Jahres- und Tätigkeitsbericht 2012/13

Transkript

1 Institut für Parasitologie der Vetsuisse-Fakultät sowie der Medizinischen Fakultät der Universität Bern (IPB) STS 175 Nationales Referenzlabor für Toxoplasmose Nationales Referenzlabor für Trichinellose Nationales Referenzlabor für Tritrichomonose Nationales Referenzlabor für Beschälseuche Nationales Referenzlabor für Neosporose Direktor: Prof. Dr. B. Gottstein Jahres- und Tätigkeitsbericht 2012/13 (Annual scientific reports in English) 20. & 21. Berichtsjahr 1 IPB: Jahresbericht 2012_13 / annual report 2012_13

2 Inhaltsverzeichnis Inhaltsverzeichnis Institutsbezeichnung und Arbeitsgruppen 3 Forschungsberichte (in Englisch) 4 Scientific Research Reports On individual major research projects; list of additional minor research projects; technical research services. a) Öffentlichkeitsarbeit 13 Publikationsliste international begutachteter Arbeiten; Buchkapitel und Presseartikel; Vorträge und Posters, internationale Partnerschaften Aus-, Fort- und Weiterbildung 19 Unterrichtsveranstaltungen für Veterinärmediziner; Unterrichtsveranstaltungen für Humanmediziner und Biologen. Interne und externe Fort- und Weiterbildungsaktivitäten; Institutsbibliothek und Unterrichtsmaterial. b) Gäste und Gastseminarien 21 Dienstleistungen / Diagnostik 22 Dienstleistungen im Sektor Veterinärparasitologie; Dienstleistungen im Sektor medizinische (Human)- Parasitologie. Akkreditierung,Qualitätsmanagement und Qualitätskontrollen. Personelles und Personalbestand 24 Akademisches Profil des Kaderpersonals, Personalbestand und Personalmutationen c) IPB-Service 27 Who is who? (solution on page 26) 2 IPB: Jahresbericht 2012_13 / annual report 2012_13

3 Institutsbezeichnung und Arbeitsgruppen INSTITUTSBEZEICHNUNG Institut für Parasitologie der Vetsuisse und der Medizinischen Fakultät der Universität Bern Länggass-Strasse Bern TEL FAX Internet URL (Sekretariat, nur nachmittags) (Leitung); (Sekretariat) ARBEITSGRUPPEN DES INSTITUTES (Personalbestand 31.Dezember 2013) Prof. Dr. Bruno Gottstein Institutsleitung Daniela Hayoz (Sekretariat) Prof. Dr. Norbert Müller Dr. med. vet. Caroline Frey Dr. med. vet. Brigitte Hentrich Dr. Gertrud Rosenberg Liliane Krähenbühl Caroline Müller Ha Thu Trang Nguyen Didier Hirt Ursula Kurath Bernadette Merkle Dienstleistung & angewandte Forschung Prof. Dr. Andrew Hemphill PD Dr. Joachim Müller Dr. Markus Spiliotis Dr. Stefan Kunz Dr. Mahbubur Rahman Dr. Cornelia Spycher Dr. Britta Stadelmann Dr. Denise Zysset-Burri Denise Aeschbacher Vreni Balmer Ghalia Boubaker IsabeL Hostettler Junhua Wang Pablo Winzer A. Läderach (ITPA) N. Vukovic (AD) Grundlagenforschung Hausdienst IPB: Jahresbericht 2012_13 / annual report 2012_113 3

4 Forschungsberichte / scientific reports SCIENTIFIC RESEARCH REPORTS 2012/13 List of research grants / projects active in 2012/13 (N) = new project generated during 2012/13; (P) = new prolongation of project during 2012/13 Swiss National Science Foundation SNSF Schweizerischer Nationalfonds SNF 1) ECHINOCOCCUS MULTILOCULARIS AND ALVEOLAR ECHINOCOCCOSIS: THE MOLECULAR STRUGGLE (P) Gottstein B, Hemphill A Swiss National Science Foundation No A /1 Project period: Project budget: CHF ) ECHINOCOCCUS MULTILOCULARIS AND ALVEOLAR ECHINOCOCCOSIS: IMMUNOMODULATORY MECHANISMS AS POTENTIAL TARGET FOR IMMUNO- AND CHEMOTHERAPEUTICAL INTERVENTION (N) Gottstein B, Hemphill A Swiss National Science Foundation No A /1 Project period: Project budget: CHF ) NEOSPORA CANINUM AND NEOSPOROSIS: STUDIES IN THE MOUSE MODEL (P) Hemphill A Swiss National Science Foundation No A /1 PROJECT PERIOD: PROJECT BUDGET: CHF ) OF MICE AND SHEEP: PREVENTION AND TREATMENT OF NEOSPORA CANINUM INFECTIONS (N) Hemphill A Swiss National Science Foundation No PROJECT PERIOD: PROJECT BUDGET: CHF ) FUNCTIONAL ANALYSIS OF NITROREDUCTASES FROM THE INTESTINAL PROTOZOAN PARASITE GIARDIA LAMBLIA. Müller N, Müller J. Swiss National Science Foundation No A /1 Project period: Project budget: CHF ) KISS AND FLUSH A MINIMIZED ENDOCYTIC SYSTEM IN THE PROTOZOAN GIARDIA LAMBLIA Spycher C Swiss National Science Foundation, Ambizione, No. PZ00P3_132120/1 Project period: Project budget: CHF IPB: Jahresbericht 2012_13 / annual report 2012_13

5 Forschungsberichte / scientific reports Federal Office for Civil Protection Bundesamt für Bevölkerungsschutz 1) ANALYSE DER PATHOGENITÄTSMECHANISMEN IN NAEGLERIA FOWLERI (P) Gottstein B, Müller N Project No Project period: , Prolongated: Project budget: CHF CHF Programme Interreg IV A France-Suisse 1) ISOTOPECHINO (P) Gottstein B, Hemphill A, Müller J Project period: Project budget: CHF &Kt. Bern CHF &Kt. Waadt CHF Research support by Foundations / Others Forschungsbeiträge durch Stiftungen / Andere 1) Bordier Affinity Products, Crissier CHF November ) Bordier Affinity Products, Crissier CHF November ) Gottfried und Julia Bangerter-Rhyner-Stiftung A. Hemphill: CHF June 2012 CHF June ) Novartis Stiftung für medizinisch-biologische Forschung Nr. 12A02 The endophilin-like protein EmP29 in Echinococcus multilocularis metacestodes A. Hemphill CHF Mai 2012 Industry Industrie: 1) Bayer HealthCare AG, Leverkusen/Germany PRÜFUNG NEUER WIRKSTOFFE ZUR IN VITRO BEHANDLUNG VON NEOSPORA CANINUM, TOXOPLASMA GONDII UND BESNOITIA BESNOITI Gottstein B Project period: Project budget: CHF IPB: Jahresbericht 2012_13 / annual report 2012_113 5

