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OIE Collaborating Centre Reports Activities in 2011 Title of Collaborating Centre: Address of Collaborating Centre: Zoonoses in Europe Friedrich-Loeffler-Institut Federal Research Institute for Animal Health Headquarters Südufer 10 17493 Greifswald – Insel Riems GERMANY Tel.: +49(0)38351 71102 Fax: +49(0) 38351 71151 e-mail address: website: [email protected] www.fli.bund.de Name of Director of Institute (Responsible Official): Prof. Dr Dr h.c. Thomas C. Mettenleiter (President of the FLI) Name (including Title and Position) of Head of the Collaborating Centre (formally OIE Contact Point): Prof. Dr Dr h.c. Thomas C. Mettenleiter (President of the FLI) Name (including Title and Position) of writer of this report (if different from above) Dr Jens Schell (Research Coordination) Annual reports of OIE Reference Centres, 2011 1 Zoonoses in Europe Summary of activities specifically related to the mandate of OIE Collaborating Centres 1. Activities as a centre of research, expertise, standardisation and dissemination of techniques within the remit of the mandate given by the OIE Centre of research: The Friedrich-Loeffler-Institut (FLI) is a higher independent Federal authority with the legal task to perform research on infectious diseases of animals. It houses all national reference laboratories (NRL) for notifiable infectious diseases of animals in Germany as well as seven OIE reference laboratories (OIE-RL) of which five concern zoonoses: Avian Influenza, Newcastle Disease, Bovine Herpesvirus-1 Infection, Brucellosis, Chlamydiosis, Glanders, and Rabies. FLI is also the national authority to give market authorization for diagnostic tests for infectious diseases in animals and participates in epidemiological investigations of animal disease outbreaks. FLI is involved in several national and international research projects. It is a leading partner in EPIZONE, a FP6 NoE with 19 partners on epizootic disease diagnosis and control. Furthermore, it is involved in large EU FP7 consortia dealing with swine influenza (FluPig), avian influenza (EMPERIE), or (Re-) Emerging Infectious Diseases (EDENext, ANTIGONE). FLI is a member of the European Network for Imported Viral Diseases (ENIVD) on diagnosis of "imported", rare and emerging viral infections. On the national level it co-ordinates the National Research Platform for Zoonoses. This network bundles the national research activities in the field of zoonotic infectious diseases. Viral Diseases: Crimean-Congo-Haemorrhagic-Fever (CCHF): A CCHFV seroepidemiological study in small and large ruminants in twelve southeastern European states is carried out. For this purpose FLI scientisis visited their counterparts in Albania, Macedonia and Turkey. Samples and techniques were exchanged with Ukraine, Kosovarian and Bulgarian scientists. Filoviruses (Ebola/Marburg): Participation in a ring trial exercise to standardize the diagnostic methodology (qRT-PCR) for filoviruses. Hantaviruses: Development of diagnostic tools for serological and molecular biological detection of hantavirus infections in rodents and other small mammals was continued. Influenza A: Avian Influenza (AI): An OIE Twinning Project with the Egyptian NRL for AI at Giza, Egypt, ceased in 2011. Apart from training activities several research papers were compiled in the frame of this project. Research focussed on problems with vaccination caused by the endemic situation of HPAI in Egypt. A close-meshed surveillance of H5N1 HPAIV is mandatory for an early warning of the rise of antigenic drift variants. The presence of such variants requires adaptation of the vaccine. Vaccination needs to be continued to suppress the amount of circulating virus at the human animal interface. However, human infections in Egypt continue to be registered. Details are available from the final twinning project report. Pandemic Influenza A/H1N1pdm: In the course of a short term project passive surveillance of influenza virus infections in populations of domestic swine in the Northwest of Germany was conducted. According to reports from the medical sister institute of the FLI in Germany, the Robert-Koch