Zoonoses in Europe
OIE Collaborating Centre Reports
Activities in 2013
Title of Collaborating Centre: / Zoonoses in EuropeAddress of Collaborating Centre: / 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: / 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)
Summary of activities specifically related to the mandate of
OIE Collaborating Centres
ToR: To provide services to the OIE, in particular within the region, in the designated specialty, in support of the implementation of OIE policies and, where required, seek for collaboration with OIE Reference Laboratories
ToR: To identify and maintain existing expertise, in particular within its region
1. Activities as a centre of research, expertise, standardisation and dissemination of techniques within the remit of the mandate given by the OIE
Disease control
Epidemiology, surveillance, risk assessment, modelling
Training, capacity building
Zoonoses
Wildlife
Avian diseases
Aquatic animal diseases
Animal welfare
Diagnosis, biotechnology and laboratory
Veterinary medicinal products
Vaccines
Food safety
Feed safety
Other (Name the category: )
Remark: FLI’s main area of expertise regarding the OIE mandate is “Zoonoses”, however expertise exists also concerning the other marked areas.
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 runs a Collaborating Centre of the WHO for rabies. Furthermore it is designated as a reference centre of the UN Food and Agriculture Organization (FAO) for influenza in animals and Newcastle disease as well as classical swine fever. 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.
Zoonooses (Viral Infections)Title of activity / Scope
Animal Influenza
incl. A/H1N1pdm: / Passive surveillance of influenza virus infections in populations of domestic swine in the Northwest of Germany revealed cocirculation of at least for stable lineages including a reassortant between A/H1N1pdm and enzootic porcine influenza viruses (H1pdmN2). Increasing antigenic diversity among circulating strains was evident (Harder et al., 2013).
Avian Influenza (AI): / Research on new vaccines targeting the highly pathogenic avian influenza virus (HPAIV) epizootic in Egypt have been taken up. Work of further characterization of recent circulating HPAIV H5N1 and H9N2 AIV strains from Egypt is ongoing.
Pathogenesis studies in chickens, pigeons and ferrets with Chinese origin A/H7N9 virus have been conducted in comparison to low pathogenic European origin H7N7 AIV indicating that also European H7 LP viruses harbour some potential to infect mammalian species.
Crimean-Congo-Haemorrhagic-Fever (CCHF) / Recombinant N-Proteins of CCHFV strains coming from European, Asian and African origin were produced and diagnostic assays for the detection of specific antibodies in small and large ruminants have been developed and validated using reference sera. Comparative studies including the in-house ELISA, the commercial Vectorbest CCHF ELISA, which is based on inactivated full virus antigens, and a commercial immunofluorescence assay have been performed.
Cattle sera from Macedonia and Albania were analysed and antibody prevalences were observed. Further CCHFV antibody testing of sera from additional Balkan countries is foreseen. Moreover, collaboration with partners in Mauritania, Sierra Leone, Cameroon and DR Congo was initiated. A total of 6000 domestic animal sera were received and analysis of these sera has started.
In addition to working on the serological test quantitative RT-PCRSs for the detection of CCHFV-RNA were further optimized.
Japan-Encephalitis: / Immunization of rats and mice with a human vaccine and recombinant glycoprotein (E protein) for generation of monoclonal antibodies. The antibodies will be selected for specific binding to JEV with minimal cross reactivity to viruses from the same serogroup (e.g. West Nile virus, Tick-borne encephalitis virus etc.). Development of ELISA based serological diagnostic systems for differentiation of JEV infection from infections with other Flaviviruses.
Nipah/Hendravirus infections: / The qRT-PCR assays for the diagnosis of Henipa virus infections were validated and can be applied for the analysis of diagnostic samples.
Polyclonal and monoclonal antibodies against functional proteins of the hendra virus were generated. A second validation study of an indirect in-house antibody ELISA based on the N-protein is ongoing using a larger set of samples including sera from pigs experimentally challenged with the Nipah virus.
Orthopoxvirus infections / Several cases of orthopoxvirus infections in animals were diagnosed and 3 different cowpox viruses (CPXV) could be isolated and characterized (2 isolates from Alpacas and 1 isolate from a cat).
More than 2300 samples from wild rodents (voles) were screened for orthopoxvirus-specific DNA.
A method for full-length sequencing of CPXV was established using next generation sequencing. More than 10 different cowpox virus isolates from different animal species were full-length sequenced.
Parapoxvirus infections / Several cases of parapox virus infections (Orf) could be confirmed.