6 Forschungsberichte / scientific reports 2) Intervet International, Boxmeer, The Netherlands PREVENTION OF NEOSPORA CANINUM INFECTION. Hemphill A Project period: Project budget: ) TI Pharma PHOSPHODIESTRASE INHIBITORS FOR THE TREATMENT OF NEGLECTED TROPICAL DISEASES Hemphill A Project period: Project budget: ) MegaCor Diagnostik GmbH, Austria VALIDATION FASTEST NEOSPORA CANINUM Hentrich B Project period: Project budget: SCIENTIFIC REPORTS ON MAIN RESEARCH PROJECTS 2012/13 ECHINOCOCCOSIS Prof. Dr. B. Gottstein, Prof. Dr. A. Hemphill, Prof. Dr. N. Müller, Dr. M. Spiliotis, G. Boubaker, PD Dr. J. Müller Cooperative investigations with: Dr. M. Siles-Lucas, Salamanca; Dr. D. Vuitton, Dr. L. Millon, Dr. F. Grenouillet, Dr. O. Blagosklonov, Dr. J. Knapp, Besançon; Prof. P. Deplazes, Dr. P. Torgerson, Zürich; Prof. B. Mühllhaupt, Zürich; Prof. K. Brehm, Würzburg; Prof. J. Keiser, Swiss Tropical Institute, Basel; Prof. D. Boykin, Atlanta, Prof. J. Vennerstroem, Omaha; PD Dr. Carsten Vock, Vienna. Humans represent an aberrant intermediate host for the fox tapeworm Echinococcus multilocularis, while small rodents (mice) function as natural intermediate hosts. Infection in humans can cause alveolar echinococcosis (AE), a severe disease with fatal outcome if not treated appropriately. Immunologiocal studies: We have previously shown that E. multilocularis infection affected peritoneal DCs such as to remain in an immature or resting state as well, characterized by low expression of costimulatory molecules and MHC class II (I-a) molecules. A concomitant high level of TGF-β-expression classifies these DCs within cells with suppressive features. Together, impairment of MØ and DC maturation and antigen presentation indicated that a successful E. multilocularis infection induces an anergix c status of immunity in the host. Subsequent experiments at the T cell level documented that regulatory T cells (Tregs) interfere in the complex immunological host response to infection. Indeed, a subpopulation of regulatory CD4+CD25+ T cells isolated from E. multilocularis-infected mice reduced ConA-driven proliferation of CD4+ T cells. The high expression levels of Foxp3 mrna by CD4+ and CD8+ T cells suggested that subpopulations of regulatory CD4+Foxp3+ and CD8+Foxp3+ T cells were involved in negatively modulating the immune responses of E. multilocularis-infected mice. Our most recent data demonstrated now that a novel CD4+CD25+ Treg effector molecule FGL2 contributes to the outcome of E. multilocularis infection by promoting Treg cell functions (Wang et al., submitted); they 6 IPB: Jahresbericht 2012_13 / annual report 2012_13

7 Forschungsberichte / scientific reports give evidence for a role of IL-17 in FGL2 regulation, and suggest that targeting FGL2 could be used for the development of novel treatment approaches in this parasitic disease. Small scale in vitro culture system for E. multilocularis primary cells: Echinococcus multilocularis is the laboratory model organism for cestodes with human relevance, because its genome is sequenced and more important, in vitro systems for culturing different larval stages and extracted primary cells are established. Until now, primary cells could only be cultured in the presence of feeder cells rendering the system non useful to screen for influences of bioactive molecules without feeder cell derived side effects. To circumvent this obstacle, we developed an axenic feeder cell free small scale (96-/384-well) culture system for primary E. multilocularis cells that was tested on its ability to screen large drug-candidate compilations or to detect growth factor derived effects on development, proliferation and/or survival. Characterization of the Echinococcus antigen P29: The antigen P29 was firstly described from E. granulosus as a serological marker to follow-up the infection status after surgery on young patients. It was shown, that the P29-antibody titer decreases in between 6-12 months on cured patients and stayed stable on non-cured patients. However, beside of the DNA/protein sequence and first experiments of its diagnostic usability nothing was known about P29. We characterized and analyzed the p29 bioinformatically and diagnostically for the E. granulosus complex and E. multilocularis. P29 is present as a single copy gene and between the different species one recombinantly expressed P29 can be used to detect infections with both, E. granulosus complex and E. multilocularis. When used as a vaccine on mice, P29 reduces parasite load when applied as a protective vaccine prior infection and additionally in an immunotherapeutic approach when applied during a manifested E. multilocularis infection. Deeper characterization on its biological importance for E. multilocularis showed, that P29 is highly important for survival of the parasite, since knock-down via RNA interference resulted in a death-phenotype. By Yeast- Two-Hybrid screen a possible interaction partner (cytoplasmatic malate dehydrogenase) was identified. Especially for the biological characterization of P29 further experiments still have to be performed. Recombinantly expressed E. multilocularis antibodies: A monoclonal antibody (anti Em2/G11) recognizing the carbohydrate structure (part of the native Em2 antigen) in the laminated layer of E. multilocularis was isolated by Peter Deplazes in 19xx. We cloned the antigen specific heavy- and lightchain of this antibody and constructed a single chain antibody (scg11-3) to be expressed in E. coli. During the following experiments we realized, that scg11-3 was working with similar specificity compared to the native monoclonal antibody, but the yield of functional scg11-3 was very low in E. coli extracts. Therefore, we designed a spacer-linked G11-3 diabody (G11-3-dia) that is now functional and expressed with high yields in mammalian Hek cells. Recombinant proteins can be easily mutated, for example additional tags can be added or single amino acids can be changed. Now, G11-3-dia is used to establish (i) a cleanup system for native Em2 antigen and (ii) a sandwich Elisa for the detection of Em2-antigen in body liquids of infected mice or human patients. Future plans involve (iii) the linking of radioactive or chemical substances to be used for directed treatment of Alveolar Echinococcosis and imaging techniques to localize E. multilocularis lesions in mice and human patients. Additionally, (iv) peptide sequences stimulating the immune system of infected mice will be added to direct the hosts immune response closely to the infection herd. Taken together, G11-3- dia will be applied in antigen-cleanup, diagnostic/localization/follow-up and treatment purposes. Experimental chemotherapy studies: The current chemotherapy of alveolar echinococcosis (AE) relies on albendazole and mebendazole, which are presently used for the treatment of non-surgical cases, and as a supplementary treatment prior and post-surgery. During AE, metacestodes proliferate asexually and infiltrate the neighbouring tissue, forming a complex mass consisting of peripherally proliferating metacestodes and centrally located necrotic tissue, all intermingled with connective tissue and immune cells, which is difficult to treat. Novel chemotherapeutical options should be developed to cure AE and CE. One approach to speed up drug discovery for AE is to find new uses for already approved drugs, or compounds and compound classes that are already in development This process, called drug repositioning, has been widely used in the field of rare and neglected diseases such as AE, where the interests of the pharma companies are not compatible with the expected financial benefits in drug development. Our laboratory has applied E. multilocularis cell-based and metacestode-based drug screening assays employing compound libraries from different commercial and non-commerical sources (FDA, MMV-malariabox, several academic collaborators) and identified several mefloquin-, artemisininand pentamidine-derivatives, as well as ruthenium-based organometallic compounds, which show interesting capacities to kill the metacestodes in vitro. In vivo studies in the mouse model demonstrated the efficacy of oral application of the anti-malarial compound mefloquin, which prompted us to further IPB: Jahresbericht 2012_13 / annual report 2012_113 7

