler using a double labeled probe and we are currently validating this approach.
from L.major and L.tropica. The heterogeneity of L.major in Iran has confirmed in this study.In view of the fact that isoenzyme characterization has recommended as a reliable and precise method for species and strain identification of isolates, obtained data in last decade is very helpful for research investigations in the filed of leishmaniasis in Iran. Key words: Leishmaniasis, zoonotic, anthroponotic, isoenzyme
SY02/03-04
SY02/04-01
MICROSATELLITES CHARACTERIZATION OF LEISHMANIA ISOLATED STRAINS
PHLEBOTOMINE SANDFLIES AND LEISHMANIA: COEVOLUTION AND CAPTURES
REALE S.(1), GRAVINO A.E.(2), MANNA L.(2), LUPO T.(1), PIAZZA M.(1), MIGLIAZZO A.(1), VITALE F.(1) (1) Istituto Zooprofilattico sperimentale della Sicilia, PALERMO, ITALY ; (2) Università Federico II°, NAPOLI, ITALY
DEPAQUIT J. USC AFSSA « VECPAR » - Faculté de Pharmacie - Université de Reims Champagne-Ardenne, REIMS, FRANCE The relationships between Leishmania and their exclusive vectors: the Phlebotomine sandflies (Diptera: Psychodidae) are discussed.This reflexion is based 1) on known links between Leishmania and sandflies (like the common transmission of L. infantum by the Phlebotomus belonging to the subgenus Larroussius and its capture by Lutzomyia longipalpis in America) 2) on biogeography and paleobiogeography of parasites and vectors 3) on paleontological data 4) interactions between Leishmania and sandflies including vectorial competence and capacity.
Most part of Leishmania infantum strains circulating in Mediterranean area, are belonging to zymodeme MON-1, despite their very wide geographical distribution. Different approach are been developed to improve the discrimination Leishmania genus using biochemical and molecular methods. Microsatellite are tandem repeats DNA sequences distributed abundantly in the eukaryotic genomes that may reveal important genetic polymorphisms at sub-species level. In the daily activity of Italian National Reference Centre for Leishmaniasis (C.Re.Na.L.) of Palermo, a high number of Leishmania strains are isolated from infected biological samples. In this work the authors describe a new epidemiological tool to study the genetic differences trough the isolated Leishmania parasite. We analyzed different repetitive DNA regions, previously described by other authors, to investigate as the microsatellite patterns observed within the strains, could be correlated with the geographic distribution. Depending by the annealing temperature three group of PCR tests were performed for each sample, to generate labelled amplicones. Finally the products were analized by 3130 Applied Biosystems genetic analyzer and the results were elaborate by Gene Mapper software (AB), to associate the patterns to each zymodema. Some evaluations as these data could be applied in epidemiologic and pathogenetic studies are in progress.
SY02/04-02
CHARACTERISATION OF TRYPANOSOMES FROM AUSTRALIAN MARSUPIALS
AVERIS S.(1), THOMPSON A.(1), LYMBERY A.(1), WAYNE A.(2), MORRIS K.(2), SMITH A.(1) (1) Murdoch University, PERTH, AUSTRALIA ; (2) Department of Environment and Conservation, PERTH, AUSTRALIA Trypanosomes are widespread blood parasites of vertebrates. Some species such as Trypanosoma brucei and T. cruzi cause disease in humans and/ or livestock. Very little is known about trypanosomes found in Australian marsupials. In this study we screened a range of Australian native mammals to determine the presence of trypanosomes by PCR targeting the 18S rRNA gene. Blood samples were obtained from 81 animals: 4 Shark Bay mice (Pseudomys fieldi); 30 boodies (Bettongia lesueur); 24 chuditch (Dasyurus geoffroii), 7 brush-tailed possums (Trichosurus vulpecula); 6 woylies (Bettongia penicillata); 2 western barred bandicoots (Perameles bougainville); 3 banded hare wallabies (Lagostrophus fasciatus); 2 western mice (Pseudomys occidentalis) and 9 bilbies (Macrotis lagotis) from various geographical locations in Western Australia. We found that 2 Shark Bay mice; 2 boodies; one chuditch; 1 brush-tailed possum and 4 woylies were positive for trypanosomes. The trypanosomes isolated from the chuditch were most closely related to T. bennetti, previously isolated from an American kestrel, and those isolated from two separate woylie populations placed on a branch of their own within a clade containing other trypanosomes from Australian vertebrates. The phylogenetic relationship of the potentially novel trypanosomes is discussed.
SY02/03-05
BIOCHEMICAL CHARACTERIZATION OF CAUSATIVE AGENTS OF CUTANEOUS LEISHMANIASIS (CL) IN LAST DECADE AND CURRENT STATUS OF CL IN IRAN
HATAM G.R.(1), HABIBI P.(1), HOSSEINI S.M.H.(2) (1) Shiraz University of Medical Sciences, SHIRAZ, ISLAMIC REPUBLIC OF IRAN ; (2) Razi Institute, SHIRAZ, ISLAMIC REPUBLIC OF IRAN Isolates of Leishmania from patients with cutaneous leishmaniasis were obtained from different parts of Iran and characterized by isoenzyme electerophoresis using polyacrylamide gel and cellulose acetate as matrix. Six superior enzyme systems were used mainly for isolated organisms included malate dehydrogenase, nucleoside hydrolase1&2, phosphoglucomutase, glucose phosphate isamerase and 6 hosphoglucanate dehydrogenase. In suspicious conditions excess systems were used which the most important of them were glucose 6 phosphate dehydrogenase and estrase. Relative migrations of isoenzyme bands of each isolates were compared with World health organization reference strains. This study has established some old foci and has introduced a lot of new foci of ZCL in central, southwest, south, southeast, west, and northeast of Iran. Several zymodemes were obtained
SY02/04-03
EPIDEMIOLOGICAL STUDIES LEISHMANIOSIS IN THE PROVINCE LLEIDA (CATALONIA, NE OF SPAIN)
ON OF
BALLART C.(1), BARÓN S.(2), GUERRERO I.(1), ALCOVER M.(1), PORTÚS M.(1), GÁLLEGO-CULLERÉ M.(1) (1) Universidad de Barcelona, BARCELONA, SPAIN ; (2) 37
Universidad de Granada, GRANADA, SPAIN
GENOTYPES OF LEISHMANIA SPP. USING PCR-RFLP ASSAYS IN CLINICAL SAMPLES OF PATIENTS&RESERVOIRS IN TURKEY
The knowledge of the epidemiological situation of leishmaniosis in Spain is fragmentary due to different reasons: it is not obligatory to declare human cases in all the Spanish autonomous communities, leishmaniosis is not recognized as a major health problem, there are not official statistics on canine leishmaniosis and there are few entomological data concerning sand flies vectors. This lack of knowledge is especially highlighted in the north, including the province of Lleida (12 administrative divisions, AD) where we have undertaken retrospective and prospective surveys. A review of official data about human leishmaniosis detected an incidence rate higher than expected (0-4.9 annual incidence/100,000 inhabitants). Entomological surveys conducted in July 2006 allowed us to detect the two proved vectors of leishmaniosis in Spain: Phlebotomus ariasi present in 9/12 AD, with a density of 2.3 specimens/m2 and representing the 2.6% of the collection and P. perniciosus present in 10/12 AD, with a density of 10.2 specimens/m2 and representing the 11.6% of the collection. Their presence is conditioned by the altitude. For the first time an isolate of Leishmania (MCAN/ES/2007/BCN-759) from the area has been identified by MLEE belonging to the zymodeme MON-1 of Leishmania infantum. Work supported by projects AGL2004-06909-C02-01/GAN (Ministerio de Ciencia y Tecnología, Spain) and 400764/GOCECT-2003-010 (6th Framework Programme, EU)
GUNES K.(1), OZENSOY TOZ S.(1), ERTABAKLAR H.(2), ERTUG S.(2), OZBEL Y.(1) (1) Ege University Medical School, IZMIR, TURKEY ; (2) Adnan Menderes University Medical School Dept. of Parasitology, AYDIN, TURKEY Leishmaniasis is an infection that caused by the parasites belonging to the genus Leishmania. The clinical forms of the disease can be summarized in three topics: Visceral (VL), cutaneous (CL) and mucocutaneous (MCL) leishmaniasis. The first two of these forms are seen in Turkey and the reservoirs are the dog and the man respectively. In our study, 37 Leishmania isolates that have been cultivated in the NNN medium since 1996 were analyzed by using molecular tools comparing the restriction patterns of the DNA bands belonging to the ribosomal ITS region amplified by PCR method. By using one of the restriction nucleases, Taq I, different restriction patterns were obtained between the isolates from VL and CL, and also CL isolates from Sanliurfa and Ege Region. There were different patterns in three of dog isolates while rest of the VL isolates that originated from the man and the dog were similar. In addition to this, RFLP patterns of 14 Sanliurfa isolates out of 21 CL isolates were almost totally homogenous except one. The causative agents were confirmed as Leishmania infantum for VL, and Leishmania tropica for CL in these regions in Turkey. But the RFLP patterns of the isolates obtained from the patients with CL in Sanliurfa were not the same as the isolates from Ege Region except one in comparison by the genotype. The analyses of the molecular results together with izoenzyme typing will be supportive for the epidemiological aspect.
SY02/04-04
THE IMPACT OF AGRICULTURE DEVELOPMENT IN THE SPREAD OF VISCERAL LEISHMANIASIS IN TUNISIA
AOUN K.(1), JEDDI F.(1), AMRI F.(2), ZOUARI B.(3), BOURATBINE A.(1) (1) Institut Pasteur de Tunis, TUNIS, TUNISIA ; (2) Service de Pédiatrie-Hôpital de Kairouan, KAIROUAN, TUNISIA ; (3) Section de Médecine préventive-Faculté de Médecine de Tunis, TUNIS, TUNISIA
SY03/01-02
STUDIES ON THE BIOLOGY OF MALARIA VECTOR (ANOPHELES LABRANCHIAE) IN THE TONGA LAKE
Until the late 70’, cases of Visceral leishmaniasis (VL) were restricted in Tunisia to the Northern part of the country. From the 80’, a geographical spread of the disease to many areas of the Center is observed. It is also associated with a dramatically increase of the incidence of cases. Thus, annual incidence raised up from 5-10 cases before the 70’ to an average of 99,6 cases in this last decade. The mean yearly incidence rate in children less than 5 years, the most concerned by VL in Tunisia, has reached 9,6/100000 during the period 1996-2006, which represents one of the highest rates in Mediterranean countries. The districts of the Center, rarely involved before the 80’, register actually more than 40% of all cases. The geographical spread and the increase of the number of VL cases are mainly linked to the recent mobilization of the hydraulic resources in the arid areas of the center by the promotion of wells and dams construction as well as hills lakes implementation. In fact, in spite of their benefits on the improvement of irrigation, agriculture and farming, thesis water plans induce environmental changes that are generally auspicious to sandflies and dogs, the former as vectors and the later as reservoir of the causative agent Leishmania infantum. As a matter of fact, a positive correlation is found between the distribution of hydraulic restraints and the incidence of VL (r’=0,47, p90º and incomplete in certain caryophyllideans. With respect to the number of plates of intercentriolar bodies, there are 1–3 in the caryophyllideans and 3–7 in the trypanorhynchs. A major difference between the groups is in the number of axonemes, with one in the Caryophyllidea and two in the Trypanorhyncha. The phylogenetical implications of such differences with respect to other cestode orders will be also among the points addressed.
SY08/02-01
INDUCTION AND EXECUTION OF APOPTOSISLIKE CELL DEATH IN PLASMODIUM BERGHEI OOKINETES
HURD H., ARAMBAGE S., ALI M. Keele University, KEELE, UNITED KINGDOM
Ookinete development in vivo and in vitro is associated with a huge death rate, with parasites showing multiple features of apoptosis-like cell death including nuclear chromatin condensation, DNA fragmentation and activation of caspaselike molecules. The proportion of parasites exhibiting these markers increases with time. The mode of death is, however, contentious as components of molecular pathways involved in metazoan apoptosis are missing from the Plasmodium genome. Inhibitor studies suggest the caspase-like molecule detected using fluorescent markers is a clan CD cysteine protease rather than belonging to clan CA. Three members of this family, the metacaspases (MCP), are present in P. berghei. However, PbMCP1 and PbMCP2 knockout parasites did not differ from wild type ookinetes in the proportion exhibiting apoptotic markers (45% at 22h post-incubation). Proteomic approaches have been initiated to identify the caspase-like molecule. Although a proportion of ookinetes exhibit loss of mitochondrial membrane potential this does not appear to be associated with the onset of an apoptosis-like pathway. In vitro, apoptosis-like death is induced by increased levels of nitric oxide, but not LDOPA and nitric oxide donors in a blood meal inhibit the formation of ookinetes in the mosquito midgut. These results contribute to our understanding of the morphological and cellular pathways involved in the death of ookinetes in the midgut lumen.
SY08/01-05
COMPARATIVE TEM STUDIES ON THE INFECTIVE ONCOSPHERES OF SEVEN SELECTED SPECIES OF CYCLOPHYLLDEAN CESTODES OF MEDICAL AND VETERINARY IMPORTANCE.
SWIDERSKI Z.(1), MLOCICKI D.(1), GRYTNER-ZIECINA B.(2) (1) Institute of Parasitology Polish Academy of Sciences; Medical University of Warsaw, WARSAW, POLAND ; (2) Medical Univeristy of Warsaw, WARSAW, POLAND The ultrastructure of the infective oncospheres of seven species belonging to three families of cyclophylldean cestodes was examined as follows: : [1] Taeniidae, 2 species: Echinococcus granulosus and E. multilocularis; [2] Anoplocephalidae, 4 species: Inermicapsifer madagascariensis, Anoplocephaloides dentata, Gallegoides arfaai and Mosgovoyia ctenoides; and [3] Hymenolepididae: Hymenolepis diminuta. The infective oncospheres of these seven species were compared with each other with respect to the following five cell types: (1) a bi-nucleate subtegumental cell; (2) a bi- nucleate penetretion gland; (3) two nerve cells; (4) numerous somatic cells (=myocytons of both somatic and hook musculature); (5) a variable number of germinative cells (about 8-20), which have an important role in metacestode or cercoid formation. The comparison of the number of oncospheral cells by means of TEM suggest that the progressive reduction in their number represents a general trend in cestode evolution,as one of their ontogenic adaptatations to a parasitic way of life.
SY08/02-02
HOST-CELL APOPTOSIS IN TAENIA SOLIUM INDUCED BRAIN GRANULOMAS IN NATURALLY INFECTED PIGS
SIKASUNGE C.S.(1), PHIRI I.K.(1), JOHANSEN M.V.(2), WILLINGHAMIII A.L.(3), LEIFSSON P.S.(4) (1) Department of Paraclinical Studies, School of Veterinary Medicine, UNIVERSITY OF ZAMBIA, P.O.BOX 32379, LUSAKA, ZAMBIA ; (2) DBL - Centre for Health Research and Development, FACULTY OF LIFE SCIENCES, UNIVERSITY OF COPENHAGEN, THORVALDSENSVEJ 57, DK-1870 FREDERIKSBERG C, DENMARK ; (3) WHO/FAO Collaborating Centre for Parasitic Zoonoses, Faculty of Life Sciences, UNIVERSITY OF COPENHAGEN, DYRLæGEVEJ 100, 1870 FREDERIKSBERG C, DENMARK ; (4) Department of Veterinary Pathobiology, Faculty of Life Sciences, UNIVERSITY OF COPENHAGEN, RIDEBANEVEJ 3, 1870 FREDERIKSBERG C,
SY08/01-06
PRIMARY CELL CULTURE CHARACTERIZATION FROM ECHINOCOCCUS GRANULOSUS LARVAL STAGE
ALBANI C.M., ELISSONDO M.C., CHISARI A., DENEGRI G.M. Universidad Nacional de Mar del Plata, MAR DEL PLATA, ARGENTINA The ability to cultivate helminth cells in vitro or to establish helminth cell lines is very difficult and only few cases have been described in the literature. The aim of this work was to characterize a primary cell culture from Echinococcus 63
DENMARK
The increase in malaria burden is certainly caused by the resistance of the parasite to drugs and by the high fatality rate carries by cerebral malaria despite chemotherapy. In both case, research on apoptosis could pave the way of new options to resolve a part of this terrific problem. First, we demonstrated that Plasmodium apoptosis could be triggered by quinolines and that differences in apoptosis capability among clones could be related to drug resistance. The relationship between loss of apoptosis and CQ resistance could provide the opportunity to develop a different lead drug targeting metacaspase regulation. Second, one must admit that the therapeutic efficacy of new antimalarial drugs is not associated with improved survival, suggesting a role for adjunctive therapies in the early phase of the disease. We are now using erythropoietin for a clinical trial in endemic area to demonstrate the concept of neuroprotection during cerebral malaria. Taken together, these experiments provide more evidence that deciphering the apoptosis mechanisms and the regulation of life-or-death decision in Plasmodium and human brain will drive the way from research lab to endemic areas in a confidently hurried manner.
To assess whether apoptosis occurs in pig brain granulomas due to T. solium cysticerci, brain tissues from 30 pigs naturally infected with T. solium cysticercosis were evaluated by terminal deoxynucleotidyl transferase-end labeling (TUNEL) staining. In addition, tissues were stained with CD3 marker to identify T lymphocytes. Examination of TUNEL stained tissues showed apoptotic cells in early lesions that contained live or viable cysticerci. Apoptotic cells were primarily found interspersed with normal cell types, and were mostly located in the inflammatory infiltrate. Late or advanced granulomas with disintegrated scolices did not show TUNEL positive cells. CD3+ cells were found in both early and advanced lesions and apoptosis mainly colocalized with CD3+ T lymphocytes. This suggests that these cells are constantly undergoing apoptosis and thus die as soon as they arrive at site of infection. Apoptosis indeed may be one way by which T. solium cysticerci down-regulate the host’s cellular immune response in early cysticercosis. Therefore, further research is needed to establish if other cells besides T- lymphocytes are also a target for destruction by cysticerci in early cysticercosis as well as studies to assess if cysteine protease is expressed by viable cysticerci in situ.
SY08/03-01
HOW THE TOXO1 LOCUS TOXOPLASMOSIS OUTCOME
SY08/02-03
FEATURES OF PLASMODIUM FALCIPARUM APOPTOSIS AND ROLE OF THE PFMCA1 METACASPASE PROTEIN
DIRECTS
CAVAILLÈS P.(1), BISANZ C.(1), PAPAPIETRO O.(2), FOURNIÉ G.J.(2), CESBRON-DELAUW M.F.(1) (1) Adaptation et Pathogénie des Microorganismes, UMR 5163, Institut Jean Roget, GRENOBLE, FRANCE ; (2) INSERM U563 CPTP, TOULOUSE, FRANCE
MESLIN B., BEAVOGUI A.H., BARNADAS C., KAISER K., PICOT S. UCBL, LYON, FRANCE
Toxoplasmosis remains a healthcare problem in pregnant women and immunocompromised patients. Like humans, rats usually develop a subclinical infection and the LEW rat exhibits a peculiar refractoriness to T.gondii infection (no antibody response and no cyst). This is a unique model of total resistance to toxoplasmosis that enabled genetic studies between LEW (resistant) and BN (susceptible) rats. Linkage analysis in (LEW/BN) F2 rats identified a single locus located on chromosome 10 that controls toxoplasmosis outcome, we named Toxo1. Positional cloning using reciprocal congenic lines LEW.BNc10 and BN.LEWc10 narrowed down Toxo1 locus to a 1.5 Mb region including 39 genes. The role played by macrophages in the immune defense against T. gondii led us to investigate both penetration and proliferation of the parasite in rat peritoneal macrophages in vitro. Whereas there is no difference in the invasion rate, Toxo1 of Lew origin inhibits 90% of Toxoplasma proliferation in macrophages. Beside the fact that Toxo1 controls parasite proliferation, additional functional studies have highlighted the role of Toxo1 in the control of cell. Based on these results, we identified 2 major candidate genes in Toxo1 that could be responsible of these phenotypes and are now working on the role of these genes in the outcome of toxoplasmosis.
The ability to undergo apoptosis, previously thought to be exclusive to multicellular organisms, has been demonstrated in unicellular parasites. Concerning Plasmodium, causing agent of malaria, apoptosis figures as a key component in life cycle success. Here we report evidence for Plasmodium falciparum erythrocytic stage apoptosis upon chloroquine treatment. Cell condensation has been detected with the presence of DNA fragmentation (TUNEL-positive nuclei) and similar mitochondrial depolarization patterns have been observed after incubation with chloroquine and CCCP a well-known depolarizing reagent (JC1 staining). Concerning apoptosis pathway, metacaspases have been described as caspase-like proteins in plants and several unicellular organisms. Three metacaspases like proteins have already been annotated in P. falciparum genome Pf13_0289, PF14_0363 and Pf14_0160 (PfMCA1, PfMCA2 and PfMCA3 respectively). Sequences analysis of PfMCA1 revealed presence of a catalytic caspase domains and a putative caspase recruitment domain (CARD) belonging to the death domain (DD) superfamily. To investigate molecular characterization, PfMCA1 was cloned and expressed in several heterologous systems to examine its processing and enzymatic activity. It has been shown a prodomain cleavage according to the common caspase maturation processing, and a metacaspase-like activity indicating PfMCA1 could be involved in P. falciparum apoptosis.
SY08/02-04
APOPTOSIS AND MALARIA: FROM BENCH TO BED
PICOT S., MESLIN B., BEAVOGUI A.H., BIENVENU A.L. Hospices Civils de Lyon, Malaria Research Unit, EA 4170, Université Claude Bernard, LYON, FRANCE 64
SY08/03-02
SY08/03-04
IDENTIFICATION OF ANOPHELES GAMBIAE GENES THAT AFFECT PLASMODIUM INFECTION
PROTEOMIC ANALYSIS OF LOW ABUNDANCE PROTEINS INVOLVED IN HOST-PARASITIC INTERACTION IN SHEEP SELECTED FOR RESISTANCE AND/OR SUSCEPTIBILITY TO TRICHOSTRONGYLUS COLUBRIFORMIS
JARAMILLO GUTIERREZ G.(1), BRANDT .S.(2), KUMAR S.(3), MOLINA-CRUZ A.(3), SCHNEIDER D.(2), BARILLAS-MURY C.(3) (1) UNIVERSITE LIBRE DE BRUXELLES, GOSSELIES, BELGIUM ; (2) STANDFORD UNIVERSITY, CALIFORNIA, UNITED STATES ; (3) NATIONAL INSTITUTE OF HEALTH, ROCKVILLE, UNITED STATES
BINOS S.(1), BUCK C.(1), ANDRONICOS N.(2), WINDON R.(2), MCDONAGH M.(1) (1) DPI Victoria, Bioscience Research Division, MELBOURNE, AUSTRALIA ; (2) CSIRO Livestock Industries, ARMIDALE, NSW, AUSTRALIA
Malaria parasites must complete a complex developmental cycle in Anopheles mosquito before transmission to a vertebrate host. Mosquitoes vary greatly in their ability to transmit malaria, but the molecular basis for these differences is poorly understood. Previous studies indicate that in vitro cultured ookinetes develop to mature oocysts when injected into Drosophila. A genetic screen identified several mutations that significantly reduced or increased Drosophila’s ability to support oocyst development. In the present study, we have identified homologues for six selected genes in Anopheles gambiae, and confirmed their expression in adult females. dsRNA-mediated knockdown of five of these genes affected Plasmodium berghei infectivity in the mosquito. None of the identified genes have been previously implicated in innate immune responses or interactions with Plasmodium. Moreover, four of these selected genes also affect An. gambiae infection with the human malaria parasite P. falciparum. We conclude that the Drosophila-Plasmodium model was successful in identifying unexpected genes that affect P. berghei and P. falciparum development in the mosquito using a genome-wide mutagenesis strategy.
Gastrointestinal nematodes remain the major cause of losses in productivity for the sheep meat and wool industries. Increased resistance of nematodes to current anthelmintics has resulted in the need to develop alternate parasite control strategies, including breeding for resistance. The plasma proteome has enormous biochemical diversity containing proteins of wide dynamic range in concentration. Proteins of interest which play a key physiological role in host-parasite interaction and which potential aid with resistance to parasitic challenge are usually present in plasma as low abundant proteins (LAP). To enhance detection and quantification therefore for these biomarkers of resistance, it is necessary to deplete plasma of proteins in high and medium abundance (HMAP) that constitute more than 95% of the total plasma proteome. In this study a plasma subproteome of sheep selected for resistance or susceptibility to infection with the nematode Trichostrongylus colubriformis have been quantitatively compared by I-TRAQ labelling following HMAP depletion and Liquid Chromatography/Mass Spectrometry (LC/MS). We anticipate that by identifying differences in the proteins expressed in these animals during critical times of infection/challenge we will enhance our understanding of host-parasite interplay. Ultimately this knowledge may also assist with selecting sheep for resistance to gastrointestinal nematodes.
SY08/03-03
FREP 4: A MODEL OF A TREMATODERESPONSIVE POLYPEPTIDE IN BIOMPHALARIA GLABRATA HEMOLYMPH
SY08/03-05
ADEMA C.M., ZHANG S.M., LOKER E.S. University of New Mexico, ALBUQUERQUE, UNITED STATES
MOLECULAR CHARACTERISATION OF TOTAL AND DIFFERENTIALLY EXPRESSED SECRETOME OF TRYPANOSOMA CONGOLENSE: IDENTIFICATION OF NOVEL VIRULENCE AND/OR PATHOGENIC FACTORS.
Fibrinogen-related proteins (FREPs) are produced by the snail Biomphalaria glabrata following infection with digenetic trematodes, most prominently following exposure to Echinostosma paraensei. Of the known FREPs, the one most consistently upregulated following infection is FREP4. Recombinant FREP4 has been produced and used to generate antibodies that specifically bind FREP4 in profiles of snail blood, a finding confirmed using proteomics. Using this antibody, we show that FREP4 increases sharply in abundance in snail blood following exposure, and that this band quickly broadens to encompass a range of molecular weights. The breadth of this band is not a simple function of glycosylation. As some FREP4 sequences examined to date are variable, and given that Southern analysis indicates that FREP4 is represented by only 1-3 coding sequences, the possibility is being investigated that FREP4 is diversified, potentially in hemocyte populations expanding as a consequence of infection. Four BAC clones containing FREP4 have been found and are being investigated for associated regulatory sequences, and our use of a >1152 feature oligo-based microarray supports earlier observations that FREP4 is the most consistently and strongly upregulated of all the known FREPs. FREP4 can provide basic insights into the nature of the invertebrate response to parasites. This study is supported by NIH grants RO1 AI24340 (ESL), AI52363 (CMA) and AI67686 (SMZ).
HOLZMULLER P.(1), GRÉBAUT P.(2), BRIZARD J.P.(2), SEVENO M.(3), DEMETTRE E.(3), JOUIN P.(3), VINCENDEAU P.(4), BENGALY Z.(5), BOULANGÉ A.(1), CUNY G.(2) (1) CIRAD, MONTPELLIER, FRANCE ; (2) IRD, MONTPELLIER, FRANCE ; (3) IGF, MONTPELLIER, FRANCE ; (4) Université Bordeaux 2, BORDEAUX, FRANCE ; (5) CIRDES, BOBODIOULASSO, BURKINA FASO
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Animal trypanosomosis is one of the most severe constraints to agricultural development in Sub-Saharan Africa and is also an important disease of livestock in Latin America and Asia. The causative agents are various species of protozoan parasites belonging to the genus Trypanosoma, among which T. congolense and T. evansi are the major pathogenic species. The extracellular position of the trypanosomes implies to consider both the parasite and its excreted-secreted factors (secretome) in the course of both parasite multiplication in the host and the physiopathological processes. The postgenomic era stimulated the development of new techniques and bioinformatics tools to identify the locations, functions and interactions of the gene products in living organisms. These advances in proteomics led us to propose both a global and a comparative approach of the secretome of T. congolense and
and influences reproductive success of the analyzed S.mansoni populations. For the direct analysis of the epigenotype of S.mansoni we developed a native chromatin immunoprecipitation protocol that requires a minimum amount of cells, and we demonstrate what chromatin modifications take place during the development of the parasite.
T. evansi. T. congolense clones of differing virulence and pathogenicity were used as a model to improve the proteomic approach to highlight novel molecules related to virulence and pathogenicity. The molecular identification of differentially expressed molecules correlated with either the virulence process or exhibiting potential properties to induce pathogenic dysregulation are promising to define new potential molecular targets for improved field diagnosis and new strategies of interference with the infectious process of animal trypanosomosis.
SY08/04-03
AN EXPERIMENTAL APPROACH FOR THE SCREENING OF TRYPANOSOMATID CONSERVED SECRETED PROTEINS
SY08/04-01
CORRALES R.M., MATHIEU-DAUDÉ F., GARCIA D., BRENIÈRE S.F., SERENO D. Institut de Recherche pour le Developpement, MONTPELLIER, FRANCE
MOLECULAR DETERMINATION OF GAMETOCYTOGENESIS IN PLASMODIUM FALCIPARUM
WITKOWSKI B.(1), BENOIT-VICAL F.(1), DEYMIER C.(2), SERTORIO M.(2), BERRY A.(2) (1) LCC CNRS, TOULOUSE, FRANCE ; (2) Service de Parasitologie CHU Rangueil, TOULOUSE, FRANCE
Among the most important protozoan parasitic species causing disease in humans, Leishmania major, Trypanosoma cruzi and Trypanosoma brucei have had their genomes sequenced recently. In order to complete their life cycle, encompassing an insect vector and a vertebrate host, these pathogens have developed various strategies to shape their environment, modulate the host immune response and invade the target cell. Materials secreted by the parasite play pivotal roles in these processes and may therefore represent targets for vaccines or rational drug design. In order to identify new trypanosomatid conserved secreted proteins through the classical secretory pathway; we combined a web-based bioinformatic screening approach with a functional test that takes advantage of the facility to genetically transform Leishmania. Thirteen hypothetical conserved proteins in trypanosomatids were selected by bioinformatic screening of the three genome databases as those potentially secreted via the endoplasmic reticulum. Transfection of these putative secreted proteins in Leishmania and detection of the tagged proteins in the extracellular medium confirmed the computer predictions in about 25% of the cases. Thus, the bioinformatic method, combined with the functional test, provides a reliable and accurate method for the identification of secreted proteins involved in the classical secretory pathway of trypanosomatids.
