OIE Reference Laboratory Reports s15

OIE Reference Laboratory Reports Activities in 2011

Name of disease (or topic) for Rabies which you are a designated OIE Reference Laboratory:

Address of laboratory: Institute for Epidemiology, Friedrich-Loeffler-Institute - Federal Research Institute for Animal Health, Seestrasse 55, D-16868 Wusterhausen, GERMANY

Tel.: +49 33979 80186

Fax: +49 33979 80200

e-mail address: [email protected]

website: www.fli.bund.de

www.who-rabies-bulletin.org

Name (including Title and Dr Thomas Müller, senior scientist Position) of Head of Laboratory (Responsible Official):

Name(including Title and Drs Thomas Müller, Conrad Freuling Position) of OIE Reference Expert:

Name (including Title and Dr Thomas Müller Position) of writer of this report (if different from above):

Annual reports of OIE Reference Centres, 2011 1 Rabies

Part I: Summary of general activities related to the disease

1. Test(s) in use/or available for the specified disease/topic at your laboratory

Test For Specificity Total

Fluorescent antibody test Antigen N protein, all genotypes 1032

RTCIT Virus isolation all genotypes 48

RT-PCR RNA all genotypes 553

Sequencing Strain characterization N-P region, full length genome 100

RFFIT Virus neutralising RABV, EBLV-1, EBLV-2 376 Antibodies

FAVN Virus neutralising RABV 14 Antibodies

Virus titration Oral vaccines for wildlife / 65 Quality assessment

2. Production and distribution of diagnostic reagents

Amount supplied nationally Amount supplied to other Type of reagent (including for own use) countries

Panel of 10 anti-N-monoclonal - 100 ml antibodies

anti-G-monoclonal antibodies - 10 ml

BHK21 cells, murine neuroblastoma cells for virus propagation and isolates of European Bat Lyssavirus (EBLV) type -1 and -2 is supplied to other OIE Members.

Part II: Activities specifically related to the mandate of OIE Reference Laboratories

3. International harmonisation and standardisation of methods for diagnostic testing or the production and testing of vaccines

a) Establishment and maintenance of a network with other OIE Reference Laboratories designated for the same pathogen or disease and organisation of regular inter-laboratory proficiency testing to ensure comparability of results

 The OIE Reference Laboratory at FLI organised an international analytical and diagnostic ring trial for lyssavirus (RABV, EBLV-1, -2) real-time RT-PCRs, in which 16 European laboratories including two OIE Reference Laboratories for Rabies participated.

 The OIE Reference Laboratory participated (i) in the annual proficiency testing regarding quality assessment of rabies serological testing for dogs and cats within the frame of the EU pet travel

2 Annual reports of OIE Reference Centres, 2011 Rabies

scheme, (ii) the 2010 inter-laboratory comparison test on FAT, RTCIT and RT-PCR and (iii) the inter-laboratory comparison test on the detection of tetracycline (as a biomarker) for monitoring of ORV campaigns as organised by the EU-RL for Rabies, ANSES, Malzeville, France.

b) Organisation of inter-laboratory proficiency testing with laboratories other than OIE Reference Laboratories for the same pathogens and diseases to ensure equivalence of results

 The OIE Reference Laboratory at FLI organised an international analytical and diagnostic ring trial for lyssavirus (RABV, EBLV-1, -2) real-time RT-PCRs, in which 16 European laboratories including two OIE Reference Laboratories for Rabies participated.

 The OIE Reference Laboratory participated in a collaborative study on quality issues of anti-rabies conjugates for use in FAT as organised by the EU-RL for Rabies, ANSES, Malzeville, France.

 The OIE Reference Laboratory at FLI joint international co-operation on harmonising methods for quality control of vaccines and participated in an inter-laboratory comparison test on titration of the oral rabies virus vaccine SAD Bern currently used in ORV campaigns in Europe organised by the EU-RL for Rabies, ANSES, Malzeville, France.