8 Forschungsberichte / scientific reports investigate a range of mefloquine-derivatives. Promising in vivo effects were also obtained with a pentamidin derivative, DB1127, however, only when the drug was applied intraperitoneally. In addition, investigations on potential targets of selected drugs in Echinococcus are being carried out in order to explore their potential mechanism of action in the metacestode stage. NEOSPOROSIS, TOXOPLASMOSIS & BESNOITIOSIS Prof. Dr. A. Hemphill, Prof. Dr. B. Gottstein, Prof Dr. N. Müller, PD Dr. J. Müller, Dr. C. Frey, Dr. B. Hentrich, Dr. Karim Debache, Dr. Ferial Alaeddine, Dr. C. Guionaud, Thierry Monney, Fabienne Barna, Michelle Schorer, Pablo Winzer Collaborative investigations with: Prof. S. Leib, Dr. D. Grangirard, Bern; Prof. Alexandre Leitao, Lisbon, Portugal; Dr. Helder Cortes, Evora, Portugal; Prof. Luis Ortega-Mora, Madrid, Spain; PD Dr. M. Hässig, Prof. D Boykin, Atlanta, USA; Prof. Chad Stephens, Augusta, USA; PD Dr. Carsten Vock, Vienna, Austria, Prof. Wes VanVoorhis, Seattle, USA; Dr. Kajode Ojo, Seattle, USA. Vaccines against neosporosis: from mice to sheep: Neospora caninum is an obligatory intracellular apicomplexan parasite and causes abortion and stillbirth in cattle, and neuromuscular disease in dogs. The worldwide impact on the cattle industry is significant. Options for intervention are selective culling, vaccination and chemotherapy, and the latter two require research to identify suitable targets for preventive or therapeutic intervention. A major focus has been the development of vaccines against neosporosis, based on targets that are involved in adhesion and entry into host cells by the invasive stages of the parasite, and these studies are carried out in mice as a model for proof of concept. Thus, we hypothesize that antigens involved in host cell entry mechanisms represent good candidates for the development of a vaccine against N. caninum infection. Based on our investigations so far, we could largely confirm this hypothesis in a non-pregnant mouse model, where we could show that a series of antigens associated with either the surface of the parasites or organelles that mediate invasion actually confer protection against challenge infection upon vaccination. However, further investigations in the fetal infection model showed, that pregnancy alters the immune status of mice in such a way that the protective properties of vaccine candidates are substantially diminished. We therefore started to investigate the effects of physically linking recombinant antigens to distinct Toll-like receptor ligands in order to improve immunity and increase protection. In cooperation with partners in Madrid we are now in the process of developing an experimental N. caninum infection model in sheep to mimic the situation in cattle more accurately (but at a lower cost than in cattle), and for future applications of this model in vaccination and chemotherapy. Drugs against N. caninum and other apicomplexans: in vitro and in vivo chemotherapy studies: An immunological approach alone might not be sufficient to efficiently combat this parasitic infection, and we have therefore also explored potential chemotherapeutical tools for treatment. Prospectively, this strategy could combine the two options: (i) application of a preventive vaccine to minimize infection and (ii) application of an efficacious compound that eliminates residual parasites that had achieved infection despite of vaccination. We have studied a number of compound classes that could be potentially useful for chemotherapeutical use in Neospora-infected animals. These include miltefosine, thiazolide compounds such as nitazoxanide, dicationic pentamidine derivatives such as DB750 and DB745, ruthenium-based organometallic compounds, and calcium-dependent protein kinase (CDPK) inhibitors. Especially CDPK inhibitors exhibited outstanding activity against N. caninum infection in mice in both non-pregnant and pregnant animals. Bovine besnoitiosis: Bovine besnoitiosis (disease caused by infection with the protozoa Besnoitia besnoiti) is emerging in Europe. Indeed, the first cases have been detected in Switzerland in Our institute contributed in the diagnosis of these cases. Furthermore, Dr. C. Frey received a personal grant from the Swiss national Science Foundation to spend a research year investigating bovine besnoitiosis in Spain, at Saluvet group, Complutense University, Madrid, Spain. 8 IPB: Jahresbericht 2012_13 / annual report 2012_13

9 Forschungsberichte / scientific reports GIARDIASIS Prof. Dr. N. Müller, PD Dr. Joachim Müller, Prof. Dr. A. Hemphill, Dr. C. Spycher, Dr. S. Kunz Cooperative investigation with: Prof. Dr. A. Hehl, Institute of Parasitology; Zürich; Dr. David Leitsch, Dept. of Specific Prophylaxis and Tropical Medicine, Medical University of Vienna, Vienna, Austria; The Pathogen Functional Genomics Research Center; J. Craig Venter Institute, Rockville, Ma., U.S.A. Chemotherapy of giardiasis: function of nitroreductases in the metabolism of nitro drugs metronidazole and nitazoxanide: The protozoan parasite Giardia lamblia causes the intestinal disease giardiasis that may lead to acute and chronic diarrhea in humans and various animal species. Nitro drugs such as the nitroimidazole metronidazole (MET) and the nitrothiazolide nitazoxanide (NTZ) are currently used for treatment of giardiasis. We are interested in the mode of action of, and the mechanisms involved in resistance to, these nitro drugs. Using affinity chromatography, we identified a nitroreductase, GlNR1, as a NTZ-binding protein. Moreover, G. lamblia has a second nitroreductase in its genome, GlNR2. While G. lamblia trophozoites overexpressing GlNR1 exhibited increased susceptibility to nitro drugs, trophozoites overexpressing GlNR2 were less susceptible to these drugs as compared to control trophozoites. The biological function(s) of GlNR1, and GlNR2, are not known so far. Assuming that the presence of nitroreductases must have selective advantages for the parasite, we are currently investigating if GlNR1 and GlNR2 are involved in the reduction of toxic nitrous compounds thus preventing nitrosative stress. Such compounds are: (i) nitric oxide (NO) produced either by endogenous processes (from L-arginine), or by the host, and (ii) nitroso or hydroxylamine intermediates present in the digestive tract. Accordingly, we are investigating both on the mrna (by reverse transcription PCR) and protein (by immunoblot) level if GlNR1 and GlNR2 are up-regulated in Giardia cultures upon exposure of the cells to different nitro compounds and compounds that influence endogenous NO levels (NO donors, L-arginine). Nitrosative stress conditions can also be achieved by co-cultivating parasites with NO-producing intestinal epithelial cells. Our project also includes an antisense RNA gene silencing approach in order to reduce the GlNR1 and GlNR2 expression levels in G. lamblia. Here, we make use of various constructs providing low to high level gene expression and test if transgenic knockdown parasites exhibit a characteristic growth phenotype in response to nitro drugs and natural nitrosative agents. In summary, our planned studies on GlNR1 and GlNR2 highlight the biological function(s) of nitroreductases in G. lamblia and provide important information about their role as drug targets thus allowing rational screening approaches for the identification of novel compounds suitable for treatment of giardiasis and related parasitic diseases. Validating the phosphodiesterase of Giardia lamblia as new drug target: There is a superb combination of evidence validating cyclic nucleotide phosphodiesterases (PDEs) as meaningful drug targets in protozoan parasites. PDEs hydrolyze the second messengers camp and cgmp to its inactive 5 -monophosphate forms and thus are key players in cyclic nucleotide signaling. In different protozoan parasites such as trypanosomes, Leishmania and malaria-causing Plasmodium, cyclic nucleotide signaling plays a pivotal role in vital cellular functions such as cell differentiation or cell cycle control. Interfering in the camp signaling cascade by inhibiting PDEs may be lethal for parasites as was demonstrated in Trypanosoma brucei, the causative agent of the fatal sleeping sickness. There is ample evidence that the same principal works also in Giardia lamblia. In Giardia, camp has been suggested to be important in cell differentiation (excystation) and parasite attachment to intestinal cells, thus the PDE could play a vital role in the infection biology of this parasite. Our preliminary experiments revealed that some of the PDE inhibitors identified so far have a potential to kill Giardia cells. Strikingly, these compounds are also efficient inhibitors of the Giardia PDE. These results lead us to the hypothesis that the Giardia PDE is essential for the survival of the organism and represents an important drug target. In our ongoing studies, we aim to validate the PDE as novel drug target by applying a pharmacogenetic approach. We will genetically decrease and increase PDE expression levels in G. lamblia trophozoites. The so generated transgenic strains will be tested for phenotypically changes including lethal effects - and for altered susceptibility against PDE inhibitors. These principal studies will be complemented by further experiments, which shall help to characterize the function of the PDE in G. lamblia. IPB: Jahresbericht 2012_13 / annual report 2012_113 9