The gametocytes, sexual blood-stages of P. falciparum, represent the parasite stage that can be transmitted to the mosquito vector during the infected bloodmeal. Whereas microscopic determination of gametocytogenesis is long, painful and poorly discriminating, here we provide a molecular methodology to predict in vitro gametocytogenesis potency of various Plasmodium strains. Quantification, trough RT-PCR, of four specific sexual stage genes: Psf16, Psf25, Pfg27 and S 18S rRNA were investigated on in vitro culture of Plasmodium falciparum strains. Our work shows that the early overexpression of the only Pfs 16 onto the first 72h of parasite culture is correlated with the capacity to produce in vitro mature gametocytes (14 days later). These results were validated by microscopic method. This protocol is a fast and efficient tool to predict sexual stages production and in fine to study transmission potency of the parasites in various conditions and especially in presence of antimalarial compounds.
SY08/04-02
THE ROLE OF EPIGENETICS IN THE GENERATION OF PHENOTYPIC VARIABILITY IN HOST-PARASITE INTERACTIONS: THE BIOMPHALARIA GLABRATA/ SCHISTOSOMA MANSONI MODEL
SY08/04-04
HIGHLY POLYMORPHIC MICROSATELLITES IN NEOSPORA CANINUM
BAZZOCCHI C.(1), CERUTTI M.C.(1), KRAMER L.(2), GRANDI G.(2), SASSERA D.(1), BANDI C.(1), FERRI E.(1), GENCHI C.(1) (1) Facoltà di medicina Veterinaria; University of Milan, MILANO, ITALY ; (2) Facoltà di Medicina Veterinaria, University of Parma, PARMA, ITALY
COSSEAU C., MITTA G., AZZI A., GRUNAU C. Université de Perpignan, PERPIGNAN, FRANCE
Epigenetics is the science of heritable but modifiable information. We believe that the reason for the existence of such an alternative information transmitting system lies in the evolutionary advantage that provide epigenetic mechanisms. Theoretical considerations support a model in which epigenetic variability increases phenotypic variability (i.e. the number of different phenotypes in a population) and allows populations to explore the adaptive landscape and change their fitness prior to modifications of the genotype. In this model, epigenetic variability is induced by environmental changes. Phenotypic variability is particular important for rapidly evolving biological systems under high selective pressure such as parasites and their hosts. These systems such as the human blood fluke S.mansoni and its intermediate host, the fresh-water snail B.glabrata, are therefore models of choice for the study of epigenetic processes that create phenotypic variations. We show here for the first time that direct modulation of the epigenetic variability increases indeed the phenotypic variability
Neospora caninum is a coccidian parasite that was first associated with neuromuscular disorders in dogs and soon after recognized as a major cause of infectious abortion in cattle worldwide, causing important economic losses. N. caninum is transmitted horizontally via ingestion of bradyzoites or oocysts and across the placenta. Several strains of N. caninum have been isolated from cattle and dogs and, based on the worldwide distribution of the parasite and its potential capacity for sexual reproduction, a large genetic polymorphism in N. caninum is expected and preliminary demonstrated. Consequently, the purposes of this study were: 1) to identify, using a microsatellites approach, a method for the evaluation of the genetic intraspecies diversity of this parasite; 2) to understand the epidemiology and pathogenicity of N. caninum in field analysis. Sequences of N. caninum derived from expressed sequence tags of Nc-1 tachyzoite and Nc-Liv 66
between resistant alleles of P. falciparum parasites that share ancestry and those that have arisen independently. Little information is available to assess the evolutionary origin of P. vivax drug resistant parasites. Our study was conducted in Madagascar between 2006 and 2007. 160 P. vivax isolates were collected from eight sentinel sites. Pvdhfr and dhps genes were amplified and sequenced. We scanned 10 kb flanking P. vivax dhfr and dhps genes and chose 5 microsatellite markers around dhfr and 3 around dhps which were amplified and sequenced too. 71% percent of our isolates were of mutant type in the dhfr gene (single to triple mutant alleles) and 16% in the dhps gene (single mutant allele). Microsatellites were found to be polymorphic with 2 to 8 alleles each, mean values of 5. Expected heterozygosity at each locus was calculated. We didn’t find an association between pvdhfr and pvdhps mutations and the flanking microsatellite alleles in our isolates, even in triple dhfr mutant genotypes. Mutant genotypes seem to have arisen from multiple origins. The role of microsatellite markers in the monitoring of drug resistant P. vivax parasites should be further explored.
tachyzoite cDNA libraries have been screened for microsatellite identification. Microsatellites were analysed with GeneMapper program using a ABI Prism 310 Genetic Analyser. Three microsatellite markers exhibited high polymorphism within 10 N. caninum isolates showing from 7 to 8 alleles each. These three markers have then been used for identifications of N. caninum strains in different cattle farms in the North of Italy.
SY08/04-05
GENOTYPING OF ECHINOCOCCUS GRANULOSUS SENSU LATO: OVERVIEW OF THE MITOCHONDRIAL COX1 AND NAD1 POLYMORPHISM AND EVALUATION OF THE MULTILOCUS MICROSATELLITE EMSB
MAILLARD S.(1), MA S.(2), HAAG K.L.(3), GOTTSTEIN B.(4), RADONJIC I.(5), BENCHIKH-ELFEGOUN M.C.(6), KNAAP J.(1), PIARROUX R.(1) (1) Laboratoire de Parasitologie et de Mycologie, BESANCON, FRANCE ; (2) Institute of Parasitology, Medical College of Qinghai University, XINING CITY, CHINA ; (3) Universidade Federal do Rio Grande do Sul, PORTO ALEGRE, BRAZIL ; (4) Institute of Parasitology, University of Bern, BERN, SWITZERLAND ; (5) Institute of Microbiology and Immunology, University of Belgrade Faculty of Medicine, BELGRADE, SERBIA AND MONTENEGRO ; (6) Laboratory of Parasitology, University of Mentouri, CONSTANTINE, ALGERIA
SY09/01-02
NEW GENES OF HARD TICK IXODES RICINUS, INVOLVED IN TICK INNATE IMMUNITY: MLDOMAIN CONTAINING PROTEIN, DER-P2LIKE ALLERGEN AND TICK RECEPTOR FOR OSPA (TROSPA).
HORACKOVA J.(1), RUDENKO N.(2), GOLOVCHENKO M.(2), GRUBHOFFER L.(2) (1) Faculty of Sciences, University of South Bohemia, CESKE BUDEJOVICE, CZECH REPUBLIC ; (2) Biology Centre AS CR, CESKE BUDEJOVICE, CZECH REPUBLIC
Previously composed by 10 strains (G1-G10), the taxonomy of Echinococcus granulosus sensu lato has been progressively revised. The genetic polymorphism of this cestoda is currently assessed with mitochondrial markers. However, their relative homogeneity within a given strain limits their use for epidemiological investigation. Furthermore, the cross-fertilizing processes can not be dectected in this approach. The tandemly repeated multilocus microsatellite EmsB has recently proved its contribution in three epidemiological studies of E. multilocularis. In order to evaluate its potential usefulness for E. granulosus s.l., we studied 160 hydatid cysts collected in Algeria, Mauritania, Ethiopia, Romania, Serbia, China and Brazil, from various intermediate hosts (sheep, cattle, pigs, dromedaries and human patients). With the mitochondrial coxI and nadI markers, we distinguished five known genotypes, as well as some microvariants. They were clustered in three groups, E. granulosus sensu stricto, E. ortleppi and E. canadensis. The microsatellite EmsB showed a higher polymorphism and discriminated the cysts according their geographical origin. In addition, one of the brazilian cyst exhibited an hybrid profile suggesting genetic exchanges between two different species (E. granulosus s.s. and E. ortleppi).
Tick Ixodes ricinus is the vector of many pathogens in Europe. Expression of many tick genes is regulated by blood feeding or pathogen invasion. New genes encoding for proteins that are involved in tick innate immunity are presented below. Genes encoding two proteins from ML (MD-2-lipid recognition) domain protein family, characterized by presence of N-terminal signal peptide and six conserved cysteine residues, were unknown in hard ticks until now. Der-p2-like allergen and MLdomain containing protein have putative binding sites for lipids. Expression of genes is strongly induced by blood intake, which can indirectly confirm the participation of these proteins in lipid transport or in recognition of lipidic molecules on the surface of the pathogens. Protein products of these intronless genes belong to the group II of ML domain protein family. Expression of the gene for tick receptor for OspA (TROSPA) was induced in I. ricinus by infection with Borrelia burgdorferi. Analysis of the genomic sequence showed that this gene contains 2 exons (404 bp and 93 bp long) and one phase 2 intron (479 bp long). The genomic organization of I. ricinus TROSPA was similar to one of I. scapularis with small difference in the length of the first exon: 404 bp in I. ricinus and 392 bp in I. scapularis, respectively. The nucleotide substitutions that caused the amino acid changes and several deletions in I. scapularis gene were observed in comparison to I. ricinus gene.
SY08/04-06
MULTIPLE ORIGIN OF PLASMODIUM VIVAX DHFR AND DHPS MUTANT ISOLATES FROM MADAGASCAR
BARNADAS C.(1), TICHIT M.(2), BOUCHIER C.(2), PICOT S.(1), MÉNARD D.(3) (1) EA4170, LYON, FRANCE ; (2) Plate forme génomique 1; Institut Pasteur, PARIS, FRANCE ; (3) Unité de recherche sur le paludisme; Institut Pasteur de Madagascar, ANTANANARIVO, MADAGASCAR Genetic basis of resistance to sulphadoxine pyriméthamine is point mutations in genes encoding the target enzymes of the drug (DHFR and DHPS). Microsatellite markers located in the flanking regions of these genes have been used to distinguish 67
exposed to MLCr and ConAAg pulsed DCs. There was no significant change in the CD4+CD25+FoxP3 cell population compared to control cultures. Results obtained in this work suggest the existence of more balanced Th1/Th2 response to TS antigens instead of exclusive predominance of Th2 immune response.
SY09/01-03
TICK(S) DEFENCE PROTEINS: SIMILARITY, SITE OF EXPRESSION AND STAGE-SPECIFIC EXPRESSION.
MATEJOVSKA T.(1), RUDENKO N.(2), GOLOVCHENKO M.(2), GRUBHOFFER L.(2) (1) Faculty of Sciences, University of South Bohemia, CESKE BUDEJOVICE, CZECH REPUBLIC ; (2) Biological Centre AV CR, CESKE BUDEJOVICE, CZECH REPUBLIC
SY09/01-05
DIFFERENTIAL EFFECT OF DENDRITIC CELL MATURATION BY EXCRETORY/SECRETORY ANTIGENS DERIVED FROM DIFFERENT HELMINTH SPECIES
Ixodes ricinus is the principal European vector of severe human diseases such as Lyme disease and Tick-borne encephalitis. Immune responses in ticks involve proteins of defence reactions or pathogen pattern recognition. Defensins are effectors of tick immune system. They are induced by bloodfeeding or pathogen invasion and are known for their antimicrobial activity against bacteria, fungi and protozoan parasites. Defensins were identified in Amblyomma americanum (1 isoform), A. hebraeum (2), Boophilus microplus (1), Dermacentor variabilis (2), Hyalomma longicornis (2), I. scapularis (1), Ornithodoros moubata (4). Some species expressed more than one defensin isoform. Recently, we showed that 2 isoforms of I. ricinus defensin are involved in its immunity. Def1 is expressed in midgut, while Def2 - in midgut, ovary, malphigian tubes and salivary glands. Both peptides were induced by blood feeding in adult females, nymphs and larvae. They differ in 3 amino acids and show the intron/exon genomic structure. We found defensin peptides in D. reticulatus (1), D. marginatus (1), O. tartakovskyi (1), O. papillipes (2), O. puertoricensis (2) and O. rastratus (2). While mature part of newly identified defensins was conserved among the species, the prepropeptide was rather heterogeneous. The diversity of defensins from different tick species might explain the competence of I. ricinus as a vector of B. burgdorferi and TBEV in comparison to nonvector ticks.
LANGELAAR M.(1), ARANZAMENDI C.(1), FRANSSEN F.(1), VAN DER GIESSEN J.(1), VAN DIE I.(2), RUTTEN V.(3), PINELLI E.(1) (1) RIVM, BILTHOVEN, NETHERLANDS ; (2) VU Medical Center, AMSTERDAM, NETHERLANDS ; (3) Faculty of veterinary medicine, UTRECHT, NETHERLANDS During chronic infections with helminths a Th2 type of immune response and a regulatory network are generally induced. These responses can benefit parasite survival and can also affect the host’s immune response to other pathologies such as allergies. Depending on the helminth species, protection against and enhancement of allergic manifestations have both been reported. The present study aims at determining the effect of antigens derived from different helminth species on APC maturation in vitro. For this purpose, we used excretory/secretory (E/S) antigens derived from the roundworms Toxocara canis and Trichinella spiralis. As antigen presenting cells (APC) murine bone marrow derived dendritic cells (DC) were used. These cells were incubated with the helminth E/S antigens, alone or in combination with different TLR ligands. The cytokine production and the expression of various surface markers were measured. Results indicate different effects of helminth antigens on APC maturation. While helminth antigens alone do not significantly affect the production of the various cytokines or expression of surface markers, Trichinella spiralis, but not Toxocara E/S antigen suppresses maturation of immature DC induced by E.coli LPS. Our results indicate that the outcome of DC maturation can be affected differently depending on the helminth species. These results could explain the different findings on the association between helminth infection and allergy.
SY09/01-04
THE ROLE OF DENDRITIC CELLS IN PRESENTATION OF DIFFERENT TRICHINELLA SPIRALIS ANTIGENS AND SUBSEQUENT T CELL POLARIZATION
ILIC N.(1), HEPWORTH M.(2), GRUDEN-MOVSESIJAN A.(1), SOFRONIC-MILOSAVLJEVIC L.(1), GRENCIS R.(2) (1) Institute for the Application of Nuclear Energy - INEP, BELGRADE, SERBIA AND MONTENEGRO ; (2) Faculty of Life Science, University of Manchester, MANCHESTER, UNITED KINGDOM
SY09/02-03
IN VIVO PROTECTIVE ROLE OF IMMUNOGLOBULIN E AND EOSINOPHILS IN MURINE TRICHINELLOSIS AS SHOWN BY DOWN REGULATING TH2 RESPONSES
The role of dendritic cells (DCs) in generation and maintaining immune response to Trichinella spiralis (TS) has not been completely elucidated. In this study we utilized antigens from all three life stages of TS (muscle larvae antigens: crude – MLCr; Concanavalin A selected components of MLCr – ConAAg; excretory-secretory products – ES L1; deglicosilated fraction of ES L1; excretory-secretory products of adult parasites – AdES; or, a soluble extract of newborn larvae – NBLAg) to prime DCs of C57BL/6 mice. Incomplete maturation of DCs was observed since MHC II was moderately up-regulated in response to all antigens except NBL and MLCr. Major up-regulation of ICAM1 but not expression of CD80 and CD86 co-stimulatory molecules were observed with all TS antigens. Elevated levels of IL-10, IL-12p70, IL-6 and TNFalfa were produced by all antigen stimulated DCs. Naive CD4+ T cells, primed with TS antigen pulsed DCs, had increased production of IL-4, IL-9, IL-10, IL-13 and IFNgama. Decreased production of IL-6 and TNFalfa was detected in all antigen primed T cell cultures, except for those
DEL PRETE G.(1), CHIUMIENTO L.(2), AMEDEI A.(1), PIAZZA M.(2), CAPONI L.(2), D'ELIOS M.(1), CODOLO G.(3), DE BERNARD M.(3), MASETTI M.(2), BRUSCHI F.(2) (1) UNIVERSITA' DI FIRENZE, FLORENCE, ITALY ; (2) UNIVERSITA' DI PISA, PISA, ITALY ; (3) UNIVERSITA' DI PADOVA, PADOVA, ITALY AIM: In vivo evaluate the relations between blood eosinophils (Eo), total IgE (tIgE), Trichinella spiralis (Ts) specific IgE (TsIgE), plasma Creatine Kinase (CK) and aldolase levels and worm burden in both infected with Ts untreated (C) and Th2down regulated (with the recombinant Neutrophil Activating Protein from Helicobacter pylori (HP-NAP) Ts infected mice (T). METHODS: BALB/c female mice were infected with Ts larvae and half received intraperitoneal (i.p.) PBS, whereas the rest of infected animals received 10 g of HP-NAP i.p. on day 10 and 68
is controlled almost exclusively by ivermectin, which prevents pathology by targeting microfilariae. However, this reliance on a single control tool has led to interest in vaccination as a potentially complementary strategy. Eight recombinant Onchocerca proteins, selected on the basis of protective efficacy against infective larvae in rodent models and recognition by sera from putatively immune humans, were used together to immunise cattle exposed to natural transmission of O. ochengi. After 22 months, vaccination had not affected the burden of adult parasites, but patent infection (microfilaridermia) was reduced by 42% (P = .015). Serological responses were mounted to all eight proteins, and the ratio of IgG1:IgG2 was apparently modulated by the adjuvant selected for antigen delivery. As microfilariae appear to have been targeted by this vaccine despite an intention to protect against infective larvae, the development of an anti-microfilarial vaccine deserves specific attention.
28 post infection. Blood total leukocyte and differential counts, tIgE, Ts-IgE and CK and aldolase levels were evaluated at different days post infection (d.p.i.). Parasite burden as muscle larvae (ML) obtained by artificial digestion of each single mouse carcass was estimated at 56 d.p.i. RESULTS: The ML burden was lower (p < 0.02) in C than in T. In both groups there was an inverse relationship between ML burden and tIgE levels (r – 0.617, p = 0.0013 and r -0.678, p = 0.0001, respectively), Ts-IgE levels (r -0.529, p < 0.01) and more significantly in T (r -0.601, p < 0.001) or Eo on day 42 (r -0.390, p = 0.0592 and r -0.803, p = 0.0001). No significant relation was observed between worm burden and plasma CK or aldolase levels, in both groups. CONCLUSIONS: This study provides evidence for a partial protective effect of Th2 responses in Ts infection, more evident in Th2 down regulated animals.
SY09/02-04 SY09/03-02
NATURALLY ACQUIRED ANTIBODIES TO THE VARO VARIANT FROM P. FALCIPARUM PFEMP1 PROTEIN IN BENINESE CHILDREN
EVALUATION OF PROTECTIVE EFFECT OF RECOMBINANT PROTEIN OF EIMERIA USED AS AN ADJUVANT AGAINST CHRONICAL TOXOPLASMOSIS IN MICE
LOKOSSOU A.G.(1), VIGAN-WOMAS I.(2), GUILLOTTE M.(2), MERCEREAU-PUIJALON O.(2), MIGOT-NABIAS F.(1) (1) IRD, COTONOU, BENIN ; (2) Institut Pasteur, PARIS, FRANCE
HEDHLI D., MÉVÉLEC M.N., DIMIER-POISSON I. UFR sciences pharmaceutiques, TOURS, FRANCE
The Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is an important virulence factor on the surface of infected erythrocytes. Naturally acquired antibodies to PfEMP1 expressed by parasites causing severe malaria are suggested to be protective and of major interest for vaccine development. The VarO serotype, derived from the Palo Alto line, has strong properties of rosetting adhesion (mainly mediated by the DBL1a domain), demonstrated in Saimiri sciureus monkeys, and reproduced in in vitro infected human erythrocytes. Antibodies directed to VarO DBL1a domain reverse rosetting in vitro, but information on the human antibody reactivity to VarO expressing parasites is lacking. Specific IgG and subclasses to recombinant proteins of VarO domains (DBL1a, DBL2bC2 and CIDRg) were determined in Beninese children (mean age 4.5 years) including 65 and 37 children with severe and uncomplicated malaria, respectively, and 52 asymptomatic carriers of P. falciparum. IgG responders were older than others (all P < 0.0001) with prevalence rates ranging from 23 to 54%. DBL1a generated higher IgG levels than other proteins, whatever the clinical presentation of malaria (all P < 0.0001). Asymptomatic children had higher IgG levels to all proteins than symptomatic ones (all P < 0.0001). These results established the existence of a clinically-related antibody reactivity to VarO in humans in natural conditions of exposure to P. falciparum parasites.
The important impact of Toxoplasma gondii in human and veterinary medecine leads us to characterize some immunological mechanisms involved in host resistance to infection within the framework of a vaccinal strategie. In this work we studied the effect of the Toxoplasma gondii RH total antigen (TAg) co-delivered with recombinant antigen of Eimeria (rEA) (a profilin like protein and an inducer of protozoantargeted innate immunity) as an adjuvant in CBA/J mice against chronical stage of toxoplasmosis. TAg was administrated three times with or without rEA, by nasal or intraperitoneal routes. Results shows that mice immunized with TAg-rEA present significant higher IgG titer and IL-12 rate in their sera then mice immunized with TAg alone. In vitro we determine an upregulation of specific IL-2 and IFN- by spleen cells stimulated with TAg. Investigation of protection against the chronic phase of infection shows a significant decrease in the number of tissue cysts from the TAg-rEA immunized group (71% and 55% of reduction when immunization by intraperitoneal and nasal routes respectively) compared with non immunized group. Mice immunized by TAg alone present less protection rates (54% and 1% of reduction when immunization by intraperitoneal and nasal routes respectively) compared with control group. Our data demonstrated that protection was improved by associating TAg and rEA for further developement of rEA as an adjuvant candidate.
SY09/03-01
SY09/03-03
FIRST FIELD TRIAL OF A RECOMBINANT VACCINE AGAINST ONCHOCERCIASIS: EVALUATION IN CATTLE EXPOSED TO ONCHOCERCA OCHENGI
CD8+ BUT NOT CD4+ T CELLS MEDIATE ANTITOXOPLASMA GONDII IMMUNITY IN A CHRONIC MURINE MODEL BY IMMUNIZATION WITH PULSED DENDRITIC CELLS
MAKEPEACE B.(1), JENSEN S.A.(1), LANEY S.(2), NFON C.(1), NJONGMETA L.(1), TANYA V.(3), WILLIAMS S.(2), BIANCO A.(1), TREES A.(1) (1) Liverpool School of Tropical Medicine, LIVERPOOL, UNITED KINGDOM ; (2) Smith College, NORTHAMPTON, MA, UNITED STATES ; (3) Institut de Recherche Agricole pour le Développement, NGAOUNDÉRÉ, CAMEROON
GUITON R., ZAGANI R., DIMIER-POISSON I. UFR des Sciences Pharmaceutiques, TOURS, FRANCE
Onchocerca volvulus (the aetiological agent of River Blindness) 69
Toxoplasma gondii, an obligate intracellular protozoan, is the causative agent of toxoplasmosis. However, an effective vaccine for this disease still needed. Dendritic cells trigger early and specific immune responses in murine toxoplasmosis. Indeed, they synthesize IL-12 and activate a protective Th1
response. Vaccination with pulsed dendritic cells is very efficient, but the interacting lymphocytic effectors have not been determined. Therefore, we carried out in vivo depletion of the two major lymphocyte populations, CD4+ or CD8+, and studied the subsequent cellular immune response and protective mechanisms induced. Our findings suggested that CD4+ T lymphocytes only play a minor role in Th1 cytokine secretion, Th2 cytokine negative regulation and are not involved in the protection of mice. By contrast, the increasing number of intracerebral cysts and depletion of CD8+ T cells were strongly correlated, revealing a prominent role for CD8+ T cells in the protection of mice. We also found that splenic CD8+ T lymphocytes induce IFN- and IL-10 production whereas CD8+ T lymphocytes from mesenteric lymph nodes induce IFN- production but inhibit IL-10 production. We concluded that CD8+ cells are the main effectors following dendritic cell vaccination.
vaccine development against malaria. Present on both merozoite and sporozoite stage, AMA1 is a critical invasion molecule of the apicomplexans. Despite strong animal data supporting the vaccine candidacy of AMA1, the genetic diversity of the parasite vis-à-vis its AMA1 sequence is a cause of concern to vaccinologists. About 10% of AMA1 amino acids are polymorphic and immune response to AMA1 is highly straindependent. Our lab is mapping the strain-specific and crossreactive inhibitory epitopes of AMA1. Using chimeric proteins and an in vitro merozoite growth inhibition assay (GIA), we have recently mapped the Antigenic Escape Residues (AER) of AMA1 (Dutta et al. Proc Natl Acad Sci USA. 2007; 104:1248893). The AER are polymorphic sites, that localize within inhibitory epitopes of AMA1, and mutations at the AER confer escape from invasion inhibitory antibodies. Chimeric proteins containing AER from two different strains induce antibodies that cause high level inhibition of both the target strains. AER map can be used to predict the spectrum of efficacy of a vaccine during Phase 2 trials and for formulating a multi-allelic cocktail vaccine. Using chimeric proteins in ELISA and GIA, we are now mapping the cross-reactive inhibitory epitopes of AMA1. In the future, we hope to use the cross-reactive and strain-specific epitope maps to design a Pan-reactive AMA1 vaccine.
SY09/03-04
THE CELLULAR RESPONSE OF SHEEP VACCINATED WITH A RECOMBINANT FASCIOLA HEPATICA PHOSPHOGLYCERATE KINASE (PGK) AND INFECTED WITH FLUKE METACERCARIAE.
KOZAK M., JEDLINA-PANASIUK L., WESOLOWSKA A., JAROS S., WEDRYCHOWICZ H. W. Stefanski Institute of Parasitology PAS, WARSAW, POLAND
SY10/01-03
INTRA-INDIVIDUAL VARIATION IN THE NUCLEAR RDNA IN SIBLING SPECIES OF THE ANISAKIS SIMPLEX COMPLEX (NEMATODA: ANISAKIDAE)
The aim of this study was to determine the cellular response of sheep after immunization with a novel candidate vaccine antigen, PGK. Male merino lambs were vaccinated intramuscularly with 100g of PGK cDNA and then boosted twice subcutaneously with 100 g of recombinant PGK protein. Bupivacaine and montanide were used as adjuvants, respectively. Control animals were injected with the adjuvant alone. Vaccinated and control lambs were challenged orally with 250 metacercariae, a month after the last immunization. Flow cytometry was used to investigate cell phenotypes in the blood of experimental animals. The overall immune response in both groups was dominated by a Th2 type. However in the vaccinated group, the level of CD8+ cells was higher compared to non-vaccinated one. Furthermore, there was an increase in the number of B cells, which was most visible after the second and third immunization. The percentage of cells exhibiting MHC II receptors was also elevated in the vaccinated group compared to the controls. Moreover, from the 2nd week after the challenge infection, there was a higher eosinophil response in immunized animals. These results indicate that vaccination may have stimulated a shift of the immune response towards a protective one but further research must be conducted to investigate the influence of other immunization protocols on vaccine efficacy. Research funded by the EC project DELIVER, contract No. FOOD-CT-2005-023025
D'AMELIO S.(1), BUSI M.(1), FARJALLAH S.(2), INGROSSO S.(1), CAVALLERO S.(1), PAGGI L.(1) (1) Sapienza University of Rome, ROME, ITALY ; (2) Institut Supérieur de Biotechnologie de Monastir, MONASTIR, TUNISIA Anisakis simplex s.l. comprises at least three sibling species (A. simplex s.s., A. pegreffii and A. simplex C), differing in their genetic structure and in geographic distributions and host preferences. PCR-RFLP analysis of nuclear rDNA spacers proved fruitful for the identification of sibling and morphospecies in the genus Anisakis. Previous studies conducted on larvae from fishes from Iberian and Tunisian waters revealed the existence of specimens showing heterozygote genotypes, where alleles of A. simplex s.s. and A. pegreffii were both represented. It has been suggested that these specimens may represent hybrid forms between the two species. Alternatively, intra-individual variation may involve the incomplete efficiency of concerted evolution or the persistence of ancestral polymorphisms in the rDNA spacers after speciation. To investigate whether or not heterozygote individuals may represent hybrid forms, specimens showing the RFLP pattern of A. simplex s.s., A. pegreffii and heterozygotes were analysed comparing the ITS region with two mitochondrial genes (cox2 and rrnS) and one single copy nuclear gene (EF-1). The analysis of the two mtDNA genes confirms the ability of the rDNA markers to correctly discriminate between the two species, when showing the parental alleles. The analysis of the EF-1 gene, highly polymorphic, indicates that this region is potentially reliable to unravel the nature of heterozygote individuals.
SY09/03-05
OVERCOMING ANTIGENIC DIVERSITY OF THE MALARIA VACCINE CANDIDATE APICAL MEMBRANE ANTIGEN-1.
DUTTA S.(1), SPRING M.(1), BATCHELOR A.(2), CLAYTON J.(1) (1) Division of Malaria Vaccine Development, Walter Reed Army Institute of Research, SILVER SPRING, MARYLAND, UNITED STATES ; (2) School of Pharmacy, University of Maryland, BALTIMORE, MARYLAND, UNITED STATES
SY10/01-04
LUNGWORM TRANSMISSION IN PORPOISES AND SEALS: MOLECULAR TOOLS DETERMINE VERTEBRATE INTERMEDIATE HOSTS
LEHNERT
Apical Membrane Antigen-1 (AMA1) is an important target for 70
K.(1),
SAMSON-HIMMELSTJERNA
G.V.(2),
WOHLSEIN P.(3), SCHAUDIEN D.(3), SIEBERT U.(1) (1) Research and Technology Center, Christian-AlbrechtsUniversität zu Kiel, 25761 BUESUM, GERMANY ; (2) Institute for Parasitology, Stiftung Tieraerztliche Hochschule Hannover, 30559 HANNOVER, GERMANY ; (3) Institute for Pathology, Stiftung Tieraerztliche Hochschule Hannover, HANNOVER, GERMANY
SY10/01-06
BIODIVERSITY OF PARASITES OF CORAL REEF FISH IN THE LAGOON OF NEW CALEDONIA (SOUTH PACIFIC): 10,000 SPECIES, OR MORE?
