♦ 04 July 2016 ♦   Central Veterinary Institute, part of Wageningen University and Research Centres – Lelystad       Dutch National Reference Laboratory The Central Veterinary Institute (CVI) collaborates with public and private partners to safeguard animal and public health. CVI contributes to the prevention, eradication and control of animal infectious diseases through research, diagnostics and consultancy. By doing so, CVI helps to guarantee international trade and to preserve the international top position of the Dutch livestock industry. In 1904 we started as the National Serum Institute in Rotterdam. In 1959 this institute merged with the Governmental veterinary research institute (situated in Amsterdam) into the CVI. CVI is the national reference institute for notifiable animal diseases in the Netherlands. For these diseases CVI has an important and decisive task to determine the final diagnosis.   Rabies in the Netherlands In the period from 1875 to 1923 several laws were installed to eradicate dog as well as sylvatic rabies in the Netherlands, leading to the official OIE rabies free status in 1923. In the period from 1974-1988 the free status was lost however, due to a reintroduction of sylvatic rabies through foxes from the east through Belgium and Germany. After 1988 the country regained it OIE free status but by then was confronted with a new phenomenon, European bat lyssavirus. Since 1987 a nationwide surveillance system was set up, and since then, over 5000 bats have been tested for EBLV, resulting in 377 cases of EBLV-1 in serotine bats and 5 cases of EBLV-2 in pond bats. Since that time the only classical rabies cases were import cases. The last incident occurred in 2012 when a 8,5-week-old puppy was imported from Morocco. A day after arrival in Amsterdam the puppy showed aggressive behaviour and two days later suspicion of rabies arose and was confirmed by CVI. Over forty people were given post-exposure-prophylaxis and several contact animals were euthanized for rabies testing. Picture: EBLV-1 and EBLV-2 cases in the Netherlands from 2004 to 2016   Laboratory and staff Rabies diagnostics at CVI is performed by the Diagnostic Unit. All suspect cases, bats as well as larger mammals (mostly foxes and dogs) are tested by FAT and real-time RT-PCR. The FAT is ISO17025 accredited, as well as the EBLV-1 real-time PCR. Other PCR tests such as the RABV and EBLV-2 real-time PCR and several conventional PCR are extensively validated and run under ISO9001 standards. The Fluorescence Antibody neutralisation test (FAVN) is used for determination of antibody titres in pets as well as humans. These tests are performed by a team of technicians and staff to authorize results. Since CVI has a 24/7 duty to perform rabies diagnostics in cases of direct contact of humans with suspect animals such as bats or sometimes foxes we maintain a minimum of eight people per team. Besides the diagnostics tasks, CVI has limited resources for rabies research. Besides phylogenetic analyses of all the positive bat cases and a study on molecular species determination we have been involved in several studies aiming at further analysis of bats in search of other potential zoonotic viruses. In 2012 we co-authored a paper on bat Coronaviruses in the Netherlands and to date we are involved in a study on the Dutch ABC islands (Aruba, Bonaire and Curacao).                     Contacts E.A. (Bart) Kooi, Ph.D. Projectleader Rabies and Biosafety Officer Central Veterinary Institute of Wageningen UR Houtribweg 39, 8221 RA Lelystad e-mail: bart.kooi@wur.nl Internet: www.cvi.wur.   Presentation of Dutch NRL.pdf

♦ 04 July 2016 ♦ The annual online activity questionnaire was submitted to all National Reference Laboratories (NRLs) early 2016 to collect and collate data on methods used and results of tests carried out in the Community in the frame of rabies control programmes (Commission regulations (EU) N° 737/2008 and N°415/2013). This document reviews the 2015 analysis performed in 27 NRLs from the European Union and in 7 NRLs from third countries involved in a rabies control programme. The report is available n the private part of the EURL for rabies website for private members only.  

