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OIE Reference Laboratory Reports Activities in 2010 Name of disease (or topic) for which you are a designated OIE Reference Laboratory: Address of laboratory Bee diseases Chemisches Veterinäruntersuchungsamt Freiburg P.O.B. 100462, D-79123 Freiburg, Germany Tel.: (+49-761) 150.21.75 Fax: (+49-761) 150.22.99 e-mail address: [email protected] website: Name of Head of Laboratory (Responsible Official): Dr Wolfgang Ritter Name of OIE Reference Expert: Dr Wolfgang Ritter Name of writer of this report (if different from above): Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 1 Bee diseases 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 Disease Test Positive Negative Total Acariosis Microscopic 1 87 88 American foulbrood Culture tech. 264 1428 1692 RT-PCR1 30 10 40 European foulbrood Culture tech 16 13 29 Nosemosis Microscopic 29 64 93 Varroosis Macroscopic 115 16 131 Tropilaelaps spp. Macroscopic 0 115 115 Small hive beatle Macroscopic 0 112 112 Acute Bee Paralysis Virus (ABPV) RT-PCR2 16 84 100 Chronical Bee Paralysis Virus (CBPV) RT-PCR1 0 12 12 Deformed wing virus (DWV) RT-PCR2 68 32 100 Kashmir bee virus (KBV) RT-PCR1, 2 0 10 10 Sac brood virus (SBV) RT-PCR1, 2 6 94 100 1Complete body; 2Heads only Diagnosis of bee diseases refers to parasites, bacteria, fungi and viruses as agents As parasites are recognised the ectoparasitic mites Varroa destructor and Acarapis woodi as well as the microsporidium Nosema spp. All of them are diagnosed exclusively by macroscopic or microscopic examinations and identified on the basis of their morphological qualities. Bacterial diseases mainly occur in the bee brood. Paenibacillus larvae (formerly called Bacillus larvae), the agent of American Foulbrood, and Melissococcus pluton, the agent of European Foulbrood, are cultivated on special nutrient media and then identified under the microscope. After being cultivated on a nutrient medium, the agents of the most important fungus diseases, Ascophaera apis and Aspergillus flavus, can easily be distinguished by microscopy. Some of the agents can be analyzed by PCR technique. Bee viruses can be identified by PCR. The respective tests were performed in the frequency shown in the table. 2. Production and distribution of diagnostic reagents For the diagnosis of European Foulbrood and American Foulbrood, strains of P.l. and M.p., respectively, were given to different diagnostic laboratories as reference strains. On the national level another institute in our order produces and distributes standards for detection of P.l. in honey. The diagnosis of P.l. and various viruses was performed for different countries. 2 Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 Bee diseases 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 The CVUA Freiburg as national reference laboratory for notifiable bee diseases continued the interlaboratory test with further testing institutions in different countries. An international interlaboratory test for different OIE members has been prepared for the year 2011. 4. Preparation and supply of international reference standards for diagnostic tests or vaccines In cooperation with other institutes different reference materials were created. 5. Research and development of new procedures for diagnosis and control Paenibacillus larvae bacteremia in humans In a collaborative research program with the University Hospital Freiburg, Germany, P. larvae was detected in sterile compartments of 5 patients with clinical and laboratory evidence of infection. Given the fact that all patients were injection drug users, the mode of infection was thought to be intravenous administration of contaminated methadone, resulting in P. larvae bacteremia. Our hypothesis is supported by the isolation of P. larvae from honey or honey-containing methadone provided to 2 patients. Recently, several Paenibacillus species have been reported to cause bacteremic infections in humans. Several aspects provide strong evidence for a genuine P. larvae bacteremia. First, the present cases were observed over a period of several years, and detection of P. larvae thus occurred in different charges of blood culture bottles, which argues against pseudobacteremia. Second, isolation of P. larvae was reported independently by two microbiology laboratories, making contamination highly unlikely. Third, in patient 1 isolation succeeded at different times and in samples of different compartments. Moreover, the detection of P. larvae in honey-prepared methadone and honey strongly suggests genuine bacteremia as a consequence of injection of contaminated material. Biochemical and molecular identification of P. larvae may be difficult and time-consuming. Misinterpretation of blood culture results because of incomplete differentiation or confusion with other gram-positive spore forming-bacteria (e.g., Bacillus species) has to be taken into consideration. Underestimation of the frequency of true P. larvae bacteremia therefore cannot be excluded. Thus, infectious disease physicians, microbiologists, and pharmacists need to be aware that injection of material contaminated with P. larvae, such as honey-prepared methadone, may cause bacteremic infection. Diagnosis of Small Hive Beatle In 2010, the year under review, diagnosing methods of different diseases were verified. The method for diagnosing the