JUSTINE J.L. Institut de Recherche pour le Développement, NOUMÉA, NEW CALEDONIA
Lungworms infect a wide range of mammals and are usually found in the respiratory tract. Most terrestrial metastrongyloids are heteroxenous using molluscs as intermediate hosts. Little is known about the biology of lungworms infecting marine mammals and their transmission in the marine environment. Lungworm disease and associated pneumonia are important in the health of wild populations and a common cause of mortality in porpoises and seals. In this study we dissected various species of benthic fish from the North Sea (n=250). Larval nematodes from the gastrointestinal tract were removed. We isolated larval DNA and amplified the ITS-2 region from their rDNA. The primers used had been designed to sequence ITS2 regions from adult specimens of all six metastrongyloid species infecting porpoises and seals in the North Sea. Histology and in-situ-hybridisation using newly developed species-specific probes were performed on intestinal tissue. ITS-2 (18S rDNA control) sequences of larvae found in dab were identified as Pseudalius inflexus (Pseudaliidae), a porpoise lungworm, and sequences from plaice were determined as Parafilaroides gymnurus (Filaroididae), a seal lungworm. Turbot from aquaculture were infected experimentally with lungworm larvae. This is the first study to determine three intermediate hosts for lungworms of these marine mammals. It elucidates the so far unknown life cycle of two metastrongyloid species parasitizing seals and porpoises.
New Caledonia has the largest lagoon in the world (25,000 km2) and a very rich fish fauna, with more than 1,600 shore fish species listed. A programme has been initiated to study biodiversity of fish parasites of New Caledonia, including monogeneans, cestodes, digeneans, nematodes, and other parasites. Examples of parasite biodiversity will be given, especially for the monogeneans. For the Diplectanidae from Epinephelinae (groupers), an astonishing biodiversity has been revealed. Almost all species have diplectanids. On a single organ, the gill, one species, the highfin grouper, Epinephelus maculatus, has 10 diplectanids, plus 2 ancyrocephalids, a total of 12 monogenean species; in addition, there are also 3 species of copepods and 1 species of isopod, giving a total of 16 gill parasite species. The malabar grouper, Epinephelus malabaricus, has 11 monogeneans. The speckled blue grouper, Epinephelus cyanopodus, has 9 monogeneans, and a total of 41 species of parasites. Many of these parasite species are strictly specific. Biodiversity of fish parasites in New Caledonia is estimated to be 10,000 species. For all the coral reefs of the world, the numbers are even higher, possibly 20,000 to 30,000 species of fish parasites. As coral reefs are threatened, these figures have consequences for the numerical evaluation of endangered species; extinction of a single fish species would sometimes result in the extinction of 10–20 parasite species.
SY10/01-05
THREE-DIMENSIONAL VISUALISATION OF THE DEVELOPMENTAL STAGES OF AN APICOMPLEXAN FISH BLOOD PARASITE IN ITS INVERTEBRATE HOST
SY10/02-02
MYXOZOAN PARASITES IN TWO IMPORTANT CANDIDATE SPECIES IN MEDITERRANEAN AQUACULTURE: STRIPED SEA BREAM, LITHOGNATHUS MORMYRUS AND SHARPSNOUT SEA BREAM, DIPLODUS PUNTAZZO
HAYES P.(1), SMIT N.(2), SEDDON A.(1), WERTHEIM D.(1), DAVIES A.(1) (1) Kingston University, LONDON, UNITED KINGDOM ; (2) University of Johannesburg, JOHANNESBURG, SOUTH AFRICA Although widely used in medicine, the use of three-dimensional (3D) visualisation in parasitology to date appears to be limited. In this study, developmental stages of a marine fish haemogregarine, Haemogregarina curvata (Apicomplexa: Adeleorina), observed in histological sections of its leech vector, and their relationship to host cells were investigated by 3D visualisation of brightfield and confocal microscopy images. 3D visualisations of confocal images, in particular, through sections of infected leech salivary cells showed developmental stages of H. curvata in some detail, providing an appreciation of the shape of individual stages, their spatial layout and the reduced cellular contents of infected host cells. Meronts located marginally within salivary gland cells were observed to have small tail-like connections to the host cell membrane; this host cell-parasite interface had not been obviously visible in twodimensional (2D) light micrographs and no records of a similar connection in apicomplexan development have been traced. We believe this to be the first account of the use of 3D visualisation to study the developmental stages of an apicomplexan parasite in its invertebrate host. The elucidation of the extent of the development of the haemogregarine within the leech salivary cells, together with the unusual connections between meronts and the host cell membrane, illustrates the potential future use of 3D visualisation in this field.
ALAMA-BERMEJO G., RAGA J.A., HOLZER A.S. Cavanilles Institute of Biodiversity and Evolutionary Biology, University of Valencia, PO BOX 22085, 46071 VALENCIA, SPAIN Myxozoans have been reported as the most important pathogens in Mediterranean aquaculture systems. Histozoic myxozoa of the genus Kudoa and Unicapsula can cause postmortem myoliquefaction in commercial fishes. A species of Unicapsula from the muscle of L. mormyrus was characterised morphologically and molecularly, and its damage to the muscle tissue was investigated to provide important information on potential future culture problems. High mortalities in cultures of D. puntazzo have been associated with a myxozoan gall bladder parasite, Ceratomyxa diplodae. This species has also been described from Diplodus annularis, Dicentrarchus labrax and Dentex dentex. In the current study the species complex C. diplodae from different hosts was characterised molecularly and developmental stages of the parasite are described by means of electron microscopy. Furthermore, to clarify whether direct transmission of the parasite is possible in culture systems, experimental infections of D. puntazzo were carried out orally, intraperitoneally and via bath treatment. 71
SY10/02-03
SY10/02-05
CHARACTERISATION, LOCALISATION AND INFECTION DYNAMICS OF THE RENAL MYXOZOANS GADIMYXA ATLANTICA AND ZSCHOKKELLA HILDAE IN CULTURED ATLANTIC COD GADUS MORHUA L. USING MOLECULAR APPROACHES
OCCURANCE AND INTENSITY OF NITZSCHIA STURIONIS (MONOGENEAN: CAPSALIDAE) IN THE STURGEONS ( CHONDROSTEI: ACIPENSERIDAE) OF SOUTH OF CASPIAN SEA
TAVAKOUL S., JALALI B., HALAJIAN A. Islamic azad university Tehran science and research branch, TEHRAN, ISLAMIC REPUBLIC OF IRAN
HOLZER A.S.(1), WOOTTEN R.(2), SOMMERVILLE C.(2) (1) Cavanilles Institute of Biodiversity and Evolutionary Biology, University of Valencia and Institute of Aquaculture, University of Stirling, (Scotland, UK), P.O. BOX 22085, 46071 VALENCIA, SPAIN ; (2) Institute of Aquaculture, University of Stirling, FK9 4LA, SCOTLAND, UNITED KINGDOM
Nitzschia sturionis is a monogenean in Capsalidae family parasitizing gills of sturgeons (Chondrostei fishes). Five sturgeon species live in south of Caspian Sea including Acipenser stellatus, Acipenser gueldenstaedti, Acipenser persicus, Acipenser nudiventris and Huso huso. In this survey, samples of four species ( including : A. stellatus, A. gueldenstaedti, A. persicus, A. nudiventris) were collected from February 2007 to April 2008 and after recording biometric characteristics, their gills were examined by standard parasitology methods. The worms were identified by parasites identification keys(Yamaguti,Bykowskaya). Standard statistical analysis on Nitzschia sturionis (mean intensity, standard deviation, prevalence, abundance and dominance) were analysed according to species, seasons, geographical location, sex, length and weight of the sturgeons and location in the gill and length of the worms. This results have shown in the tables of the article. For example the prevalences of Nitzschia sturionis according to species of sturgeon are : A. stellatus 55.5%, A. gueldenstaedti 0%, A. persicus 36.1%, A. nudiventris 0% . Also, in this study Diclybothrium armatum was separated from the gills of sturgeons.
Atlantic cod from various sites in Scotland were found infected with two myxozoan kidney parasites, G. atlantica and Z. hildae, which inhabit the renal tubules and the collecting ducts of the organ. Molecular characterisation by 18S rDNA sequencing of the myxozoans permitted phylogenetic positioning, specific primer design and the development of a PCR detection assay which allows for the specific detection and differentiation between the two parasites. Cod aged 0+ to 2+ years from three different hatcheries and one on growing facility were then tested by the PCR assay for the presence of the two myxozoan species and revealed important differences with regard to infection dynamics, levels and seasonality of infection. Furthermore, the use of the specific primers in a rapid in situ hybridisation protocol identified specific microhabitats for the two myxozoans in the excretory system of the kidney of the infected cod. The results of this study are discussed with regard to differences between the cod culture systems and the biology of myxozoan life cycles.
SY10/02-06
ULTRASTRUCTURE OF THE DIGESTIVE SYSTEM OF MACROGYRODACTYLUS CONGOLENSIS PRUDHOE, 1957, A MONGENEAN SKIN PARASITE OF THE NILE CATFISH CLARIAS GARIEPINUS
SY10/02-04
THE IMPACT OF THE COPEPOD PARASITE, LERNAEOCERA BRANCHIALIS, ON CULTURED JUVENILE COD, GADUS MORHUA.
SOMMERVILLE C., SMITH J.L., WOOTTEN R., TILDESLEY A., MAXWELL L. Institute of Aquaculture, University of Stirling, FK9 4LA, STIRLING, SCOTLAND, UNITED KINGDOM
EL-NAGGAR M.(1), KEARN G.(2), ARAFA S.(3) (1) Mansoura University, Faculty of Science, MANSOURA, EGYPT ; (2) University of East Anglia, School of Biological Sciences, NORWICH, UNITED KINGDOM ; (3) Mansoura University, Faculty of Science, Zoology Department, MANSOURA, EGYPT
The cod is a new species for mariculture where wild cod fisheries have declined. Juveniles are stocked in net pens in inshore sites where natural infection of gadoid fish with L. branchialis takes place. L. branchialis is a pennellid copepod parasite, widely distributed in the North Atlantic. It has a two host life cycle, settling on the gills of the gadoid as an adult female. The female penetrates the branchial artery and undergoes metamorphosis as it extends the cephalic and thoracic segments in the contra blood flow. The head terminates within the ventral aorta or the bulbus arteriosus. The main feature of the pathogenesis of the invasion process is an organised clot enveloping the cephalothoracic region of the parasite. We report the maturation of the lesion over the lifespan of the parasite. Morphological studies of the endoparasitic elements of the adult female soma revealed the appendages associated with the pathology. To assess the impact on growth performance, cultured juvenile cod were experimentally infected and the growth rate compared with uninfected fish. Whereas there was no significant difference in the Specific Growth Rates (SGR) of infected and uninfected cod, the number of parasites significantly influenced the SGR. There was little effect on growth at the lower infection level of 1-2 parasites but 3-5 parasites results in impaired performance.
Gyrodactylid monogeneans are potential pathogens of freshwater and marine fishes. Macrogyrodactylus congolensis live on the skin and fins of Clarias gariepinus. The aim of the present study was to illustrate the ultrastructure of the digestive system of M. congolensis. The pharynx consists of an anterior muscular and a posterior mainly glandular regions. The anterior region possesses 6 pharyngeal papillae while the posterior region has glandular syncytium and radial muscle fibres. The glandular syncytium possesses spherical, electron – dense granules. Oesophageal glands contain abundant, small, electron-dense secretory bodies. The intestine consists of a short median tube and two long , blind caeca. Both the tube and caeca have an uninterrupted syncytial gastrodermis resting on a fibrous basal matrix and possesses many lamellae, different kinds of vacuoles (V1,V2 and V3), many tubular structures, lipid –like droplets and melanin –like particles, The possible functions of these structures were discussed.
SY10/03-02
TRANSMISSION ECOLOGY OF POLYSTOMA 72
GALLIENI (PLATHYHELMINTHES, MONOGENEA), AN OPPORTUNIST PARASITE OF HYLA MERIDIONALIS (ANURA, HYLIDAE)
LAMELLODISCUS (MONOGENEA, DIPLECTANIDAE) PARASITES OF SEVERAL SPARID FISH HOSTS: TRUE GENERALISTS OR ONGOING SPECIATIONS?
BADETS M., VERNEAU O. Université Via Domitia, PERPIGNAN, FRANCE
POISOT T., DESDEVISES Y. Université Pierre et Marie Curie, Observatoire Océanographique de Banyuls, BANYULS CEDEX, FRANCE
The Monogenea is a highly diversified group of ecto parasites with a direct life cycle. They infest mainly gills and skin of actinopterygian and chondrichthyan fishes and to a lesser extent internal cavities of freshwater sarcopterygians. Among them, the Polystomatidae is the single family recovered from tetrapods like turtles and amphibians. Whereas 21 genera are recorded within this family, Polystoma is the only genus to perform two distinct life cycles depending of the physiological stage of its amphibian host. It is indeed either completed on gills of tadpoles or inside the urinary bladder of frogs leading to two different phenotypes adapted to maximize transmission efficiency in regards to host ecological stages. We explored this duality across the stripless tree frog Hyla meridionalis and its specific parasite Polystoma gallieni during two years in a temporary pond in South of France. Results obtained from field data and through experimental infestations revealed fine-tuned adaptations and numerous trade-off between parasite reproductive output and span of life upon which this opportunist strategy lies. Parasite-parasite as well as hostparasite interactions are highlighted to explain success of parasite transmission which leads ultimately to the infestation of more than 95% of young metamorphosed frogs in wild populations. Our results complete the ecological forces and processes shaping polystomatids’ life history traits trough evolution.
In spite of their short generation time, direct life cycle and high host specificity, most monogeneans parasite species show no profile of cospeciation with their fish hosts. It can therefore be hypothesized that monogeneans are able to perform numerous and frequent host switches (increasing their host range), shortly followed by speciation events to maintain their observed level of specificity. We investigated this question in the system formed by Lamellodiscus and their sparids hosts, because of the existence within the Lamellodiscus genus of several generalists species (harbouring up to 6 host species). From 8 hosts species, we sampled 120 individuals from generalist parasite species, and conducted a morphometric analysis on the attachment organ, called the haptor. Parasites were genotyped by sequencing their ITS1 and Cox1 genes. Morphometric and molecular datasets were compared to find out if generalists parasites form groups according to their hosts. This analysis allowed us to discriminate between true generalists (species showing no differenciation on their different hosts), species displaying phenotypic plasticity (morphometric variation with no genotypic differenciation), and ongoing speciations. Furthermore, the taxonomic status of several Lamellodiscus species recently described from morphological criteria was assessed on the basis of molecular data.
SY10/03-03
LINK BETWEEN INTRASPECIFIC VARIABILITY AND HOST SPECIFICITY IN LAMELLODISCUS MONOGENEANS
SY10/04-01
PARASITES AS SELECTIVE FORCES IN ECOLOGICAL DIVERSIFICATION - EXAMPLES FROM STICKLEBACKS
DESDEVISES Y.(1), KACI-CHAOUCH T.(1), VERNEAU O.(2) (1) Université Pierre et Marie Curie, Laboratoire Arago, BANYULS-SUR-MER, FRANCE ; (2) Université de Perpignan Via Domitia, PERPIGNAN, FRANCE
KALBE M.(1), EIZAGUIRRE C.(1), WEGNER K.M.(2) (1) Max Planck Institute for Evolutionary Biology, PLOEN, GERMANY ; (2) Institute of Integrative Biology, ETH-Zentrum, ZURICH, SWITZERLAND
We investigated whether host specificity is linked to interindividual variability within several species of Lamellodiscus monogeneans, fish gill ectoparasites of the Sparidae. Fish were sampled in the northeastern part of the Mediterranean Sea, and four specialist parasite species (using one single host species), and three generalist species (using several host species) were collected. We assessed intraspecific variability from morphological and molecular data. Morphometric variability of the attachment organ (i.e. the opisthohaptor) was estimated from measurements of several sclerified parts. Genetic variability was calculated based on comparisons of ribosomal DNA sequences of the first internal transcribed spacer (ITS1). Morphometric variances in the specialist versus generalist species were compared via principal component analysis and F-tests, and uncorrected genetic distances (pdistances) were estimated within species. We showed that the inter-individual variances of morphometric characters, as well as genetic distances, are clearly greater within generalist species than within specialists. These findings suggest that a relative increase in morphological and molecular variability promotes the ability to colonise new host species in monogeneans. This supports the hypothesis that intraspecific variability could be a potential determinant of host specificity.
The Red Queen Hypothesis describes the relationship between hosts and parasites as an evolutionary arms race, in which both partners continuously have to adapt to each other in order to maintain their own fitness. Three-spined sticklebacks (Gasterosteus aculeatus) are ideal vertebrate model organisms to study the evolution of hostparasite interactions on a small geographical scale. In northern Germany, it has been shown that sticklebacks e.g. from rivers and lakes can be regarded as genetically distinct ecotypes, which also harbour different habitat-specific macroparasites. We comparatively monitored stickleback populations and their parasite fauna in a river and a lake. During the observation period of 7 consecutive years/generations, neutral genetic markers (microsatellites) showed less differentiation than the immunologically relevant MHC class IIB genes in all three habitats, indicating that selection on these genes, which are responsible for specific parasite recognition and immunological memory, outweighs genetic drift. Additionally, as an ecological/genetical intersection, we included a small ditch, where sticklebacks of a lake genotype encounter river-like ecological conditions. These fish were intermediate, in terms of abundance of some parasite species, as well as in a number of morphological characteristics. This indicates that habitatspecific parasites play an important role in ecological diversification of host populations.
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SY10/04-04 SY10/04-02
PARASITES OF APOLLONIA MELANOSTOMA (NEOGOBIUS MELANOSTOMUS) (PALLAS) AND NEOGOBIUS KESSLERI (GÜNTHER, 1861) (GOBIIDAE:OSTEICHTHYES) FROM THE DANUBE RIVER IN AUSTRIA
WHAT CAN WE LEARN FROM PARASITES ABOUT SWORDFISH (X. GLADIUS) STOCK ASSESSMENT AND HEALTH STATE?
DAMIANO S.(1), MATTIUCCI S.(2), GARCIA A.(3), SANTOS M.N.(3), NASCETTI G.(1) (1) DECOS - Tuscia University, VITERBO, ITALY ; (2) DSSPSapienza University of Rome, ROME, ITALY ; (3) IPIMARCRIPSUL, OLHÁO, PORTUGAL
MÜHLEGGER J.M., JIRSA F., KONECNY R., FRANK C. University of Vienna, VIENNA, AUSTRIA
The two invasive fish species, the round goby Apollonia melanostoma syn.Neogobius melanostomus (Pallas) and the bighead goby Neogobius kessleri (Günther, 1861) have established a firm population in Austrian waters during the last 15 years. As there has been no record of the parasite community from these populations, a total of 79 specimens of A. melanostoma and 12 specimens of N. kessleri have been examined for parasites between May and October 2007 from three different sampling sites from the Danube River in Austria. In total twelve parasite taxa could be recovered. The protozoans Trichodina sp., and Ichtyophthirius multifilis from the gills and skin, two crustacean species Paraergasilus brevidigitus, Ergasilus sieboldii, from the gills and the two monogenean genera Gyrodactylus sp., Dactylogyrus sp. from the skin and gills as well, all occurring in low prevalence and intensities. Furthermore cystacanths of the acanthocephalan Acanthocephalus lucii could be recovered from the body cavity. Metacercaria of the digenean species Diplostomum spaethaceum and Thylodelphys clavata were found in the lens of the eye and the vitreous humour, respectively. The two digeneans Nicolla skrjabini and Bunodera nodulosa could be recovered from the intestine. Most striking seems the finding of metacercaria of the digenean Bucephalus polymorphus encysted in the skin and fins with prevalence up to 78 %, which is the first record of this parasite in Austrian waters.
The use of parasites as biological tags in population dynamics of marine fish proved to be a useful tool for discriminating stocks and to evaluate fish health state. The swordfish, Xiphias gladius, as a high commercial value seafood, represents a big challenge for the fishing industry. Population genetics studies demonstrated that the Mediterranean swordfish stock, separated from those of north and south Atlantic, presents a sensible genetic variability reduction. Host-populations showing low genetic variability levels are prone to infection by pathogens (“Red Queen Hypothesis”, Van Valen, 1973). 22 metazoan parasite taxa were collected from 523 X. gladius, fished in nine sampling areas throughout Atlantic Ocean and Mediterranean Sea. Among the metazoan identified, anisakid nematodes of the genera Anisakis and Hysterothylacium, identified to species level by genetic markers, emerged to be useful biological tags for the distinction between the Mediterranean and the Atlantic swordfish stocks. High infection levels in the Mediterranean stock has been evidenced by the highly pathogen copepod Pennella spp., while opposite trend has been observed in the Atlantic stock.
SY10/04-03
TREMATODE COMMUNITY STRUCTURE IN VALVATA MACROSTOMA WITH A SPECIAL REFERENCE TO TRANSMISSION PATTERNS OF ICHTHYOCOTYLURUS PILEATUS
SY10/04-05
HOST INTRODUCTION AND PARASITES: A CASE STUDY ON THE PARASITE COMMUNITY OF THREE FISH SPECIES IN THE HAWAIIAN ISLANDS
FALTYNKOVA A., VALTONEN E.T., KARVONEN A. University of Jyväskylä, JYVÄSKYLÄ, FINLAND
We examined the spatiotemporal structure of the trematode community in the hermaphroditic prosobranch snail Valvata macrostoma by collecting a total of 1,103 snails monthly from two sites (near shore, 1-2 m deep; and 70 m offshore, 5-6 m deep) in Lake Konnevesi, Central Finland, in May-October 2007. In 51 % (n=557) of the snails we found a community of 10 trematode species with cercariae of 7 and metacercariae of 5 species. The system was characterised by high spatial variation in infection of individual species with high infection rates in the shallow (all 10 species) and dominance of only one species (Ichthyocotylurus pileatus) in the deeper site. The total prevalence of infection changed with season peaking in July (79 %), and being higher in early summer (64 %) than in autumn (8-32 %), which suggests that the infections overwinter. We suggest that these spatial differences in species composition may emerge through site-preferences of bird definitive hosts, whereas the temporal patterns are likely to reflect seasonality of parasite transmission and dynamics of the snail population. In a detailed study on the transmission patterns of I. pileatus to perch (Perca fluviatilis), we found that the transmission took place in July and August and was correlated with the prevalence in snails. Despite of observed temporal (July-August) and spatial (deeper site only) restrictions, the transmission resulted in 100 % prevalence in juvenile (0+) fish.
VIGNON M., SASAL P. Laboratoire Biologie et Ecologie Tropicale et Méditerranéenne, PERPIGNAN, FRANCE
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Peacock groupers (Cephalopholis argus), blacktail and bluestripe snappers (Lutjanus fulvus and L. kasmira) were intentionally introduced to Hawaiian archipelago from French Polynesia 50 years ago to enhance local fisheries. Following introduction, these species spread rapidly, became extremely abundant and caused controversial issues. The entire metazoan parasite community of these three species was compared between their native ranges (both Society archipelago and Marquesas Islands, French Polynesia) and their introduced range (Hawaii). Polynesian and Hawaiian snappers are infected with a highly diversified parasite community (up to 30 species), suggesting that parasite species were introduced with their Lutjanid hosts. We discuss the potential threat of these exotic parasites to the native fish community in Hawaii. On the other hand, while Polynesian groupers have a diverse parasite community, they exhibit a depauperate ensemble of parasites in Hawaii (they lost parasites common in Polynesia). None of the fish acquired new parasites present in Hawaiian waters. This study provides the first complete parasitological survey for a wild introduced fish between native and introduced range in a coral reef ecosystem and provides opportunity to gain insight into the role of parasites in determining the success of host
invasions. Considering the homogenisation of biota in marine ecosystems, our study may also have implications for biological conservation
These include genes implicated in the eel’s somatic growth (the Insulin-like Growth Factor 1 and the Thyroid Hormone Receptor beta), osmoregulation (the Na+/K+ATPase beta 1 and the Aquaporin 3) and hematopoiesis (the Hemoglobin alpha-chain). Our results showed an absence of effect on genes involved in fish growth. However, the parasite may have an effect on the osmoregulation and hematopoiesis. We also noted a differential impact of male and female parasites on the expression of some genes, perhaps owing to the sexual dimorphism in body size of the parasite.
SY10/04-06
MONITORING OF TREMATODE COMMUNITIES (CERCARIAE) IN LYMNAEA STAGNALIS (GASTROPODA, PULMONATA) BY MARKRECAPTURE METHOD
SOLDANOVA M.(1), FALTYNKOVA A.(2) (1) Institute of Parasitology, Biology Centre, Czech Academy of Sciences, Faculty of Science, University of South Bohemia in Ceske Budejovice, 31 Branisovska, 370 05, CESKE BUDEJOVICE, CZECH REPUBLIC ; (2) Institute of Parasitology, Biology Centre, Czech Academy of Sciences, 31 Branisovska, 370 05, CESKE BUDEJOVICE, CZECH REPUBLIC
SY10/05-04
HEAVY METALS IN THE TAPEWORM CARYOPHYLLAEUS LATICEPS (PALLAS 1781) AND ITS HOST THE NASE CHONDROSTOMA NASUS (L. 1758) FROM TWO AUSTRIAN RIVERS: BIOINDICATIVE ASPECTS
The intimate association between trematodes and molluscs denotes them as suitable model systems for parasite-host interaction studies. We studied the trematode infra- and component community in the great pond snail Lymnaea stagnalis using mark-recapture. We collected the snails in three-week intervals from ponds in areas of Jindichv Hradec and Tebo in South Bohemia, Czech Republic, examined them for cercarial shedding, measured and marked their shells and released them to their original ponds. We recorded a high infection rate as well as recapture-rate, more than thousand individuals of snails were recaptured, most of them once and a few even up to five times. The trematode component community in L. stagnalis consisted of 13 species; cercariae dominant throughout the season were those of Plagiorchis elegans and Opisthioglyphe ranae (Plagiorchiidae). We found a considerably high number of multiple infections and frequent consecutive changes of infections within individual snails. An explanation of this phenomenon is rather difficult as our method provides only indirect evidence of interspecific interactions and does not include snail dissection. Recaptures of infected overwintered snails indicate that trematodes survive winter in first intermediate host. Mean size as well as survival rate of infected snails was significantly higher compared to uninfected snails suggesting that trematode infections alter the life-history of their hosts.
JIRSA F., STOJANOVIC A., LEODOLTER-DVORAK KRACHLER R. University of Vienna, WIEN, AUSTRIA
During parasitological examinations of the predominantly herbivorous fish species nase Chondrostoma nasus (L.1758) from different rivers in Austria a well established population of the tapeworm Caryophyllaeus laticeps (Pallas, 1781) was detected in the intestine of the fish from two sites. One of the sites is reported unpolluted, the other one polluted with cadmium, lead and zinc. The concentrations of these metals in the tissue of the gills, muscle, intestine and liver of the fish and in the parasite were determined by inductively coupled plasma - optical emission spectrometry (ICP-OES). Both the nase and C. laticeps show a bioindicative ability for metal pollution in rivers. Cadmium was found mainly in the liver with maximum concentrations of 1.57 g/g dry weight from the unpolluted site and 5.58 g/g from the polluted site. Lead was found mainly in the intestinal wall and liver in concentrations between 0.09 and 4.05 g/g, and 0.26 and 1.94 g/g, respectively. In the samples from the polluted site it also could be detected in the gills (mean 1.38 g/g). Zinc could be detected in all fish tissues, the highest concentrations occurred in the liver, showing no significant differences between the two sites. The parasite C. laticeps shows even higher capacities for metal accumulation: cadmium was detected in concentrations up to 5.1 times higher, lead up to 9.7 and zinc up to 3.0 times higher in the tapeworm compared to the fish liver.
SY10/05-03
CHANGES IN GENE EXPRESSION IN EUROPEAN EELS (ANGUILLA ANGUILLA) INDUCED BY INFECTION WITH SWIMBLADDER NEMATODES (ANGUILLICOLA CRASSUS)
FAZIO G., MONÉ H., DA SILVA C., SIMON-LEVERT G., ALLIENNE J.F., LECOMTE-FINIGER R., SASAL P. UMR 5244 CNRS-EPHE-UPVD, Biologie et Ecologie Tropicale et Méditerranéenne, PERPIGNAN, FRANCE The European eel, Anguilla anguilla, is considered an endangered species that is under pressure from many factors. Among these factors, the introduced parasite, Anguillicola crassus, is thought to play an important role in the decline of eel populations. These nematodes have been shown to negatively affect many fitness-related traits in eels (e.g growth, osmoregulation, stress tolerance). Nevertheless, there has been little work on the way by which the host-parasite interaction influences the molecular regulation of these key physiological processes. We analyzed, experimentally, the effect of this nematode on the expression of genes involved in the physiology of European eels during their continental life.
M.,
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SY10/05-05
SY11/01-02
PARASITES, HEAVY METALS AND SANITARY RISK BOUND TO THE CONSUMPTION OF A PRIMITIVE FISH (PROTOPTERUS ANNECTENS) IN IVORY COAST.
POPULATION STRUCTURE OF TOXOPLASMA GONDII IN GABON
MERCIER A.(1), NGOUBANGOYE B.(2), SALLE B.(2), JAFFRE (2), AJZENBERG D.(1), DARDÉ M.L.(1) (1) Faculté de Médecine, EA3174 NETEC, CNR/CRB Toxoplasmose, LIMOGES, FRANCE ; (2) CIRMF Centre International de Recherche de Franceville/ Centre de Primatologie, FRANCEVILLE, GABON
CISSE M., GNAYORO M.M.A., OUATTARA A., GOURENE G. Université d'Abobo-Adjamé, ABIDJAN, IVORY COAST
The Protopteridae constitute the current representatives of the Subclass Dipneusta which is the most archaic group of living osseous fishes. The origin of this family goes back up to Devonian (355 - 408 million years). At present, there is only a single species in Australia (Neoceratodus forsteri), one in Amazonia (Lepidosiren paradoxa) and only one in West Africa (Protopterus annectens). P. annectens is often captured in Ivory Coast in the hydroagricultural small lakes of the North (Dams of Solomougou, Guemou, Katiali, Sambakaha, Nabingué), in lakes (Kossou, Taabo, Buyo) and in the plains of inundation of the river Bandama where it constitutes a source of protein appreciated by the rustic communities. The plain of inundation of the region of Oumé is the meeting place of raw waste water coming from the city and waters of the river Bandama. In this place many of specimens of P. annectens are daily captured by the local residents. After taxonomic description of their parasites, we discuss the sanitary risk, the place of Protopterus annectens in the zoological scale and the relationship with evolution of the accommodated parasites. In order to appreciate toxicological implications, heavy metals (Cd, Cu, Zn, Pb, and Fe) in the water, the sediments and the fish were considered.