10 Forschungsberichte / scientific reports Investigation of the protein-secretory machinery in Giardia lamblia: Giardia lamblia has a minimized organelle repertoire, and most strikingly lacks a classical stacked Golgi apparatus. Nevertheless, Giardia trophozoites constitutively secrete variant surface proteins, and dramatically increase the volume of protein secretion during differentiation to cysts. Eukaryotic cells have evolved an elaborate system for quality control (QC) of protein folding and capacity in the endoplasmic reticulum (ER). Upon ER-overload, an unfolded protein response (UPR) is triggered on transcriptional/translational level aiming at alleviating ER stress. In Giardia, a minimized secretory machinery and absence of glycan-dependent QC suggests that a genetically conserved UPR (or functional equivalent) to cope with insults to the secretory system has been eliminated. We tested this hypothesis of UPR elimination by profiling the transcriptional response during induced ER-folding stress. We showed that on the contrary, ER-folding stress triggers a stressor-specific, ER-directed response with upregulation of only ~ 30 genes, with different kinetics and scope compared with the UPR of other eukaryotes. Computational genomics revealed conserved cisacting motifs in upstream regions of responder genes capable of stressor-specific gene regulation in transfected cells. Interestingly, the sensors/transducers of folding stress, well conserved in model eukaryotes, are absent in Giardia suggesting the presence of a novel version of this essential eukaryotic function. NAEGLERIA FOWLERI Prof. Dr. N. Müller, Dr. Denise Zysset-Burri, Prof. Dr. B. Gottstein Cooperative investigation with Drs. N. Schürch, M. Wittwer (Spiez), Prof. Dr. S. L. Leib (Bern); Drs. A. Kiderlen, A. Äbischer, Robert-Koch-Institute, Berlin, Gemany Identification of potential pathogenicity factors of the free-living amoeba Naegleria fowleri by genomic, transcriptomic and proteomic approaches: Naegleria species are free-living amoebae found worldwide in soil and warm fresh water. Although around 30 species have been described, Naegleria fowleri is the only pathogen that causes primary amoebic meningoencephalitis (PAM) in humans. PAM is a disease of the central nervous system that mainly affects healthy children and young adults with a history of recreational activity in lakes, pools and hot springs. Infection occurs when water contaminated by N. fowleri enters the noses of swimmers and the amoebae reach the brain through the olfactory nerve tract. Several days after infection, the patients suffer from headache, fever, vomiting, nausea and behavioral abnormalities. Because most of the infected persons fail to be diagnosed rapidly, they die one to two weeks after exposure to the infectious water source. Despite extensive research over the last decades, the determinants for this acute and fast progressing nature of PAM are still unknown. Thus, the overall aim of my thesis was to provide further insights into the pathogenesis of N. fowleri. To achieve this aim, we established a novel pathogenicity model for N. fowleri based on different compositions of axenic growth media. We showed that the presence of liver hydrolysate in the medium resulted in rapidly proliferating trophozoites with a smaller cell diameter and a higher pathogenic potential in mice. Based on this model, we performed proteomic approaches comparing highly with weakly pathogenic trophozoites to identify proteins and cellular locations with a potential role in the pathogenesis of N. fowleri. As scaffold for the identification of differentially expressed proteins, we de novo sequenced the 30-Mb genome as well as the transcriptome of N. fowleri. Using these data, we found that the extent of relationship of N. fowleri and its non-pathogenic relative Naegleria gruberi, whose genome has been sequenced in 2010, is comparable to that between Trypanosoma brucei and Trypansosoma cruzi. Furthermore, we proposed a low similarity between the coding sequences of these two Naegleria species. Through the comparative proteomic approaches, we identified 22 novel proteins with a potential role in the pathogenesis of N. fowleri. Via cellular component analysis of these potential pathogenicity factors, the membrane of the amoeba was identified as a key location where pathogenic processes may occur. These processes may involve actin-dependent vesicular trafficking mechanisms. The results of the project provide the basis for further examination using reverse genetic approaches to demonstrate the involvement of the identified candidate proteins in the pathogenesis of PAM. Furthermore, proteins with a confirmed role in pathogenic processes will serve as a basis for subsequent pharmacological studies to identify novel drug targets of N. fowleri 10 IPB: Jahresbericht 2012_13 / annual report 2012_13

11 Forschungsberichte / scientific reports BOVINE AND FELINE TRITRICHOMONOSIS Dr. C. Frey, Prof. Dr. B. Gottstein, Prof. Dr. N. Müller, PD Dr. J. Müller, K. Reinmann, C. Bernasconi Cooperative study with: Dr. Louis Ortega (Saluvet group, UCM, Spain), Dres. Carlos Campero & Marcelo Fort (INTA, Argentina), Prof. Jan Slapeta (University of Sydney, Australia), Dr. Klaus Henning (FLI, Germany), Dr. David Leitsch (Medical University, Vienna), Prof. Michael Hess (University of Veterinary Medicine, Vienna). Tritrichomonas foetus is an economically important venereal disease in cattle. The parasite is an OIE list B pathogen, and bulls used for artificial insemination in Switzerland have to be closely monitored for T. foetus. With these measures, Switzerland has become free from tritrichomoniasis. In the late 1990ies reports were released about T. foetus causing large bowel diarrhea in cats in the USA. We developed a PCR to be used for the diagnosis of cats affected by T. foetus. Furthermore, we successfully applied the culture system used in bulls for a similar diagnostic approach in cats. Subsequently, cats with a history of diarrhea were screened with both methods. We found a very high prevalence of over 25% in this population at risk. As it is very important to know whether the parasites of cattle and of cats are taxonomically identical, we identified two genetic markers, ITS-2 and elongation factor 1alpha, and could show that T. foetus isolates from cats and cattle are very closely related, but display genetic differences in both loci. In collaboration with an Australian research group, a wider panel of 10 genomic markers was developed and applied to various bovine and feline isolates. These in-depth analyses confirmed that feline and bovine isolates indeed are genetically distinct. A large-scale survey targeting naturally mating bulls in Switzerland could not find bovine T. foetus in any of the samples, thus suggesting that bulls in Switzerland are free from T. foetus. Our genetic and epidemiological studies thus suggested that feline and bovine T. foetus do not interfere in natural cycles. OTHER PROJECTS 2012/13 (Projects & cooperations funded / supported by extramural grants) ASSESSMENT OF RECOMBINANT ANTIGENS FROM FASCIOLA HEPATICA FOR THE IMMUNODIAGNOSIS OF HUMAN AND ANIMAL FASCIOLOSIS. Prof. Dr. M. Spiliotis, Ghalia Boubaker, Marianne Schneeberger, Cristina Huber, Prof. Dr. B. Gottstein Cooperative study with: PD Dr. M. Doherr (VPHI) and Prof. Dr. H. Cortes (Evora University). To improve the serodiagnosis of human fasciolosis caused by Fasciola hepatica, we comparatively evaluated the accuracy of two different enzyme-linked immunosorbent assays (ELISAs) based on the use of recombinant F. hepatica saposin-like protein-2 antigen (recsap2). Although the F. hepatica E/S antigen exhibited a slightly higher diagnostic sensitivity, the higher specificity performance of recsap2 renders this antigen very suitable for application in low endemic areas, especially when coupled to an easy and standardized production facility as compared to the relatively complex production procedure for an E/S antigen. Conclusively, the recsap2-elisa can be used as a routine individual serodiagnostic test for human fasciolosis. IMMUNOLOGY AND GENETICS OF EQUINE RECURRENT AIRWAY OBSTRUCTION AND DEFENCE AGAINST NEMATODES (SNF) Prof. Dr. V. Gerber (main applicant), Horse Clinics Prof. Dr. B. Gottstein, Prof. Dr. T. Leeb, Prof. Dr. E. Marti Schalch (other applicants) The hygiene-hypothesis postulates an inverse relationship between certain parasitic and other infectious diseases and measures of atopy and asthma in humans. Recurrent airway obstruction (RAO; IPB: Jahresbericht 2012_13 / annual report 2012_113 11