Institute, three human infections with swine-derived influenza viruses occurred in 2011. Direct contact between infected swine and patients was confirmed in all cases. Results are shared with other European partners in the FP7 ESNIP3 project. New orthobunyavirus in cattle (“Schmallenberg virus”): Analysis at the FLI identified a novel orthobunyavirus of the Simbu serogroup, the “Schmallenberg virus” in cattle and sheep. A newly developed real-time RT-PCR was transfered to the laboratories in other European countries. There is severe concern about the congenital defects Schmallenberg virus induced in newborn lambs, goats and calves. Although some members of the Simbu serogroup, e.g. Oropouche virus, are zoonotic, the risk for humans is assessed to be very low or negligible, as the Schmallenberg virus is closely related to Shamonda virus and so far no clinical signs in humans have been reported. Nevertheless, clinical and serological surveillance in humans should be performed in the regions with positive animals to update the risk assessments. Nipah / Hendra: The molecular (qRT-PCR) and serological (ELISA) assays for the diagnosis of Henipa virus infections were established Orthopoxviruses: In 2011 further cowpox virus (CPXV) strains from different animal species were isolated and characterized. 2 Annual reports of OIE Reference Centres, 2011 Zoonoses in Europe Rabies: Participation in the annual proficiency testing regarding quality assessment of rabies serological testing for dogs and cats within the frame of the EU pet travel scheme as well as in inter-laboratory comparison test on FAT, RTCIT and RT-PCR and on the detection of tetracycline (as a biomarker) for monitoring of ORV campaigns as organized by the CRL for Rabies. Organisation of an international analytical and diagnostic ring trial for lyssavirus (RABV, EBLV-1, -2) specific real-time RT-PCRs, in which 16 European laboratories including two OIE-RL for Rabies participated. A collaborative study on quality issues of anti-rabies conjugates for use in FAT and an international co-operation on harmonising methods for quality control of vaccines currently used in ORV campaigns in Europe was joined. We also have continued to establish closer links with other OIE-RLs and WHO CCs for rabies. A novel diagnostic qRT-PCR assay for the detection of Bokeloh Bat Lyssavirus (BBLV) was developed and validated. Rift Valley fever (RVF): In the frame of Arbozoonet project (FP7) intense collaborations are ongoing with the OIE-RL for CCHF and RVF at the Pasteur institute, Paris as well as NICD Johannesburg, South Africa. FLI developed monoclonal antibodies to N- as well as G1 and G2 proteins and a G1-based ELISA. This ELISA was validated on almost 2000 ruminant sera from SA. Viral Equine Encephalitis (VEEV, WEEV, EEEV): Organization of a ring trial to standardize the diagnostic methodology (qRT-PCR). Participants were EU NRL in other EU member states (UK, B, F etc.). A collaboration is ongoing with Aphis, Ames, USA (OIE-RL) to develop and validate novel ELISAs using recombinant instead of full virus antigens. West Nile Fever (WNF): Serological (ELISA, immunofluorescence assays, PRNT, micro-neutralisation test) and molecular biological methods (different quantitative realtime-PCR) were further validated for the detection of West-Nile-Virus (WNV) infections in animals. Collaborations with the NRLs of France, Austria, Cyprus, Poland, Switzerland and Romania are ongoing. New Realtime PCR protocols for WNV were developed and evaluated. A vivid exchange of techniques and information to evaluate WNV diagnostic assays exists with the newly established EU-CRL for equine diseases, ANSES, Paris, France. Molecular and serological techniques, viruses and samples (serum, paraffin blocks) were supplied by/to the NRLs of Austria and Poland. An exchange of serum samples for the determination of WNV antibody titres with the NRL in Romania took place and interactions are growing. Likewise are connections to the Greek NRL for WNV. Bacterial Diseases: Bovine tuberculosis/Paratuberculosis: Ongoing research projects “M. bovis in the wildlife-livestock-human interface in East and Southern Africa (DFG)” and “Integrated