Rift-Valley fever: / In 2013 the monitoring studies from a RVFV outbreak in Mauretania 2010 were finished and showed the involvement especially of small ruminants and camels in RVFV infection and transmission The studies were supported by a novel Gn based indirect ELISA. In addition, a set of 45 monoclonal antibodies against structural proteins of RVFV were generated and characterized. They will be used and provided for several approaches to enable and facilitate antigen detection assays (competitive ELISAs, indirect immunofluorescence, immunohistochemistry). The previous work demonstrates the preparedness for monitoring studies in Europe and will alleviate further studies on vector competence and susceptibility of European lifestock to RVFV in close collaboration with Dr. Noël Tordo (OIE Reference Centre for Viral Haemorrhagic Fevers, Paris).
Schmallenberg virus infection / More than 2000 samples of domestic ruminants and wild life were tested for SBV-antibodies and virus. Semen samples of bulls were screened for SBV-RNA. Selected SBV-strains were full-length sequenced and phylogenetically typed. PCR-systems for the diagnosis of SBV especially in semen samples were developed and validated including 2 international ring trials.
More than 15 000 midges were screened for SBV-RNA.
There are members of the Simbu-serogroup which are known to be zoonotic, however, for SBV it is confirmed that no zoonotic potential exists.
Usutu virus infection / The molecular biological and serological monitoring in wild birds was continued in close collaboration with different German bird clinics. Investigation of dead wild birds in close collaboration with the local state veterinary laboratories and with the Bernhard-Nocht-Institute for Tropical Medicine in Hamburg to detect USUV infected birds.
Viral Equine Encephalitis (VEEV, WEEV, EEEV): / Establishment of hybridoma cell lines kindly provided by J. Roehrig, CDC, and inclusion of produced monoclonal antibodies in the ongoing development of new ELISA based diagnostic systems. Further characterisation of these antibodies and investigation of their ability to bind to recombinant glycoproteins. Production of recombinant alphaviral glycoproteins in eucaryontic cells (BHK-21) for the use of glycosylated proteins in diagnostic sytems
West Nile Fever: / Further development of different diagnostic tools for molecular biological detection (Pan-Flavi-PCR, microarray) of WNV infections in birds and horses. Different micro-neutralisation tests as a method to detect the antibody cross reactivities between the Flavivirus family members were performed. Virological and serological monitoring studies of wild birds in the last year. Differential diagnostic analysis on request of third parties.
Rabies / The OIE-RL leads a core group of the Partners for Rabies Prevention (PRP) dedicated to develop a Blueprint for Bat Rabies Prevention.
The OIE RL participated (i) in the 2013 annual proficiency testing regarding quality assessment of rabies serological testing for dogs and cats within the frame of the EU pet travel scheme, (ii) the 2013 inter-laboratory comparison test on FAT, RTCIT, conventional and RT-PCR, and (iii) an inter-laboratory comparison among the network of OIE-RL for rabies of the direct rapid immunohistochemical test (dRIT) for diagnosis of rabies.
Zoonooses (Bacterial Infections)
Title of activity / Scope
Bovine tuberculoses, Para tuberculoses / Research is focussing on improved and novel strategies for diagnosis and control of diseases (mycobacterial infections?).
In the frame of the project “Mycobacterium bovis in the wildlife-livestock-human interface in East and Southern Africa.” (DFG Program: African network building for the control of neglected zoonoses) mycobacterial from livestock and wildlife (113 from Tanzania, 44 from Kenya” were sequence-analyzed for species identification samples
Ongoing research project are “Integrated control of neglected zoonoses: improving human health and animal production through scientific innovation and public engagement” (EU FP7 – 221948; ICONZ) and “Development of novel diagnostic strategies for the ante-mortem immunodiagnosis of bovine tuberculosis and Johne´s disease (EMIDA ERA-Net, MYCOBACTDIAGNOSIS)”.
Brucelloses / 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. The laboratory carried out diagnostic testing for several OIE Member Countries (Spain, Egypt, Chile). Training courses for serological and molecular diagnostic techniques were performed for UAE, Pakistan and Egypt.
Chlamydioses / Protocols and technical expertise of the RL's standard real-time PCR assays and DNA microarray tests were provided to laboratories in the Netherlands, Poland, Croatia, Ukraine and France. Researchers from the Netherlands, Poland and Ukraine were received in the laboratory for short technical visits. Chromosomal DNA aliquots from reference strains of Chlamydia spp. were shipped to labs in 8 different OIE member states (Ireland, Spain, France, the Netherlands, Ukraine, Poland and Australia). A total of 889 diagnostic tests on avian chlamydiosis) and 443 on ovine chlamydiosis were conducted to examine field samples sent in from other OIE countries.
EHEC / Ongoing research project “Occurrence of EHEC O104:H4 genes in cattle herds from Germany and Spain” (EU ANTIGONE).
Protocol and technical expertise of the reference laboratory standard STEC colony hybridization assay were provided to the laboratories in Spain (SaBio-IREC (CSIC-UCLM-JCCM), Ciudad Real, Spain; VISAVET Health Surveillance Centre, Universidad Complutense, Madrid, Spain).