Small Hive Beetle in the bee colony developed by us (Schäfer et al, 2009) proved to be efficient and easily applicable, both for veterinarians and animal owners. Therefore, this method is recommended for general practice. Diagnosis of Paenibacillus larvae in Debris of bee colonies The examinations on the evidence of Paenibacillus larvae, the cause of American Foulbrood, were extended to the analysis of faeces of bee colonies. A method was developed at the Institute in Dole (CSR). According to first results, this method could be suitable especially for the examination of bee colonies in winter cluster. A final conclusion, however, will only be possible after termination and evaluation of the complete field test next year. Pathogenesis of European Foulbrood in Germany and Switzerland The examinations on different variations of the pathogenesis of European Foulbrood have been continued. Whereas this disease has become more and more epidemic in Switzerland, it is regionally restricted in Germany and only rarely spread within neighboring apiaries. As flight routes of the bees from both areas are overlapping, different types of pathogens can be excluded as a reason. According to the results achieved until now, management method and hive types could have an influence on the epidemic process. The examinations are going to be continued and intensified, especially along the borderline of both countries. Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 3 Bee diseases 6. Collection, analysis and dissemination of epizootiological data relevant to international disease control Data on bee losses in the EU and other parts of the world were collected. 7. Provision of consultant expertise to OIE or to OIE Members The OIE expert Dr W. Ritter participated at various hearings of experts at the Veterinary Board of the European Union. 8. Provision of scientific and technical training to personnel from other OIE Members The OIE expert, Dr Ritter, participated in the establishment of diagnostic methods at various laboratories. 9. Provision of diagnostic testing facilities to other OIE Members The Service was requested from Austria, Italy, Switzerland, Czech Republic and Turkey on an individual level (institutes, companies, organisations). 10. Organisation of international scientific meetings on behalf of OIE or other international bodies In 2010, the year under review, the 42 th International Congress of Apimondia at Buenos Aires (Argentine) was prepared. More than 4000 participants are expected in harvest 2011. Among other topics, a total number of four symposiums are planned about different topics of bee health. In the week before the congress, an OIE symposium will be held. The diagnosis and control of Varoosis and the American Foulbrood will be the focal points. They are organized in cooperation with local scientists and collaborators of the veterinary authorities. In cooperation with an NGO and by order of the European Commission, the author has been offering training courses for veterinarians working in the civil service. During the one week’s course at Prague (Czech Republic) the basic facts about diagnosis and control of the most important bee diseases as well as the most important official and international regulations are taught. In the afternoons, staff members of the Czech Institute at Dole will demonstrate the contexts in practice. During the year under review, two courses with 45 participants each from different countries of the European Union and neighboring countries were realized. Another two courses are planned for the following year. 11. Participation in international scientific collaborative studies COLOSS/COST Action As official delegate of the Federal Republic of Germany, the author participated at different activities of the Colloss Project funded by the European Union (COST) since 2009. 12. 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 Ritter W. Different statements and presentations on bee losses at different meetings. 4 Scientific publications in peer-reviewed journals Conte Y., Marion E., Ritter W. Varroa mites and honey bee health: can Varroa explain part of the colony losses? Apidologie 41 (2010) 3353-363* Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 Bee diseases Rieg S., Bauer T.M., Peyerl-Hoffmann G., Held J., Ritter W., Wagner D., Kern W.V., Serr A. 2010. Paenibacillus larvae Bacteremia in Injection Drug Users. Emerging Infectious Diseases (16) 3: 487-489 Genersch E., von der Ohe W., Kaatz H., Schroeder A., Otten Ch., Büchler R., Berg S., Ritter W., Mühlen W., Gsider S., Meixner M. Liebig G., Rosenkranz P. 2010. The German bee monitporing project: a long term study to understand periodically high wintrer losses of honey bee colonies. Apidologie: 41:332-352. Schaefer, M., Ritter, W., Pettis, J.S., Teal, P.E., Neumann, P. 2010. The role of infestations with Aethina tumida and Varroa destructor. J. Econ. Entomol. 1o3 (1):10-16 Other communications Several statements, expert opinion and consultancy reports 13. Inscription of diagnostic kits on the OIE Register i) Did you participate in expert panels for the validation of candidate kits for inscription on the OIE Register? If yes, for which kits? None ii) Did you submit to the OIE candidate kits for inscription on the OIE Register? If yes, for which kits? None _______________ Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 5