control of neglected zoonoses (EU FP7; ICONZ)”. Participation in a conference organized by the CRL for Bovine Tuberculosis to initiate harmonization of diagnostic methods in Europe. Brucellosis: A diagnostic multiplex PCR microarray assay to detect and differentiate Brucella spp. was developed and a project is running to evaluate the use of Raman spectroscopy for direct diagnosis of Brucella ssp. DNA positive control material for PCR and reference sera for brucellosis and B. ovis as well as B. ovis, B. canis and Yersinia enterocolitica O9 antigens have been provided to national and international laboratories. Chlamydiosis: Cell cultured samples as well as chromosomal DNA from reference and field strains were provided to OIE-member states. In research and development emphasis was laid on optimisation of existing assays. Protocols and technical expertise of real-time PCR assays and DNA microarray tests have been provided to laboratories in Argentina, Croatia, Hong Kong, Italy, the Netherlands, Switzerland, and Ukraine. Partners from Croatia sent a series of samples for re-examination in order to ensure their accreditation status. Dourine: Camel sera from CVRL, Dubai, UAE, were tested for the presence of antibodies for confirmatory purpose and the results were transmitted. Glanders: A new Westernblot assay showing the same sensitivity but higher specificity than the CFT was evaluated with field sera. A project is running to evaluate the use of Raman spectroscopy for direct diagnosis of B. mallei in samples. B.mallei - positive and negative control serum; DNA and LPS from B. mallei and B. pseudomallei strains was prepared and provided to other laboratories. Tularemia: Several hundred sera of red foxes (Vulpes vulpes) and wild boars (Sus scrofa) were screened serologically as sentinel animals for tularemia. Ticks were collected in areas with enzootic tularemia and tested for Francisella tularensis using PCR assays. Q fever: Improvement and standardization of methods for diagnosis of Q fever. At present, FLI scientists are collaborating with institutes from Poland, the Netherlands, France, Moldova, Belarus, and Kenya. Parasites - Echinococcosis: Establishment and publication of a mathematical model for analysing the spatial and temporal distribution of Echinococcus multilocularis. Studies on the spread and behaviour of the raccoon dog (Nyctereutes procyonoides) as a new definitive host of E. multilocularis in eastern and central Europe. Validation Annual reports of OIE Reference Centres, 2011 3 Zoonoses in Europe of molecular techniques for diagnosis and for the differentiation of Echinococcus spp. samples and for the distinction of Echinococcus spp. from Taenia spp. eggs. Prions - TSE: Ongoing close collaborations with NRLs of other European countries. BSE or scrapie positive samples and reference materials (e.g. fresh brainstem material or paraffin-embedded fixed tissue) have been supplied. Special emphasis was put on the collaboration with the Polish NRL. Moreover, FLI provided ring trial samples to the Canadian NRL to support their quality control system. 2. Proposal or development of any procedure that will facilitate harmonisation of international regulations applicable to the surveillance and control of animal diseases, food safety or animal welfare Viral Diseases: Rabies: Contribution to the “Blueprint for Rabies Prevention and Control (http://www.rabiesblueprint.com/)” The OIE-RL leads a core group of the Partners for Rabies Prevention (PRP) dedicated to develop a Blueprint for Red Fox Rabies Prevention. TBE: Participation in WHO meetings to discuss and approve the WHO Position Paper and the WHO Background Paper: “Vaccines against tick-borne encephalitis”. Bacterial Diseases: Brucellosis: Participation in the EU Brucellosis Bovine Milk iELISA Proficiency Ring-trial and the CFT harmonisation (organized by CRL for Brucellosis). FLI also participated in the standardisation of Fluorescence Polarisation Assay (FPA) (organized by OIE-RL Argentina) Chlamydiosis: The introduction of an alternative diagnostic reference test for chlamydial infections to replace cell culture is proposed. This procedure includes a validated real-time