12 Forschungsberichte / scientific reports horse asthma) is a major health problem of stabled horses. Two large half-sibling horse families and large numbers of unrelated RAO cases are used to study if a genetic adaptation to strongylid nematode parasites may predispose horses living in an unnatural environment (hay-feeding; frequent deworming) to RAO. In order to study the immunological response to antigens relevant to RAO and intestinal strongylid parasitism, peripheral blood mononuclear cells are stimulated with hay dust extract, Aspergillus fumigatus extract, extract from small strongylids and recombinant strongylid antigens. In both of the families, RAOaffected animals and controls are compared. MICROPET FOR THE DEVELOPMENT OF NEW IMAGING MARKERS IN AE (INTERREGIV) Prof. Dr. B. Gottstein, Prof. O. Blagosklonov (main applicants), C. Porot, J. Wang, Prof. Y. Seimbille, D. Camporese, Dr. J. Knapp, Prof. D. Vuitton, S. Germain [18F]- fluorodeoxyglucose (FDG) is a validated tracer of AE lesions; however, it does not directly reflect parasite viability but rather peri-parasitic host inflammatory processes. The ideal tracer should be able to assess the course of AE upon direct uptake by the metacestode through its metabolic activity. We designed a radiolabelling protocol which could be assessed for activity in host and parasite cell cultures in vitro. We compared the uptake of FDG and of some other PET tracers ([[18F]-fluorotyrosine (FET), [18F]- fluorothymidine (FLT), [18F]-fluorometylcholine (FMC), and sodium [18F]-fluorine (NaF)) by in vitro studies on human hepatocytes, human leukocytes and in vitro-cultivated metacestode vesicles. Preliminary results showed that leukocytes were the greediest for all fluorinated tracers but mainly for FDG. FLT appeared to be the best promising candidate for AE functional imaging because we observed very high FLT-uptake in parasite vesicles and no FLT-uptake in human leukocytes. In order to confirm FLT potential in vivo, we elaborated a PET imaging protocol for appropriate murine AE primary and secondary infection studies. World-wide first PET images of AE in animals were acquired after intraperitoneal metacestode injection; FDG-uptake was studied at advanced infection time points. Experiments confirmed that FDG distribution in PET reflects inflammatory reactions in the peri-parasitic metacestode tumour-like tissue. Other smaller projects... - Equinella-Meldesystem zur Erfassung infektiöser Erkrankungen von Pferden in der Schweiz. Dr. C. Frey (IPB); Dr. F. Wohlfender (VPHI). - Oxyurenproblem bei Klammeraffen des Zoo s Basel: (Masterarbeit Stéphanie Borel). Prof. B. Gottstein (IPB), Dr. B. Hentrich (IPB), Dr. S. Hobi (Zoo Basel), Prof. R. Gasser (Melbourne University) - Validierung des Trichin-L, Trichinella Antigen Test Kit der Firma BIO-RAD für den Nachweis von Trichinella Antigenen im Schwein: Validierungsstudie für die Zulassung als Diagnostischen Test durch das BLV. Dr. B. Hentrich (IPB). - The effect of commercial anthelmintic Herb-All WORM on early Ascaris suum migration. (Dissertation Salomé Siegenthaler). Prof. W. Zimmermann, Prof. B. Gottstein, Dr. B Hentrich, Prof. M. Doherr. 12 IPB: Jahresbericht 2012_13 / annual report 2012_13

13 Öffentlichkeitsarbeit TECHNICAL SERVICES IN RESEARCH ELECTRON MICROSCOPY Preparative methods for basic electron microscopical techniques and immunogold-labeling are available. TEM access is ensured at the Institute of Anatomy at the Medical Faculty, University of Bern, SEM is carried out at the Electron Microscope Laboratory of the Department of Chemistry, University of Bern. PUBLIKATIONEN 2012/13 Peer-reviewed scientific articles (2012) Barnes TS, Deplazes P, Gottstein B, Jenkins DJ, Mathis A, Siles-Lucas M, Torgerson PR, Ziadinov I, Heath DD: Challenges for diagnosis and control of cystic hydatid disease. Acta Trop. 123: 1-7 (2012). Burri DC, Gottstein B, Zumkehr B, Hemphill A, Schürch N, Wittwer M, Müller N: Development of a high- versus low-pathogenicity model of the free-living amoeba Naegleria fowleri. Microbiology. 158: (2012) Debache K, Hemphill A. Effects of miltefosine treatment in fibroblast cell cultures and in mice experimentally infected with Neospora caninum tachyzoites. Parasitology 139: (2012). Debache K, Hemphill A. Intra-cisternal vaccination induces high-level protection against Neospora caninum infection in mice. Vaccine 30: (2012). Eichhorn L, Zimmermann W, Gottstein B, Frey CF, Doherr MG, Zeeh F: Erstmalier Nachweis von Lungenwürmern bei Freilandschweinen in der Schweiz. Schweiz. Arch. Tierheilk. 154: (2012). Frey CF, Berger-Schoch AE, Hermann DC, Schares G, Müller N, Bernet D, Doherr MG, Gottstein B: Vorkommen und Genotypen von Toxoplasma gondii in der Muskulatur von Schaf, Rind und Schwein sowie im Katzenkot in der Schweiz. Schweiz. Arch. Tierheilk. 154: (2012). García-Lunar P, Ortega-Mora LM, Schares G, Gollnick NS, Jacquiet P, Grisez C, Prevot F, Frey CF, Gottstein B, Álvarez-García G: An inter-laboratory comparative study of serological tools employed in the diagnosis of Besnoitia besnoiti infection in bovines. Transboundary and Emerging Diseases, 60: (2012). Jenkins EJ, Peregrine AS, Hill JE, Somers C, Gesy K, Barnes B, Gottstein B, Polley L: Detection of European strain of Echinococcus multilocularis in North America [letter]. Emerg Infect Dis [serial on the internet] June [date cited]. Keutgens A, Simoni P, Detrembleur N, Frippiat F, Giot JB, Spirlet F, Aghazarian S, Descy J, Meex C, Huynen P, Melin P, Müller N, Gottstein B, Carlier Y, Hayette MP: Fatal alveolar echinococcosis of the lumbar spine. J Clin Microbiol. 51: (2012). IPB: Jahresbericht 2012_13 / annual report 2012_113 13