4. Preparation and supply of international reference standards for diagnostic tests or vaccines

see point 2

The anti-N-MAb panel can be used for virus typing of lyssaviruses and is able to distinguish between virus variants circulating in different host species. The panel is supplied upon request to national and international laboratories in a lyophilised form.

5. Research and development of new procedures for diagnosis and control

The OIE Reference Laboratory at the FLI in collaboration with the OIE Reference Laboratory at the AHVLA, UK, the Technical University of Denmark, National Veterinary Institute, Lindholm, Denmark and the Central Veterinary Institute, Lelystad, The Netherlands further validated a TaqMan© real-time RT-PCR developed for EBLV-1 and 2 considering OIE recommendation and artificial labelled positive and internal controls developed. Furthermore, a TaqMan© real-time RT-PCR for the detection of the newly discovered Bokeloh Bat Lyssavirus (BBLV) was developed that can be used in active bat rabies surveillance.

Test protocols for RABV-specific real-time RT-PCRs and other RT-qPCR applied in various European NRLs for Rabies were compared regarding specificity and sensitivity.

In collaboration with an industrial partner research on the development of a new anti-N-MAb based FITC-labelled anti-rabies conjugate was finished and the product validated against commercially available ones. The new anti-N- MAb based FITC-labelled anti-rabies conjugate has an improved sensitivity and sensitivity and is characterised by a very low or absent background staining.

The OIE Reference Laboratory is a collaborator in a RAPIDD (Research and Policy in Infectious Disease Dynamics) Working Group on Bat Infection Dynamics funded through the US government with the focus on modelling EBLV-1 and EBLV-2 infections in host reservoirs.

Antigenenic sites of the newly discovered Bokeloh Bat Lyssavirus (BBLV) were characterized and compared with other lyssavirus species. Experimental (in mice) and serological studies using a panel of 100 sera from human vaccines were conducted to determine pathogenicity of BBLV and cross-neutralisation with RABV specific antibodies.

Eighteen inactivated Human Rabies Vaccines were biochemically characterized and their virus strains genetically identified.

Susceptibility of free-roaming raccoons (Procyon lotor) from the German founder population for RABV (dog- mediated, fox-mediated and raccoon-mediated strain, EBLV-1) was experimentally studied.

The OIE Reference Laboratory leads a core group of the Partners for Rabies Prevention (PRP) dedicated to develop a Blueprint for Red Fox Rabies Prevention.

6. Collection, analysis and dissemination of epizootiological data relevant to international disease control

Edition of 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 2011, the issues 03/2010, 04/2010, 1/2011 and 02/2011 were edited and distributed throughout the world. The following Miscellaneous Articles were published in the issues mentioned above: (i) Passive and Targeted Surveillance for European Bat Lyssavirus in Swiss Bats, (ii) Development of a Blueprint for Rabies Prevention and Control, (iii) Rabies outbreak in Małopolskie voivodship in Poland, (iv) Third European workshop for rabies and implementation of the EU-RL website, (v) The European Union strategy for external cooperation with neighbouring countries on rabies control, (vi) Bat rabies surveillance in Finland, (vii) Human Rabies in a Romanian boy – an ante mortem case study and (viii) Review of the analysis related to rabies diagnosis and follow-up of oral vaccination performed in NRLs in 2010. To enhance the quality of rabies surveillance data, an online submission of rabies surveillance data, e.g. positives and negatives, to the European rabies database of the RBE was favoured. Access and use of the RBE website was analysed using tracking software.

In order to assess ORV programs in Western, Central and Eastern Europe we collected data for each country for the time period 1978-2010 on (i) size and location of vaccination areas, (ii) oral rabies vaccine virus strain used, (iii) bait density used, and (iv) modalities of bait distribution for each individual campaign. Spatial information data was subsequently digitized and geo-referenced and entered into the European (ArcGIS) database for further analysis. The influence of (1) the initial epidemiological situation (rabies incidence) at the beginning of ORV campaigns, (2) the size of the affected area and (3) the length of borderline to neighbouring rabies endemic regions on the duration and costs of rabies elimination was analysed.

7. Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the pathogen and the disease concerned

The OIE Reference Laboratory at FLI A has implemented a documented quality management system and is accredited according to norm ISO/IEC 17025. The usual contents of the quality manual follow the outline of the ISO/IEC 17025 standard.

Rabies diagnostic and research is conducted under biosafety level 3** and 3, respectively.

8. Provision of consultant expertise to OIE or to OIE Member Countries

The following consultant expertise was provided by rabies experts of the OIE Reference Laboratory at the FLI:  the 10th meeting of the South East African Rabies Group, Maputo, Mozambique, January 2011  Rabies Oral Vaccination Monitoring Workshop, Pristina, Kosovo, February, 2011  the meeting of Partners for Rabies Prevention, Banna, Italy, May 2011 and San Juan, Puerto Rico, USA, October 2011  the EU task force meeting to assess the oral vaccination programmes of foxes against rabies in Bulgaria, Sofia, March 2011  annual meeting with the candidate laboratory and provincial veterinary laboratories in the frame of the OIE Laboratory Twinning Project with Turkey, March 2011  International Bohol Rabies Conference, Bohol, Philippines, June 2011  OIE Global conference on Rabies control: Towards Sustainable Prevention at the Source, Incheon, Korea, September 2011  National Zoonosis Symposium, Berlin, Germany, October 2011  22nd International Rabies in the Americas Conference, San Juan, Puerto Rico, October 2011

 RAPIDD meeting, Cambridge, UK, October 2011  Extraordinary meeting in relation with the rabies situation in the common border area between Bulgaria, Greece and the Former Yugoslav Republic of Macedonia, Sofia, Bulgaria, December 2011  Contribution to a quantitative risk assessment on the change in likelihood of rabies introduction into the United Kingdom as a consequence of adopting the existing harmonised Community rules for the non- commercial movement of pet animals (client DEFRA - upon request of the OIE Member)  Contribution to the development of the “Blueprint for Rabies Prevention and Control (http://www.rabiesblueprint.com/)”  Leadership position in developing a “Red Fox Rabies Blueprint” for the Partners for rabies Prevention

9. Provision of scientific and technical training to personnel from other OIE Member Countries

Training of scientific and technical staff: In 2011 we have hosted colleagues from other OIE member countries including one person from Iran and one from South Africa. The training included basic epidemiology and 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).

Also, training in epidemiology, implementation of adequate rabies surveillance and epidemiological analysis was provided for one colleague from Iran. A staff member of the OIE reference laboratory is supervisor for a PhD thesis of a Latvian colleague on evaluation of ORV campaigns in Latvia.

Witihin the frame of an African “Joint Master’s Programme in Transboundary Animal Disease Management” (MTADM) project 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, staff members of the OIE Reference Laboratory acted as supervisors for a master student from the Addis Ababa University, Ethiopia.

The OIE Reference Laboratory conducted advisory service within the frame of the OIE Laboratory Twinning Project with Turkey

10. Provision of diagnostic testing facilities to other OIE Member Countries

Diagnostic facilities were provided to the American Armed Forces stationed in Germany for confirmatory testing and sequencing of rabies virus isolates from Iraq and Afghanistan. We also have continued establishing closer links with other OIE reference laboratories for rabies and WHO CCs for rabies, e.g the AHVLA, UK, Pasteur Institute Paris France, and CDC, USA.

11. Organisation of international scientific meetings on behalf of OIE or other international bodies

The OIE reference laboratory for rabies at the FLI was member of the scientific committee of the OIE Global conference on Rabies control: Towards Sustainable Prevention at the Source, Incheon, Korea, September 09-11, 2011.

12. Participation in international scientific collaborative studies

The OIE Reference Laboratory at FLI continued work within a WHO co-ordinated research project on research and development on a murine monoclonal antibody (MAbs) cocktail to replace HRIG and ERIG for rabies post- exposure prophylaxis (PEP) in humans and conducted in vitro studies of plant derived murine MAbs.

