Zoonoses in Europe
Activities in 2009
Zoonoses in Europe
Prof. Dr Thomas C. Mettenleiter
Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health
Headquarters, Südufer 10, 17493 Greifswald – Isle of Riems, GERMANY
Tel.: (+49-38351) 71.02, Fax: (+49-38351) 71.51
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 for notifiable infectious diseases of animals. It is also the national authority to give market authorization for tests for diagnosis of infectious diseases in animals and participates in epidemiological investigations of animal disease outbreaks
The FLI is a leading member of the network of excellence 'Epizone' funded by the European Union which brings together 20 institutes to coordinate research on epizootic diseases of animals. On the national level it coordinates together with the university of Münster and the Telematikplattform für Medizinische Forschungsnetze e.V. (TMF) the National Research Platform for Zoonoses. This network bundles the research activities in the field of zoonotic infectious diseases. Development of flexible and sustainable solutions to strengthen research, prevention, and therapy of zoonotic infectious diseases are its main objectives.
Bovine tuberculosis/Paratuberculosis: Research project (Deutsche Forschungsgemeinschaft [German Research Foundation] TA 668/1-1): Mycobacterium bovis in the wildlife-livestock-human interface in East and Southern Africa. African network building for the control of neglected zoonoses.Research project (EU FP7 – 221948; ICONZ): Integrated control of neglected zoonoses: improving human health and animal production through scientific innovation and public engagement.
Participation in the final meeting of the EU Coordination Action VENoMYC (Veterinary Network of Laboratories Researching into Improved Diagnosis and Epidemiology of Mycobacterial Diseases) in Turin, 17th-20th June, 2009. Summing up of the workshop “Laboratory diagnosis of Mycobacterium spp.”.Participation in the meeting of the EU Coordination Action VENoMYC (Veterinary Network of Laboratories Researching into Improved Diagnosis and Epidemiology of Mycobacterial Diseases) in Madrid, 23rd – 25th March, 2009. VNTR/MIRUs and DVR spoligotyping for M. bovis typing - Results of the VNTR-DVR Spoligotyping Ring Trial”.
Participation in the meeting of the EU Community Reference Laboratory for Bovine Tuberculosis in Madrid, 5th – 6th November, 2009
Brucellosis:A second international ring trial for brucella genotyping with Multilocus Variable Number of Tandem Repeat Analysis (MLVA) was organized by our lab together with Italian colleagues (Istituto Zooprofilattico Sperimentale "G. Caporale", Teramo). 12 DNA samples were sent out to 15 laboratories in 11 countries and results from 12 labs were received.
Chlamydiae:The Reference Laboratory for Chlamydiosis initiated the drafting of a review article on recent advances in the laboratory diagnosis of chlamydia infections, which was published in 2009 (Sachse et al. Vet. Microbiol. 135, 2-21; see list of publications). This paper contains numerous recommendations and comments, all of which represent the common position of the leading European researchers in the field of veterinary chlamydiology
Protocols and technical expertise of real-time PCR detection and DNA microarray tests have been provided to laboratories in the Netherlands, Switzerland, France, Ireland, Sweden and Argentina. These methods are being used by diagnostic labs in a number of countries, and some of them have the potential to become standard procedures.
Echinococcosis:Identification of the red-necked wallaby (Macropus rufogriseus) as an intermediate host of Echinococcus multilocularis. Studies on the spread and behaviour of the raccoon dog (Nyctereutes procyonoides) as a new definitive host of Echinococcus multilocularis in eastern and central Europe. After accreditation of the laboratory according to ISO 17025:2005, the laboratory validated molecular techniques for diagnosis and for the differentiation of Echinococcus spp.samples and for the distinction of Echinococcus spp. from Taenia spp. eggs. Contribution to a new edition of the WHO/OIE Manual on Echinococcosis.
Glanders: A protocol of the Western Blot technique for the serological diagnostic was evaluated and compared with other diagnostic techniques.
Hantavirus: FLI continued the development of diagnostic tools for serological and molecular biological detection of hantavirus infections in rodents and other small mammals. These techniques were developed in close collaboration with the Bundeswehr Institute of Microbiology, Munich, the Bernhard-Nocht-Institute for Tropical Medicine, Hamburg, the Institute of Virology, Charité, Berlin and the Institute of Biotechnology in Vilnius, Lithuania.