14 Öffentlichkeitsarbeit Knapp J, Staebler S, Bart JM, Stien A, Yoccoz NG, Drögemüller C, Gottstein B, Deplazes P: Echinococcus multilocularis in Svalbard, Norway: Microsatellite genotyping to investigate the origin of a highly focal contamination. Infect Genet Evol. 12: (2012). Küster T, Zumkehr B, Hermann C, Theurillat R, Thormann W, Gottstein B, Hemphill A: Voluntary ingestion of antiparasitic drugs emuslified in honey represents an alternative to gavage in mice. Journal of the American Association for Laboratory Animal Science 51: 1-5, Küster T, Lense N, Barna F, Hemphill A, Kindermann MK, Heinicke JW, Vock CA. A new promising application for highly cytotoxic metal compounds: η6-arene ruthenium(ii) phosphite complexes for the treatment of alveolar echinococcosis. J Med Chem 55: (2012). Lesser M, Braun U, Deplazes P, Gottstein B, Hilbe M, Basso W: Erste Fälle von Besnoitiose bei Rindern in der Schweiz. Schw. Arch. Tierheilk. 154: (2012). Lobsiger L, Frey CF, Müller N, Rosenberg G, Schweizer T, Gottstein B: Breitet sich die bovine kutane Leishmaniose in der Schweiz aus? Schw. Arch. Tierheilk. 154: (2012). Marreros N, Frey CF, Willisch CS, Signer C, Ryser-Degiorgis MP: Baseline coprological data in apparently healthy Alpine ibex (Capra ibex ibex) from two Swiss colonies. Vet. Parasitol. 186: (2012). Monney T, Debache K, Grandgirard D, Leib SL, Hemphill A. Vaccination with the recombinant chimeric antigen recncmic3-1-r induces a non-protective Th2-type immune response in the pregnant mouse model for N. caninum infection. Vaccine 30: (2012) Peregrine AS, Jenkins EJ, Barnes B, Johnson S, Polley L, Barker IK, De Wolf B, Gottstein B: Alveolar hydatid disease (Echinococcus multilocularis) in the liver of a Canadian dog in British Columbia, a newly endemic region. Can Vet J 53 : (2012). Reinmann K, Müller N, Kuhnert P, Campero CM, Leitsch D, Hess M, Henning K, Fort M, Müller J, Gottstein B, Frey CF: Tritrichomonas foetus isolates from cats and cattle show minor genetic differences in unrelated loci ITS-2 and EF-1α. Vet Parasitol. 185(2-4): (2012). Schorer M, Debache K, Barna F, Monney T, Müller J, Boykin D, Stephens CE, Hemphill A. Di-cationic arylimidamides act against Neospora caninum tachyzoites by interference in membrane structure and nucleolar integrity and are active against challenge infection in mice. Int I Parasitol Drugs Drug Res 2: (2012). Slapeta J., Müller N., Stack C.M., Walker, G, Lew-Tabor A., Tachezy J. Frey C.F: Comparative analysis of Tritrichomonas foetus (Riedmuller, 1928) cat genotype, T. foetus (Riedmuller, 1928) cattle genotype and Tritrichomonas suis (Davaine, 1875) at 10 DNA loci. Int. J. Parasitol. 42: (2012). Schweiger A., Grimm F., Tanner I., Müllhaupt B., Bertogg K. Müller, N., Deplazes P. Infection 40: (2012). Sidler D, Brockmann A, Mueller J, Nachbur U, Corazza N, Renzulli P, Hemphill A, Brunner T. Thiazolide-induced apoptosis in colorectal cancer cells is mediated via the Jun kinase-bim axis and reveals glutathione-s-transferase P1 as Achilles'heel. Oncogene. 31: (2012). Spiliotis M. Inverse fusion PCR cloning. PLoS One doi: /journal.pone (2012). Takumi K, Hegglin D, Deplazes P, Gottstein B, Teunis P, van der Giessen J: Mapping the increasing risk of human alveolar echinococcosis in Limburg, The Netherlands. Epidemiol Infect. 140: (2012). 14 IPB: Jahresbericht 2012_13 / annual report 2012_13

15 Öffentlichkeitsarbeit Peer-reviewed scientific articles (2013) Alaeddine F, Hemphill A, Debache K, Guionaud C. Molecular cloning and characterization of NcROP2Fam-1, a member of the ROP2 family of rhoptry proteins in Neospora caninum that is targeted by antibodies neutralizing host cell invasion in vitro. Parasitology. 140: (2013). Barna F, Debache K, Vock CA, Küster T, Hemphill A. In vitro effects of novel ruthenium complexes in Neospora caninum and Toxoplasma gondii tachyzoites. Antimicrob Agents Chemother 57: (2013). Berzina I, Müller N, Krudewig C, Silaghi C., Matise I., Welle M. Histopathological changes in the skin associated with persistent canine granulocytic anaplasmosis. J. Vet. Int. Med. 27: Boubaker G, Macchiaroli N, Prada L, Cucher MA, Rosenzvit MC, Ziadinov I, Deplazes P, Saarma U, Babba H, Gottstein B, Spiliotis M: A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex. PLoS Negl Trop Dis 7(1): e2017. doi: /journal.pntd (2013). Debache K, Hemphill A. Differential effects of intranasal vaccination with recombinant NcPDI in different mouse models of Neospora caninum infection. Parasite Immunol. 35: (2013). de Marval F, Gottstein B, Weber M, Wicht B: Imported diphyllobothriasis in Switzerland: molecular methods to define a clinical case of Diphyllobothrium infection as Diphyllobothrium dendriticum, August Eurosurveillance, Volume 18, Issue 3, 17 January (2013). Frey CF, Gutiérrez-Expósito D, Ortega-Mora LM, Benavides J, Marcén JM, Castillo JA, Casasús I, Sanz A, García-Lunar P, Esteban-Gil A, Alvarez-García G: Chronic bovine besnoitiosis: Intra-organ parasite distribution, parasite loads and parasite-associated lesions in subclinical cases. Vet Parasitol. 197: (2013). Keutgens A, Simoni P, Detrembleur N, Frippiat F, Giot JB, Spirlet F, Aghazarian S, Descy J, Meex C, Huynen P, Melin P, Müller N, Gottstein B, Carlier Y, Hayette MP: Fatal alveolar echinococcosis of the lumbar spine. J. Clin. Microbiol. doi: /jcm (2013) Küster T, Hermann C, Hemphill A, Gottstein B, Spiliotis M: Subcutaneous infection model facilitates treatment assessment of secondary Alveolar echinococcosis in mice. PLoS Negl Trop Dis. 23;7(5):e2235 (2013). Küster T, Kriegel N, Boykin DW, Stephens CE, Hemphill A. In vitro and in vivo activities of dicationic diguanidino compounds against Echinococcus multilocularis metacestodes. Antimicrob Agents. Chemother 57: (2013). Lanz S, Gerber V, Marti E, Rettmer H, Klukowska-Rötzler J, Gottstein B, Matthews JB, Pirie S, Hamza E: Effect of hay dust extract and cyathostomin antigen stimulation on cytokine expression by PBMC in horses with recurrent airway obstruction. Vet Immunol Immunopathol. 155: (2013). Maksimov P, Zerweck J, Dubey JP, Pantchev N, Frey CF, Maksimov A, Reimer U, Schutkowski M, Hosseininejad M, Ziller M, Conraths FJ, Schares G: Serotyping of Toxoplasma gondii in cats (Felis domesticus) reveals predominance of type II infections in Germany. PLoS One. 8(11):e80213 (2013). Mols-Vorstermans T, Hemphill A, Monney T, Schaap D, Boerhout E. Differential Effects on Survival, Humoral Immune Responses and Brain Lesions in Inbred BALB/C, CBA/CA, and C57BL/6 Mice Experimentally Infected with Neospora caninum Tachyzoites. ISRN Parasitology Article ID , Monney T, Grandgirard D, Leib SL, Hemphill A. Use of a Th1 Stimulator Adjuvant for Vaccination against Neospora caninum Infection in the Pregnant Mouse Model. Pathogens 2: (2013). IPB: Jahresbericht 2012_13 / annual report 2012_113 15