♦ 01 July 2016 ♦ The inter-laboratory test for rabies diagnosis is now over. A total of 54 laboratories, including 28 non-EUMS laboratories participated to the 2016 session of the test. This year the test was composed of a panel of 10 samples to be tested with the chosen technique(s) (FAT, RTCIT, Conventional and/or Real Time RT-PCR) and a corresponding online technical questionnaire for each tested technique. The report should be available in autumn 2016. In the meantime, participants will receive an online satisfaction questionnaire regarding this test.  

♦ 16 June 2016 ♦ Despite the unfortunate Alsatian weather of the day, the annual Workshop for Rabies was a frank success. Forty-five rabies European scientists attended the meeting in the European Capital City of Strasbourg, leading to constructive exchanges on rabies. Additionally to the representative of the European Commission, Pedro Rosado Martin (G5 Unit of EC DG Santé), the EURL team was pleased to welcome for the first time speakers from the Food and Agriculture Organization of the United Nations (FAO) and from the French Directorate General for Food (DGAl). The analysis of the results of inter-laboratory tests for rabies diagnosis organized in 2015 by the EURL for rabies was, as usual, presented with an open discussion on the subject. The EURL for rabies also presented on one hand a performance study of disinfectants used in the context of molecular biology techniques, and on the other hand the progress of the study on the evaluation methods of Real Time RT-PCR. The Balkan region was in the spotlight this year, and several laboratories attended the meeting (Albania, Bosnia and Herzegovina, Republic of Kosovo, Former Yugoslav Republic of Macedonia and Serbia) as well as other eastward countries such as Moldova, Ukraine and Turkey. Each of them presented both the rabies situation and the rabies programmes in progress in their country. Presentations are available on the EURL for rabies website for private members only. Next workshop for rabies will be organized in June 2017 in Spain, together with the meeting of the EURL for rabies serology.  

♦ 05 July 2016 ♦ [Taken form the homepage of the Euroreference website] “The first issue of Euroreference (Journal of Reference) was launched in June 2009 as a free online AFSSA publication in both French and English. The original aim was to facilitate the dissemination of information about refe¬rence activities among stakeholders both at national and European levels. The topics covered included all scientific and technical aspects of analytical methods and their role in surveillance plans and reference-related regulatory issues. As a common forum for members of reference laboratory networks and all the players concerned, it was geared to National and European Reference Laboratories, animal health and food safety diagnostic laboratories, Ministries and decision-makers, directors of institutions, etc. The AFSSA/ ANSES Editorial board was rapidly joined by representatives of CODA-CER¬VA (Belgium), ISS (Italy) then PIWET (Poland).” The new of EuroReference is “co-edited by CODA-CERVA (Belgium), ANSES and SCL (France), BfR and FLI (Germany), ISS, IZSAM and IZSLER (Italy), PIWET (Poland), APHA and FERA (UK) in addition to the EPPO.” A .pdf version of EuroReference can be downloaded on the website.  

♦ 01 June 2016 ♦ Forty-five participants from the EC Member States and eastern bordering countries as well as international institutions such as FAO, the European Commission and the French Ministry of Agriculture (DGAl) will attend next workshop for for rabies that will be held on 16th June in Strasbourg, France. The provisional agenda has been sent to all participants and is also available on the EURL website (private part only). It includes a discussion on the results of the inter-laboratory tests of rabies diagnosis organised by the EURL in 2015, some key points of the rabies laboratory activities, and an overview of the significant rabies events that occurred in 2015 as well as a focus on the rabies situation in the Balkans and Eastward countries.  

♦ 30 May 2016 ♦ Samples for this year Inter-laboratory test for rabies were sent on Monday 30 May. The test is composed of a panel of 10 samples to be tested with the chosen technique(s) (FAT, RTCIT, Conventional and/or Real Time RT-PCR) as well as a corresponding online technical questionnaire for each tested technique. Results from participating laboratories have to be sent to the EURL for rabies and online technical questionnaires should be answered before 30 June 2016. A satisfaction questionnaire will be sent after the test to all participants. The report on this test should be issued in October 2016.  