Furthermore, we lead a working group of the Partners for Rabies Prevention dedicated to develop a “Blueprint for Red Fox Rabies Prevention”. The Blueprint is expected to be approved in May 2012.

13. Publication and dissemination of information relevant to the work of OIE (including list of scientific publications, internet publishing activities, presentations at international conferences)

 Presentations at international conferences and meetings

o MÜLLER T, DIETZSCHOLD B, ERTL H, FOOKS AR, FREULING C, FEHLNER-GARDINER C, KLIEMT J, MESLIN FX, RUPPRECHT CE, TORDO N, WANDELER AI, KIENY MP. Current status of research on murine monoclonal antibodies for PEP. 10th meeting of the South East African Rabies Group (SEARG), 25-28 January 2011, Maputo, Mozambique

o FREULING C, MÜLLER T. Rabies oral vaccines and their quality insurance. Rabies Oral Vaccination Monitoring Workshop, 12 February 2011, Pristina, Kosovo

o MÜLLER T, FREULING C, General principals of monitoring of Oral Vaccination Campaigns Rabies Oral Vaccination Monitoring Workshop, Pristina, Kosovo, 12.02.2011

o MÜLLER T, FREULING C, VOS A. Europe – Fox Rabies Prevention Blueprint - Operational part update, Meeting of the Partners for Rabies Prevention, 04-06 May 2011, Banna, Italy

o MÜLLER T, FREULING C, DEMETRIOU P. Global Projects – European Union (EU) and Surrounding Areas. Meeting of the Partners for Rabies Prevention, 04-06 May 2011, Banna, Italy

o RUPPRECHT CE, FOOKS AR, MÜLLER T. “A Road Ahead?” Rabies Research Advances and Needs in the 21st Century. International Bohol Rabies Conference, 13-15 June 2011, Bohol, Philippines

o MÜLLER T, FREULING CM. Case Report (3) – Europe (focus on wildlife) Rabies elimination in Europe – a success story. Global Conference on Rabies Control 7-9 September 2011, Incheon–Seoul (Republic of Korea)

o MÜLLER T, HOFFMANN B, HÖPER D, BEER M, TEIFKE J, FINKE S, METTENLEITER TC, FREULING C. Bat rabies in Germany - discovery of a novel lyssavirus in a Natterer’s bat (Myotis nattereri). 4th EURL workshop of the EU Rabies National Reference Laboratories, 21 September 2011, Nancy, France

o MÜLLER T, HOFFMANN B, HÖPER D, BEER M, TEIFKE J, FINKE S, METTENLEITER TC, FREULING C. Bat rabies in Germany - discovery of a novel lyssavirus in a Natterer’s bat (Myotis nattereri). National Zoonosis Symposium, 06-07 October 2011, Berlin, Germany

o FREULING CM, FUHRMANN L, AYLAN O, UN H, FOOKS AR, MCELHINNEY L, JOHNSON N, MÜLLER T. Molecular epidemiology of rabies in the Middle East and Central Asia with special emphasis on Iraq and Afghanistan. National Zoonosis Symposium, 06-07 October 2011, Berlin, Germany

o HANKE D, HOFFMANN B, SCOTT T, FREULING C, MÜLLER T, NEL L, BEER M, HÖPER D. Lyssavirus whole genome sequencing. National Symposium on Zoonosis Research, 06-07 October 2011, Berlin, Germany

o SCHATZ J, HEIDECKE D, WIBBELT G, DOLCH D, POTT-DÖRFER B, OHLENDORF B, ECCARD JA, MÜHLE RU, FREULING C, MÜLLER T. Passive surveillance for bat lyssaviruses in indigenous bat species in Germany (1998-2010). National Symposium on Zoonosis Research, 06-07 October 2011, Berlin, Germany

o MÜLLER, T, FREULING CM, VOS A. Chapter on wildlife rabies in the Canine Rabies Blueprint. Meeting of the Partners for Rabies Prevention, San Juan, Puerto Rico, 16 October 2011