16 Öffentlichkeitsarbeit Müller J., Schildknecht P., Müller N. Metabolism of nitro drugs metronidazole and nitazoxanide in Giardia lamblia: characterization of a novel nitroreductase (GlNR2). J. Antimicrob. Chemother. 68: (2013). Schaarschmidt D, Gilli U, Gottstein B, Marreros N, Kuhnert P, Daeppen JA, Rosenberg G, Hirt D, Frey CF: Questing Dermacentor reticulatus harbouring Babesia canis DNA associated with outbreaks of canine babesiosis in the Swiss Midlands. Ticks Tick Borne Dis. 4: (2013). Spekker K, Leineweber M, Degrandi D, Ince V, Brunder S, Schmidt SK, Stuhlsatz S, Howard JC, Schares G, Degistirici O, Meisel R, Sorg RV, Seissler J, Hemphill A, Pfeffer K, Däubener W. Antimicrobial effects of murine mesenchymal stromal cells directed against Toxoplasma gondii and Neospora caninum: role of immunity-related GTPases (IRGs) and guanylate-binding proteins (GBPs). Med Microbiol Immunol 202: (2013). Zangger H., Ronet C., Desponds C., Kuhlmann F.M., Robinson J., Hartley M.A., Prevel F., Castiglioni P., Pratlong F., Bastien P., Müller N., Parmentier L., Saravia N.G., Beverley S.M., Fasel N. Detection of Leishmania RNA virus in Leishmania parasites. PLoS Negl. Trop. Dis. 7, e2006. doi: /journal.pntd (2013). Peer-reviewed Reviews and book-chapters (2012) Frey CF, Müller N: Tritrichomonas - Systematics of an enigmatic genus Mol. Cell. Probes 26: (2012). Peer-reviewed Reviews and book-chapters (2013) Alvarez-Garcia G, Frey CF, Ortega-Mora LM, Schares G: A century of bovine besnoitiosis: an unknown disease re-emerging in Europe. Trends Parasitol. 29: (2013). Hemphill A, Müller N, Müller J. Thiazolides, a novel class of anti-infective drugs, effective against viruses, bacteria, intracellular and extracellular protozoan parasites and proliferating mammalian cells. Anti-Infective Agents 11, (2013). Hemphill A, Debache K, Monney T, Schorer M, Guionaud C, Alaeddine F, Mueller N, Mueller J. Proteins mediating the Neospora caninum-host cell interaction as targets for vaccination. Front. Biosci. (Elite Ed). 5, (2013) Müller J, Hemphill A. New approaches for the identification of drug targets in protozoan parasites. Int. Rev. Cell. Mol. Biol. 301, (2013). Müller J, Hemphill A. In vitro culture systems for the study of apicomplexan parasites in farm animals. Int. J. Parasitol. 43, (2013). Soeiro MN, Werbovetz K, Boykin DW, Wilson WD, Wang MZ, Hemphill A. Novel amidines and analogues as promising agents against intracellular parasites: a systematic review. Parasitology. 140, (2013). 16 IPB: Jahresbericht 2012_13 / annual report 2012_13

17 Öffentlichkeitsarbeit VORTRÄGE & POSTER (P) 20102/ Cretu CM, Staniceanu F, Voinea L, Mihailescu P, Pop M, Malis S, Popa LG, Stanescu D, Coroiu Z, Gottstein B: Human dirofilariasis: emergent diseases in Romania? EMOP, Cluj, Romania, July Frey CF: Durchfall bei älteren Kälbern bekannte und unbekanntere Erreger. Vetsuisse Nutztierabend, Bern, 13. März Frey CF: Koproskopische Diagnostik parasitärer Erreger Möglichkeiten unter Praxisbedingungen. Kleinwiederkäuertagung, Zürich, 16. Februar Frey CF: Behandlungsschema Parasitologie therapeutische und prophylaktische Massnahmen. Kleinwiederkäuertagung, Zürich, 16. Februar Gottstein B, Hässig M: Concomitatnt parameters promoting Neospora-induces abortion in cattle? ApiCowplexa-meeting, Lisboa, October 25-28, Gottstein B: Echinococcus multilocularis und alveoläre Echinokokkose. Weiterbildungsseminar für Tierärztlich-technisches Personal KTK. 30. April Gottstein B: Echinococcuss multilocularis infection in patients with AIDS or suffering from other immunodeficiencies. Int. Meeting on HIV and associated infectious diseases., Würzburg, January 19-20, Gottstein B: Visceral Larva Migrans. 22nd ECCMID, London, March 31 April 3, Gottstein B: Wildbahnassoziierte Zoonosen: Auswahl parasitärer Erreger. XVIII. Diagnostik-Symposium, Schaan, Lichtenstein, Hemphill A. In vitro induction of stage conversion in the parasite Neospora caninum. 3R-ecopa- Swisstox, Technopark Zürich, Nov 19-20, Hentrich B: Parasitologische Labordiagnostik unter Praxisbedingungen. Graubündner Frühjahrstagung,Thusis, , Schweiz. Müller N: Drug resistance formation in the intestinal protozoan parasite Giardia lamblia. Phil Nat. Faculty, University Basle: May 7, Müller N, Nillius D., Hemphill A., Müller J.: Functional analyses of nitro drug activation and resistance formation in Giardia lamblia. V International Giardia and Cryptosporidium Conference, Wellington, New Zealand, January 31 February 3, 2012 Spiliotis M, Hirschi P, Kiss F, Largiader C, Hemphill A, Gottstein B: A chip based RNA-microarray for Echinococcus multilocularis. Annual Meeting of the SGTP, St. Gallen, June 21-22, Bernasconi C, Bodmer M, Doherr M, Janett F, Thomann A, Iten C, Hentrich B, Müller N, Gottstein B, Frey CF: Prävalenzschätzung von Tritrichomonas foetus-infektionen bei Stieren in der Schweiz. DVP Parasitology Meeting, Giessen, 8-10 Juli, IPB: Jahresbericht 2012_13 / annual report 2012_113 17

18 Öffentlichkeitsarbeit Bernasconi C, Bodmer M, Doherr M, Janett F, Thomann A, Iten C, Hentrich B, Müller N, Gottstein B, Frey CF: Tritrichomonas foetus: a prevalence study in naturally mating bulls in Switzerland. Buiatrissima, Bern/Switzerland, 30. August, Frey CF, Gutiérrez-Expósito D, Ortega-Mora LM, Benavides J, Marcén JM, Castillo JA, Casasús I, Sanz A, García-Lunar P, Álvarez-García G. Chronic bovine besnoitiosis: histopathological findings and parasite distribution and load in subclinical cases. Buiatrissima, Bern/Switzerland, 30. August Frey CF, Hentrich B: Zecken-übertragene Krankheiten beim Rind und Hund. Weiterbildung Berner Tierärzte, Langenthal, 14. November Gerber V, Dolf G, Bründler P, Nussbaumer S, Frey C, Leeb T, Gottstein B: Association between resistance to strongylid nematodes and recurrent airway obstruction (RAO) in warmblooded horses. DVP Parasitology Meeting, Giessen, 8-10 Juli, Gottstein B, Basso W: Vorkommen und Bedeutung von Toxoplasma gondii beim Schwein in der Schweiz. GST Jubiläumskongress, Bern, 5-7 Juni, Gottstein B, Müller J, Greif G, Cortes H: In vitro efficacy of Toltrazuril against Besnoitia besnoiti infection in Vero and HFF cells. 24th WAAVP Meeting, Perth, Australia, August 26-29, Gottstein B: Echinococcus multilocularis infection (alveolar echinococcosis): recent developments. Morgenfortbildung Viszerale Chirurgie und Medizin, Inselspital Bern, January 10, Gottstein B: NRL presentation on the Epidemiology of Parasitic zoonoses in Switzerland. 8th Workshop of national Reference Laboratories for Parasites, ISS, Roma, May, Gottstein B: In vitro and in vivo investigations to develop functional imaging by Positron Emission Tomography (PET) for murine and human alveolar echinococcosis. Annual SSTMP meeting, Basle, October 31, Hemphill A, Küster T, Kriegel N, Boykin DW, Stephens CE. In vitro and in vivo activities of dicationic compounds against Echinococcus multilocularis metacestodes. 24th International Conference of the World Association for the Advancement of Veterinary Parasitology. Perth, August 25-29, 2013 Hemphill A, Barna F, Debache K, Vock CA. In vitro (& in vivo) effects of novel ruthenium complexes in Neospora caninum and Toxoplasma gondii tachyzoites. 24th International Conference of the World Association for the Advancement of Veterinary Parasitology. Perth, August 25-29, Hemphill A, Barna F, Debache K, Vock CA. In vitro (& in vivo) effects of novel ruthenium complexes in Neospora caninum and Toxoplasma gondii tachyzoites. Apicowplexa 2: 2nd international meeting on apicomplexan parasites in farm animals. Kusadasi, Turkey, October 31-Novermber 2, Knapp J, Umhang G, Boue F, Hornaz V, Millon L, Gottstein B: Contamination à Echinococcus multilocularis des hôtes primates captifs et rongeurs. Congrès Soc. Franç. Parasitol., Dijon, May 15-17, Lampe K, Mätz-Rensing K, Becker T, Schrod A, Brehm K, Gottstein B, Janko C, Kaup FJ: Diagnosis and prophylaxies of alveolar echinococcosis in old world monkeys at the German primate center. Int. Conf. on Dis. of Zoo and Wild Animals, Vienna, May 8-11, Müller N. Chemotherapy of giardiasis: antagonistic function of nitroreductases in (in)activation of nitro drugs metronidazole and nitazoxanide. Annual SSTMP meeting, Basle, October 31, Müller N.: Chemotherapy of giardiasis: function of nitroreductases in the metabolism of nitro drugs metronidazole and nitazoxanide. Drug Design and Drug Development Seminar, Würzburg, Germany, April 11-13, IPB: Jahresbericht 2012_13 / annual report 2012_13