Glanders / An OIE-Twinning project with NRCE, Hisar, India was started. Training courses for serological and molecular diagnostic techniques were performed for Iraq. 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 / Various nutrient media for culture of F. tularensis subsp. holarctica were compared with polymerase chain reaction (PCR) in a study with samples from wild animals (also published).
A serological investigations of wild boars (Sus scrofa) and red foxes (Vulpes vulpes) as indicator animals for circulation of Francisella tularensis in Germany was performed and published.
German Francisella tularensis isolates from European brown hares (Lepus europaeus) were typed using genetic and phenotypic markers in a cooperation with Sweden to elucidate their phylogeographic relation (also published).
PCR assays were provided to Federal Laboratories and a ringtest was organized for the detection of F. tularensis using PCR assays with participants from Germany and Austria.
Zoonooses (Parasite Infections)
Title of activity / Scope
Dourine / Dourine control sera were provided to the Laboratoire Régional d´Analyses et de Recherches, Casablanca Morocco,
40 horse sera from Kyrgyzstan were examined for dourine antibodies
Echinococcosis / A mathematical model for analysing the spatial and temporal distribution of Echinococcus multilocularis was further developed. The spread and behaviour of the raccoon dog (Nyctereutes procyonoides) as a new definitive host of Echinococcus multilocularis in eastern and central Europe was studied. A total of 33 foxes from Brandenburg, Germany, were tested for the presence of Echinococcus multilocularis by the Intestinal Scraping Technique. The parasite was detected in 15 samples. Molecular typing techniques (EmsB microsatellite analysis; detection of the mitochondrial markers cox1, nad1 and atp6) were established to study the spatial and temporal epidemiology of the infection of foxes with the parasite.
A PhD student from Sudan was trained in study design, sampling and echinococcosis diagnosis. 33 out of 143 canine fecal samples from different regions in Sudan contained taeniid eggs, which were molecularly typed by multiplex PCR (Boubaker et. al., 2013). 535 sheep, 291 goats, 735 cattle and 430 camels were examined for hydatid cysts at slaughterhouses. While the prevalence was low in cattle (1.4%), sheep (0.3%) and goats (0.3%), a substantial proportion of camels was found infected (25.3%).
Zoonooses (Transmissible Spongiforme Encephalopathies, Prions)
Title of activity / Scope
Prions –TSE: / Pathogenesis studies and risk assessment studies focus more on atypical BSE. Close collaborations with other NRLs and with research institutes were continued. BSE or scrapie positive samples and reference materials (e.g. fresh brainstem material or paraffin-embedded fixed tissue) were supplied to cooperating partners.
ToR: To propose or develop methods and procedures that facilitate harmonisation of international standards and guidelines applicable to the designated specialty
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
Proposal title / Scope/Content / Applicable areaHarmonisation of diagnostic techniques on Echinococcosis / This study was done in the framework of APHAEA (harmonised Approaches in monitoring wildlife Population Health, And Ecology and Abundance) in collaboration with IREC, Spain, IZSTO, Italy, VetAgroSup, France, SVA Sweden, DTU, Denmark, FIWI at the University of Bern, Switzerland, the Artemis Research Institute, the Netherlands, IZSLER, Italy, and the University of Turin, Italy. / Surveillance and control of animal diseases
Food safety
Animal welfare
Real time PCR subtyping of swine influenza viruses / In order to cope with the increasing diversity of swine influenza viruses multiplex RT-qPCRs were developed to diagnose four hemagglutinin and two neuraminidase lineages including A/H1N1pdm in swine samples. / Surveillance and control of animal diseases
Food safety
Animal welfare
“Blueprint for Fox Rabies Prevention” / Completion and updating of the “Blueprint for Fox Rabies Prevention” (http://www.rabiesblueprint.com/) / Surveillance and control of animal diseases
Food safety
Animal welfare
Reliable detection method for Schmalenberg virus RNA in semen / Development and validation of PCR-system for the reliable detection of SBV-RNA in semen samples / Surveillance and control of animal diseases
Food safety
Animal welfare
In vitro production procedure of dourine antigen for CFT / The OIE-recommended method of production of Trypanosoma equiperdum whole cell antigen for diagnosis of dourine requires the infection of rats followed by harvest and purification the pathogen from blood. In vitro propagation of T. equiperdum saves animal lives and is expected to generate better standardized antigen for CFT and the opportunity to detect partial antigens of higher specificity to differentiate T. equiperdum from T.evansi. / Surveillance and control of animal diseases
Food safety
Animal welfare
ToR: To establish and maintain a network with other OIE Collaborating Centres designated for the same specialty, and should the need arise, with Collaborating Centres in other disciplines