PCR detection assay in conjunction with the ArrayTube® microarray test. The proposal is currently being discussed among the chlamydia research community. Glanders: The laboratory participated in the glanders specific international ring trials VETQAS CFT (organized by VLA Weybridge) and ANSES Glanders ring trial (organized by CRL for Glanders). Furthermore it participated in the ring trial detection of highly pathogenic agents including B.mallei/pseudomallei (organized by RKI, Germany, within the EU-project EQADeBa). Parasites - Echinococcosis: Advising the EU-CRL for Parasitoses in Italy, Rome, in designing proficiency trials on E. multilocularis at the international level. Prions - TSE: FLI collaborates closely with the EU-CRL for TSEs in Weybridge, UK and other NRLs to conduct an EU-wide batch control system for rapid tests for the mass screening of ruminant samples for BSE and scrapie. The FLI is responsible for the EU wide batch control of the TeSeE Test (BioRad). 3. Networking a) Maintenance of a network with other OIE Collaborating Centres designated for the same specialty, and FLI is closely linked with NCFAD Winnipeg, AAHL Geelong, ARS Ames, HPA Salisbury, IAH Pirbright, IVI Mittelhäusern, Laboratoire P4 Inserm Jean Mérieux Lyon, the Plum Island Animal Disease Center and the BSL4-Laboratory in Wuhan (China). In the frame of NADIR and ERINHA, FLI collaborates with the major European research institutes that are running BSL-3 and BSL-3+ (NADIR) and BSL4 (ERINHA) infrastructures. b) Should the need arise, maintenance of a network with Collaborating Centres in other disciplines – 4. Placement of expert consultants at the disposal of the OIE The President of the FLI is a member of OIE’s Scientific Commission for Animal Diseases 4 Annual reports of OIE Reference Centres, 2011 Zoonoses in Europe The FLI contributed to the Technical Consultation in Support of the OIE Emerging Pandemic Threats Program activities in Paris, France on 25 - 27 January 2011 Viral Diseases: Crimean-Congo-Haemorrhagic-Fever (CCHF): Invited expert on CCHF surveillance at the 1st Veterinary Forum for South East Europe, 3-4 February 2011, Budapest. Rabies: Consultant expertise was provided at (i) the Partners for Rabies Prevention Meetings in Italy and Puerto Rico, (ii) workshop on Rabies and Oral Vaccination Monitoring in Kosovo, (iii) the 10th SEARG Meeting in Mozambique, (iv) the EU task force meeting on assessment of ORV programmes in Bulgaria, and (v) the extraordinary meeting in relation with the rabies situation in the common border area between Bulgaria, Greece and the Former Yugoslav Republic of Macedonia, in Bulgaria. Rift Valley Fever (RVF): Invited expert at the ad hoc Workshop "Rift Valley Fever vaccine development; progress & constraints" - FAO headquarters Rome, 19-21 January 2011 Bacterial Diseases: Brucellosis: Contribution to the brucellosis chapters of the Terrestrial Manual. Consultancy and support for setting-up two specialized brucellosis laboratories in the United Arab Emirates and initiation of laboratory twinning. The Twinning project with Kosovo to strengthen control on bacterial zoonoses was successfully completed. Chlamydiosis: Update of the OIE Manual's chapter 2.3.1. Avian Chlamydiosis and review of chapter 2.7.7. Enzootic Abortion of Ewes. A fact sheet on enzootic abortion of ewes was prepared within the European DISCONTOOLS project. Glanders: Consultant expertise in handling the latest outbreaks and for eradication and control of the disease was given to Kuwait and Lebanon on request. Further expertise was provided at the Glanders Workshop in Recife, Brazil and at the conference of the CRL in Paris, France. Parasites - Echinococcosis: Contribution to a new edition of the WHO/OIE Manual on Echinococcosis. Prions - TSE: Contribution to the revisions of the BSE chapter 11.6., the scrapie chapter 14.9., and appendix 3.8.5 of the OIE Terrestrial Animal Health Standards Commission. 