Toxoplasma gondii is found worldwide, but the genotype distribution varies across the continents. A clonal population structure with 3 main lineages (types I, II, and III) was described in Europe and USA, whereas the genetic diversity is higher in South America. Genetic data concerning Toxoplasma in Africa are scarce and do not allow any genetic population analysis. Seropositive animals originating from different areas in Gabon (from 50 to 700 km apart), were analyzed for Toxoplasma isolation and genotyping: 53 chickens from 8 geographically distant locations [Libreville, Franceville, Makokou, Bakoumba, La Lopé, Lékoni and Songola (Congo)] and 45 animals from Dienga, a village near the frontier of Congo, where a high prevalence of Toxoplasma infection was previously found. Sixty-eight isolates were obtained by bioassay in mice. Five genotypes were identified using 6 low polymorphic microsatellite markers: 9 isolates had a type III genotype and 7 a genotype characterized by a defined mixture of I and III alleles, already found elsewhere in Africa (African#1); 2 isolates were variants of this genotype,11 isolates had a genotype with a mixture of alleles I, II, and III and 1 a mixture of alleles I and II.. The addition of 9 more polymorphic microsatellite markers leads to the detection of 30 genotypes. The analysis of data demonstrates a clonal epidemic population structure within each area, but a global genetic diversity higher than in France.
SY10/05-06
INFLUENCE OF SEDIMENT CONTAMINATED WITH UNTREATED PULP AND PAPER EFFLUENT ON WINTER FOUNDER, PLEURONECTES AMERICANUS: DOSE RESPONSE AND GRADIENT STUDIES
SY11/01-03
GENETIC DIVERSITY OF PLASMODIUM FALCIPARUM POPULATIONS FROM SITES ACROSS AFRICA INFERRED FROM SNP UNDER SELECTION VS. NEUTRAL SNP
KHAN R.A. Memorial University of Newfoundland, ST. JOHN'S., CANADA
This study investigated pulp and paper mill contaminated sediment on winter flounder, Pleuronnectes americanus, based on laboratory and field studies. Flounder, captured from a pristine site, were exposed in a flow-through system for 16 wk to sediment collected at 2, 5, 7 and l0 km from the outfall. A group of controls was placed in uncontaminated sediment. Mortality, lower condition factor, enlarged liver and toxicopathic lesions were significantly greater in fish submerged in sediment from 2 km than at other locations. Two ectoparasites, Trichodina jadranica, and Gyrodactylus pleuronecti, were observed only in the control group while in the digestive tract, Steringophorus furciger, was more abundant in fish in all other groups than at 2 km. Analysis of data from a previous gradient study on fish captured at various distances, 2, 5, 7 and l0 km down-current from the outfall revealed an enlarged liver, elevated detoxification enzymes and prevalence of toxicopathic lesions were significantly greater in samples taken at 2 km than at other sites. However, two metazoan parasites, S. furciger (Digenea) and Echinorhynchus gadi (Acanthocephala) in the digestive tract were more abundant at more distant locations and also in the controls. These results suggest that fish health was compromised at the site nearest to the outfall where high concentrations of toxic contaminants persisted.
MAÏGA O.(1), DJIMDÉ A.(2), FALCAO S.(1), BROUSSIER E.(1), RENARD E.(1), HOUZÉ S.(3), JEAN FRANÇOIS V.(4), DOUMBO O.(2), LE BRAS J.(3), CLAIN J.(1) (1) Université Paris Descartes - EA 209, PARIS, FRANCE ; (2) Malaria Research and Training Center, BAMAKO, MALI ; (3) Centre National de Référence du Paludisme, PARIS, FRANCE ; (4) Service Nationale de Lutte Antipaludique, PORT-AUPRINCE, HAITI History and evolution of Plasmodium falciparum can be traced through the analysis of DNA sequence variation.To analyse how P. falciparum African populations differentially evolved at medically important loci, we are analysing 32 single nucleotide polymorphisms (SNPs) in ~300 isolates from 6 populations. The study focuses on 2 groups of SNPs: 1) twelve functional SNPs in genes under the selection exerted by immune response or drug pressure (sSNPs) ; and 2) twenty SNPs located on 13 of the 14 chromosomes in genomic regions under no known selection, defined as neutral SNPs (nSNPs). The nSNPs were chosen through a bioinformatic scan of the PlasmoDB database. A simple and robust PCR-RFLP assay was set up for each of the SNPs included in the study. To examine how the populations are genetically differentiated, we will calculate the 76
F-statistics for all pairs of populations by using the population allele frequencies across the nSNPs and the sSNPs separately. Relationship between differentiation and geographical distance will be evaluated through the use of nSNPs only. The genetic differentiation revealed by nSNPs and sSNPs will be compared to detect how local selections have shaped the diversity in the different populations. Data analysis is ongoing.
The Niayes area of Senegal harbours a population of the tsetse fly G. palpalis gambiensis, despite annual rainfall of only 400 mm and a strong human encroachment. It is an important agricultural and cattle production area, with a strong presence of exotic breeds for milk production, but their costeffectiveness is threatened by trypanosomoses. A tsetse control campaign has thus been launched by the Direction de l’Elevage with the technical assistance of the International Atomic Energy Agency. Tsetse populations in the Niayes are separated from the major tsetse belt by a 70 km-long very dry area that creates a physical barrier to tsetse dispersal. Their isolation was confirmed by an assessment of the gene flow using microsatellite DNA markers (FST = 0.10 to 0.16, p 2.72, p in Niakhar) and of season (> during rainy season). Those findings support the idea that anti-mosquito saliva antibodies naturally raised by domestic animals could be a good marker of exposure to Anopheles bites and, hence, help to better understand the role of animals in malaria transmission.
In the end of 2005, a nationwide free LLINs distribution targeting most vulnerable populations was coupled with a national polio vaccination campaign, aiming to reduce morbidity and mortality among young children in Niger. It provided a unique opportunity to evaluate the impact of such an intervention on Plasmodium falciparum transmission and to gather informations about its benefits for human population, but also undesired side effects potentially occuring (e.g. insecticide resistance in vector populations). We developed a multidisciplinary approach to evaluate potential effects of the LLINs coverage on human, parasite and vector populations. Multiple parameters were followed from the 2005 transmission season (pre-LLINs period) to the two subsequent years (postLLINs period), involving specific sampling schemes and analysis methods regarding the different actors of malaria epidemiology in the field. An overview of the methodology employed with emphasis on innovative tools will be given, followed by major results obtained during the study that will be discussed relating to the epidemiologic and biogeographic contexts of the country.
SY12/02-05
SY12/03-02
INFERRING TRANSMISSION PATTERNS OF SCHISTOSOMA MANSONI USING MOLECULAR MARKERS AND PARENTAGE ANALYSIS
ANTIBODY RESPONSES TO SALIVA OF TRIATOMA INFESTANS: THEIR POTENTIAL AS EPIDEMIOLOGICAL TOOL FOR CHAGAS DISEASE SURVEILLANCE
STEINAUER M.(1), HANELT B.(1), AGOLA L.(2), MWANGI I.(2), MAINA G.(2), KARANJA D.(3), MKOJI G.(2), LOKER E.(1) (1) University of New Mexico, ALBUQUERQUE, UNITED STATES ; (2) Centre for Biotechnology Research and Development, Kenya Medical Research Institute, NAIROBI, KENYA ; (3) Vector Biology and Control Research Centre, Kenya Medical Research Institute, NAIROBI, KENYA
SCHWARZ A.(1), MEDRANO-MERCADO N.(2), COLLIN N.(3), TEIXEIRA C.R.(3), HELLING S.(4), ASSUMPÇÃO T.C.(5), MARCUS K.(4), RIBEIRO J.M.(3), VALENZUELA J.G.(3), BILLINGSLEY P.F.(6), STERNBERG J.M.(7), SCHAUB G.A.(8) (1) Group Zoology/Parasitology, Ruhr-University Bochum & School of Biological Sciences, University of Aberdeen, BOCHUM & ABERDEEN, GERMANY & UK ; (2) Laboratory of Chagas disease and Immunoparasitology, COCHABAMBA, BOLIVIA ; (3) Laboratory of Malaria and Vector Research, NIAID/NIH, ROCKVILLE, UNITED STATES ; (4) Medical Proteome Center, BOCHUM, GERMANY ; (5) Laboratory of Host-Parasite Interface, BRASILIA, BRAZIL ; (6) Sanaria Inc., ROCKVILLE, UNITED STATES ; (7) School of Biological Sciences, ABERDEEN, UNITED KINGDOM ; (8) Group Zoology/Parasitology, BOCHUM, GERMANY
Schistosomes infect 200 million people worldwide and many suffer from severe morbidity as a consequence of these parasites. Schistosoma mansoni is the primary infectious agent of schistosomiasis in western Kenya where drug treatment, HIV, and hybridization with other schistosomes possibly interact to exert selective pressures on this parasite. Our research program uses molecular and evolutionary techniques to address epidemiological issues related to this parasite in its environment. Because they live within the circulatory system and cannot be removed from a living human, only indirect inferences can be made about adult worm populations. Our approach uses microsatellite data from the offspring of human
The 'Southern Cone Initiative' was highly successful in significantly reducing populations of Triatoma infestans, the 82
main vector of Chagas disease in Latin America. To dectect re-emerging T. infestans populations at an early stage after the control programmes have finished, new methologies are required for substained, long-term monitoring of onceendemic Chagas disease regions. While feeding, triatomines induce an antibody response in their hosts. An immune response of short lived hosts, such as chickens and guinea pigs, can indicate a recent exposure to triatomid bites and be a potential measure of transmission risk. Therefore, this study analysed the antibody response of chickens and guinea pigs to the saliva of different T. infestans strains. Highly immunogenic antigens of 14, 21 and 26 kDa were recognised by all chicken sera and a 79 kDa salivary protein by all guinea pig sera after few feeding events of T. infestans. The antigens were analysed by mass spectrometry and blasted against the T. infestans salivary glands cDNA library. Out of four identified antigens, a 14.6 kDA salivary secreted protein was expressed and tested against animal sera of free-living hosts of T. infestans. This protein was recognised by all chicken and guinea pig sera and thus represents a useful immuno-epidemiological marker for the detection of low-level infestations of T. infestans for countries with Chagas disease control programmes.
GLAREOLUS
VOJTA L.(1), BECK R.(1), CURKOVIC S.(2), MRLJAK V.(3), MARINCULIC A.(1) (1) Department of Parasitology and Parasitic Diseases with Clinic, Faculty for Veterinary Medicine, University of Zagreb, ZAGREB, CROATIA ; (2) Department of Anatomy, Histology and Embryology, Faculty for Veterinary Medicine, University of Zagreb, ZAGREB, CROATIA ; (3) Clinic for Internal Diseases, Faculty for Veterinary Medicine, University of Zagreb, ZAGREB, CROATIA Wild rodents play an important role in nature as reservoir hosts for many pathogens, including some that can be transmitted to other animals. Hepatozoon sp. parasitizes many mammals, especially rodents, and its infections are dependent on tickvectors. High prevalence of Hepatozoon sp. in small wild rodents has been reported in Northern and Central America, West Africa and Europe. Due to previously non-existing data on hepatozoonosis in wild animals in Croatia, we investigated 120 small wild rodents, including 48 woodmice Apodemus sylvaticus, 37 yellow-necked mice Apodemus flavicolis, 2 striped field mice Apodemus agrarius and 33 bank voles Clethrionomys glareolus. DNA was isolated from spleen tissue and the screening PCR on the 666 bp fragment of 18S rRNA revealed 35 (29%) hepatozoon-positive rodents: 25 (75,8%) C. glareolus, 7 (14,6%) A. sylvaticus, 3 (8,1%) A. flavicolis and no A. agrarius. Positive samples were confirmed by partial sequencing of 18S rRNA gene. Sequences were blasted against Hepatozoon sp. 18S SSU rRNA available at GenBank. The alignments have revealed 33 Hepatozoon sp. sequences identical to the isolates AY600625 and AY600626 from Spanish bank voles and 2 sequences with the highest homology to H. ayorgbor isolate EF157822 from phyton. Obtained results are in agreement with investigations in other European countries, which showed that Hepatozoon infections are especially common in the bank voles.
SY12/03-03
INCIDENCE RATE AND ENVIRONMENTAL DETERMINANTS OF CANINE VISCERAL LEISHMANIASIS IN ALPES-MARITIMES, FRANCE.
PIN-DIOP R.(1), VALERIO L.(2) (1) Laboratoire Vétérinaire Départemental des Alpes-Maritimes, SOPHIA-ANTIPOLIS, FRANCE ; (2) Faculté de Médecine, Université de Nice-Sophia Antipolis, NICE, FRANCE Leishmaniasis affects approximately 2 million new patients each year. Due to a flagellate protozoa of the genus Leishmania, the disease is transmitted by the bite of phlebotomine sand flies. Visceral leishmaniasis (VL) is a widespread zoonosis, due to systemic dissemination of the parasite. The district of AlpesMaritimes, in southern France, is an endemic area where dogs are at the same time the reservoirs and the main hosts of L. infantum. The purpose of this study was to assess the effects of environmental factors on canine VL incidence. The cases of VL in dogs, detected by serological analyses between 2000 and 2007, were used to assess disease incidence in 139 out of the 163 communes of Alpes-Maritimes. Environmental features suspected to influence vector density and disease transmission were assigned to each commune in a Geographical Information System. Univariate and multivariate statistical analyses were performed at the commune level, to search for associations between the 2000-2007 mean incidence and landcover, altitude, slope and aspect. Depending on the year, the median incidence rate varied from 16.6 per 1000 to 6.5 per 1000, with an overall declining trend. VL was found to be strongly associated with slope, non continuous urban area, wild medi scrubland and transitional woodland-shrub. The results suggest that environmental patterns can be used to assess VL transmission risk in Alpes-Maritimes and in similar ecological areas.
SY12/03-05
FIRST MOLECULAR CHARACTERIZATION OF CANINE HEPATOZOONOSIS IN CROATIA USING A SCREENING PCR AND SEQUENCING METHODOLOGIES
VOJTA L.(1), BECK R.(1), CURKOVIC S.(2), ZIVICNJAK T.(1), MRLJAK V.(3), MARINCULIC A.(1) (1) Department of Parasitology and Parasitic Diseases with Clinic, Faculty for Veterinary Medicine, University of Zagreb, ZAGREB, CROATIA ; (2) Department of Anatomy, Histology and Embryology, Faculty for Veterinary Medicine, University of Zagreb, ZAGREB, CROATIA ; (3) Clinic for Internal Diseases, Faculty for Veterinary Medicine, University of Zagreb, ZAGREB, CROATIA Apicomplexan blood parasites of the genus Hepatozoon have been described from a wide range of mammals and other vertebrates. Sexual stage with sporogony takes place in different blood-sucking arthropods, and asexual phase in endothelial cells of various organs or parenchymal cells of the liver. Up to now two Hepatozoon species that infect canids, including domestic dogs, have been described: H. canis, a relatively widespread protozoan, and H. americanum, which seems to be present only in North America. H. canis has been reported in Asian, African and South American dogs, but although widespread among European wild canids and rodents, it was found only in few dogs in Spain and Greece. We investigated a population of 924 apparently asymptomatic dogs from different Croatian regions. DNA was isolated from canine blood and the screening PCR on the 666 bp fragment of 18S
SY12/03-04
INFECTION WITH HEPATOZOON IN THE SMALL CROATIAN RODENTS APODEMUS SYLVATICUS, APODEMUS FLAVICOLIS, APODEMUS AGRARIUS AND CLETHRIONOMYS 83
rRNA revealed 108 (11,8%) positive dogs. Positive samples were confirmed by partial sequencing of 18S rRNA gene. The consensus sequences, derived from various sequence data sets, were blasted against 18S SSU rRNAs of Hepatozoon sp. available at GenBank. The alignments have revealed 106 H. canis and 2 H. sp. sequences. Among H. canis isolates we found small heterogeneity, while H. sp. isolates were identical to the Spanish isolate AY600625 from Clethrionomys glareolus. Obtained results indicate higher prevalence and significance of hepatozoonosis in Croatia than previously believed.
Infection status was assessed using a copro-antigen ELISA and copro-PCR. This paper develops a temporal model that describes transmission to dogs and which accounts for uncertainty in dog infection status. Parameter uncertainty is assessed using Bayesian simulation techniques. The model was used to assess between village differences in infection pressure that might arise from heterogeneity in ecological conditions. The role of ecological variation on infection rates in dogs is discussed.
SY12/04-04
ECHINOCOCCUS MULTILOCULARIS, AN EMERGING PARASITE IN THE MOST WESTERN EUROPEAN BORDER LINE.
SY12/04-02
PREDICTIVE MAPPING OF HOST ASSEMBLAGES: AN EXAMPLE WITH SMALL MAMMALS IN WESTERN CHINA
VAN DER GIESSEN J.(1), VERVAEKE M.(2), DE VRIES A.(1), BROCHIER L.(3), CHU M.(1), LOSSON B.(4), TEUNIS P.(1), TAKUMI K.(1) (1) RIVM, BILTHOVEN, NETHERLANDS ; (2) University of Antwerpen, ANTWERPEN, BELGIUM ; (3) Scientific Institute of Public Health, BRUSSELS, BELGIUM ; (4) Université de Liège, LIÈGE, BELGIUM
VANISCOTTE A., PLEYDELL D., RAOUL F., GIRAUDOUX P. University of Franche-comté, BESANÇON, FRANCE
Linking host spatial distributions to environmental variables can provide key information for understanding and predicting the transmission of a parasite in space. When a large number of potential intermediate hosts are shown to co-occur within a diversity of habitats, community level modelling helps to summarise such complex data sets by defining groups of species/sites, i.e assemblages. We built a predictive model for the niches of small mammal assemblages including potential Echinococcus multilocularis intermediate hosts, in two areas of western China (Sichuan) by a three step modelling procedure. First, eight assemblages were defined using a multinomial logistic model associated with a redundancy reduction procedure. Then, niches of these assemblages were modelled w.r.t. the environmental space of each sampled area using a Multiple Adaptive Regression Spline. Elevation and band 6 of a Landsat Enhanced Thematic Mapper image (which correlates with land surface temperature) were the main factors relating to assemblage distributions in both study areas. The importance of vegetation indices, which are correlated to the amount of vegetation, differed between the two locations. Finally, the model providing the lowest predictive classification error was chosen to map assemblage occurrence probabilities beyond the sampled locations. The predictive error component induced by extrapolation of model predictions on non trained locations is discussed.
We analyzed red fox data from an area close to the westernmost margin of E. multilocularis habitat in Europe, Belgium and a neighboring province (Limburg) in The Netherlands (NL), with the aim of studying the emergence of the parasite in this area. Spatial coordinates of the locations of infected and uninfected foxes have been determined by GPS. The spatial distribution of the prevalence of infection among sampled foxes has been modeled as an ellipsoidal gradient, demonstrating increasing prevalence in southeastern direction and E. multilocularis infection could be shown to have a continuous distribution across national borders. A mathematical model describing the parasite population dynamics both in time and in space was fitted to the worm burdens of foxes sampled between 1996 and 2006 in the NL. We found a strong indication that the parasite’s reproduction number R0 is greater than 1 and that the parasite is spreading to a wider region in Limburg. Now, the infected foxes can also be found close to the city of Maastricht. Based on the R0 derived from the mathematical model of the parasite’s transmission, we analysed the effect of public health measures aimed to eradicate the infection. Increased infection pressure of E. multilocularis in north-western Europe is most likely to occur at present. Although sofar, human cases have not been found in the Netherlands, we assume that this will change in the near future.
SY12/04-03
SPATIO-TEMPORAL VARIATION IN ECHINOCOCCUS MULTILOCULARIS INFECTIONS IN TIBETAN DOGS
SY12/04-05
IMPROVING A SPATIAL MODEL FOR ECHINOCOCCUS MULTILOCULARIS TRANSMISSION STUDY
PLEYDELL D.(1), MOSS J.E.(2), BUDKE C.M.(3), QIU J.M.(4), CHEN X.W.(4), WANG Q.(4), CRAIG P.S.(2), GIRAUDOUX P.(1) (1) University of Franche-Comté, BESANÇON, FRANCE ; (2) University of Salford, GREATER MANCHESTER, UNITED KINGDOM ; (3) Texas A&M University, COLLEGE STATION, UNITED STATES ; (4) Institute of Parasitic Disease Control of Sichuan Provincial CDC, CHENGDU, CHINA
QUINTAINE T.(1), POULLE M.L.(1), THULKE H.H.(2) (1) Centre de Recherche et de Formation en Eco-éthologie (2C2A-CERFE) & Laboratoire de Parasitologie-Mycologie, EA 3800, IFR 53, Université de Reims Champagne Ardenne, BOULT-AUX-BOIS & REIMS, FRANCE ; (2) Department of Ecological Modelling, Helmholtz Centre for Environmental Research, UFZ, LEIPZIG, GERMANY
The Qinghai-Tibetan plateau has been described as a large meta-stable transmission focus for Echinococcus multilocularis. Infection rates in canid definitive host populations vary as a function of the availability of infected small mammal intermediate hosts which in turn varies as a function of complex interactions within a set of environmental variables. The infection status of dogs was monitored in 13 villages from Serxu County, NW Sichuan, China over a three year period. Each dog was dosed with praziquantel at the start of the study.
In Europe, the cycle of Echinococcus multilocularis (E.m.) includes mainly red fox (Vulpes vulpes) as definitive host and grassland voles as intermediate hosts. It implies a free eggs stage dispersed in the environment with fox faeces. Ecological factors therefore strongly influence parasite transmission, acting both on hosts and parasite. Our aim was to evaluate the 84
influence of ecological factors on the E.m. transmission by using the “Echi” model developed by Hansen et al. (2004). Echi is both a grid based and an individual based model, with grid corresponding to rodent distribution and individuals to foxes. Foxes move on the grid and modify it by hunting and defecating. The influence of ecological factors on E.m. transmission is assessed by testing the capability of different scenarios to reproduce the observed ranges of fox and intermediate host prevalence. We first improved the input of the models by collecting field data on fox prevalence, fox density, intermediate hosts and fox faeces distribution among six habitats. We then tested the effect of the intermediate hosts and fox faeces distribution on E.m. transmission. Potential use of the model in the identification of such key factors of E.m. transmission is discussed. Hansen F. et al., 2004. Processes leading to a spatial aggregation of Echinococcus multilocularis in its natural intermediate host Microtus arvalis. Int. J. Parasitology 34, 3744
TRANSMISSION: ECHINOCOCCUS MULTILOCULARIS AS A STUDY CASE
RAOUL F., AIME B., LAMBERT J., N'GUYEN VIET H., GIRAUDOUX P. University of Franche-Comté, BESANCON, FRANCE Predator-prey interaction is a key factor determining transmission intensity of trophically transmitted parasite. However, few studies document the relationship between prey availability, prey consumption by predators, and its consequence on transmission. We investigated how the variation of density of two small mammal E. multilocularis intermediate hosts (IH) in the field, on a local scale (n x 10km2), impact fox diet and subsequently parasite transmission intensity in foxes. Ten study sites were sampled in Franche-Comté (France) in winter from 1995 to 1999. IH population densities were estimated using index methods. Fox faeces were collected in the field to estimate (i) the occurrence of IH in the diet, and (ii) the infection status of the fox population (coproantigen detection). Mixed-effects models were used to account for site pseudo-replication. Results suggest no correlation between M. arvalis density in the field and its occurrence in the diet, and a significant positive correlation between A. terrestris density and its occurrence in the diet. The median infection level measured in faeces, as well as the high infection values were significantly correlated to the biomass of A. terrestris only. This study suggests an opportunistic behaviour of foxes towards A. terrestris, and a preference (or a better accessibility to) for M. arvalis. Here, A. terrestris, the bigger IH species, seemed to play a dominant role in transmission.
SY12/04-06
REMOTE SENSING, LANDSCAPE EPIDEMIOLOGY AND RISK ASSESSMENT IN THE ONCHOCERCIASIS AMAZONIAN FOCUS OF SOUTHERN VENEZUELA
BOTTO ABELLA C.(1), SUÀREZ L.(2), VILLAMIZAR N.(3), CORTÉS J.(3), CORONEL P.(3), ESCALONA E.(1), GRILLET M.E.(2) (1) Instituto de Medicina Tropical / Universidad Central de Venezuela & Centro Amazónico de Investigación y Control de Enfermedades Tropicales Servicio Autónomo, CARACAS & PUERTO AYACUCHO, VENEZUELA ; (2) Instituto de Zoología Tropical / Universidad Central de Venezuela, CARACAS, VENEZUELA ; (3) Centro Amazónico de Investigación y Control de Enfermedades Tropicales Servicio Autónomo, PUERTO AYACUCHO, VENEZUELA
SY12/05-03
A QUANTITATIVE EVALUATION OF ECHINOPARYPHIUM ACONIATUM TREMATODA PARASITE POPULATION IN CHANY LAKE ECOSYSTEM, WESTERN SIBERIA, RUSSIA
Remote sensing, geographical information systems (GIS) and landscape epidemiology approach is helping to understand spatial distribution of vector borne diseases and to assess the risk of infection. Onchocerciasis, a filarial infection transmitted by Simulium flies, comprise an important challenge to public health for the Yanomami population in the Amazonian Venezuelan focus. The objective of this work is to assess the influence of landscape and different environmental variables on the risk of onchocerciasis infection in the Upper Orinoco basin. Pretreatment prevalence of infection was assessed in 63 out of 131 yanomami communities by skin biopsy that was performed in 2856 individuals and environmental variables were described for each community with the aid of GIS. Principal component analysis was used to characterize the habitat of hyperendemic onchocerciasis. Multiple regression analysis showed that high prevalence of infection is associated with high degree of steepness of the slope ground and low river order. A river factor negatively associated with prevalence of infection was identified. Discriminant analysis demonstrated that hyperendemic onchocerciasis is mostly prevalent in mountainous and plateau landscapes, associated with rivers of low order and volcanic substrate. These variables allowed to correctly classify > 92% of communities as highly endemic. This information is relevant for epidemiological mapping in remote rainforest areas.
YURLOVA N. Institute of Animal Systematics and Ecology, Siberian Branch of Russian Academy of Sciences, NOVOSIBIRSK, RUSSIAN FEDERATION Long-term (1990-2005) changes in abundance of free living cercariae (cercarial flow) of E. aconiatum, common trematoda species in Chany Lake ecosystem (Novosibirsk region, Russia) were quantified. The changes in abundance of metacercariae we also quantified to reducing relatively to a snail host density (individual /sq.m). Vegetated shallow zone of Chany Lake bay was selected as a control area for evaluation of parasite (cercariae and metacercariae) population (individual/sq.m). Prevalence and intensiy of E. aconiatum we quantified in all potential hosts: 3 snail species of the first intermediate host and 17 snail species of the second ones; the density of snails infected with parthenitae and metacercariae was calculated. The daily productivity of cercariae, their abundance and biomass were estimated also. The results demonstrated that cercarial productivity of E.aconiatum in the abundant obligatory snail host Lymnaea stagnalis population averaged over 61 thousand/day (range 9000 and 3, 2 million/day). Thus trematode agents comprise a substantial biomass in this ecosystem. The long-term changes in cercarial and metacercarial abundance are characterized by significant cyclic fluctuation with cycle duration being 4-7 years and correlate with changes in population sizes and densities of intermediate hosts, which
SY12/05-02
PREY DENSITY, PREDATOR-PREY INTERACTION AND CESTODE 85
are mediated by cyclic alterations in water levels in the lake. This study was financed by Russian Fund of Basic Research 0704-01416.
with the highest V.I., covering 20% of the explored surface. At farm level, the proportion of pasture perimeter with V.I.=5 was significantly related to the B. divergens seroprevalence: the farms with less than 20% of their pasture perimeter with V.I.=5 showed low seroprevalences. When above 20%, the farms had a significantly higher risk of high seroprevalences.
SY12/05-04
HUMAN AND ANIMAL SEROPREVALENCE OF TOXOPLASMA GONDII IN GABON : COMPARISON BETWEEN A RURAL AND A URBAN AREA
SY12/05-06
FLEATICKRISK : A CLIMATIC MODEL DEVELOPED TO MONITOR AND PREDICT THE ACTIVITY AND THE DENSITY OF 3 TICKS SPECIES AND THE CAT FLEA IN FRANCE.
MERCIER A.(1), NGOUBANGOYE B.(2), SALLE B.(2), BISVIGOU U.(3), AKUE J.P.(2), DARDÉ M.L.(1) (1) Faculté de Médecine, EA3174 NETEC, CNR/CRB Toxoplasmose, LIMOGES, FRANCE ; (2) CIRMF Centre International de Rcherche de Franceville/ Centre de Primatologie, FRANCEVILLE, GABON ; (3) CIRMF Centre International de Recherche de Franceville, FRANCEVILLE, GABON
BEUGNET F.(1), LOUKOS H.(2), CHALVET-MONFRAY K.(3), BICOUT D.(3) (1) MERIAL, LYON, FRANCE ; (2) CLIMPACT - Paris VI, PARIS, FRANCE ; (3) ENVL, LYON, FRANCE The aim was to develop a geographical and climatological model to survey the activity and density of arthropods. The model integrates data from 100 ground stations providing 4 measures of temperature and humidity per day, linked with satellite analysis in real time and also vegetal coverage (biotopes : urban areas, sub-urbans, rural, forest). The published biological parameters of Ixodes ricinus, Dermacentor reticulatus, Rhipicephalus sanguineus and Ctenocephalides felis were transformed in matrix. The activity was defined by the capacity of infesting host. The mathematical modelling permitted to create 2 index : a weekly activity index (ranging from 0 to 100) and a cumulative index (from 0 to 1000). The cumulative index is calculated over 10 weeks, with an adjustment linked to the past season. A 7 days predictive activity index has been put in place using the daily meteo predictions. The spatial resolution is 27 km. Due to the importance of tick biotopes, a local adaptation of the results must be done by veterinarians. The accuracy of the data was studied with sentinel vet clinics as well as comparison with published surveys. A full year validation survey is ongoing with a collaboration with the French army health services and the collect of ticks iin 15 sites. The model and its maps are available for vets on www.FleaTickRisk.com. It helps to explain the rise of ectoparasites and the peaks of vector borne diseases like canine babesiosis.