Influenza: Avian Influenza: Research into avian influenza virology in 2009 focussed on molecular tools enhancing diagnostic capabilities. Studies on the production of recombinant hemagglutinin antigen tailored for use in subtype-specific ELISAs have been successfully finished. Promising results have also been obtained for a genetically modified porcine cell line suitable to promote the replication of a wide variety of avian influenza viruses of low pathogenicity independent of supplementation of the culture medium with trypsin. In addition, commercial kits for real-time RT LAMP analysis of subtype H5 and H7 viruses have been evaluated; while the LAMP technology offers advantages particularly for on-site use, the currently available kits were found significantly less sensitive than the gold standard method, real-time RT PCR.
An OIE twinning project between the National Laboratory for Veterinary Quality Control on Poultry Production (NLQP), Institute for Animal Health Research (ARI), Cairo, Egypt, and the Friedrich-Loeffler-Institute (FLI), Isle of Riems, Germany continued. Ongoing collaborations within OFFLU, the OIE/FAO network of expertise on Avian influenza.
Influenza:New Influenza A/H1N1 “Swineflu”: Research projects in 2009 focussed on disease pathogenesis and transmissibility studies in pigs. Current activities evaluate commercially available vaccines against influenza virus infections in pigs for their potential to protect from virus infection, clinical signs, and/or virus shedding.
Newcastle Disease: Research was focussed on NDV in pigeons. Beside molecular characterization and pathotyping of currently circulating NDV isolates from pigeons a study on prevalence of Avian Paramyxovirus in wood pigeons (Columba palumbus) was conducted. In addition investigations on immune response of turkeys after NDV-vaccination were initiated.
The OIE twinning project between the National Laboratory for Veterinary Quality Control on Poultry Production (NLQP), Institute for Animal Health Research (ARI), Cairo, Egypt, and the Friedrich-Loeffler-Institute (FLI), Isle of Riems, Germany was ongoing. The program is focused on collaboration for AI and ND diagnosis. This year a total of seven scientists visited the laboratory for training purposes.Besides national activities for federal agencies the Laboratory provided assistance for NDV – diagnostic for laboratories from the United Emirates and Ethiopia.
Orthopoxviruses:In 2009, several human infection clusters with cowpox virus (CPXV), transmitted by so-called pet rats, occurred in Germany and France. Therefore, in a series of animal experiments, pet rats, Wistar rats, and cattle were infected with a CPXV strain isolated from an infected pet rat. Reference materials from the infected animals and pathogenesis data were collected. Infection of Wistar rats was established as a suitable infection model for the recent CPXV.
Rabies: 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 quarterly. In 2009, the issues 03/2008, 04/2008, 1/2009 and 02/2009 were edited and distributed throughout the world. Data are free to download and available from our website (
Salmonellosis in cattle:Molecular and epidemiological analysis of SalmonellaDublin infections in cattle in Germany. Epidemiological analysis of bovine salmonellosis according to the data in National Animal Disease Reporting System in Germany.
TSE: The National TSE Reference Laboratory at the FLI has been in close collaboration with the NRLs of the other European countries, especially with those of EU members. This is reflected in numerous EU-funded research projects that have been joined during the last year(s). In the context of these activities FLI has supplied other TSE NRLs and research groups with several dozens of BSE or scrapie positive samples and reference materials (e.g. fresh brainstem material or paraffin-embedded fixed tissue). A special emphasis was put on the collaboration with the Polish NRL regarding the biochemical characterization of the first notified atypical scrapie cases in Poland. Moreover, the German NRL has been in close contact with the Canadian NRL over the last years and has supplied this laboratory with ring trial samples in order to support their quality control system.
Tularemia:The National Tularemia Reference Laboratory at the FLI has been in contact with the NRLs of France and Switzerland as well as other laboratories in Austria, China, Poland and Spain that are involved in the detection of biological agents. An exchange of strains, techniques and information to evaluate diagnostic and typing tools took place.