19 Aus-, Fort-, und Weiterbildung Peregrine A, Jenkins EJ, Gesy K, Kerr M, Scott S, Barnes B, Skelding A, Gottstein B: Emergence of Echinococcus multilocularis in dogs in Canada? 24th WAAVP Meeting, Perth, Australia, August 26-29, Porot C, Wang J, Germain S, Seimbille Y, Camporese D, Knapp J, Vuitton DA, Blagosklonov O, Gottstein B: In vitro and in vivo investigations to develop functional imaging by Positron Emission Tomography (PET) for murine and human alveolar echinococcosis. 24th WAAVP Meeting, Perth, Australia, August 26-29, Schaarschmidt D, Gilli U, Gottstein B, Marreros N, Kuhnert P, Daeppen JA, Rosenberg G, Hirt D, Frey CF. Questing Dermacentor reticulatus harbouring Babesia canis DNA and associated with outbreaks of canine babesiosis in the Swiss Midlands.Annual meeting of the Society for Veterinary Epidemiology and Preventive Medicine (SVEPM), Madrid, Spain, May 20-22, Skelding A, Brooks A, Stalker M, Mercer N, de Villa E, Gottstein B, Peregrine AS: Hepatic alveolar echinococcosis in a dog in Ontario, Canada. AAVP Annual Meeting, Chicago, July 20-23, Skelding, A., Brooks, A., Stalker, M., Mercer, N., de Villa, E., Gottstein, B., Peregrine, A.S. (2013) Hepatic alveolar echinococcosis in a dog in Ontario, Canada. In: Proceedings of the 58th Annual Meeting of the American Association of Veterinary Parasitologists, 20th-23rd July 2013, Chicago, Illinois, U.S.A., Abstract #69, page 87. Spiliotis M, Stadelmann B, Gottstein B, Hemphill A: Validation and application of a cellular large-scale drug-screening assay for the identification of novel chemotherapies against human alveolar echinococcosis. Drug Design and Development Seminar, Würzburg, April Wang J, Huber C, Gottstein B: The novel CD4+CD25+ regulatory T cell effector molecule Fibrinogenlike Protein 2 (FGL2) contributes to the control of murine alveolar echinococcosis. 25th Swiss Immunology Meeting, Schloss Wolfsberg, April 2-4, Wang J, Porot C, Germain S, Seimbille Y, Camporese D, Knapp J, Vuitton DA, Blagosklonov O, Gottstein B: In vitro and in vivo investigations to develop functional imaging by Positron Emission Tomography (PET) for murine and human alveolar echinococcosis. Annual Meeting of the SGTP, Basel, October 31, Zysset-Burri D, Müller N, Gottstein B, Schürch N, Wittwer M: Genomic, transcriptomic and proteomic identification of pathogenicity factors from Naegleria fowleri. FLAM July 14-19, AUS-, FORT- UND WEITERBILDUNG AUSBILDUNG Veterinärparasitologie (HS: Herbstsemester; FS: Frühjahrsemester) VETSUISSE - CURRICULUM Gottstein B: Oekologie mit Beispielen aus der Parasitologie. [8 Stunden, 1. Studienjahr]. (FS) Gottstein B, zusammen mit Frey C, Hentrich B, Sager H: Integralunterricht (Vorlesung und Kurs/Praktikum) "Veterinärmedizinische Parasitologie im NOZ ( Nicht-organzentrierter Unterricht ) sowie in verschiedenen Organblöcken gemäss neuem Vetsuisse Studienplan. (HS und FS). Gottstein B, zusammen mit Frey C, Hentrich B und Sager H: "Klinische Parasitologie (Kern- und Mantelunterrichts) gemäss neuem Vetsuisse Studienplan [4. Vetsuisse Studienjahr]. (HS und FS). IPB: Jahresbericht 2012_13 / annual report 2012_113 19

20 Aus-, Fort-, und Weiterbildung Frey C, zusammen mit Hentrich B. und Gottstein B: Parasitologische Fälle. Blockstudium gemäss neuem Vetsuisse Studienplan [5. Vetsuisse Studienjahr]. (HS und FS). AUSBILDUNG - Medizinische Parasitologie (Human- und Veterinärparasitologie) Ganze Veranstaltung (3 ECTS) Müller N, Hemphill A, Stadelmann B, Frey C, Gottstein B: Medizinische Parasitologie und tropische Parasitosen, 3 Stunden/Woche. Für Biologen (Block M9P) und Mediziner (HS). Gottstein B, Müller, N., B. Hentrich, G. Rosenberg: Diagnostische Methoden im Bereich Parasitologie; einwöchiger Blockkurs Teilpensum Müller N: Vorlesung im Rahmen der Zell- und Molekularbiologie, Block M5, 4 Stunden FORT- UND WEITERBILDUNG Veterinär- und Humanparasitologie Für die institutsinterne Fort- und Weiterbildung wurden regelmässig Seminare / Journal-Clubs sowie wöchentliche diagnostische Kolloquien abgehalten. Das Institut hat 2012/13 folgende Weiterbildungsveranstaltungen organisiert und durchgeführt: - Trichinella im Fleisch. Kurs und Praktikum für Tierärzte, Laborpersonal und Fleischkontrolleure. Hentrich B, Gottstein B, Rosenberg G, Bern Gastvorträge: Dr. Kasten Nöckler, BFR Berlin; Dr. Gerd Bünger, Vetlabor, Deutschland Habilitationen, Dissertationen und Diplom/Master-Arbeiten, Abschlüsse im 2012/13 - Christoph Bernasconi, Dr. med. vet. : Supervisors: C. Frey, B. Hentrich, B. Gottstein - Marianne Schneeberger (Masterarbeit, vet.-med.): Production and assessment of diagnostic recombinant SAP-2-antigen from Fasciola hepatica. Supervisors: B. Gottstein, G. Boubaker, M. Spiliotis - Denise Zysset-Burri (PhD-Arbeit): Identification of potential pathogenicity factors of the freeliving amoeba Naegleria fowleri by genomic, transcriptomic and proteomic approaches Supervisor N. Müller, B. Gottstein (IPB); M. Wittwer (Spiez Laboratory) 20 IPB: Jahresbericht 2012_13 / annual report 2012_13