Toxoplasma gondii is found in humans and animals worldwide, but the prevalence varies between different populations according to climatic characteristics, presence of felids and way of life. Our study takes place in Gabon. The seroprevalence of Toxoplasma infection in humans and animals was evaluated in Dienga, a village near the frontier of Congo. A Modified Agglutination Test (MAT) was used to detect anti-Toxoplasma antibodies in 346 humans, 168 chickens, 94 goats, 44 sheep, 77 dogs, 32 cats, 45 rats (Ratus ratus) and 40 wild rodents. A high prevalence found in humans (74,5%) and variable in domestic and peridomestic animals (50.3% for the chickens, 84% for the goats, 97.7% for the sheep, 60.3% for the dogs, 81.3% for the cats, 2.2% rats and 2.5% wild rodents) demonstrates an intense circulation of Toxoplasma in this rural area of Gabon. SY12/05-05
A VEGETATION INDEX QUALIFYING PASTURE EDGES IS RELATED TO TICK DENSITY AND REFLECTS THE RISK OF HIGH BABESIA DIVERGENS SEROPREVALENCE IN DAIRY CATTLE HERDS
AGOULON A.(1), MALANDRIN L.(2), LEPIGEON F.(1), VENISSE M.(1), BONNET S.(3), BECKER C.(1), HOCH T.(2), L'HOSTIS M.(1), BASTIAN S.(1), PLANTARD O.(2), BEAUDEAU F.(1) (1) Ecole Nationale Vétérinaire, NANTES, FRANCE ; (2) INRA, NANTES, FRANCE ; (3) INRA, MAISONS-ALFORT, FRANCE Babesia divergens, transmitted by the tick Ixodes ricinus, is the main agent of bovine babesiosis in France. This Apicomplexa is often present in asymptomatic carriers, with a large panel of seroprevalence among herds (0 to 87% in our study), regardless of clinical cases which are scarce. Although numerous factors (including vegetation type) are known to influence tick density, no risk factor of carrying B. divergens has been yet identified. Our study aimed to explore a vegetation index as an indirect marker of the within-herd B. divergens seroprevalence. In spring 2007, in 20 dairy cattle herds of Western France, all dairy cows were blood sampled and IFI serology was individually performed to measure the B. divergens seroprevalence. In addition, I. ricinus nymphs were collected by drag sampling along transects on the vegetation of each pasture perimeter for each farm. Tick density was significantly related to a Vegetation Index (V.I., ranging from 1 to 5) taking into account the abundance of trees or bushes at the pasture edge: most ticks (54%) were found in transects
SY13/01-02
MOBILE BIOLOGY LABORATORY FOR ANTIMALARIAL DRUG SUSCEPTIBILITY EVALUATION IN THE FIELD
SINOU V.(1), DESBORDES M.(1), DESPLANS J.(1), NGUYEN V.D.(2), NGYUEN T.T.H.(2), GAZIN P.(3), PARZY D.(1) (1) IMTSSA, MARSEILLE ARMÉES, FRANCE ; (2) Centre d'Analyse et de Recherche des Médicaments Militaire, HANOI, VIET NAM ; (3) Université de la Méditerranée, EA 864, MARSEILLE, FRANCE A mobile laboratory was develop to primarily assess the antimalarial drug susceptibility of Plasmodium falciparum in endemic areas where no well-equipped laboratory is available. The main elements of this laboratory include i) a trigaz incubator chamber supplied with CO2 and N2 compressed in two 5 L steel cylinders allowing two weeks of culture, ii) a class II biological safety cabinet, i) a centrifuge, iii) a microscope and iv) a portable ELISA plate reader. All were integrated into two compartimented cases weighting 250 kg (volume = 2 m3) whom the lid can be used as a working surface. The mobile laboratory was used during 2006-2007 in the Binh Phuoc province of Vietnam, on the Campuchia-Vietnam border. 86
By these years 160 fresh isolates of P. falciparum were collected and 60 were cultured using 96-pre-coated well microculture plate. The evaluation of parasite growth was by a colorimetric method based on the measurement of the levels of parasite histidine-rich protein II. A total of 45 P. falciparum isolates were successfully tested for their susceptibility to dihydroartemisinin, quinine, chloroquine and doxycycline. In conclusion, the mobile laboratory allows to identify the drug resistance status of Plasmodium infections in the field just like in the lab. In the future, this laboratory will meet the particular demands of developing countries, such as organizing and providing efficace response in face of epidemic in remote regions.
hybridization. We detect Pneumocystis in both non invasive samples, such as oropharyngeal washing or induced sputum, and invasive bronchoalveolar lavage samples. Interestingly, we found an excellent correlation between PCR results on bronchoalveolar lavage fluids and oropharyngeal washing in infected patients. The specificity of the mtLSUrRNA TD-PCR assay on oropharyngeal washing was 100% and the sensitivity was 90%. In patients with negative microscopic test, when Touch Down - PCR results where positive in bronchoalveolar lavage fluids and negative in oropharyngeal washing, the absence of PcP was clinically confirmed and colonization was concluded. In this kind of patients, nested - PCR on OW was usually positive.
SY13/01-03
SY13/01-05
OOCYST DETECTION OF TOXOPLASMA IN WATER: AN EXAMPLE OF DETECTION FROM WATER IN CHAMPAGNE-ARDENNE, FRANCE.
VILLENA I. Laboratoire FRANCE
parasitologie,
CHU
Maison
Blanche,
HUMAN-VECTOR IMMUNE INTERACTIONS DURING MALARIA : IDENTIFICATION OF AN IMMUNOLOGICAL MARKER OF EXPOSURE TO ANOPHELES GAMBIAE BITES BASED ON A PEPTIDE DESIGN OF THE GSG6 SALIVARY PROTEIN
REIMS,
POINSIGNON A.(1), CORNELIE S.(1), MESTRES-SIMON M.(2), LANFRANCOTTI A.(2), ROSSIGNOL M.(1), BOULANGER D.(1), CISSE B.(3), SOKHNA C.(4), ARCÁ B.(2), SIMONDON F.(1), REMOUE F.(4) (1) Institut de Recherche pour le Développement, MONTPELLIER, FRANCE ; (2) University Federico II, NAPLES, ITALY ; (3) Université Cheikh Anta Diop, DAKAR, SENEGAL ; (4) Institut de Recherche pour le Développement, DAKAR, SENEGAL
Toxoplasma gondii is a protozoan parasite capable of infecting a large variety of hosts. Several recent outbreaks of toxoplasmosis were related to drinking water. We propose a strategy for Toxoplasma oocysts detection, based on a multirisk waterborne parasitic approach including Giardia and Cryptosporidium recovery. Our strategy involves three steps: concentration and filtration of the water to recover small numbers of Toxoplasma oocysts, elution and purification on a sucrose density gradient, and detection based on PCR and mouse inoculation to determine the presence and the infectivity of recovered oocysts. This strategy was applied to environmental water samples. Among the interpretable samples, we detected Toxoplasma DNA in several samples, including Raw Surface Water, whose environmental matrices may be contaminated by soil washing after rainfall, Underground Water samples chosen by public health officials because of frequent pathogen recovery and occasionally in Public Drinking Water. None of the samples were positive by bioassay. Mouse bioassay is still the reference method to detect viable oocysts, but technics is too long to be purposed as an efficiency method. We need tools for rapid detection of oocysts in water, like them proposed for detection of Giardia cysts and Cryptosporidium oocysts by immunomagnetic separation and immunofluorescence detection. So that, development of monoclonal antibodies against T. gondii oocyst wall is required.
The evaluation of human antibody response to arthropod salivary proteins could be an epidemiological indicator of exposure to vector bites. In the objective to increase specificity of malaria exposure, we identified the SG6 salivary protein as Anopheles genus specific and antigenic in children living in a malaria endemic area. The objective of the present study was to evaluate if the IgG response to the An. gambiae gSG6 protein could be an immunological marker of exposure specific to An. gambiae bites. To optimize the specificity and reproducibility, we designed five gSG6-based peptides sequences by a bio-informatic approach, taking into consideration i) their potential antigenic properties and ii) their absence of cross-reactivity with proteins sequences of other arthropods/organisms. The five gSG6-peptides showed different antigenic properties with gSG6-P1 and gSG6-P2 exhibiting the higher antigenicity. However, a significant increase of the specific IgG response during the rainy season and a positive association between IgG level and the level of exposure to An. gambiae bites was only significant for gSG6P1. This step by step approach suggests that gSG6-P1 could be an optimal candidate as marker for evaluating exposure to An. gambiae bites. This marker could be employed as geographical indicator to map the risk of malaria and it could also represent a direct criterion of efficacy in the evaluation of vector control strategies.
SY13/01-04
CLINICAL VALUE OF CURRENT PCR FOR DETECTION OF PNEUMOCYSTIS JIROVECII IN INVASIVE VERSUS NON-INVASIVE CLINICAL SAMPLES
DURAND-JOLY I. Institute Pasteur of Lille, LILLE, FRANCE
Diagnosis of Pneumocystis pneumonia (PcP) is conventionally based on detection of the opportunistic fungus Pneumocystis jirovecii in lower respiratory tract samples by using microscopy. Molecular diagnosis of PcP is usually more sensitive. For more than ten years, in the Lille University hospital, we used this sampling associated with PCR for PcP diagnosis. In order to decrease the risk of cross-contamination, we used a Touch Down - single round PCR targeting the mitochondrial large subunit ribosomal RNA region, followed by oligoblotting
SY14/01-03
ETHICS AND RESEARCH METHODS
GAZZOLA S.(1), LÉVÊQUE L.(1), CHABÉ M.(2), DEI-CAS E.(3), NUROCK V.(1) (1) Department of Philosophy, Lille 3 University, VILLENEUVE D'ASCQ, FRANCE ; (2) Parasitology-Mycology Service, Pharmaceutical Sciences Faculty, EA3609, Lille 2 University, & IFR-142 Lille Pasteur Institute, LILLE, FRANCE ; (3) 87
Parasitology-Mycology Service, Microbiology Department, EA3609 Faculty of Medicine, Lille 2 University, University Hospital Centre & IFR-142 Lille Pasteur Institute, LILLE, FRANCE
SY14/02-02
RESEARCH ETHICS AND DECISION-MAKING PROCESS
LÉVÊQUE L.(1), GAZZOLA S.(1), DELHAES L.(2), DEI-CAS E.(2), NUROCK V.(1) (1) Department of Philosophy, Lille 3 University, VILLENEUVE D'ASCQ, FRANCE ; (2) Parasitology-Mycology Service, Microbiology Department, EA3609 Faculty of Medicine, Lille 2 University, University Hospital Centre & IFR-142 Lille Pasteur Institute, LILLE, FRANCE
Biomedical researcher meets the gap between paradigm of biology and observational data. Indeed, properties of living are not forcedly covered by current theoretical frameworks. Most biological data result from reduction. However nonlinear dynamics of living generate often complex behaviours or emergent properties uneasily got by reduction ways. Researcher is thus faced to the emergence – reduction dilemma, a debate with ethical involvements that lead parasitology researchers to bioethical questioning. Limits of reductionism lead to think about new ways for approaching the living. It is hypothesized that researcher standing back his practice gives himself a sort of freedom area. Here he is able to exert critical thinking in order to clarify epistemological concerns related with research strategy. Ethical questioning is legitimate because the strategies which are the subject of an epistemological choice, have impact on living’s representations. The choice is influenced by the will of reaching the most accurate living’s representation, researcher career, and political and economic constraints. The researcher becomes therefore an ethical subject in an ethical space. New possibilities, not only epistemological constraints, become available. This ethical space is the moment of a choice that contributes crucially to the living’s representation. Thus, ethics questions the link between scientific validity and social value of the living’s knowledge.
Decision-making process studies target mainly the medical world. The research context is increasingly complex. Therefore, questioning about the decision-making processes that underlies all research has growing importance. Researcher is caught in an actor’s game that exceed the doctor-patient relationship. Pharmaceutical industry, associations, health authority, ethics committees, scientific community, social world drive directly or indirectly his research, a process that is consciously or not lived by it. Stakes and outcomes of current or specific situations of either conflict or cooperation need to be understood. Likewise, how the researcher reconciles values, expectations or rules in its research? Cognitive or targeted purpose, thematic priorities, choice of method, responsibility, reputation, productivity, justice, common good intervene in its decision, but not all reconcilable. We have therefore to clarify tension mechanisms, understand complementary or opposite interactions among the multiple partners, which drive or limit the researcher decision ability. Here, we aim at showing that research strategies have an impact on representations of society and its mores, as well as the ethical dimension of research. It will be highlighted that exchange among the involved partners confers ethical value to the decision. The present analysis aims at clarifying underlying mechanisms which only rarely are the object of researcher's reflection.
SY14/02-01
CURRENT BIOETHICAL PARASITOLOGY
ISSUES
IN
BOURY D.(1), DEI-CAS E.(2) (1) Centre of Medical Ethics Catholic University of Lille, EA4031 Catholic University of Lille & Lille 2 University, LILLE, FRANCE ; (2) Parasitology-Mycology Service, Microbiology Department, EA3609 Faculty of Medicine, Lille 2 University Hospital Centre & IFR-142 Lille Pasteur Institute, LILLE, FRANCE
SY14/02-03
HOW PARASITOLOGY IS TAUGHT IN MEDICAL FACULTIES IN EUROPE? PARASITOLOGY LOST?
BRUSCHI F. UNIVERSITA' DI PISA, PISA, ITALY
In parasitology, for many reasons, the range of questions that can be ascribed to bioethics remains, to a large extent, unexplored. Yet parasitic diseases, often included among the neglected diseases, constitute the most common infections among the poorest billion people, causing about 534,000 deaths a year and leading to long-term infirmities and poverty. So, the number of public health control programs increases. Programs setting-up entails ethical consequences. In this paper, we analyse the bibliography to suggest three main issues which epitomize ethical stakes in parasitology: accounting the complexity of the field of intervention (with economic, social and cultural involvements), putting the principle of justice into practice (by acknowledgment of communities as actors) and managing the tensions between divergent scientific approaches of research. Involved in producing knowledge and mastering technological tools, parasitologists, researchers or clinicians, are called on to assume a specific responsibility, which does not simply consist in respecting statutory text or ethical rules. They can help citizens, as well as political and institutional heads to have a better understanding of the complex mechanisms of parasitic endemic diseases, the foreseeable consequences and risks of decisions made, to shed qualified light on public health issues and to transmit their values to students whose training they are in charge of.
The study of Parasitology and parasitic diseases is still necessary and will always be important to the practice of modern medicine, even in industrialised countries, especially when considering how it is easy now to travel from the north to the south on the world and vice-versa in just a matter of hours. Despite of these facts, however, many medical schools throughout the western world give this subject little attention in their teaching programs, if any at all. The results of a survey in different medical faculties in Europe, after the administration of a dedicated questionnaire are presented and compared with those obtained ten years ago in a similar manner. In particular, the situation in France, Germany, Italy and Poland shows the decrease of Parasitology Departments in many Faculties, as well as the reduction of the number of hours dedicated to this discipline in independent courses. European situation is compared with that of North-, South America, China and South East Asia. Some suggestions are given to improve the scenario.
SY15/01-02
FOLLOW UP OF PLASMODIUM FALCIPARUM GENOTYPE AND ANTI-VAR2CSA IMMUNE 88
RESPONSE SENEGAL.
DURING
PREGNANCY
Schistosoma mansoni and PbA affected the development of CM: infection with 50 cercaria, 7 weeks prior to PbA infection, reduced CM from 65% to 33%, while infection with 100 cercaria prevented CM altogether. Our results indicate that administration of immunomodulators reduces CM. This treatment extends the time window available for conventional anti-plasmodial drug treatment and therefore may increase cure rates.
IN
GUITARD J.(1), COTTRELL G.(1), MOULOPO MAGNOUHA N.(1), SALANTI A.(2), LI T.(1), SOW S.(3), DELORON P.(1), TUIKUE NDAM N.(1) (1) IRD, PARIS, FRANCE ; (2) University of Copenhagen, COPENHAGEN, DENMARK ; (3) Thiadiaye Hospital, THIADIAYE, SENEGAL Pregnancy associated malaria is responsible for morbidity in area of P. falciparum endemicity. Pregnant women develop protective anti-VSA IgG when infected. Their major target is var2csa encoded PfEMP1. Parasites expressing distinct variants of VAR2CSA seem to infect primigravidae or multigravidae. The aim of this study was to determine the dynamic of P. falciparum genotypes during pregnancy and to study the associated acquisition of anti-VAR2CSA immune response. To study the dynamic of P. falciparum population during pregnancy, msp2 genotyping and analysis of the var2csa DBL5epsilon sequence were performed. ELISA was used to measure the level of VAR2CSA DBL5epsilon specific IgG and subclasses and their levels were analyzed in relation to the infection. The results highlight the selection of some parasite variants that were able to persist over several weeks and found at delivery in the placenta. Differential evolution of anti-VAR2CSA IgG3 were observed. Significant increase of anti-DBL5epsilon IgG3 between enrolment and delivery was associated with uninfected placenta at delivery in primigravidae while this was related to an infection of the placenta in multigravidae. These results support the hypothesis that parasites infecting primigravidae and multigravidae expressed different VAR2CSA variants and highlight the fact that VAR2CSA possesses immunogenic epitopes that can be of major interest for any vaccination strategy.
SY15/01-04
IL-13 INDUCES EXPRESSION OF CD36 RECEPTOR IN HUMAN MONOCYTES THROUGH PPAR ACTIVATION AND CONTRIBUTES TO THE ENHANCEMENT OF PLASMODIUMPARASITIZED ERYTHROCYTES PHAGOCYTOSIS
OLAGNIER D.(1), COSTE A.(1), BENOIT-VICAL F.(2), BALARD P.(1), PIPY B.(1), BERRY A.(1) (1) EA2405, IFR31, Institut Louis Bugnard, TOULOUSE, FRANCE ; (2) LCC, CNRS, UPR8241, TOULOUSE, FRANCE Severe and fatal malaria is associated with the failure of host defences to control parasite replication, excessive secretion of Th1 cytokine such as TNF- and sequestration of parasitized erythrocytes (PE). CD36 has been presented as a major sequestration receptor able to bind PE via the interaction with the parasitic protein PfEMP1. However, CD36 receptor is also a component of the pattern recognition receptors on monocytes that recognizes PE and induces a non-specific phagocytosis process. It has been shown that agonists of the nuclear transcription factor PPAR induced an increase in CD36 mediated phagocytosis of PE. In this work, we have demonstrated that CD36 expression was induced in human monocytes following exposure to IL-13, a Th2 cytokine via the activation of PPAR. This signalling pathway involved phospholipase A2 activation and production of 15d-PGJ2, an endogenous PPAR ligand. Then, we have shown that CD36 and PPAR were involved in IL-13 mediated phagocytosis of PE by human monocytes. These results reveal a novel role for IL13, a non inflammatory cytokine, which is able to enhance the clearance of PE. Specific up-regulation of CD36 on monocytes may represent a novel strategy in falciparum malaria therapy.
SY15/01-03
IMMUNE INTERVENTION FOR TREATMENT OF CEREBRAL MALARIA
WAKNINE J.H.(1), GOLD D.(2), FLESCHER E.(2), MCQUILLAN J.(3), HUNT N.H.(3), BARENHOLZ Y.(4), BARHUM K.(4), ANVIR Y.(4), GOLENSER J.(1) (1) The Hebrew University of Jerusalem, JERUSALEM, ISRAEL ; (2) Department of Human Microbiology, Tel Aviv University, TEL AVIV, ISRAEL ; (3) Molecular Immunopathology Unit, Institute for Biomedical Research, University of Sydney, NSW, SYDNEY, AUSTRALIA ; (4) Biochemistry (Laboratory of Membrane and Liposome Research), The Hebrew University of Jerusalem-Hadassah Medical School, JERUSALEM, ISRAEL Cerebral malaria is mainly due to overproduction of Th1 cytokines and underproduction of others; e.g. during infection in IL-10 knockout mice, there was greater parasite sequestration, cerebral edema and hemorrhage compared with the wild type mice. Treatment of mice infected by P. berghei ANKA (PbA) with curcumin (2x50mg/kg/day/gavage, 0-5 day p.i.) prevented CM and delayed death by 10 days. There was no difference in parasitemia between experimental and control groups. Thus, curcumin alters the outcome, corresponding to immunopathogenesis. -methasone hemisuccinate (BMH), is a glucocorticoid prodrug used for its anti-inflammatory activity. Treatment with 20mg/kg BMH decreased the incidence of CM from 90% to 67%. Administration of BMH encapsulated into sterically stabilized nanoliposomes delayed symptoms and reduced CM to 18%. BMH was not effective against P. falciparum in vitro, hinting at an in vivo immunological mechanism. Concomitant infections in malaria-endemic areas may lead to altered T cell responses. Coinfection of mice with
SY15/01-05
PROGRESS WITH MALARIA ELIMINATION IN THE WHO EUROPEAN REGION
EJOV M.(1), KURDOVA-MINTCHEVA R.(2) (1) WHO EURO, COPENHAGEN, DENMARK ; (2) National Centre of Infectious and Parasitic Diseases, SOFIA, BULGARIA
89
In the recent years some clear success in malaria control is being achieved in the WHO European region. From 1999 to 2007, the reported number of malaria cases declined from 90712 to 1069. The incidence of malaria in all affected countries has been brought down to such levels that interruption of transmission of malaria has become a feasible objective. At present, autochthonous malaria continued to pose a challenge in 6 out of the 52 Member States of the Region, namely Azerbaijan, Georgia, Kyrgyzstan, Tajikistan, Turkey and Uzbekistan. Taking into account the results achieved by 2007, it is likely that P. vivax malaria will be eliminated in Armenia and Turkmenistan by 2010, and Tajikistan will be free from falciparum malaria by 2010. In areas and countries where malaria had been eliminated, attention is given to maintaining the malaria-free status. Interrupting malaria transmission by 2015 and eliminating the disease within all affected countries of
the Region is the ultimate goal of the new regional strategy. All malaria-affected countries of the Region endorsed the Tashkent Declaration “The Move from Malaria Control to Elimination, 2005”, which has supported and facilitated their decisions to undertake the new elimination effort. Elimination of malaria will contribute to strengthening the health system realizing the wide future expectations in terms of industry, trade and tourism in countries presently affected by malaria.
that are causing major economic losses to sheep industry throughout the world. Amino acid sequence characterization of nAChR predicted subunits showed characteristic features of neurotransmitter-gated ion-channel such as an extracellular dicysteine loop and the presence of four transmembrane domains. Phylogenetic relationship analyses of the nAChR subunit family revealed high evolutionary conservation among nematode species. In addition, four genes encoding close homologues of unc-29 C. elegans gene were identified providing evidences that nAChR subunit repertoire in trichostrongylids may be more diversified than the one of C. elegans.
SY15/02-02
NEMATODE TARGETED TREATMENTS IN GOATS UNDER TEMPERATE CLIMATE USING BODY CONDITION SCORING
SY15/02-04
PARAUD C., PORS I., CHARTIER C. AFSSA, NIORT, FRANCE
RECONSTITUTING SENSITIVE ACHR XENOPUS OOCYTES
Leaving susceptible worms in refugia by treating only a proportion of animals within a flock may be a way to manage anthelmintic resistance in small ruminants. A study was conducted in 3 French flocks to test the efficiency of body condition scoring as a criteria related to nematode infection. All the goats present the day of sample (193, 154 and 182 respectively for farms 1, 2 and 3) were individually sampled for faeces and scored for body condition. Milk production and lactation number were recorded. The mean egg excretions were 901, 874 and 1947 eggs per gram respectively for farms 1, 2 and 3. A significant negative correlation was observed between BCS and FEC in farm 3. The sensitivity and specificity of the following indicators: BCS (treatment of the goats having BCS lower than 2), milk production (treatment of the 25% highest producing goats), lactation number (treatment of the primiparous) and of their combinations were calculated for this farm, using as gold standards individual FECs 1000 or 1500 epg. Regarding BCS, 43% of the goats had a lumbar score lower than 2, the sensivity was 54 and 52 %, respectively for 1000 and 1500 epg. The combination of the 3 indicators (69% of the goats treated) gave the best sensitivity (80 and 83%, respectively for 1000 and 1500 epg). Body condition scoring may be useful when infection is at a high level but combination of indicators highly improve the detection of the highest infected goats.
BOULIN T.(1), GIELEN M.(1), RICHMOND J.(2), PAOLETTI P.(1), BESSEREAU J.L.(1) (1) ENS Paris, PARIS, FRANCE ; (2) University of Illinois, CHICAGO, UNITED STATES The levamisole receptor (LevR) is an ionotropic acetylcholine receptor activated by the antihelmintic drug levamisole. LevR is one of the major drug targets for treatment of parasitic nematode infections in livestock. It is also the best characterized AChR expressed in C. elegans. However, its basic pharmacological properties remain largely unknown due to the lack of a reliable heterologous expression system. We have succeeded in getting robust expression of the LevR by using information derived from C. elegans genetics. In C. elegans, at least eight genes are necessary to produce LevR: 5 genes encode AChR subunits and 3 genes encode proteins involved in intracellular trafficking and protein maturation. We have co-injected the corresponding cRNAs into Xenopus oocytes and observed robust expression of the LevR. Strikingly, each of the 8 genes is required for LevR expression in Xenopus oocytes. Acetylcholine elicits currents in the A range, with fast activation kinetics and no desensitization. Similarly, robust responses are obtained for levamisole, which behaves as a partial agonist. d-Tubocurarine, MLA and hexamethonium, antagonists of AChR, inhibit the currents. Acetylcholine and levamisole EC50 calculated in dose-response experiments are 25 M and 10 M, respectively. Our expression system could serve as the first in vitro system to test new pharmacological agents, and it could be adapted to express the LevR of parasitic nematode.
SY15/02-03
LEVAMISOLE-SENSITIVE NICOTINIC ACETYLCHOLINE RECEPTOR SUBUNIT GENE INVESTIGATION IN THE TRICHOSTRONGYLINA NEMATODES HAEMONCHUS CONTORTUS, TELADORSAGIA CIRCUMCINCTA AND TRICHOSTRONGYLUS COLUBRIFORMIS
SY15/02-05
EOSINOPHILS IN HAEMONCHUS CONTORTUSINFECTED RESISTANT AND SUSCEPTIBLE BREEDS OF SHEEP: ABOMASAL TISSUE RECRUITMENT AND IN VITRO FUNCTIONAL STATE
NEVEU C., CHARVET C., FAUVIN A., CORTET J., CABARET J. INRA, NOUZILLY, FRANCE
The nicotinic acetylcholine receptor (nAChR) is an important determinant of signal transmission at the neuromuscular junction. Nematode muscle nAChRs are selectively targeted by many drugs such as the anthelmintic family agents imidazothiazoles (e.g., levamisole) currently used against gastrointestinal parasitic nematodes of small ruminants. In the free-living nematode Caenorhabditis elegans, the levamisolesensitive nAChR is composed of five multi-transmembrane spanning subunits encoded by unc-29, lev-1, unc-63, unc-38 and lev-8 genes. Here we have identified and sequenced unc29, lev-1, unc-63 and unc-38 orthologs isolated from three contortus, trichostrongylid nematodes (Haemonchus Teladorsagia circumcincta and Trichostrongylus colubriformis)
THE LEVAMISOLEOF NEMATODES IN
JACQUIET P.(1), LACROUX C.(1), TEREFE G.(2) (1) UMR INRA/ENVT 1225, Ecole Nationale Vétérinaire, TOULOUSE, FRANCE ; (2) Faculty of Veterinary Medicine, DEBRE ZEIT, ETHIOPIA Introduction and objectives: Eosinophils are considered key elements in the response against helminth infections. Recently, a lethal effect of sheep eosinophils was demonstrated on Haemonchus contortus infective larvae in vitro. The present study was initiated to assess i) the Il-5 mRNA gene expression in the infected mucosa and in the draining lymph node, ii) the blood eosinophilia, iii) the recruitment of eosinophils in the 90
abomasal tissue and iv) the in vitro larval killing ability of blood eosinophils during a primary H. contortus infection in two breeds of sheep: the Martinik Black Belly sheep (resistant breed) and the INRA 401 sheep (susceptible breed). Results: Il5 mRNA gene expression, blood eosinophilia and tissue recruitment of eosinophils were similar in the two breeds before the experimental infection and four days after but substantially higher in the resistant breed than in the susceptible breed 15 days after. However, no significant difference was observed in the in vitro larval killing potential of eosinophils between the two breeds. In addition, Martinik Black Belly sheep harboured ten times less worms than INRA 401 sheep. Conclusion: Eosinophilic responses have been observed in both breeds during the two first weeks of a primary infection. The intensity of this response seemed to be quantitatively higher in the resistant breed; however the intrinsic effectiveness of eosinophils was similar in the two breeds.
when used in combination with Glibenclamide, an ABC transporters blocker. Interestingly, Glibenclamide accumulates in an intracellular tubular system located at the anterior end of the cell, the nucleus and along the parasite. This tubular system is labeled by the acidic marker LysoTracker-red, but not by the nuclear marker 4',6-diamidino-2-phenylindole, the mitochondrial marker MitoTracker-red, or the endocytic marker FM 4-64. These results demonstrate that ABC transporters inhibited by Glibenclamide are located at the membrane of intracellular organelles from Leishmania major. These organelles, probably associated with the lysosomal multivesicular system, may be involved in sequestering Glucantime from the cytosol. The inhibition of the ABC transporters located at the membranes of these organelles increases significantly the effective concentration of Glucantime and its leishmanicidal activity. Since combination therapy has been addressed as a way to decrease the cytotoxicity of individual drugs, the duration of therapy, and the potential development of resistance the herein described results could thus be of fundamental impact for leishmaniasis therapy.