♦ 04 April 2016 ♦ Latvian National Reference Laboratory Institute of Food Safety, Animal Health and Environment BIOR (Institute BIOR) - Rīga, Latvia The Institute of Food Safety, Animal Health and Environment BIOR (Institute BIOR) is the National Reference Laboratory for rabies in Latvia. The Institute BIOR consists of three laboratories situated in Riga (Animal Diseases Diagnostic Laboratory, Laboratory of Food and Environmental Investigations and Medical Microbiology Laboratory) and three Regional laboratories. Two Divisions of Animal Diseases Diagnostic Laboratory (ADDL) deal with rabies diagnostics and Monitoring of ORV: Virology Division and Pathology Division. Rabies team Staff present on 01.04.2016: From the left: Daina Čavare (M.Sc.), Ņina Baranovska, Irina Kaļiņičenko (M.Sc.), Ludmila Soboļeva. On the background: Ieva Rodze(M.Sc.), Zita Muižniece (Dr.biol.) and Svetlana Cvetkova (M.Sc.). Staff missing from the photo: Marianna Dobroštana (PhD student), Veronika Buboviča (Dr.biol.), Jūlija Trofimova (M.Sc.), Marija Bazanova (veterinarian) and Inga Pigiņka-Vjačeslavova (PhD student). The head of ADDL is Ieva Rodze who coordinates all work for rabies diagnostic in the Laboratory and provides for cooperation with Food and Veterinary Service on rabies surveillance. Rabies specific diagnostics are carried out mainly in Virology Division. Head of Division is Marianna Dobroštana. In these activities six senior experts and two laboratory technicians from Virology Division are involved. Two senior experts from Pathology Division perform necroscopy and tetracycline detection in teeth. Rabies technique Laboratory techniques used for diagnostic tests in frame of passive surveillance is FAT, RTCIT and conventional hn-RT-PCR (P.R. Heaton et al., 1997), virus genotype sequence analysis. Serological tests used for vaccination control are FAVN and ELISA. ORV campaign control is performed by bait titration, serological tests by ELISA (sero-conversion detected by Ab-ELISA) and luminescent microscopy (for tetracycline detection in teeth). The laboratory is accredited according to ISO 17025 and all rabies laboratory techniques used in BIOR have been accredited. Latvia rabies status Latvia has been approved free of rabies since 2015, but vaccination is still carried out close to Latvia’s border with Russia and Belorussia. Now our main task is monitoring of efficiency of ORV, therefore we examine about 1200 samples (from foxes and racoon dogs) per year. In frame of passive surveillance we examine 200 to 300 samples for virus detection and about 200 serological tests for pet international movement per year. Since 2013 research work has been started for investigation of rabies status in bat population, that is the subject of our colleague Marianna Dobroštana’s PhD studies. Contact Responsible expert - Dr.Zita Muižniece: zita.muizniece@bior.lv Virology Division Animal Diseases Diagnostic Laboratory Institute of Food Safety, Animal Health and Environment BIOR Lejupes iela 3, Rīga, LV-1076, LATVIA Website: www.bior.lv   Document in .pdf version  

♦ 29 February 2016 ♦ Every year, the EURL is in charge of elaborating a technical report detailing all rabies activities undertaken and their outputs in the frame of the regulations No 737/2008 and 415/2013. The issue for 2016 (report on year 2015) has just been released and is now online on the EURL for rabies website. Consultation is for members of the EURL network only (restricted access).  

♦ 01 September 2016 ♦ From 1st September 2016, a new model of animal health certificate for pets movements into an EU Member State will have to be used by EU approved Laboratories, in order to avoid further forgery laboratory reports on the results of the rabies antibody titration test following the Commission Implementing Regulation (EU) 2016/561 of 11 April 2016 amending Annex IV to Implementing Regulation (EU) No 577/2013 as regards the model of animal health certificate for dogs, cats and ferrets moved into a Member State from a territory or a third country for non-commercial purposes.