West Nile Fever:The FLI has developed and validated serological (ELISA, Immunofluorescence-assays, micro-neutralisation test) and molecularbiological methods (different quantitative realtime-PCR) for detection of West-Nile-Virus (WNV) infections in animals. These techniques were developed in close collaboration with the Robert-Koch-Institute in Berlin, the Bernhard-Nocht-Institute for Tropical Medicine in Hamburg and the Paul-Ehrlich-Institute in Langen.
The National West-Nile-Virus Reference Laboratory at the FLI has been in contact with the NRLs of France, Austria,Cyprus, Poland, Switzerland and Romania. Realtime PCR protocols for WNV which were developed by FLI and evaluated against published methods were supplied to the Swiss WNV NRL in Bern. With the newly established EU-CRL for equine diseases, AFSSA, Paris, France a vivid exchange of techniques and information to evaluate a sensitive WNV diagnostic is has been continued. With the NRL in Austria and Poland we exchanged PCR-protocols, requirements for the safety level 3 laboratories and exchange of information for a better monitoring study to use as an early-warning system. Furthermore we carry out an intensive advisory service for the Central Veterinary Service of Cyprus. First efforts for an exchange of serum samples with WNV-antibodies and virus isolates with the arbovirus NRL in Romania were undertaken.
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
Bovine tuberculosis/Paratuberculosis: Development of a novel probe based two-target real-time PCR system for the detection of DNA of members of the M. tuberculosis complex (MTC), including M. bovis and M. caprae, in tissue samples from cattle and transfer of the protocol to the regional diagnostic laboratories in Germany.
Chlamydiae: RL Chlamydiosis proposes the introduction of an alternative diagnostic reference test for chlamydial infections to replace cell culture. 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.
Echinococcosis:Proficiency testing was prepared to compare the performance of different diagnostic techniques in various laboratories at the international level.
Glanders: In the frame of an EU-Project, the laboratory took part in two international ring trials for antigen detection of highly infectious agents, including B. mallei and B. pseudomallei together with 21 other European laboratories. Furthermore, the laboratory took part in the Glanders CFT QAU Proficiency Testing organized by the VLA, Weybridge.
Influenza:New Influenza A/H1N1 “Swineflu”:Animal experiments also investigated viral dissemination in the infected organism in order to verify food safety.
Q fever: The EFSA project "Development of harmonised schemes for monitoring and reporting of rabies and Q fever in animals in the European Union" has been continued.
Rabies: The FLI together with other 4 European rabies laboratories participated in a project funded by the European Food Safety Authority (EFSA) on the development of harmonised schemes for monitoring and reporting of rabies and Q fever in animals in the European Union. Within this project currently existing recommendations on rabies surveillance of international organisations were reconsidered and based on scientific data the introduction of a situation-based rabies surveillance both for terrestrial (classical) rabies as well as bat rabies suggested.
Salmonellosis in cattle:Further studies on improving the method for the bacterial detection of salmonellae according to the national German “Regulation on protection against salmonellosis in cattle” and comparing with the method according to ISO 6579 Annex D. As result of these activities, the existing method for bacterial detection of salmonellae according to the national German “Regulation on protection against salmonellosis in cattle” was replaced by method according to ISO 6579 Annex D.
TSE: In close collaboration with the Community Reference Laboratory for TSEs in Weybridge, UK and other NRLs, FLI scientists were involved in the establishment of an EU-wide batch control system for rapid tests for the mass screening of ruminant samples for BSE and scrapie. A major progress has been achieved in this goal, as an EU-wide batch control system has already been established for four different BSE rapid tests (IDEXX HerdChek Test, PrioSTRIP Test, Prionics Western Test, BetaPrion BSE EIA Test Kit / ajRoboscreen). The German NRL is responsible for the EU wide batch control for the TeSeE Test (BioRad), which is currently under preparation and the start of the EU wide batch control for this test is anticipated for the first quarter of 2010. Moreover, FLI has contributed to the revisions of the BSE chapter 11.6., the scrapie chapter 14.9., and appendix3.8.5 of the OIE Terrestrial Animal Health Standards Commission.
Tularemia:In order to obtain an overview of the prevalence of tularemia in Europe we propose that sentinel animals (e.g. wild boars) that are tested for other diseases on a routinely basis should be screened serologically. Serological tests for this purpose need to be evaluated and standardized.