SY15/03-02
MECHANISM OF ACTION OF MILTEFOSINE IN LEISHMANIA DONOVANI: MEMBRANE AFFINITY AND EFFECT ON PHOSPHOLIPID METABOLISM
SY15/03-04
CHEMOSENSITIVITY TO ANTIMONY OF LEISHMANIA INFANTUM ISOLATED FROM DOGS AND SANDFLIES IN THE REGION OF MONTPELLIER (SOUTH OF FRANCE)
RAKOTOMANGA M., BEN BRAHIM M., SAINT-PIERRE M., GODOY R., LIBONG D., CHAMINADE P., LOISEAU P.M. Université Paris-Sud 11, CHÂTENAY-MALABRY, FRANCE
OURY B.(1), DIABATÉ M.(1), GAZANION E.(1), VERGNES B.(1), GARCIA D.(1), PRATLONG F.(2), BASTIEN P.(2), SERENO D.(1) (1) IRD (Institut de Recherche pour le Développement), UR16 «Caractérisation et Contrôle des Populations de Vecteurs», MONTPELLIER, FRANCE ; (2) Université Montpellier 1 et Centre Hospitalier Universitaire de Montpellier, Laboratoire de Parasitologie-Mycologie, Centre National de Référence des Leishmania, MONTPELLIER, FRANCE
Miltefosine (hexadecylphosphocholine, HePC) is the first drug orally active for the treatment of visceral leishmaniasis. Taking into account of its lipid-like structure, we demonstrated its affinity for membranes and particularly for sterols suggesting an affinity for lipid rafts. Thus, sterol-depleted Leishmania donovani promastigotes by using a beta-methylcyclodextrin or a cholesterol-oxidase pre-treatment were less sensitive to HePC. Raft probably are the main domains for HePC recruitement at the level of the outer leaflet of the plasma membrane. Once HePC is internalized by the LdMT transporter described by the Gamarro group (Granada, Spain), HePC acts on the lipid metabolism by decreasing the phosphatidylcholine (PC) content of the plasma membrane whereas the phosphatidylethanolamine (PE) content is enhanced suggesting a partial inhibition of the PE-N-methyltransferase. The enhancement of lysophosphatidylcholine (LPC) content could be ascribed to a phospholipase A2 activation. References : - Rakotomanga M., Blanc S., Gaudin K., Chaminade P., Loiseau P.M., 2007. Antimicrobial Agents and Chemotherapy, 51 : 1425-1430. - Rakotomanga M., Loiseau P.M., Saint-Pierre-Chazalet M., 2004. Hexadecylphosphocholine interactions with lipid monolayers. Biochimica et Biophysica Acta, 1661: 212-218.
In the region of Montpellier (South of France), the treatment of canine leishmaniosis has been mainly based on the use of antimonials for several decades. Strains undergo therefore a high drug pressure which is likely to favour the emergence of chemoresistant strains on this long-range. To evaluate the sensitivity level to antimonials, the 50% inhibitory concentrations (IC50) of Potassium antimony tartrate (Sb(III) or trivalent antimony) and Sodium stibogluconate (Sb(V) or pentavalent antimony) were determined for promastigotes and intramacrophage amastigotes, respectively. Most of strains under study were isolated from dogs, some of them from sandflies in a limited focus of transmission between 1974 and 2006. In vitro tests showed a variability of chemosensitivities. IC50s of more sensitive promastigotes ranged from 5 g/mL to 10 g/mL while IC50s of less sensitive strains were three- to five-fold higher. Interestingly less sensitive strains have emerged as early as the first decade of using antimony for treatment of canine leishmaniosis. Results question the mode of emergence of chemoresistant strains and consequently the risk of their transmission to humans.
SY15/03-03
GLIBENCLAMIDE-RESPONSIVE ABC TRANSPORTERS IN LEISHMANIA: THEIR ROLE IN GLUCANTIME DISPOSITION AND ACTIVITY
SY15/03-05
BACTERIA VERSUS PARASITES: LACTOBACILLUS RHAMNOSUS CELL-FREE SUPERNATANT INHIBIT THE SPORULATION OF EIMERIA OOCYSTS IN VITRO
PADRON NIEVES M., DIAZ E., MACHUCA C., PONTE-SUCRE A. Universidad Central de Venezuela, CARACAS, VENEZUELA
MOLAN A.L., DE S., THOMAS D. Institute of Food, Nutrition and Human Health, Massey University, PALMERSTON NORTH, NEW ZEALAND
One cause of chemotherapeutic failure in anti-infective therapies is the increased movement of drugs across membranes through ATP-binding cassette (ABC) transporters. Herein, we show that Glucantime´s efficacy to decrease the infection rate of Leishmania-infected macrophages is enhanced
Avian coccidiosis, caused by infection with Eimeria species, is 91
considered to be one of the most important diseases of domestic poultry worldwide. The purpose of this study was to determine if the probiotic bacterium, Lactobacillus rhamnosus, generates compounds that affect the sporulation of the oocysts of three species of Eimeria using an in vitro assay. L. rhamnosus was grown in Mann-Rogosa-Sharpe broth for 24 h at 37 C anaerobically and the cells were removed from the broth by centrifugation to get the cell-free supernatant (CFS). The oocysts were incubated with undiluted or diluted (1:2-1:64 in water) CFS for 72 h at 25-29 C. Following incubation sporulated and non-sporulated oocysts were counted and percent sporulation determined. In control incubations containing oocysts of E. tenella, E. acervulina and E. maxima, 82.5-86% of the oocysts sporulated. In incubations containing diluted (1:16) CFS, 25%, 29% and 36% of the oocysts of these 3 species sporulated, respectively, and this corresponds to 70%, 67% and 57% inhibition in sporulation. (P1000; P=4,4-90,5), C. concavum met. (I=3->1000; P=83,3-95,5), D. minutus (I=1-300; P=6,8-93,3). Detected parasites caused the invasive diseases in wild flounder and also are potentially hazardous to health of flounder at its cultivation. Parasite species diversity of flounder in the Azov Sea was similar to that in the Black Sea (20-21 species), but it was smaller than in the Baltic Sea (22-25 species) and in the North Sea (25-30 species).
Cystic Echinococcosis is endemic in Argentina. To study the disease and to design control programs is necessary (among other things) the characterization of the genotypes which differ in some epidemiological features. As a consequence of the diverse geographic and climatic conditions the characterization is important in every Argentinean region. Central region of the country presents an intense farm activity where there are several types of intermediary hosts allowing the establishment of more than one epidemiological chain. The aim of this work was to determine the genotypes of Echinococcus granulosus in cattle, pig, sheep and human from Central Region of Argentina. A total of 142 isolates were recovered. DNA extraction was carried out by standard protocols. PCRs with primers for regions of the CO1 and ND1 mitochondrial genes were performed. Sixty five isolates of cattle origin belonged to the G1 genotype (common sheep strain), 35 isolates from sheep belonged to the same genotype and also 5 isolates from humans. Of a whole of 37 isolates from pigs, 17 belonged to G7 (pig strain) and the rest to G1. Within each genotype several micro variants were recorded: 3 for CO1 and 4 for ND1. This report contributes to the characterization of the E. granulosus genotypes that exist in the region and also to the knowledge of the disease epidemiology for the Central region of Argentina.
P-186
AEDES AEGYPTI POPULATION GENETICS USING SINGLE NUCLEOTIDE POLYMORPHISM
PADUAN K.S., RIBOLLA P.E.M. Universidade Estadual Paulista UNESP, BOTUCATU, BRAZIL
The mosquito Aedes aegypti is an important vector of several debilitating or lethal human arboviruses, including yellow fever and dengue. It was probably introduced into Brazil on ships 164
lineages in Northern States by allopatric fragmentation from the suture zone, and various degrees of gene flow and restricted colonization between these three major clades (Southern, Northern, and the suture zone). The lineages derived from NCA correlate well with climate derived hierarchical classification.
P-189
DETECTION AND CHARACTERIZATION OF GIARDIA AND CRYPTOSPORIDIUM IN HUNGARIAN RAW, SURFACE AND SEWAGE WATER SAMPLES
PLUTZER J.(1), KARANIS P.(2), DOMOKOS K.(1), TÖRÖKNÉ A.(1), MÀRIALIGETI K.(3) (1) National Institute of Environmental Health, BUDAPEST, HUNGARY ; (2) National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, OBIHIRO, JAPAN ; (3) Eötvös Loránd University, Department of Microbiology, BUDAPEST, HUNGARY
P-191
SURVEY OF GENETIC POLYMORPHISM IN ECHINOCOCCUS GRANULOSUS IN CENTRALEASTERN EUROPE, WITH DETECTION OF THE TWO G7 LINES AND THREE G1 VARIANTS
SNABEL V.(1), NAKAO M.(2), SEVCOVA D.(1), HUETTNER M.(3), ROMIG T.(3), SZÉNÀSI Z.(4), CIELECKA D.(5), GEORGESCU S.O.(6), SALAMATIN R.(5), EMETS A.(7), ITO A.(2), KUZMINA T.(8), DUBINSKY P.(1) (1) Parasitological Institute SAS, KOSICE, SLOVAKIA ; (2) Department of Parasitology, Asahikawa Medical College, ASAHIKAWA, JAPAN ; (3) Department of Parasitology, University of Hohenheim, STUTTGART, GERMANY ; (4) Department of Parasitology, National Center for Epidemiology, BUDAPEST, HUNGARY ; (5) Department of General Biology and Parasitology, Medical University of Warsaw, WARSAW, POLAND ; (6) 81st Surgical Clinic, University Hospital St. Spiridon, IASI, ROMANIA ; (7) Agricultural University, SUMY, UKRAINE ; (8) Institute of Zoology, NAS of Ukraine, KYIV, UKRAINE
We investigated the prevalence of Giardia and Cryptosporidium species and analysed the genotypes in 36 water samples collected from different water sources and various geographic areas in Hungary. Samples were collected from drinking water and sewage treatment plants and from the recreation area of Lake Balaton. The (oo)cysts were purified according to the USEPA 1623 method and they were detected by Immuno Fluorescence Test (IFT). Genomic DNA was extracted from all samples and then the GDH target gene for Giardia and the SSU rDNA for both Giardia and for Cryptosporidium species were amplified by PCR. 24 out of 36 samples (67%) were Giardia positive and 15 (42%) were Cryptosporidium positive by IFT. PCR confirmed that 13 out of 36 samples (36%) were Giardia positive and 10 (28%) Cryptosporidium positive. 12 Giardia and 2 Cryptosporidium PCR products were successfully sequenced. In seven samples G. lamblia Assemblage A and in one sample Assemblage B, in 4 cases Assemblage A and B have been found. In one sample C. parvum and in another separate sample C. meleagridis was detected. Sequence analysis revealed a new subtype of G. duodenalis - complex, clustered close to the Assemblage A group. This study provides the first report on simultaneous detection and genotyping of G. lamblia and Cryptosporidium species from water supplies in Hungary. Further investigations are necessary to define the public health significance of these findings.
To examine genotype diversity and intrastrain variability in Echinococcus granulosus complex in central/eastern Europe, 28 isolates from five countries were analyzed in fragments of mitochondrial and nuclear genes (cox1, nad1, ATP6, ActII). Of these, 21 isolates originated from 11 localities in Slovakia (19 isolates - pig host, 2 isolates - human), 2 isolates from northeastern Ukraine (Sumy region) from pig, 1 isolate from western Ukraine (human), 1 isolate from southeastern Hungary (human, Szeghalom, Békés county), 2 isolates from northeastern Romania (human - Tutova-Vaslui, Romanesti Iasi counties), 1 isolate from Poland (human). All isolates from Slovakia, Ukraine, and Poland possessed the G7 genotype (E. canadensis). Noteworthy, the two lines within G7 were detected in the examined sample; the former line (haplotype G7b) was associated with isolates from northeastern Ukraine (2 isolates) and 1 isolate from eastern Slovakia, the latter line (haplotype G7a) associated with remaining 20 Slovak isolates, Polish isolate and isolate from western Ukraine. The lines differed in two nucleotide substitutions in the cox1; the Slovak isolate with the G7a structure has manifested also third, additional mutation. Both Romanian isolates and Hungarian isolate (human host) exhibited the G1 genotype (Echinococcus granulosus sensu stricto). The finding of G1 in Hungary has subtly modified boundary of the known occurrence of this strain in Europe towards north.
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BIOGEOGRAPHIC ADAPTATION OF IXODES SCAPULARIS (ACARI : IXODIDAE) : GIS MODELS REVEAL POPULATION STRUCTURING AND COHERENCE WITH GENETIC LINEAGES.
ESTRADA-PEñA A.(1), MANGOLD A.(2) (1) University of Zaragoza, ZARAGOZA, SPAIN ; (2) INTA, RAFAELA (SANTA FE), ARGENTINA Climate niches used by populations of Ixodes scapularis (Ixodidae) are evaluated using a hierarchical classification of climate derived ecoregions. Analysis shows a structuring of populations associated to significantly different ecoregions. Populations in southern USA (core area of species) are enclosed by a large suture zone running from the North Atlantic States into mid Southwestern. Niche-based distribution models are created using a maximum-entropy algorithm. The amount of overlap between predicted distribution ranges is evaluated against a probability distribution of null overlap. Only populations within Southern States overlapped (ecological interchangeability), while the other populations are highly restricted to their climate niches. Populations are compared with 12S and 16S rDNA sequences, looking for coherence between climate classification and genetically derived phylogenies. Nested clade analysis (NCA) is used to determine lineages from available 12S and 16S haplotypes. This reveals a major split of northern and southern lineages, a separation of
P-192
MORPHOLOGICAL AND ISOENZYMATIC ANALYSIS OF SERGENTOMYIA MINUTA POPULATIONS OF MOROCCO
BOUSSAA S.(1), BOUMEZZOUGH A.(2), SIEBOLD B.(1), ALVESPIRES C.(3), PESSON B.(1) (1) Faculté de Pharmacie, ILLKIRCH, FRANCE ; (2) Faculté des Sciences, MARRAKECH, MOROCCO ; (3) Instituto de Hygiene e Medicina Tropical, LISBON, PORTUGAL Sandflies of the genus Sergentomyia are distributed throughout the Old World. There is no evidence that they transmit pathogens to humans but recently, the presence of Leishmania 165
donovani DNA in Phlebotomus and Sergentomyia species caught in Indian kala-azar patients dwellings suggest the possible involvement of multiple sandfly species in kala-azar transmission (Mukherjee et al., 1997). In France, Toscana virus RNA was detected in Sergentomyia minuta (Charrel et al., 2006). The number of female cibarial teeth is the only morphological character separating S. minuta into two subspecies: about 40 teeth for S. minuta minuta European subspecies and 70 for S. minuta parroti from North Africa. In contrast, previous studies in western (Rioux et al., 1975b) and in eastern Mediterranean basin (Léger et al., 1979; Pesson et al., 1984) showed that S. minuta is a single species with a continuous variation (cline) of this character. In the aim to verify this hypothesis, morphological and isoenzymatic analysis of wild populations of S. minuta parroti from Morocco and of S. minuta minuta from continental Europa was carried out. Morphological result confirmed the cline hypothesis, in contrast, genetic variability showed a separation between northern and southern Mediterranean basin populations.
P-194
GENOTYPES OF PLASMODIUM FALCIPARUM IMPORTED TO POLAND
MYJAK P., STEMPNAKOWSKA K., PIETKIEWICZ H., NAHORSKI W., SZOSTAKOWSKA B. Medical University of Gdansk,Inter-Faculty Institute of Maritime and Tropical Medicine, Chair of Tropical Medicine and Parasitology, GDYNIA, POLAND Pathogenesis of Plasmodium falciparum is connected with i. a. three high polymorphic genes: msp1,msp2 and glurp encoding surface proteins of the parasite. The proteins play an important role in penetration of P. falciparum into red blood cells. The aim of the study was to estimate what genotypes of P. falciparum were imported to Poland from malarial endemic areas as well as if the infections were associated with single or few different genotypes. Therefore, 94 blood samples were taken from Poles that came back from malarial areas (mainly Africa). Infections of P. falciparum were confirmed with microscopic investigations and PCR reactions. Genotyping was performed with the use of nested PCR as described by Snounou et al. (Trans. R. Soc. Trop. Med. Hyg. 1999, 93:360). Positive results of PCR reaction (at least in one group of the investigated genes) were noted in 82 samples. Msp2 was the most often amplified gen. Our results showed that in about 50% of patients occurred infection caused by few different genotypes of P. falciparum. The number of obtained different alleles ranged from 1 (RO3 family of the msp1gene) to 17 ( in glurp gene).The mean number of detected alleles (for one explored sample) were highest (1.8) for MAD20 family of the msp1gene, lower (1.42) in FC27 family of the msp2 gene, and lowest (1) in R033 family of msp1gene.
P-193
GENETIC VARIATION IN ECHINOCOCCUS MULTILOCULARIS FROM EUROPE ASSESSED BY DNA SEQUENCING
SNABEL V.(1), NAKAO M.(2), D´AMELIO S.(3), BUSI M.(3), SEVCOVA D.(1), ROMIG T.(4), BAGRADE G.(5), GOTTSTEIN B.(6), SRÉTER T.(7), BOUCHER J.M.(8), ITO A.(2), DUBINSKY P.(1) (1) Parasitological Institute SAS, KOSICE, SLOVAKIA ; (2) Department of Parasitology, Asahikawa Medical College, ASAHIKAWA, JAPAN ; (3) Department of Sciences of Public Health, Sapienza University of Rome, ROME, ITALY ; (4) Department of Parasitology, University of Hohenheim, STUTTGART, GERMANY ; (5) State Agency "Natural History Museum of Latvia", RIGA, LATVIA ; (6) Institute of Parasitology, University of Bern, BERN, SWITZERLAND ; (7) Veterinary Diagnostic Directorate, Central Agricultural Office, BUDAPEST, HUNGARY ; (8) AFSSA Nancy, France, NANCY, FRANCE
P-195
PROFOUND POPULATION STRUCTURE IN THE PHILIPPINE BULBUL IS NOT REFLECTED IN ITS HAEMOPROTEUS AVIAN MALARIA PARASITE
SILVA ITURRIZA A., KETMAIER V., TIEDEMANN R. Unit of Evolutionary Biology/Systematic Zoology, Institute of Biochemistry and Biology, University, POTSDAM, GERMANY
To examine genetic variability in Echinococcus multilocularis from Europe, isolates from a contiguous territory of 8 countries (Slovakia, Germany, Hungary, Poland, Latvia, Switzerland, France, Austria) were sequenced in five mitochondrial and nuclear genes (cox1, nad1, 12S rRNA, ATP6, ActII). Fixed differences in isolates attributable to the major European variant compared with isolates from North America and Japan were manifested by 1-4 nucleotide substitutions (in 3 positions in cox1 and atp6, 2 positions in nad1, 1 position in 12S rRNA). In nad1 and ATP6 genes, the major European variant shared sequences with isolates from north-central China (examined by Yang), which differ from an American/Japanese form. Several additional genetic forms were detected in European sample, among which an Austrian human isolate has differed most strikingly, displaying 10 mutations in four genes compared with remaining European isolates. 4 isolates from the Feldstetten locality (southern Germany) exhibited 5 mutations in three genes, 1 Latvian isolate displayed the form similar to that from Feldstetten, and 1 Slovak, 1 Hungarian and 1 French isolate have manifested one mutation. Given the amount and nature of mutations in examined European isolates, contribution of genetic factors to the recent expansion in parasite distribution in Europe is not likely based on gathered data. Acknowledgements: The study was supported by projects APVV-51-027605 and VEGA 2/7186/27.
Here we present a comparative phylogeographie study on the Philippine Bulbul (Hypsipetes philippinus), and its blood Haemoproteus malaria parasites. We used molecular data to ask whether the phylogeographic patterns in this insular hostparasite system might reflect co-evolutionary trajectories. To assess the pattern of genetic structuring of the Philippine Bulbul across 6 islands of the Central Philippine archipelago we used fragments of three mitochondrial genes. Birds were screened for the occurrence of parasites by species-specific PCRs of the mitochondrial cytochrome b gene. The phylogeography of the Philippine Bulbul mostly reflects the geographical origin of samples. 48 % of the analysed birds turned out to be infected with Haemoproteus (57 % with multiple infections). The parasite phylogeography reveals very little geographical structure, suggesting extensive gene flow among locations. While movements of birds among islands seem to be very rare, we found several instances of cooccurrence of evolutionary divergent haplotypes in the parasite. We conclude that historical processes have played a major role in shaping the host phylogeography, while ongoing population phenomena, possibly mediated by the invertebrate host, are predominant in the parasite. These contrasting patterns may account for the lack of an evident co-evolution in the system.
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Xth European Multicolloquium Of Parasitology Paris - France August 24th-28th, 2008
Poster session #2
167
investigations describing T. gondii oocysts detection in environmental soil samples with rapid molecular detection methods. The results of our findings showed that soil contaminated with T. gondii oocysts may create a real threat for human health. The proven presence of T. gondii oocysts in the sand-pits creates a real risk of contracting primary toxoplasmosis by children playing in contaminated sand. This work was supported by MES grant N°2P05D00729.
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COMPARISON DOT BLOT AND CULTURE METHODS TO DETECTION LEISHMANIA ANTIGEN WITHIN NATURALLY INFECTED SANDFLIES IN ABARDEJ, IRAN.
NEKOUIE H. Pasteur, TEHRAN, ISLAMIC REPUBLIC OF IRAN
Leishmaniasis, a vector borne disease caused by obligate intramacrophage protozoa, is characterized by diversity and complexity. Of the 88 endemic countries, 26 are in the old world including Iran. Zoonotic cuntaneous Leishmaniasis (ZCL) is a major health problem in many rural areas and Phlebotomus papatasi is a well known vector of Leishmania major the causative a gent of cutaneous Leishmaniasis in Iran. We collected phlebotomus by funnel traps and CDC miniature light trop. Funnels were placed on rodent burrows and also light trop at early nights. The blood Fed females Papatasi were selected and immersed in 70% alcohol for remove surface bacteria. The specimens were transferred to 0.5 ml PBS and rushed their stomach. One drop of this suspension transferred to NNN culture media contain 200 IU penicillin and were incubated at 24C° , and one drop fixed in Nitrocellulose paper for DotBlot assay. 130 blood fed females papatasi of suspension were identified by this tow method. Of specimens 25% were positive in culture media, 41% were positive by dot immunoblot . We can detect 10 μl of leishmania antigen by Dot Blot. Dot blot assay is considered a very discriminatory system for detecting Leishmania infection in fielded study and screening test. The simplicity, high sensibility and high specificity of the dot assay should make it useful for field studies.
P-198
RISK MAPPING OF THE VISCERAL LEISHMANIASIS INCIDENCE AND ROLE OF THE LOCAL ENVIRONMENTAL VARIATIONS IN AHAR AND KALAYBAR DISTRICTS, NORTH WEST OF IRAN
MAZLOUMI GAVGANI A.(1), HELALI H.(2), ABDOLLAHI H.(3), KOOSHA A.(3), GHAZANCHAEI A.(4), BADAKHSHAN H.(3), DASTGIRI S.(5), DAVIES C.(6) (1) Tabriz University of Medical Siences, TABRIZ, ISLAMIC REPUBLIC OF IRAN ; (2) Faculty of civil engineering, University of Tabriz, TABRIZ, ISLAMIC REPUBLIC OF IRAN ; (3) CDC of the Ministry of Health and Tabriz University of Medical Sciences, TABRIZ, ISLAMIC REPUBLIC OF IRAN ; (4) Infectious and Tropical diseases research center , Tabriz University of Medical Sciences, TABRIZ, ISLAMIC REPUBLIC OF IRAN ; (5) Epidemiology dept, Faculty of Medicine, Tabriz University of Medical Sciences, TABRIZ, ISLAMIC REPUBLIC OF IRAN ; (6) Dept of Infectious & Tropical Diseases, London school of hygiene and Tropical Medicine, UK, LONDON, UNITED KINGDOM Introduction VL is a vector-borne disease highly influenced by environmental factors. VL is an increasing and spreading public health problem in Iran, especially in the north-west of country. This study was developed for mapping the distribution and occurrence of VL in North West Iran in relation to different affecting parameters. Material and Methods ArcView GIS ver 3.3 (ESRI, Inc) were used to extract and map for environmental variables of villages in kalaybar districts including climate variation , topographic variation, socioeconomical impact, life style and villages environments effect, individual factors. Disease data were combined with census data to calculate disease incidence rates at varying spatial scales, including the level of the district, sub-district and village. Results Our results showed that distance from the river, rainfall, Temperature and low altitude, dog abundance and density, nomadic condition are the main variables associated with the distribution and incidence of VL in Kalaybar and Ahar. The results of the present work show a correlation between distributions of human and canine leishmaniasis cases in the kalaybar districts. Discussion We discussed the finding of this study with comparison of the other similar surveys, and found some similarities on sevral parameters. Findings of this study are relevant to the planning of effective control strategies for VL in north-west Iran. key words: GIS, leishmani, Iran
P-197
DETECTION OF TOXOPLASMA GONDII OOCYSTS IN ENVIRONMENTAL SOIL SAMPLES (SAND-PITS)
LASS A.(1), PIETKIEWICZ H.(1), SZOSTAKOWSKA B.(1), MYJAK P.(1), DUMETRE A.(2), MODZELEWSKA E.(1) (1) Medical University of Gdansk, Inter-Faculty Institute of Maritime and Tropical Medicine, Department of Tropical Parasitology, GDYNIA, POLAND ; (2) Laboratory of Parasitology, Mycology, Hygiene and Zoology, Faculty of Pharmacy, 27 BD JEAN MOULIN, 13385 MARSEILLE CEDEX, FRANCE Infections caused by Toxoplasma gondii are prevalent in humans and animals throughout the world. Soil contaminated with T. gondii oocysts may be one of the main sources of infection in human. There is poor number of reports concerning T. gondii oocyst detection in soil. Therefore, the aim of this study was to estimate occurrence of T. gondii oocysts in soil and determine the genotype of the parasites. A total number of 84 soil samples were taken from different sand-pits located in the area of the Tri-City (northern Poland). Oocysts were recovered using flotation method. Then, for specific T. gondii detection PCR reactions were performed with the use of primers targeting T. gondii B1 gene and repetitive element (REP). Positive samples were genotyping by means of restriction analysis of the appropriate T. gondii SAG2 gene fragments. The presence of T. gondii DNA was noted in 13 samples. Detected parasites were classified as genotypes: SAG2 type I and SAG2 type II. This is one of the first 168
there were no records on occurrence of DR in animals in POL, we are not sure that the 4 cases of dirofilariasis described are the domestic infections. However, the presence of parasites, even those acquired abroad, in Polish patients is another argument that confirms that human dirofilariasis expands in Europe.
P-199
PARASITES OF NON-INDIGENOUS FISH SPECIES IN THE MEDITERRANEAN SEA: TWO CASE REPORTS
MERELLA P.(1), FARJALLAH S.(2), PAIS A.(3), FOLLESA M.C.(4), GARIPPA G.(1) (1) Università di Sassari, Dipartimento di Biologia Animale, SASSARI, ITALY ; (2) Institut Supérieur de Biotechnologie de Monastir, Unité de Recherche: Génétique, Biodiversité et Valorisation des Bioressources, MONASTIR, TUNISIA ; (3) Università di Sassari, Dipartimento di Scienze Zootecniche, SASSARI, ITALY ; (4) Università di Cagliari, Dipartimento di Biologia Animale ed Ecologia, CAGLIARI, ITALY
P-201
TRENDS IN THE INCIDENCE OF PARASITIC INFECTIONS IN THE CZECH REPUBLIC OVER 1990-2007
TOLAROVA V.(1), PYSOVA I.(2), NOHYNKOVA E.(2) (1) Regional Institute of Public Health, Department of Parasitology and National Reference Laboratory for Diagnostics of Intestinal Parasitoses, PRAGUE, CZECH REPUBLIC ; (2) Teaching Hospital Na Bulovce, Department of Tropical Medicine and National Reference Laboratory for Diagnostics of Tropical Parasitic Diseases, PRAGUE, CZECH REPUBLIC
Biological invasions are considered global phenomena that may threaten biodiversity. In particular, the entry of non-indigenous species (NIS) in the Mediterranean Sea, from both the Atlantic and the Indo-Pacific, is by now a well documented phenomenon. According to CIESM, in 2007 fish NIS in the Mediterranean Sea included 33 Atlantic and 65 Indo-Pacific species. Host-parasite interactions play a key role for the success of NIS. In fact, the absence of a co-evolutionary equilibrium between hosts and parasites, and consequently the chance of NIS to enter only with a sub-set of natural parasites (i.e. the enemy release hypothesis, ERH), may result in a fitness advantage. The parasites of 2 fish NIS from the Mediterranean, the Lessepsian Fistularia commersonii (7 specimens) and the Atlantic Gaidropsarus granti (1 specimen), were examined. The parasite assemblages of both fish showed a relatively high richness and were mainly characterized by native generalist species, but also included some specific Allolepidapedon fistulariae and parasite (i.e. Neoallolepidapedon hawaiiense in F. commersonii, and Anisakis simplex s.s. in G. granti). The results suggested that both NIS entered the Mediterranean as adults, and indicated that NIS may carry part of natural parasites and/or acquire native parasites in the new habitat, suggesting caution in the uncritical acceptance of the ERH. Research supported by Fondazione Banco di Sardegna 2006
Here we report the results of the cooperation among two National Reference Laboratories and 75 field diagnostic laboratories to monitor trends in the incidence of parasitic infections in the Czech Republic over the last 18 years. The analysis has been possible due to the consistency in using of the diagnostic methods over this period. Whereas, in spite of the increasing travelling all over the world, a significant decrease in cases of common intestinal parasitoses, namely of giardiasis (from 3075 cases in 1990 to 204 in 2007) and taeniasis (from 200 to 25 cases per year over the same period), was observed, the incidence of other intestinal parasitic diseases, such as schistosomiasis, diphyllobothriasis and cyclosporiasis, remained sporadic. Annual incidence of asymptomatic intestinal infections with Entamoeba histolytica/ E. dispar was nearly the same during the observed period with 40 cases per year in average (only 5% represented infection with E. histolytica). The number of imported malaria did not show any increasing tendency either in contrast to a slightly growing number of cases of visceral leishmaniasis and extraintestinal amebiasis. Interestingly, though amebic abscess is usually an imported infection from an endemic country, several cases of the disease occurred recently in patients without any travel history.
P-200
FIRST CASES OF HUMAN SUBCUTANEOUS DIROFILARIASIS (DIROFILARIA REPENS) IN POLAND
P-202
ARNOWSKA-PRYMEK H., CIELECKA D., SALAMATIN R. Medical University of Warsaw, WARSAW, POLAND
INTRODUCTION A SIMPLE AND RAPID MODIFIED BLOOD AGAR MEDIA FOR MASS CULTIVATION OF LEISHMANIA SPP.