5. Provision of scientific and technical training, within the remit of the mandate given by the OIE, to personnel from OIE Member Countries An African “Joint Master’s Programme in Transboundary Animal Disease Management” (MTADM) project was initiated to build human resource capacity by training an effective pool of professionals in cross-border animal disease control and management and by strengthening the regional network of veterinary faculties. The course was organized by Addis Ababa University in Ethiopia and the Freie Universität Berlin with strong input and later endorsement by other partner universities in Uganda and Sudan. Part of the curriculum was held at Freie Universität Berlin with seminars given also by guest lecturers including FLI-staff. Viral Diseases: Avian Influenza: Training on classical and molecular diagnostic measures was provided to colleagues from Egypt. Crimean-Congo-Haemorrhagic-Fever (CCHF): One PhD student from Albania was trained at FLI in molecular and serological diagnostic techniques for CCHF in animals. Hantavirus: A student from the Ankara University was trained in serological and molecular diagnostic methods for hantavirus detection. Rabies: Colleagues from other OIE member countries including one person from Iran and one from South Africa were trained on basic epidemiology, surveillance and standard rabies diagnostic tests in Germany. A PhD thesis of a Latvian colleague on evaluation of ORV campaigns in Latvia and a master thesis of a student from the Addis Ababa University, Ethiopia was supervised. Bacterial Diseases: Bovine tuberculosis/Paratuberculosis: Russian scientists were trained in diagnostic methods and field control methods of bovine tuberculosis and paratuberculosis of ruminants. Laboratory training of a group of Lithuanian scientists in laboratory diagnostic methods of bovine tuberculosis. Annual reports of OIE Reference Centres, 2011 5 Zoonoses in Europe Brucellosis: Four colleagues from UAE were trained in serological methods for brucellosis diagnosis and one colleague from Oman was trained in diagnostic techniques regarding brucellosis. In Kosovo, scientist and technicians were trained in brucellosis serology and PCR. Chlamydiosis: Researchers and lab diagnosticians from Ukraine, Switzerland and Peru visited the FLI for technical training and consultations on chlamydial diagnostic methods, with emphasis on molecular detection methods. Glanders: A PhD student from the University of Veterinary & Animal Science, Lahore, Pakistan evaluated the available serological methods and commercially available antigens for glanders and compared their usefullness in surveillance and control of glanders in endemic and non-endemic regions. Scientists from Lebanon, Ukraine and Hungary were trained in detection methods for glanders. On hand support was provided to the VLA Weybridge, UK in establishing the immunoblot for glanders in their laboratories. Q fever: Training in diagnostic methods for the detection of Q fever has been performed for a colleague from Moldova. Parasites - Echinococcosis: Consultation with scientists from Sudan to set up studies to assess the epidemiological situation regarding Echinococcus granulosus s.s. in the respective countries. A PhD student from Sudan was trained in study design and echinococcosis diagnosis. 6. Organisation of scientific meetings on behalf of the OIE OIE Global conference on Rabies control: Towards Sustainable Prevention at the Source Incheon, Korea, September 09-11, 2011 (Scientific committee member) 7. Coordination of scientific and technical studies in collaboration with other laboratories, organisations or collaborating centres Viral Diseases: Crimean-Congo-Haemorrhagic-Fever (CCHF): Study on the prevalence and spread of the CCHF virus and its main vector, the Hyalomma marginatum tick in Southeastern Europe is co-ordinated in the frame of EDENext project.Hantavirus: Coordination of the research network “rodent-borne pathogens”. Nipah / Hendra: Collaboration with scientists of AAHL Geelong. Rabies: In vitro studies of plant derived murine MAbs for the WHO coordinated research project on research and development on a murine monoclonal antibody (MAbs) cocktail to replace HRIG and ERIG for rabies postexposure prophylaxis (PEP) in humans were conducted. The working group of the Partners for Rabies Prevention dedicated to develop a “Blueprint for Red Fox Rabies Prevention” was lead. The Blueprint