3.Placement of expert consultants at the disposal of the OIE
Influenza:Avian Influenza: Members of the OIE Reference Laboratory for Avian Influenza at the FLI contributed to three OFFLU technical groups. Expertise on the availability of commercial AI-related diagnostics, the production and dissemination of PCR standard controls, and on use of AI vaccines in Indonesia is continuously provided. In addition, Dr. Harder represented OFFLU at a meeting of the WHO Technical Advisory Group on molecular influenza diagnosis.
Influenza:New Influenza A/H1N1 “Swineflu”: Dr. Vahlenkamp participated as expert in three meetings organized by the EU discussing and approving the ‘Working document on surveillance, monitoring and control measures for the pandemic (H1N1) 2009 influenza virus in pigs and in poultry’.
Newcastle Disease: Revison of the O.I.E. Chapter 2.3.14. NEWCASTLE DISEASE (Editor Claudio L Afonso)
Rabies:Expert opinion on the evaluation of the Rabies Platelia II ELISA test in the frame of the pet travel scheme
Other activities related to the mandate
of OIE Collaborating Centres
4.Provision of scientific and technical training, within the remit of the mandate given by the OIE, to personnel from OIE Members
Bovine tuberculosis/Paratuberculosis:Dr. Sodnomdarjaa, Director of the State Central Veterinary Laboratory, Mongolia, stayed in the FLI for consultation about diagnosis of bovine tuberculosis and paratuberculosis.
Brucellosis:One responsible veterinarian from the NRL Lithuania was trained for one week in serological methods. Two scientists from University of Veterinary & Animal Science, Lahore, Pakistan and from the NationalResearchCenter, Cairo, Egypt underwent a three-month training on laboratory diagnosis of brucellosis.
Chlamydiae:In 2009, researchers and lab diagnosticians from France, Italy, Belgium and Sweden visited the reference lab for technical training and consultations on chlamydial diagnostic methods, with emphasis on molecular detection methods.
Glanders: One PHD student from University of Veterinary & Animal Science, Lahore, Pakistan started the work in the field of “Molecular Seroepidemiology of Glanders”. Veterinarians from Brazil, Poland, Spain and UK stayed each for a two-weeks training on laboratory diagnosis of glanders.
Hantavirus: In the frame of the network “Rodent-borne pathogens” FLI has a close collaboration with a large number of groups in Germany. These collaborations are mainly driven by the exchange of materials for the surveillance of infections with hantaviruses and other zoonotic pathogens such as Ljungan virus, orthopoxviruses, tick-borne encephalitis virus, Borrelia spp., Coxiella burnetii, Leptospira spp., E. coli and other bacterial pathogens, and endoparasites. Serological and molecular biological studies on the orthopox virus prevalence in wild rodents from Germany were performed in a close collaboration with the Haartman Institute, Helsinki, Finland. For training purposes young scientists from other laboratories in Germany were involved in the hantavirus investigations.
Influenza: Avian Influenza: Within the OIE twinning project extensive training visits of staff have been organized. Training on the job has been provided at the FLI for seven colleagues from the NLQP. Further training was provided for colleagues from Bangladesh and Russia.
Newcastle Disease: A one year training program for Dr. Fufa Dawo Bari, a scientist from the university of Adis Abeba, Ethiopia was ongoing. He left the Laboratory on September 5th 2009. Issues of training comprised molecular and classical techniques of NDV diagnosis. During the second half of the training the focus was on phylogenic analysis of NDV-isolates.As part of the OIE twinning project with the NLQP, Egypt scientists were trained on classical diagnostic methods (2 week program), on production of reference materials (4 week program) and pathogenicity testing (4 weeks program).
Q fever: A training in diagnostic methods for the detection of Q fever was performed for colleagues from Spain and Poland.
Rabies:Training of scientific and technical staff: In 2009 we hosted a number of colleagues from other OIE member countries including one person from Lithuania, two persons from Ukraine, two persons from Turkey, two persons from Mongolia. The training included all standard rabies diagnostic tests, e.g. rabies diagnosis, rabies serology, follow-up investigations of oral vaccination campaigns and virus characterization (virus typing, sequencing, rabies surveillance, evaluation of ORV campaigns).