o MÜLLER TF, FREULING CM, SELHORST T, KLIEMT A, MESLIN FX. Retrospective quantitative assessment of ORV of foxes in europe between 1978 and 2010. XXII International Conference on Rabies in the Americas, 17 – 22 October 2011, San Juan, Puerto Rico

o FREULING CM, TEIFKE JP, FINKE S, MALERCZYK C, MÜLLER T. Bokeloh Bat Lyssavirus- Pathogenicity and Vaccine Cross Neutralization. XXII International Conference on Rabies in the Americas, 17 – 22 October 2011, San Juan, Puerto Rico

o MÜLLER TF, FREULING CM. Strategic consideration for rabies surveillance and planning of ORV campaigns in the triangle border area between Macedonia, Bulgaria and Greece. Extraordinary meeting in relation with the rabies situation in the common border area between Bulgaria, Greece and the Former Yugoslav Republic of Macedonia, 22 December 2011, Sofia, Bulgaria

. Scientific publications in peer-reviewed journals

 JOHNSON N, FREULING C, HORTON D, MÜLLER T, FOOKS A (2011). Importation of rabies into the European Union and Switzerland 2001-2010. Emerg Infect Dis. 2011 Apr;17(4):753-4.

 LEMBO T, ATTLAN M, BOURHY H, CLEAVELAND S, COSTA P, DEBALOGH K, DODET B, FOOKS A, HIBY E, LEANES F, MESLIN FX, MIRANDA ME, MÜLLER T, NEL L, RUPPRECHT CE, TORDO N, TUMPEY A, WANDELER A, BRIGGS DJ. (2011) Renewed Global Partnerships and Re-designed Roadmaps for Rabies Prevention and Control. Vet Med Int. 2011:923149. Epub 2011 Jun 1.

 MULATTI P, MÜLLER T, BONFANTI L, MARANGON S. (2011). 2009-2010 emergency oral rabies vaccination of foxes in Italy: identification of residual rabies foci at higher altitudes in the Alps. Epidemiol Infect. 2011 Jul 11:1-8. [Epub ahead of print]  MCELHINNEY LM, MARSTON DA, FREULING CM, CRAGG W, STANKOV S, LALOSEVIC D, MÜLLER T, FOOKS AR. (2011) Molecular diversity and evolutionary history of rabies virus strains circulating in the Balkans. J Gen Virol. Sep; 92 (Pt 9):2171-80. Epub 2011 Jun 1.

 VOS A, NUNAN C, BOLLES D, MÜLLER T, FOOKS AR, TORDO N, BAER GM. (2011). The occurrence of rabies in pre-Columbian Central America: A Historical Search. Epidemiol Infect. Oct;139(10):1445-52. Epub 2011 Jul 29

 FREULING C, WOHLSEIN P, KELLER B; MÜHLBACH E; CONRATHS FJ, TEIFKE JP, HOFFMANN B, HÖPER D, KORTHASE C, BEER M, FINKE S, METTENLEITER TC, MÜLLER T. (2011). Novel Lyssavirus Isolated from a Natterer’s bat, Germany. Emerg Infect Dis. Aug;17(8):1519-22.

 VOS A, CONZELMANN K-K, FINKE S, MÜLLER T, FOOKS AR, NEUBERT A. (2011). Immunogenicity of a rabies virus construct with a site-directed deletion in the phosphoprotein in red foxes and skunks. Special issue: recent advances in prevention and control of rabies. Adv Prev Med. 011; 898171. Epub 2011 Sep 21.