is expected to be approved in May 2012. Rift Valley fever (RVF): Collaborations are ongoing with the OIE-RL for CCHF and RVF at the Pasteur institute, Paris as well as NICD Johannesburg, South Africa. Viral Equine Encephalitis (VEEV, WEEV, EEEV): A collaboration is ongoing with Aphis, Ames, USA (OIERL) which aims at the development and validation of novel ELISAs using recombinant instead of full virus antigens. West Nile Fever (WNF): Close links exist to the EU-CRL for equine diseases at ANSES, Paris, France. Bacterial Diseases: Bovine tuberculosis/Paratuberculosis: Novel specific protein antigens of Mycobacterium avium ssp. paratuberculosis were tested in an experimental infection study on goats coordinated by the FLI. Collaborators are CVI Lelystad, The Netherlands and the Moredun Research Institute, Edinburgh, Scotland. Brucellosis: Involvement in the validation procedure for the Brucella suis International Standard Serum. A publication is in preparation. Glanders: The cooperation with laboratories at the University Lahore, Pakistan, Laboratório Nacional Agropecuário, Ministério Da Agricultura, Recife, Brazil, CVRL Dubai, UAE, and VLA Weybridge, UK was continued. 6 Annual reports of OIE Reference Centres, 2011 Zoonoses in Europe Q fever: Collaboration with the Academy of Science of Moldova, Institute of Zoology, Chisinau, Republic of Moldova: “Prevalence of Borrelia burgdorferi sensu lato and Coxiella burnetti in ticks collected in the Republic of Moldova and Germany” (supported by the German Ministry of Science and Research [BMBF]) Prions - TSE: Close collaborations with the NRLs of other European countries as well as the Canadian NRL/OIE-RL are ongoing. A ring trial was conducted for the Canadian laboratory in order to verify their diagnostic proficiency and quality control system. 8. Publication and dissemination of any information within the remit of the mandate given by the OIE that may be useful to Member Countries of the OIE The FLI is responsible for editing the Rabies Bulletin Europe (RBE), in which numbers of reported rabies cases and rabies surveillance data from all European countries are collected and published. In 2011, the issues III and IV of 2010 as well as I and II of 2011 were edited and distributed throughout the world. Data are free to download and available from our website (http://www.rbe.fli.bund.de/). In 2011 about 400 peer-reviewed papers were published by the FLI. Furthermore scientists presented their research results at numerous national and international scientific conferences and advice was given to national and European governments. Selected publications in 2011: Freuling, C., Beer, M., Conraths, F. J., Finke, S., Hoffmann, B., Keller, B., Kliemt, J., Mettenleiter, T. C., Mühlbach, E., Teifke, J. P., Wohlsein, P., and Müller, T. (2011) Novel lyssavirus in Natterer's bat, Germany. Emerging Infectious Diseases 17:1519-1522. Haybaeck, J., Heikenwalder, M., Klevenz, B., Schwarz, P., Margalith, I., Bridel, C., Mertz, K., Zirdum, E., Petsch, B., Fuchs, T. J., Stitz, L., and Aguzzi, A. (2011) Aerosols Transmit Prions to Immunocompetent and Immunodeficient Mice. PLoS Pathogens 7:e1001257. Johnson, N., Freuling, C., Horton, D., Müller, T., and Fooks, A. R. (2011) Imported Rabies, European Union and Switzerland, 2001-2010. Emerging Infectious Diseases 17:753-754. Müller, T., Hahn, E. C., Tottewitz, F., Kramer, M., Klupp, B. G., Mettenleiter, T. C., and Freuling, C. (12-82011) Pseudorabies virus in wild swine: a global perspective. Archives of Virology 156:1691-1705. Reinhold, P., Sachse, K., and Kaltenboeck, B. (2011) Chlamydiaceae in cattle: Commensals, trigger organisms, or pathogens? Veterinary Journal 189:257-267. Riehm, J. M., Tserennorov, D., Kiefer, D., Stuermer, I. W., Tomaso, H., Zöller, L., Otgonbaatar, D., and Scholz, H. C. (2011) Yersinia pestis in Small Rodents, Mongolia. Emerging Infectious Diseases 17:1320-1322. Vos, A., Nunan, C., Bolles, D., Müller, T., Fooks, A. R., Tordo, N., and Baer, G. M. (2011) The occurrence of rabies in pre-Columbian Central America: An historical search. Epidemiology and Infection 139:1445-1452. _______________ Annual reports of OIE Reference Centres, 2011 7