The area of endemic occurrence of D. repens (DR), in Europe includes mainly Mediterranean countries and until recent time it had not crossed the border drawn by Alps. However, in some regions of Central Europe (HUN, UKR, SVK, CZE) dirofilariasis becomes the autochthonic infection. In this part of Europe, the infections of dogs as well as the cases of humans, most frequently in HUN and UKR, and recently also in SVK have been recorded. In POL, we had the case of infection with DR in patient, who had been to GRC 4 years before (Cielecka et al. 2007, Wiad Parazytol 53, supl.:165) described for the first time. Since that time, i.e. in the period of 03-2007 to 03-2008 three new cases of subcutaneous dirofilariasis have been diagnosed. All four patients live in Warsaw or Warsaw suburbs. Three of them had visited the areas of endemic dirofilariasis before the appearance of subcutaneous bumps. These were travels to GRC (1), ITA and UKR (1) and ZAF (1) in the period from several months to several years before the symptoms were identified. For one person the only visit abroad was spending holidays on the river around Prague (CZE), 10 years ago. Since
HABIBI P., MOTAZEDIAN M.H. Shiraz medical school, SHIRAZ, ISLAMIC REPUBLIC OF IRAN
Introduction: Since identification of Lieshmania based not only on external characteristics, use of some internal characteristics including molecular structure of organism, like isoenzymes.It is necessary to perform these tests 1010 organisms. Invitro mass cultivation needs different rich liquid culture media with FCS (fetal calf serum) at 22°-26°are used. Thus presentation of cheep media which can guarantee the growth of parasite seems necessary. Materials and Methods: 40-sample taken from patients suspicious of cutaneous leishmaniasis were cultivated on NNN media. Ten sample growth. Four sample mass cultivation both broth and modified solid Media were use after 2-3 times cultivations in modiefied media 1010 promastigots were obtained. compared Schisodeme pattern of RFLP (Restriction fragment length 169
polymorphism) for Leishmania major Cultivated in two manners on blood agar and in Brian Heart Infusion broth with 10% fetal calf serum. Results: Amount of promastigote growing in blood agar media proved to be 6 times greater than brain heart broth media. It took 1-2 weeks growing on the media. Similar leishmania major RFLP Schisodeme pattern mass cultivated with both methods can reveal unchanged natural of the parasite Conclusion: Use of blood agar media for mass cultivation decreased contaminations, repeating cultures and Costs of material. At least it increased growing of parasites. Key Words: blood agar, leishmaniaSpp. Iran
malaria in a boy, illustrating epidemiological and clinical aspects of pediatric severe malaria. We discuss contribution of new molecular approach to malaria diagnosis, treatment choice, and close monitoring.
P-205
GENOTYPING OF CRYPTOSPORIDIUM SPECIES FROM HUMAN STOOL IN TURKEY.
USLUCA S., AKSOY U. Dokuz Eylul University, ZMIR, TURKEY
The search for Cryptosporidium spp. in patients with diarrheal disease who referred to Parasitology Laboratory in Dokuz Eylül University, Faculty of Medicine by means of microscopic and molecular techniques and species identification was aimed. The stool samples of the diarrheic patients with the Bristol stool form scale of 5, 6 and 7, were researched by Kinyoun acid-fast staining, polimerase chain reaction and restriction fragment length polymorphism assay. Cryptosporidium spp. were detected by Kinyoun acid fast staining method in 17 stool samples, these were included in the study and microscopically 43 parasite negative samples were included as controls. Using primers Cry 9 and Cry 15, Cryptosporidium spp. spesific 550 bp band was observed in 15 stool samples. Also, 6 of 43 microscopically negative stools were detected Cryptosporidium spp. positive by PCR. We used RsaI restriction enzyme in identification of species of all Cryptosporidium spp. positive by PCR. Twenty patients had 34, 106, 413 bp bands of Cryptosporidium parvum while one patient had 34, 147, 372 bp bands spesific for Cryptosporidium meleagridis. We believe molecular methods are necessary in understanding the source and routes of transmission of Cryptosporidium spp.
P-203
COMPARISON BETWEEN MULTIPLEX PCR, IMMUNOCHROMATOGRAPHIC ASSAY AND STAINING TECHNIQUE IN DIAGNOSIS OF INTESTINAL CRYPTOSPORIDIOSIS
SOLIMAN R.H.(1), EL HAMSHARY E.M.(1), EL SAYED H.F.(2), HUSSEIN E.M.(1), EL SAYED H.Z.(1) (1) Suez Canal University- Faculty of medicine- Parasitology Department, ISMAILIA, EGYPT ; (2) Suez Canal UniversityFaculty of Medicine-Pediatrics Department, ISMAILIA, EGYPT Cryptosporidiosis is an important enteric parasitic infection that is associated with significant morbidity and mortality, especially among immunosuppressed individuals in the developing world. Numerous reports have identified two distinct genotypes of C. parvum isolates: the human genotype (Cryptosporidium hominis), and the cattle genotype (Cryptosporidium parvum), which can also infect humans. The aim of this study is to compare the performance of the traditional kinyoun’s acid fast staining to immunochromatographic strip assay, with multiplex genotyping PCR (targeting the dihydrofolate reductase gene) as a gold standard in diagnosis of cryptosporidiosis. The study was performed on 76 immunocompromised patient suffering from diarrhea attending oncology, nephrology and gastroenterology departments in the Suez- Canal university hospital. The kinyoun’s acid fast staining technique achieved 79% sensitivity and 98% specificity while the immunochromatographic strip assay achieved 89% sensitivity and 100% specificity. C. parvum comprised %68.4 of Cryptosporidium positive cases while C. hominis comprised only 26.3% indicating that transmission of Cryptosporidium in this study was mainly zoonotic. Interestingly, there was one case with mixed C. parvum and C. hominis infection. The results of the study recommend the wide scale use of the immunochromatographic strip assay as an easy and reliable method for the diagnosis of cryptosporidiosis.
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IDENTIFICATION OF CRYPTOSPORIDIUM SPP. IN TWO PATIENTS (ONE IMMUNOCOMPETENT AND ONE IMMUNOCOMPROMISED) IN TURKEY BY USING PCR-RFLP
USLUCA S., AKSOY U. Dokuz Eylul University, ZMIR, TURKEY
Cryptosporidium is an intestinal protozoan found both in humans and a wide range of animals and is known as important etiologic agent of diarrheal disease in humans affecting mostly children and immunocompromised individuals. The stool samples of two patients who referred to Parasitology Department of Dokuz Eylül University, Faculty of Medicine, were stained by Kinyoun acid-fast method and oocysts of Cryptosporidium spp were observed. First patient was 8 years old girl who had diarrhea since 5 days. She had no sign of immunodeficiency. Second patient was 48 years old, male and was complaining from intermittent diarrhea. He also had HIV, Caposi sarcoma and nonhogkin lymphoma. DNA extraction from stool samples were done with DNA stool mini kit and specific bands of Cryptosporidium spp. were detected in polymerase chain reaction. In immunocompetent girl, Cryptosporidium parvum specific 34, 106, 413 bp bands were observed in restriction fragment length polymorphism whereas in immunocompromised HIV positive patient, 34, 147, 372 bp bands were seen which were spesific for C. meleagridis known to infect avian. C. parvum may infect both immunocompetent and immunocompromised patients whereas C. meleagridis mostly infects immunocompromised patients. Though we were unable to detect the real source and the routes of transmission, the fact that both parasites are widely known to infect animals,
P-204
PEDIATRIC IMPORTED MALARIA: PLACE OF MOLECULAR APPROACH IN AN USUAL DELAYED DIAGNOSIS
DELHAES L.(1), FREALLE E.(1), BERRY A.(2), DUTOIT E.(1), LECLERC F.(1), BEAUDOU J.(2), LETEURTRE S.(1), BENOITVICAL F.(2) (1) Lille Hospital, LILLE, FRANCE ; (2) Toulouse Hospital, TOULOUSE, FRANCE Malaria is a polymorphous life-threatening disease, for which children are particularly at risk since they may appear asymptomatic at presentation. We report a case of imported 170
we believe our both patients’ infections were zoonotic origined.
opens the possibility to screen for a large number of pathogens and immediate typing in a cost efficient manner within a relatively short period of time.
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P-209 Abstract withdrawn by their authors
DEVELOPMENT OF A REAL-TIME PCR FOR DIFFERENTIATION OF GENOTYPE G1 AND CLUSTER OF GENOTYPES G2/G3 OF ECHINOCOCCUS GRANULOSUS
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PFHRP2 AND PLDH ANTIGENS DETECTION FOR MONITORING ARTEMISININ-BASED COMBINATION THERAPY (ACT) EFFICACY IN UNCOMPLICATED FALCIPARUM MALARIA IN BENIN.
MAURELLI M.P.(1), CAPUANO F.(2), RINALDI L.(1), VENEZIANO V.(1), PERUGINI A.G.(2), ALFANO S.(1), FUSCO G.(2), CRINGOLI G.(1) (1) Università Degli Studi di Napoli Federico II, NAPOLI, ITALY ; (2) Istituto Zooprofilattico Sperimentale del Mezzogiorno, NAPOLI, ITALY
HOUZE S.(1), DICKO BOLI M.(2), LE BRAS J.(1), DELORON P.(3), FAUCHER J.F.(2) (1) APHP Laboratoire de Parasitologie et EA209 Université René Descartes, PARIS, FRANCE ; (2) UR010 Santé de la mère et de l'enfant en milieu tropical, COTONOU, BENIN ; (3) IRD10 Santé de la mère et de l'enfant en milieu tropical, PARIS, FRANCE
Cystic echinococcosis caused by larval stage of cestode Echinococcus granulosus is a worldwide zoonosis, constituting one of the major public health problems in many countries of the world. To day, 10 different strains of E. granulosus have been described using mitochondrial DNA sequence data (G1-G10). In E. granulosus 12S mtDNA gene, between G1 and G3 genotypes there are two fixed nucleotide substitutions at the positions 166 and 205, whereas no mutation has been observed between G2 and G3 genotypes. Using this information, a SYBR real-time PCR assay in conjunction with melting curve analysis has been developed for a rapid differentiation of the genotype G1 from the cluster of genotypes G2/G3 of E. granulosus. After analysis, the 12S mtDNA Tm for G1 samples ranged from 76.2°C to 76.6°C, while it ranged from 76.8°C to 77.2°C for the G2 and G3 samples. Even if the Tm of G1 and G2/G3 samples are bordering, Tm ranges never overlap, so a detect mean difference of Tm of 0.6°C between G1 and G2/G3 genotypes allows a fast and simple discrimination of these genotypes. This innovative method is much more convenient economically and less time-consuming than conventional utilized methods of differentiation of E. granulosus genotypes based on PCR followed by sequencing analysis, so with this protocol a large number of samples can be rapidly tested and repeatable results can be obtained within a short time, also with a notable saving of reagents.
We compared the efficacy of 2 commercial malaria rapid diagnostic tests (RDT) detecting P. falciparum histidine rich protein 2 (PfHRP2) (Immunoquick® Malaria), or panlacticodehydrogenase (p LDH) and Pf-lacticodehydrogenase (Optimal® IT) to monitoring malaria treatment. Data were evaluated as regards to thick blood smears results. In Allada (Benin), 205 children aged 6-59 months with microscopicallyconfirmed uncomplicated falciparum malaria were randomized to receive either artesunate/amodiaquine, artemether/lumefantrine or sulfadoxine-pyrimethamine. After treatment, blood samples were examined for parasites and antigens for 42 days. At baseline, PfHRP2 based test was positive in 203 children (99%), and both pLDHs tests in 204 (99.5%). During the follow-up, PfHRP2 test produce positive results in 80.5% (132/164), 70.1% (115/164), 44.2% (69/156), 26.8% (38/142), 26.4% (33/125), 4.9 % (4/81), and 5.2% (4/77) of effectively treated children (with negative thick blood smear) at days 3, 7, 14, 21, 28, 35, and 42, respectively. With both p LDH-tests, the specificity of the antigen detection was 87% (141/162) on day 3 and ranged from 92% to 100% from day 7 until day 42. The pLDH–based RDT, but not PfHRP2–based RDT could be used in non-microscope areas for monitoring the therapeutic responses to anti-malarial therapy, especially following ACT treatment.
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A MULTIPLEX MOLECULAR ASSAY FOR DETECTION AND TYPING OF GIARDIA LAMBLIA.
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INVESTIGATION OF LIPID BINDING PROTEINS OF PARASITIC HELMINTES FROM THE GENUS TRICHINELLA AND FASCIOLA
ROELFSEMA J., BRANDES S., KORTBEEK L. National Institute of Public Health, BILTHOVEN, NETHERLANDS
RADOSLAVOV G.(1), TEOFANOVA D.(1), LIEBAU E.(2), BANKOV I.(1) (1) Institute of Experimental Pathology and Parasitology, Bulgarian Academy of Sciences, SOFIA 1113, BULGARIA ; (2) Westfälische Wilhelms-Universität (WWU), MÜNSTER, GERMANY
Giardia lamblia is a unicellular parasite that can cause severe gastro-enteritis. Several strains are known, numbered A to F. Only the A and B strain seem to infect the human intestine; other strains prefer other animals. Typing the strains is important for the diagnosis, as the B type is the more disease causing strain. In addition, typing is important for epidemiological studies. Diagnosis of a Giardia lamblia infection can be achieved in several ways. The molecular diagnostic approach usually is a PCR. One or more loci can be amplified and the products can be sequenced. Sequence analysis will reveal which strain is the infecting one. The strategy of amplifying and subsequent sequencing is slow, laborious and expensive. Using the Multiplex Ligation-dependent Probe Amplification (MLPA) we can amplify various loci and type nucleotide variations that are characteristic for the Giardia types. Because the MLPA can amplify many loci in one reaction, in contrast to real-time PCR on whatever platform, this strategy 171
Lipid Binding Proteins (LBPs) are members of a widely distributed proteins which bind and transport different lipophilic ligands. They also regulate the physiological activity, metabolism and disposition of essential hydrophobic compounds like fatty acids, phospholipids, eicosanoids and retinoids which are important molecules involved in several cellular processes including gene transcription, immune responses etc. Fatty acid binding proteins (FABPs) are mostly members of cytosolic proteins but some members are secretory proteins. There is a lot of data that helminthic FABPs from E/S products cause antigenic and allergenic reactions. In various
nematodes some of them are known such as Nematode Polyprotein Allergens/Antigens (NPA). LBP from the parasitic nematode Trichinella spiralis are not known. LBP from T. spiralis larvae has been isolated, purified and N-terminal sequenced. Secondary structure showed by CD was predominantly -structural. MW of the protein without the signal peptide is 46kDa. The protein contains two homology domains and a poly-His tail. The Cys and His residues could be involved in the metal binding properties of the protein. There is not homology with any known sequences (with the exception of the ESTs of Trichuris muris and T. vulpis). The native FABP3 from Fasciola hepatica have been purified and recombinant one has been obtained in E.coli. Their fatty acid binding activity has been determined using the fluorescent fatty acid analogue DAUDA and [14C]-palmitic acid. Our future plans are to investigate the effectiveness of the native and recombinant FABP3 as possible antigen for vaccines.
Parasitology represents the principal Ambulatory activity, realizes the prevention and the cure of parasitic disease of income animals and pets, helping the animal welfare, spreading and informing owners on the problem of zoonosis. Animal species treated in percentage: birds 50%, pets 35%, ruminants 8%, horses 2%, bees and others 5%. Therapy using 1st, 2nd, 3rd generation active costituents: idrocarbure alogenate, piperazine, flumetrina, carbamate, imidacloprid, dimetridazole, econazole, sulphachinossaline, esterophosphorique, tetramisole, febantel, griseofulvine, praziquentel, mebendazole, sulphametazina, sulphatiazolo, levomisolo, ivermectina, pirantel, niclosamide, coformulanti, tiabendazolo, morantel, niclofen, rafoxanide. The growth of modern anthelminthics coincides with the growth of chemical industry. The modern anthelminthic treatments are the presupposed of healthy breedings and warranty of elevated productions, as well as protection for the health of man. The future of Parasitology is to prepare Vaccines for animals and man. The complex vital cycles with intermediate hosts and anatomic localizations (Leishmanies in macrophagocytes, pyroplasms on erytrocytes) constitue serious limitations for this realization. Parasites escape the immune system control masking their antigens, or releasing them in circle, as well as changing their superficial antigenic composition during host passage. Chronicity develops self immunitary phenomenons.
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THE IMPACT OF THE VISUAL AIDS AND MODERN TECHNOLOGY ON INCREASING THE COMMUNICATION ABILITIES DURING THE PARASITOLOGY CLASSES IN A ROMANIAN UNIVERSITY CENTRE
NEGHINA R., NEGHINA A.M., MARINCU I., IACOBICIU I. Victor Babes University of Medicine and Pharmacy, TIMISOARA, ROMANIA
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SELECTION AND PHENOTYPE CHARACTERISATION OF SITAMAQUINERESISTANT PROMASTIGOTES OF LEISHMANIA DONOVANI
Background: The improvement of the communication between students and the teaching staff requires a feed-back interaction, reciprocity and the substitution of the verbal messages with multilateral audio-visual methods and free discussions. Being a professor means not only to possess the specific knowledge but also to have the ability in transposing and expressing them didactically. Objectives: To assess the students’ favorite Parasitology teaching methods in order to improve their knowledge assimilation and the communication with their teaching supervisor. Methods: Application during the year 2006 of an original questionnaire with ten multiple-choice questions to 115 students from Victor Babes University of Medicine, Timisoara asking about their preferences regarding the teaching method (classical/modern) and other understanding facilities for the Parasitology lectures/laboratories . Results: Most of the students: preferred the power point presentation (58%), finding it helpful (51%), able to improve communication (44%) and considered it as the primary choice for a congress exposure (83%). Multimedia movies were marked to be necessary (44%) and interesting (44%). Conclusion: Learning and understanding Parasitology efficiently depends upon the modern methods and technologies used, including the presentation of multiple figures (both schematic and real images, colored) and short educational animations.
SOARES-COIMBRA E., COJEAN S., BORIES C., MARTINEZ R., LIBONG D., CHAMINADE P., LOISEAU P.M. Université Paris-Sud 11, CHÂTENAY-MALABRY, FRANCE Sitamaquine is a 8-aminoquinoline in development for the treatment of visceral leishmaniasis by oral route. In order to anticipate the possibility of sitamaquine-resistant isolates emerging in the field, we selected a Leishmania donovani promastigote line resistant to 160 M sitamaquine, named SITA-160R by continuous stepwise in vitro drug pressure. The SITA-160R line was able to infect murine peritoneal macrophages in vitro in the same manner as the wild-type line (WT) but was less infective for Balb/C mice than its parent WT clone. This line was about 5 and 3 times more resistant to sitamaquine than the WT line on promastigote and intramacrophage amastigote forms, respectively. No crossresistance with other antileishmanial agents was observed and this resistance was stable when parasites were subcultured in drug-free medium for a long time or after in vivo passage. An HPLC-tandem mass spectrometry method for sitamaquine assay was set up and allowed to study the uptake and efflux of sitamaquine from WT and SITA-160R L. donovani promastigotes. Moreover, preliminary results from a proteomic approach led to demonstrate a difference of protein expression between WT and SITA-160R parasites. References : - Duenas-Romero A.M., Loiseau P.M., Saint-Pierre-Chazalet M., 2007. Biochimica et Biophysica Acta-Biomembranes, 1768 : 246-252. - Bories C., Cojean S., Huteau F., Loiseau P.M., 2008. Biomed. Pharmacother., in press.
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1977-2007: 30 YEARS VETERINARY AMBULATORY ON PARASITOLOGY
TOSTI M. University of Perugia, PERUGIA, ITALY
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EVALUATION OF ANTI-NEMATODE POTENTIAL OF OLEANOLIC ACID GLYCOSIDES AND GLUCURONOSIDES
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PIG AS AN EXPERIMENTAL ANIMAL MODEL TO CHARACTERIZE THE PHARMACOKINETICS OF ANTIMALARIALS
DOLIGALSKA M., JOZWICKA K., GRZELAK M., MROCZEK A., SZAKIEL A., JANISZOWSKA W. Uiwesity of Warsaw, WARSAW, POLAND
SINOU V.(1), TAUDON N.(1), MOSNIER J.(1), AGLIONI C.(2), BRESSOLLE F.(3), PARZY D.(1) (1) IMTSSA, MARSEILLE ARMÉES, FRANCE ; (2) HIA LAVERAN, MARSEILLE, FRANCE ; (3) UNIVERSITE DE MONTPELLIER I, FACULTE DE PHARMACIE, MONTPELLIER, FRANCE
Oleanolic acid and/or its derivatives are present in significant amounts in many medicinal plants. We studied if plant pentacyclic oleanane-type triterpenoids would affect Heligmosomoides polygyrus development. The rate of egg embrionation, hatching and growing of larvae L1, L2 and L3 were evaluated. Oleanolic acid glycosides, i.e. glucosides (derivatives of 3-O-monoglucoside) and glucuronides (derivates of 3-O-monoglucuronide) of Calendula officinalis or Beta vulgaris and mixture of oleanolic acid and ursolic acid isolated from Vaccinum myrtillus affected the development of free living stages of the nematode. Oleanolic acid as well as its glycosides reduced the rate of L3 surviving during prolonged storage. The protein and glycoprotein patterns of infective larvae (L3) changed when larvae developed in medium containing glucuronides; four bands of glycoproteins with molecular weight 257, 96 and 17, 15 kDa vanished when nematode growth medium was supplemented with marigold or beetroot fractions. The nematode infectivity in mice was also mostly affected by glucuronides. In conclusion plant oleanane-type triterpenoids impact on the glycoprotein phenotype of parasitic nematode.
Appropriate pharmacokinetic information’s, in particular those concerning metabolism and drug interaction, are becoming increasingly important with the increased use of combination chemotherapy in the treatment of malaria. Pig is an economical animal model and has been a well-recognized experimental animal in biomedical research for many centuries. Physiological and anatomical similarities between man and pig made this animal a good model for man in many research area. In view of these considerations, we evaluated the domestic pig as a potential experimental animal model for determining the pharmacokinetics of the artemisinin derivative, artesunate (ARTS) and its active metabolite, dihydroartemisinin (DHA). Pharmacokinetic parameters of ARTS and DHA were estimated from plasma-concentration time profiles after intravenous and intramuscular administration of ARTS to pigs using empirical Bayes methodology (NONMEM). We show that the pig model mimicks some of the variables observed in human such as absorption, distribution, metabolism and elimination with intraand interindividual variabilities consistent with those usually reported in humans for artemisinin-type compounds (between 50-70%). In conclusion, the present work shows the relevance of the swine model to study the pharmacokinetics of ARTS. Also, the swine model may be a source of information to develop newer approaches for which the application of pharmacokinetic principles is fundamental.
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SIMULTANEOUS DETERMINATION OF MONODESETHYLCHLOROQUINE, CHLOROQUINE, CYCLOGUANIL AND PROGUANIL ON DRIED BLOOD SPOTS BY REVERSE-PHASE LIQUID CHROMATOGRAPHY.
HOUZÉ P.(1), HOUZÉ S.(2), SOULETIE I.(1), GOURMEL B.(1), LEBRAS J.(2) (1) Hopital St Louis, PARIS, FRANCE ; (2) Hopital Bichat, PARIS, FRANCE
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QUININE PHARMACOKINETIC IN PIG : COMPARISON OF A HIGH FIRST DOSE REGIMEN TO A NO LOADING DOSE REGIMEN
A method for simultaneous analysis of chloroquine (CQ), proguanil (PG) and their metabolites (monodesethylchloroquine: MDCQ, cygloguanil: CG) from a whole blood sample (80 L) dried on a filter paper was developed. Sample preparation included a liquid extraction from the filter paper, followed by a solid-phase extraction (C18 Bond Elut® cartridge). Separation was obtained by reversephase liquid chromatography (HPLC) using a gradient elution on an X-Terra® column; UV detection was made at 254 nm. This assay was linear between 150 and 2500 ng.mL-1 for CQ and MDCQ and 150 and 2500 ng.mL-1 for PG and CG. The lower limit of quantification was close to 50 ng.mL-1 for CQ and MDCQ and 100 ng.mL-1 for PG and CG. No chromatographic interference from endogenous compounds or other tested anti-malarial drugs was evidenced. Chromatographic separation takes about 40 min with a coefficient of variation below 10.3% for within- and betweenbatch precision. The paper sampling method was validated in ten healthy subjects treated by Savarine®. The stability of compounds and metabolites on the filter paper was evaluated at four temperatures (-20°C, +4°C, 20°C, 50°C) and for 1, 5 and 20 days. CG concentrations were not influenced by storage conditions, whereas high temperatures and prolonged storage decreased CQ and PG levels. The proposed HPLC assay is accurate, precise and cost-effective; it can be used for epidemiological studies on anti-malarial treatments.
TAUDON N.(1), SINOU V.(1), MOSNIER J.(1), MENINI W.(2), MARTELLONI M.(1), PARZY D.(1) (1) IMTSSA, MARSEILLE ARMÉES, FRANCE ; (2) HIA St Anne, TOULON, FRANCE Severe malaria is currently treated by intravenous infusion of quinine. Classically, an initial loading dose of 17 mg/kg (expressed in quinine-base) over 4 h is follows by 8mg/kg over 4h every 8h. The loading dose enable to achieve high and efficient blood levels rapidly. However, the benefit/risk ratio of its use doesn’t always clearly appears as favourable due to a narrow therapeutic index. Thus, the loading dose is still controversy. So, we developed an experimental model using the domestic pig to study the pharmacokinetics of quinine. The relevance of this model was shown in a earlier study (submitted). Quinine was administered to 3 pigs referring the therapeutic schema described above. A second group of 3 pigs received a continue administration of quinine; 32 mg/kg over 24 h followed by maintenance doses of 8 mg/kg over 8h. In both group, the treatment was over 72h and the quantity of injected quinine was equal. Quinine concentrations in plasma samples were measured by liquid chromatography on days 1 to 7. The pharmacokinetic parameters were calculated by noncompartmental method from plasma concentration-time. We show that the values of the pharmacokinetic parameters of 173
quinine in the 2 pig groups were on the same order as those achieved in human. After the continue administration protocol, the steady-state plasma concentration of quinine is achieve in 24 h. This concentration is efficient and enable a more safety monitoring of patients.
concentrations of Abz (1.35, 8.0 and 250 μM) in vitro and compared these with Abz sensitive clones. Bi-dimensional electrophoresis analysis was carried out and the spot patterns of Abz-resistant and -sensitive stocks of G. duodenalis were compared using the LC-MS/MS technique. This allowed the identification of 8 differentially expressed proteins which are constituents of: a) cytoskeletal system (alpha 2-giardin and RanBP-1), b) fermentative metabolism (NADH oxidase, a terminal oxidase) and c) energetic metabolism (triose phosphate isomerase, phosphoglycerate kinase and ornithine carbamoyltransferase). Our results suggest that resistance to Abz in Giardia may be due to the over expression of enzymes that play an important role in structural stability, prevention of oxidative stress and enhancement of energy supply. Therefore it is possible that these proteins be potential candidates as drug targets.
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TH1 IMMUNE RESPONSE OF MICE WITH ALVEOLAR ECHINOCOCCOSIS AFTER IMMUNOMODULTION WITH NONSPECIFIC IMMUNOMODULATORS AND ALBENDAZOLE THERAPY
DVOROZNAKOVA E., PORUBCOVA J. Parasitological Institute SAS, KOSICE, SLOVAKIA
Metacestode Echinococcus multilocularis has been known for its ability to modify host immune responses to ensure its survival. The influence of immunomodulators – muramyltripeptide (MTPPE/MLV) and glucan with zinc (GIZn) – applied separately or with anthelminthic albendazole (ABZ) on T lymphocytes and macrophages was examined in mice with alveolar echinococcosis. The immunomodulative effect of ABZ was found in treated mice even for 4 weeks after the end of therapy, at the same time as the parasitostatic effect of this drug was observed. Short-time increase of serum cytokine IFN-g and activated oxidative metabolism (O2-) in macrophages were noticed during the metacestode growth reduction. MTP-PE/MLV was recognised to be the strong macrophage activator. It stimulated the O2-production for 3 months, which was supported by dominant Th1 protective response - the highest serum IFN-g level. The antiparasitic effect of MTP-PE/MLV+ABZ therapy was manifested by marked reduction of cysts growth for 3 months after the end of treatment. The advantage of additive substances was confirmed in case of GIZn. GIZn+ABZ therapy resulted in high IFN-g production and dominant Th1 response for more than 3 months after the end of therapy. GIZn+ABZ have been shown the most effective treatment, the most lowered metacestode growth was observed as early as 2 weeks from the begenning of this therapy till the end of experiment (for 4 months).
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EFFICACY OF ALBENDAZOLE TO HAEMONCHUS CONTORTUS INFECTION IN MONGOLIAN GERBILS (MERIONES UNGUICULATUS) AND INFLAMMATORY CELLS DISTRIBUTION IN STOMACH
KONIGOVÁ A., HRCKOVA G., VÀRADY M., VELEBNY S., CORBA J. Parasitological Institute of the Slovak Academy of Sciences, KOSICE, SLOVAKIA Anthelmintic resistance of gastrointestinal nematodes to benzimidazole (BZ) drugs is a major problem in many countries. Mongolian gerbils have been proposed as a suitable experimental model to study host-parasite interactions and anthelmintic activity during gastrointestinal nematode infections. In the present study the establishment rate of BZsusceptible and BZ-resistant larvae of Haemonchus contortus and the effect of drug albendazole (ABZ) was examined in stomach of gerbils within 3 weeks post infection. Higher numbers of BZ-susceptible than BZ-resistant larvae were found. Treatment with ABZ had lower effect on BZ-resistant larvae (L3 and L4 stage) in comparison with BZ-susceptible larvae. Infection elicited strong inflammation in mucosal and submucosal layers of stomach, where significantly higher mast cells (MC) numbers were seen in lamina propria mucosae and submucosae after infection with BZ-susceptible larvae. No changes were observed in eosinophil counts. Therapy decreased MC numbers more in gerbil infected with BZresistant larvae and modulated cells distribution. Apoptosis was not involved in down-regulation of MC counts after BZ-resistant H. contortus infection. Lower inflammation in stomach following ABZ therapy correlated with reduced infection, probably due to the limited stimulation of immune system. Support: by grant VEGA No. 2/7189/2, grant VEGA No. 2/7188/27 and Slovak Research and Development Agency project No.RPEU-009-06.