 MÜHLDORFER K, SPECK S, KURTH A, LESNIK R, FREULING C, MÜLLER T, KRAMER-SCHADT S, WIBBELT G. (2011). Diseases and causes of death in European bats: dynamics in disease susceptibility and infection rates. PLoS One. 6(12):e29773. Epub 2011 Dec 28.  Other communications

 PANNWITZ G, FREULING C, DENZIN N, SCHAARSCHMIDT U, NIEPER H, HLINAK A, BURKHARDT S, KLOPRIES M, DEDEK J, HOFFMANN L, KRAMER M, SELHORST T, CONRATHS FJ, METTENLEITER T, MÜLLER T. (2011). A long- term serological survey on pseudorabies virus infections in wild boar in East Germany. Epidemiol Infect. Feb 15:1-11. [Epub ahead of print]

 HÖNER O, WACHTER B, GOLLER K, HOFER H, RUNYORO V, THIERER D, FYUMAGWA RD, MÜLLER T, EAST ML. (2011). IMPACT OF A PATHOGENIC BACTERIUM ON A SOCIAL CARNIVORE POPULATION J ANIM ECOL. 2012 JAN;81(1):36-46. DOI: 10.1111/J.1365-2656.2011.01873.X. EPUB 2011 JUN 1.

 MÜLLER T, HAHN EC, TOTTEWITZ F, KRAMER, M, METTENLEITER T, FREULING C. (2011). Pseudorabies virus infections in wild swine – a global perspective. Arch Virol. Oct;156(10):1691-705

 KRUCKENBERG H, MÜLLER T, FREULING C, MÜHLE RU, GLOBIG A, SCHIRRMEIER H, BUSS M, HARDER T, KRAMER M, TESKE K, WALLSCHLÄGER D, HLINAK A. (2011). Serological and virological survey and resighting of marked wild geese in Germany. Eur J Wildl Res: 57(5): 1025-1032, DOI: 10.1007/s10344-011- 0514-1

 DELERÉ Y, WICHMANN O, MÜLLER T, FREULING C, ROGGENDORF M, ROSS S. (2011). Tollwut in Deutschland: Gelöstes Problem oder versteckte Gefahr? Epidemiologisches Bulletin, 8: 57-61

 EGGERS U, SCHULZ H, SCHULZ F, FREULING C, HLINAK A, MÜHLE RU, MÜLLER T, WALLSCHLÄGER D. (2011). Möglichkeiten interdisziplinärer Forschung zur Biologie der Vögel: das „Paradebeispiel“ Weißstorch (Ciconia ciconia) Untersuchungen in der Prignitz, Brandenburg. Berichte der Vogelwarte Hiddensee. 20: 15- 26

 BENNINGER G, DROSTEN C, GOETHE R, KÖCK R, KREIENBROCK L, LUDWIG S, MERLE R, MÜLLER T, NEUBAUER H, SACHSE K, SCHLÜTER D, STRAUSS EG, WIELER LH. (2011). Zoonosen: Bekannte und neue Infektionskrankheiten - eine Herausforderung für die Forschung an der Schnittstelle von Human- und Veterinärmedizin. Deutsche Zeitschrift für Klinische Forschung 5/6, S.42-47

 MÜLLER T, FREULING C. (2011). Tollwut. ZiVet 07/2011, S. 46-48.

 FREULING CM, KLIEMT J, MÜHLBACH E, MÜLLER T. (2011). Neues Fledermaustollwutvirus aus einer Fransenfledermaus (Myotis nattereri) isoliert. Nyctalus (N.F.), Berlin 16; 3-4: 204-206

 FREULING CM, SCHATZ J, POTT-DÖRFER B, HEIDECKE D, WIBBELT G, MÜHLDÖRFER K, KLIEMT J, MÜLLER T. (2011). Untersuchungen an Fledermaustodfunden zum Vorkommen der Fledermaustollwut in Deutschland. Nyctalus (N.F.), Berlin 16; 3-4: 212-216

 MÜLLER T, SCHATZ J, FREULING C. (2011). Fledermaustollwut - ein globaler Überblick. Nyctalus (N.F.), Berlin 16; 3-4: 197-203

 MÜLLER T, OHLENDORF B, POTT-DÖRFER B , VOS A, NEUBERT L, KLIEMT J, FREULING CM. (2011). Experimentelle Studien von EBLVs in verschiedenen Wirten - ein Überblick unter besonderer Berücksichtigung der Fledermäuse. Nyctalus (N.F.), Berlin 16; 3-4: 207-211

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8 Annual reports of OIE Reference Centres, 2011