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IDENTIFICATION AND CHARACTERIZATION OF PROTEINS DIFFERENTIALLY EXPRESSED IN GIARDIA DUODENALIS CLONES RESISTANT TO ALBENDAZOLE USING A PROTEOMIC ANALYSIS
PAZ-MALDONADO L.M.T.(1), CRUZ-SOTO M.(1), ARGÜELLOGARCÍA R.(1), MENDOZA-HERNANDEZ G.(2), ORTEGAPIERRES G.(1) (1) Departamento de Genetica y Biologia Molecular, Centro de Investigación y de Estudios Avanzados-IPN, 07360 MEXICO, MEXICO ; (2) Departamento de Bioquimica, Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 MEXICO, MEXICO Albendazole (Abz) has been extensively used in the treatment of Giardiasis since this drug is effective in controlling this infection. However its inappropriate use has caused the generation of resistant organisms. Up to now the mechanisms involved in Abz resistance in G. duodenalis have not been fully characterized. In this study we have used a proteomic analysis to identify and characterize proteins which may be differentially expressed in G. duodenalis clones resistant or sensitive to Abz. This was approached using clones resistant to different
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PREVALENCE OF ANTHELMINTIC RESISTANCE IN SHEEP FARMS IN SLOVAKIA
KONIGOVÁ A., VÀRADY M., CORBA J. Parasitological Institute of the Slovak Academy of Sciences, KOSICE, SLOVAKIA A survey of prevalence of anthelmintic resistant nematodes was carried out in 25 sheep farms in Slovakia. Anaerobic sampling kits were mailed to 45 farms, those who initially responded positively to take part in the survey. Replies were received from 174
25 farms and larval development test was used for the detection of resistance to benzimidazole and ivermectin group of anthelmintics. The presence of benzimidazole resistant populations was detected on three farms. Resistance to macrocyclic lactones was not observed. Intensity of infection was low or moderate in most of the farms. Differential diagnostics of infective larvae referred to multiple infections with several parasitic genera/species. Ostertagia spp., Trichostrongylus spp. and Chabertia ovina were the dominant species (detected on 90-100 % of the farms). Haemonchus Cooperia spp., Oesophagostomum spp., contortus, Nematodirus spp., Strongylloides papillosus and Bunostomum spp. were less frequent. In the flock where resistant nematode strains have been diagnosed, we suggested the change of anthelmintic treatment and recommended the most suitable anthelmintic control and prophylaxis measures to reduce and eliminate anthelmintic resistance on these farms. The study was supported by the Slovak Research and Development Agency project No. RPEU-009-06 and Grant Agency VEGA, Grant No. 2/7189/27 of the Scientific Agency of the Slovak Academy of Sciences.
Introduction: Considering the high prevalence of Tricomonas vaginalis (TV) in women and the known side effects of Metronidazol, the focus has been concentrated on herbal therapy in order to reduce drug side- effects in the recent decades. Materials & methods: The parasite was isolated from vagina and determined directly.Methanolic extraction was performed by percolation and essential oil prepared by hydrodistilation.parasites was added to the 10 test tubes containing Trichomona Modified CPLM medium Base(TMCPLM), metonidazole, Dimetyl solfoxide(DMSO) lavandula angyustifolia extraction of (0.1,0.01,0.001 ml and essential oil with concentration of (0.1,0.01,0.001)in order to determine the effect of these concentrations in 0,1,2,3,6,12,24,48 and 72 hours. Result: Finding suggested that Trichomonas vaginalis Could be alive in TMCPLM For 48 hours,in presence of metonidazole for 2 hours and TMCPLM for 5 hours. Also,the result releaved that methanolic extract at concentration of 0.1and 0.01 were effective at the beginning of the inoculation and at concentration of 0.001 hours respectively. Conclusion: Therfore, further investigation was carried out to check the AntiTrichomonas vaginalis activity of Lavandula angyustifolia Extract & Essential oil under Invivo Condition and if having positive effect, to be used as a drug. Key words: Trichomonas vaginalis, Trichomona Modified CPLM medium Base, Lavandula angyustifolia
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DRUGS INTERACTIONS STUDIES HIGHLIGHT A SYNERGISM BETWEEN NICOTINAMIDE AND ANTIMONIALS IN LEISHMANIA INFANTUM AMASTIGOTES
GAZANION E.(1), VERGNES B.(1), GARCIA D.(1), OURY B.(1), OUAISSI A.(2), SERENO D.(1) (1) IRD, MONTPELLIER, FRANCE ; (2) INSERM, MONTPELLIER, FRANCE
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Treatment of leishmaniasis remains problematic due to drugs toxicity and emergence of drug resistant strains. Combination therapy may prevent or limit the spread of drug resistance. Since we have previously reported antileishmanial activity of nicotinamide (Nam), a precursor in NAD synthesis, we determined its capacity to be used in combination therapy. Preliminary studies performed on intracellular forms grown in axenic condition showed that Nam induced cell growth arrest at low concentrations (< 20 mM) and apoptosis for higher concentrations. Then, in vitro efficacy and drug-drug interactions of Nam with standard antileishmanial compounds were investigated. Interestingly, Nam synergizes the cell death induced by trivalent antimony (SbIII), the active form of pentavalent antimony. We tested the Nam/antimonial combination on intramacrophagic parasites and found that Nam enhanced SbIII drug potency, but was unable to synergize with pentavalent antimonial species. Further studies performed on antimony resistant strains, in the range of physiologically achievable concentration of SbIII during antimonial treatment, demonstrate that these mutants were about three fold more resistant towards Nam than wild type strains. Our screening reveals some potential common mechanisms of action and resistance between Nam and antimonials that questions the link between nutritional status of individuals during treatment and the selection of drug resistant parasites.
IN VITRO STUDIES ON ANTIACANTHAMOEBA ACTIVITY OF NEW SYNTHESIZED BENZIMIDAZOLE DERIVATIVES
PADZIK M.(1), OLDZKA G.(1), PIETRUCZUK A.(2), PIEKARCZYK P.(3), MOCICKI D.(1), KAZIMIERCZUK Z.(4) (1) Department of Medical Biology, Medical University of Warsaw, WARSAW, POLAND ; (2) Department of Pharmaceutical Microbiology, Medical University of Warsaw, WARSAW, POLAND ; (3) Clinic of Cranio-Maxillo-Facial and Oral Surgery and Implantology, Medical University of Warsaw, WARSAW, POLAND ; (4) Institute of Chemistry, Agricultural University, Medical Research Center P. Ac. Sc., WARSAW, POLAND The Acanthamoeba infections, reported with increasing frequency in people of various part of the world, despite of advances in chemotherapy, are very difficult to treatment. Particularly, cysts of the amphizoic amoebae are extremely resistant to chemical agents. We tested in vitro antiacantamoebic effect of two new synthesized bromobenzimidazole derivatives carrying a polyfluoroalkyl substituent, each in two concentrations. Status of the surviving amoebae, cyst and trophozoite number assessed on 5-day grown population of the Neff strain of A.castellanii were compared with those of the control culture. After exposition to agents, lower amounts of moving trophozoites, tendency to encystment and increase of cyst level were visible. General amoebicidal activity of the newly synthesized benzimidazole derivatives depending on a kind and concentration of the agents is very promising; further study is conduct to acquire anti-Acanthamoeba efficacy at considerably lower concentration of chemical agents tested. The authors wish to thank the Foundation for Development of Diagnostic and Therapy in Warsaw for supporting this research.
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INVESTIGATION OF ANTITRICHOMONAS VAGINALIS ACTIVITY OF METHANOL EXTRACT & ESSENTIAL OIL OF LAVANDULA ANGYUSTIFOLIA
EZATPOUR B. Razi herbal medicine research center, KHORRAM ABAD, ISLAMIC REPUBLIC OF IRAN 175
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MOLECULAR ANALYSIS OF BENZIMIDAZOLESUSCEPTIBLE AND RESISTANT HAEMONCHUS CONTORTUS ISOLATES
MODULATION OF THE IN VITRO ANTIMALARIAL EFFECTS OF ARTEMISININ BY SELECTED EXTRACTS: THE CASE OF PAPAYA LEAF DECOCTION
CUDEKOVA P., VARADY M., CERNANSKA D., CORBA J. Parasitological Institute of SAS, KOSICE, SLOVAKIA
SANNELLA A.R.(1), BILIA A.R.(2), LAPENNA S.(2), VINCIERI F.F.(2), MESSORI L.(3), MAJORI G.(1), SEVERINI C.(1) (1) Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, ROME, ITALY ; (2) Department of Pharmaceutical Sciences, University of Florence, SESTO FIORENTINO, FLORENCE, ITALY ; (3) Department of Chemistry, University of Florence, SESTO FIORENTINO, FLORENCE, ITALY
The development of species and populations of parasitic nematodes of sheep and goats with resistance to one or more anthelmintics is an increasing problem world-wide. The aim of the study was to determine of allele and genotypes frequency in different isolates of Haemonchus contortus. Nine isolates, susceptible and resistant to benzimidazoles (BZ) anthelmintics were examined in the study. The BZ resistance is linked with the mutation in -tubulin isotype 1 gene which substitute phenylalanine (Phe) to tyrosine (Tyr) at the 200 codon of the gene. An allele specific PCR technique was used to determine the resistance status in the infective larvae based on their genotypes (rr, rS, SS). Resistant isolates had increasing frequency of resistant alleles in position 200 isotype 1 betatubulin with TAC. Frequency of BZ resistant allele in resistant isolates varied from 9 to 95 % and in susceptible isolates from 4 to 6 %. In susceptible isolates the frequency of susceptible allele varied from 58 to 84 %. Correlation between allele specific PCR and in vitro egg hatch test and larval development test have also been performed. The study was supported by the Slovak Research and Development Agency project No. RPEU-009-06 and PARASOL (FOOD-CT-2005-022851) project of EU 6th Framework Programme.
Within the framework of a research project we evaluated the possible synergistic effects in malaria treatment between artemisinins, potent antimalarial agents, and a variety of plant extracts and isolated natural constituents. In the present study, a dried extract obtained by decoction of papaya leaf was tested in vitro alone and in combination with artemisinin against 3D7 Plasmodium falciparum strain. In literature there is a report on the efficacy of a crude aqueous extract of papaya leaf on mice infested with malaria parasite in vivo. 10 g of papaya leaf collected in Burundi were extracted by decoction by boiling. The decoction was liophylised and submitted the HPLC analysis to evaluate the content of polyphenols. 1.6% of total flavonoids, expressed as rutin, were characterised as trisaccharides of kaempferol and quercetin. The antimalarial effect of extract of papaya was assayed by measuring the activity of parasite LDH. The extract of papaya inhibited P. falciparum growth with IC50 values of 166.0+/-1.23 ug/ml. Isobologram analysis showed that the extract of papaya, 100 or 150 ug/ml, used in combination whit artemisinin, exert a sinergistic effect. Similar sinergistic effect was obtained using 50 ug/ml of olive leaf extract in combination with artemisinin. These antiplasmodial interactions might be conveniently exploited to determine the constituents responsible of the synergism and design new and more effective combination therapies.
P-227
THE USE OF THE TARGETED SELECTIVE TREATMENT TO CONTROL GASTROINTESTINAL INFECTION IN SELECTED FARMS IN SLOVAKIA
VARADY M., KONIGOVA A., CERNANSKA D., CUDEKOVA P., CORBA J. Parasitological Institute of SAS, KOSICE, SLOVAKIA
P-229
Parasitic nematode infections of ruminants are mainly controlled by anthelmintic treatments, however repeated usage of anthelmintics has selected for anthelmintic resistance. Targeted selective treatments (TST) have been proposed as a means of both reducing infection and selection pressure arising from anthelmintic treatments. During the 2006 and 2007 grazing seasons we selected 5 conventional sheep farms to conduct the studies. The size of the flocks varied from 250 to 450 animals. The main objective in 2006 was to examine the suitability of a parasitological indicator FEC (faecal egg counts EPG) and pathophysiological indicators such as diarrhea index – DISCO, body condition score - BODCON and live weight gain LIVGAIN and determine their impact on parasite population dynamics and host productivity. In 2007 the efficacy of TST and their impact upon production of young animals (lambs) was examined. In general, BODCON and DISCO scores did not correlate with parasitological parameters (EPG) and LIVGAIN from the parasitized lambs (non-treated) was lower compare to the treated lambs. The TST groups of lambs (based on EPG or LIVGAIN) showed higher average LIVGAIN and lower EPG values compared to the control untreated animals. The study was supported by PARASOL (FOOD-CT-2005022851) project of EU 6th Framework Programme and Grant Agency VEGA, Grant No. 2/7189/27 of the Scientific Agency of the Slovak Academy of Sciences.
A NOVEL MANAGEMENT OPTION FOR THE ROOT-KNOT NEMATODE MELOIDOGYNE MINOR
KERR R.(1), KERR R.(1), MC NIECE E.(1), MAULE A.G.(1), FLEMING C.C.(2) (1) Queen's Unversity, BELFAST, UNITED KINGDOM ; (2) Agri Food and Biosciences Institute, BELFAST, UNITED KINGDOM The root knot nematode Meloidogyne minor is causing increasing economic difficulties throughout Western Europe due to its wide host range including sports turf, pasture grass and potatoes. Infestation by the nematode results in the stunted development of turf-grass and consequently the appearance of yellow-patch disease. This study aims to examine a novel approach to the management of M. minor, using naturally derived nematicidal compounds, in particular Furfural, derived from sugar cane. This could prove to be a more environmentally friendly alternative to chemical nematicides such as Methyl Bromide and Nemacur. Recent field trials of a Furfural containing product, Biomass Sugar, in Johnstown PGA National Golf Course, Ireland, showed an average decrease of 62% in M. minor juveniles (per 100 ml of soil), over eight of its greens. The manufacturers believe this is the result of Furfural compromising the nematode cuticle, resulting in a loss of 176
hydrostatic pressure, which proves fatal to the worm. Laboratory investigations have included S.E.M. studies which show oblation and puncturing of both the nematode cuticle and egg mass. Investigations into, mortality levels, motility and population recovery, are also under investigation. This study aims to establish if this product is an effective and environmentally friendly nematode management option.
0.4-1.6 mg/ml using inhibitory zone estimation, 0.4-1.6 mg/ml using agar dilution method and 0.2-1.6 mg/ml using broth dilution method. The minimum fungicidal concentration (MFC) of the extracts ranged from 0.8-6.4 mg/ml. The results obtained suggest that E. camaldulensis has anti–dermatophyte activity.
P-232 Abstract withdrawn by their authors
P-230
SYNTHESIS AND ANTIPROTOZOAL ACTIVITY OF 4-ARYLCOUMARINS
P-233
IDENTIFICATION OF GENES INVOLVED IN LEVAMISOLE RESISTANCE IN THE PARASITIC NEMATODE HAEMONCHUS CONTORTUS USING A CDNA-AFLP APPROACH
PIERSON J.T.(1), DELMAS F.(2), HUTTER S.(2), LAGET M.(2), DUMETRE A.(2), GASQUET M.(2), AZAS N.(2), COMBES S.(1) (1) UMR-CNRS 6264 Laboratoire Chimie Provence, Universités Aix-Marseille I, II et III, Faculté des Sciences de Saint-Jérôme, Avenue Escadrille-Normandie-Niemen, 13397 MARSEILLE CEDEX 20, FRANCE ; (2) UMR-MD3 Relations Hôte-Parasite, Pharmacologie et Thérapeutique, Université Aix-Marseille II, 27 Boulevard Jean Moulin, 13385 MARSEILLE CEDEX 05, FRANCE
FAUVIN A., CHARVET C., CORTET J., CABARET J., NEVEU C. INRA, NOUZILLY, FRANCE
Levamisole is a broad-spectrum anthelmintic drug widely used to control parasitic nematodes in livestock. However, the high efficacy of levamisole against the gastrointestinal nematode Haemonchus contortus in sheep and goats has been compromised by the development of resistance in field populations. A cDNA-AFLP (cDNA-Amplified Fragment Lenght Polymorphism)-based strategy has been used to identify genes potentially involved in levamisole resistance in Haemonchus contortus. Transcript profiles of adult nematodes from two levamisole resistant and two susceptible isolates were compared. Among 17280 transcript-derived fragments (TDFs) amplified, 26 presented a polymorphic pattern between resistant and susceptible isolates. Among those candidates a TDF named HA7 was specifically expressed in the two resistant isolates and presented strong homologies with acr-8 gene: a subunit of a nicotinic acetylcholine receptor from the free living nematode Caenorhabditis elegans. Expression studies of HA7 in resistant and susceptible isolates revealed that resistant nematodes expressed a truncated isoform of the transcript.
Malaria and leishmaniasis are the most important vector-borne parasitic diseases in the world. Their therapeutic management has become problematic because of the emergence of multidrug-resistant strains of Plasmodium falciparum, and glucantime-resistant strains of Leishmania. The development of new molecules with antiprotozoal potential is therefore an ongoing challenge. We reported the synthesis and the antiprotozoal activity of a new series of twenty-five 4-arylcoumarin derivatives. Their excellent purity allowed us to consider their cytotoxicity towards human THP1 and HepG2 cells and their potential activity against the chloroquine-resistant strain W2 of P. falciparum and the reference strain 703 of Leishmania donovani (MHOM/IN/80/DD8). Concentrations of compounds inhibiting 50% of the parasite proliferation (IC50-parasites) and 50% of the human cell proliferation (IC50-human cells), and their selective index (SI = IC50-human cells / IC50-parasites) were determined using TableCurve 2D v5.0 software. Overall, these 4-arylcoumarin derivatives have no interesting activity against P. falciparum, except one with a moderate SI>5.3. However, its evaluation was not further investigated due to its low solubility. More compounds were active against L. donovani amastigotes, with SI up to 265. Further investigations suggested that these compounds acted on the phagocytosis of Leishmania promastigotes, as described elsewhere for other coumarins.
P-234
ANTIMALARIAL ACTIVIY OF NEW QUINOLINE DERIVATIVES
GARCIA SANCHEZ R.N.(1), MARTIN PEREZ-SOLORZANO B.(1), NOGAL RUIZ J.J.(1), AMADO TORRES D.F.(2), GOMEZ BARRIO A.(1), MARTINEZ FERNANDEZ A.(1), KOUZNETSOV V.(2) (1) UNIVERSIDAD COMPLUTENSE, MADRID, SPAIN ; (2) UNIVERSIDAD INDUSTRIAL DE SANTANDER, BUCARAMANGA, COLOMBIA Malaria is one of the most important infectious diseases; moreover, it is a serious public health problem mainly in the world’s poorest countries. About 20% of all childhood deaths in Africa are due to malaria. Every 30 seconds, a child dies from malaria in Africa. Antimalarial drug resistance is widespread. Currently, artemisinin-based combination therapy is the only choice for malaria treatment in those places. For this reason, searching and developing for new drugs that are cheaper and easier to synthesize is a fundamental task. We tested 33 new quinolines for in vitro antimalarial activity against Plasmodium falciparum 3D7 strain by fluorimetric microtest using SYBR®Green I. In addition, nonspecific cytotoxicity of the most active compounds was measured in murine J774 macrophages. Five compounds showed good activity, with IC50 values lower than 1 M (from 0.69 to 0.02 M). The compound FER102 was 2 times more active than cloroquine (IC50 0.04 M). None of the compounds were cytotoxic at concentrations lower than 600 M. Further studies will be carried out in order to asses the in vivo activity and to measure the ability of these compounds
P-231
ANTIFUNGAL ACTIVITY OF EUCALYPTUS CAMALDULENSIS ON DERMATOPHYTES
FARAHYAR S., FALAHATI M., OMIDI TABRIZI N., JAHANBANI F. Iran University of Medical Sciences, TEHRAN, ISLAMIC REPUBLIC OF IRAN Methanolic leaf extracts of Eucalyptus camaldulensis were investigated for in vitro antifungal activities against Microsporum gypseum, Trichophyton rubrum, Trichophyton schoenleinii, Trichophyton mentagrophytes and Epidermophyton floccosum. The studies were carried out using broth dilution method, agar dilution method and inhibitory zone estimation. The effects of the plant extract were compared with those of griseofulvin. Eucalyptus camaldulensis showed antifungal activity against all the dermatophytes tested with minimum inhibitory concentration (MIC) values ranging from 177
to inhibit the heme biocrystallization process.
sera of 68 out of 71 filariasis patients showing an overall sensitivity of 95.8% (91.7% sensitivity for microfilaraemia group and 100% sensitivity for elephantiasis group). All negative control sera were negative for CFA, while 3 patients out of the other parasite group were negative for CFA giving an overall specificity of 96.4%. These findings suggest that (9F/10B) MAb and (5F/6H) MAb could be used as a reliable diagnostic indicator for the activity of human filariasis and as a cure monitor particularly in control programs for endemic areas.
P-235
EFFECT OF GENETIC SELECTION FOR MASTITIS RESISTANCE IN THE LACAUNE BREED OF SHEEP ON THE RESPONSE TO HAEMONCHUS CONTORTUS INFECTION
TRAORE I.(1), PFEIFFER H.(1), PREVOT F.(1), GRISEZ C.(1), BERGEAUD J.P.(1), RUPP R.(2), FOUCRAS G.(1), JACQUIET P.(1) (1) UMR INRA/ENVT 1225, Ecole Nationale Vétérinaire, TOULOUSE, FRANCE ; (2) INRA, UR631, Station d'Amélioration Génétique des Animaux, CASTANET-TOLOSAN, FRANCE
P-237
EFFECT OF AGRICULTURAL ACTIVITIES ON PREVALENCE RATES, CLINICAL AND PRESUMPTIVE MALARIA EPISODES IN CENTRAL CÕTE D'IVOIRE
Introduction and objectives: Mastitis cause significant reductions in quality and quantity of milk production in dairy sheep. Two divergent lines of Lacaune breed of sheep were selected on the somatic cell counts (SCC) in milk: a mastitis susceptible line with high number of SCC (SCC+) and a mastitis resistant line with low number of SCC (SCC-). As the origin of mastitis is mainly microbial (bacteria), resistance to this disease is mediated by a Th1 type adaptive immune response. Conversely, the resistance to the nematode Haemonchus contortus is governed by a Th2 type immune response. Therefore, the present study was initiated to evaluate the effect of selection for mastitis resistance on sheep’s responses to experimental infection with H. contortus. Results: No significant difference was registered between the two lines of sheep for faecal egg excretions, worm’s establishment and development, blood eosinophil counts, packed cell volumes, serum pepsinogen values and serum antibody measurements during three successive experimental infections. However, some immune parameters were different between the two lines as the CD4/CD8 ratios, the percentages of B lymphocytes in blood and in the draining lymph node, both higher in the mastitis resistant line. Conclusion: In this study, we demonstrated that selection for mastitis resistance on the basis of somatic cell counts did not affect the establishment, development and fecundity of H. contortus.
KOUDOU G.B. Centre Suisse de Recherches Scientifiques en Côte d'Ivoire, ABIDJAN, IVORY COAST Agricultural activities, among other factors, can influence the transmission of malaria. In two villages of central Côte d’Ivoire (Tiémélékro and Zatta) with distinctively different ecoagricultural characteristics, we assessed Plasmodium prevalence rates, fever and clinically-confirmed malaria episodes among 171, 598, 714 and 245, 689, 795 children aged 15 years by carrying out repeated cross-sectional surveys, in Zatta and Tiemelekro, respectively. Additionally, presumptive malaria cases were monitored in health centres for a 3-year period. In Tiémélékro, we observed a decrease in malaria prevalence rates from 2002 (86.1%) to 2005 (60.4%), which might be explained by changes in agricultural activities from subsistence farming to cash crop production. In Zatta, where an irrigated rice perimeter is located in close proximity to human habitations, malaria prevalence rates in 2003 (58.4%) were significantly lower than in 2002 (85.4%) and 2005 (66.0%), which coincided with the interruption of irrigated rice farming in 2003/2004. Although malaria transmission differed by an order of magnitude in the two villages in 2003, there was no statistically significant difference between the proportions of severe malaria episodes. Our study underscores the complex relationship between malaria transmission, prevalence rate and the dynamics of malaria episodes. A better understanding of this relationship can facilitate the starting of a control strategy.
P-236
IMMUNODIAGNOSIS OF HUMAN WUCHERERIA BANCROFTI INFECTION USING A PAIR OF MONOCLONAL ANTIBODIES AGAINST WORM ANTIGEN
MANSOUR W.(1), SALAH F.(1), RABIA I.(2), HENDAWY M.(2), EL GAMAL N.(3), EL BASSIOUNY A.(1), DEMERDASH Z.(1) (1) Theodor Bilharz Research Institute, Immunology Department, CAIRO, EGYPT ; (2) Theodor Bilharz Research Institute, Parasitology Department, CAIRO, EGYPT ; (3) Tropical Medicine, Faculty of Medicine, Zagazig University, CAIRO, EGYPT
P-238
FREQUENCY OF CUTANEOUS LEISHMANIASIS IN PATIENTS REFERRED TO PASTEUR INSTITUTE OF IRAN
FARAHMAND M., ASSMAR M., FARZANENEJAD Z., PIAZAK N. Pasteur Institute of Iran, TEHRAN, ISLAMIC REPUBLIC OF IRAN
This study was designed to prepare monoclonal antibodies (MAbs) against filarial worm antigen (FWA) with immunodiagnostic potential for human filariasis. From a panel of anti-filarial antigen MAbs; a pair of MAbs (9F/10B & 5F/ 6H), highly reactive with filarial antigen and showing no cross reactivity against other parasites antigens were selected and characterized. The pair was found to be of IgG1 subclass. The assay reached a lower detection limit of 125 ng/ml of filarial antigen. CFA levels were measured in serum samples from 71 filariasis patients (47 with microfilaraemia and 24 with elephantiasis), 45 patients with other parasites including schistosomiasis, fascioliasis and echinococcosis and 39 healthy individuals as negative control. CFA levels were detected in
Leishmaniasis is endemic in 88-countries throughout the world , with 1.5 to 2 million newly cases each year. Cutaneous leishmaniasis (CL) is still considered as an important health problem in many parts of the world especially the Mediterranean regions, Africa and almost all countries of the Middle East. Although this disease dose not result in death, but because of long lasting lesions, cosmetic problems, great expenses of treatment, length of cycle and side effects of the available drugs, it has created many problems . In the present analytical study the frequency of leishmanial and non leishmanial skin lesions were assessed. Initial variable included age, sex, occupation, place of residence 178
and number of lesions that all were recorded in an information data. Patients with ulcerative skin lesions were referring to the department of parasitology; Pasteur Institute of Iran was examined for CL by using direct and culture methods. Leishman body was observed in 43.4% out of 322 patients by direct smear and 45.34% by cultivation. In this study some cases of none healing and recidivans leishmaniasis was observed. Most of patients have traveled to endemic areas for CL in Iran. The results indicates that although CL is endemic in many parts of Iran ,but the frequency of non leishmanial skin lesion is very high and diagnostic test should be used before to commence the treatment. Key words: Iran, cutaneous leishmaniasis,
needed. In this study we compared the Direct Agglutination Test(DAT),Dipstick rK39and Immunofluorescent -Antibody Test (IFA)in the diagnosis of Iranian Visceral Leishmaniasis. In total sera from 29 subjects (parasitologically confirmed patients) were tested. Sensitivity of the Tests were as follows : DAT 92.3% , Dipstick rK39: 85.7%,IFA 91.7% .Specificity of the Tests were as follows : DAT 86.7% , Dipstick rK39:93.3%,IFA 93.3% . The Positive Prediction Value (PPD) of DAT 85.7%, Dipstick rK39 :92.3%,IFA 91.7% . The Negative Prediction Value (NPV) of DAT 92.9%, Dipstick rK39 :87.5% and IFA 93.3%.A significant correlation (high degree of agreement )was observed between all Tests. We conclude that these tests could be used for screening suspect patients in endemic areas. The DAT is higher sensitivity than others and the Dipstick rK39 is higher PPV, cheaper and easier than others and could be used widely.
P-239
ALLELIC POLYMORPHISM OF THE MEROZOITE SURFACE PROTEIN-1GENE OF PLASMODIUM FALCIPARUM AND CLINICAL CONDITIONS IN IRAN
P-241
SIGNS AND SYMPTOMS OF PATIENTS WITH BLASTOCYSTIS HOMINIS INFECTION
YOUSOFIDARANI H., IMANI R., PAT M. Shahrekord University of Medical Sciences, SHAHREKORD, ISLAMIC REPUBLIC OF IRAN
KESHAVARZ H.(1), HEIDARI A.(2) (1) School of Public Health, Tehran University of Medical Sciences, TEHRAN, ISLAMIC REPUBLIC OF IRAN ; (2) Department of Medical Sciences, Ministry of Sciences (GPEF), TEHRAN, ISLAMIC REPUBLIC OF IRAN
Background: Blastocystosis is the infection caused by Blastocystosis hominis which is inhabited in human intestine. Common probable clinical complaints in infected patients include abdominal discomfort, bloating, cramping, diarrhea and vomiting. Regarding to controversy about manifestation of this parasite, the aim of this study was to investigate clinical and laboratory findings associated with Blastocystosis hominis infection. Material and methods: This is a descriptive study carried out on patients referred to medical diagnosis labs in Shahrekord, Iran. In 60 patients who were positive for Blastocystosis hominis infection based on microscopic examination of stool specimens, signs and symptoms were investigated. Results: The clinical signs and symptoms were abdominal pain, diarrhea, constipation, decreased appetite, weight loss, fever, distention and tenderness. Mean fecal WBC count was 2.5 and Guardia lambilia was detected only in one case. Conclusion: The most common symptoms in Blastocystosis hominis infected patients were abdominal pain, diarrhea and weight loss. Key Words: Blastocystis hominis, signs and symptoms
Malaria is a major public health problem and is associated with 300 – 500 million cases worldwide. The block2 merozoite surface protein (MSP)-1 is Polymorphic and 3 distinct allelic families have been described as Mad20, K1 and Ro33. We investigated the Polymorphic nature of (MSP)-1 gene and clinical condition of falciparum malaria subjects in endemic area, south east, of Iran. . In 2006, National Malaria Control programmer reported 15909 malaria cases. From which nearly more than 83% were autochthonous. This study involved 80 Plasmodium falciparum patients attending randomly to the local malaria clinics and health centers. 20 patients had more sever clinical signs such as decrease of conscious, restlessness and repeated vomiting. Diagnosis of Plasmodium falciparum was confirmed by light microscopy on thick blood smears. Parasitemia in patients ranged from 500 to 30000 parasites/mm3 .The nested polymerase chain reaction was used for allelic family determination of P.falciparum isolates. K1, Mad20 and Ro33 allelic families were observed in 33, 25 and 2 uncomplicated falciparum malaria cases respectively but, in malaria patients with more sever clinical signs, k1, Mad20 and Ro33 were found in 7, 6 and 7 subjects respectively. There was significant correlation between clinical conditions and allelic family (MSP)-1 gene (P
