Nosemosis is a microsporidian disease causing mortality and weakening of honey bee colonies, especially in the event of co-exposure to other sources of stress. As a result, the disease is regulated in some countries. Reliable and harmonised diagnosis is crucial to ensure the quality of surveillance and research results. For this reason, the first European Interlaboratory Comparison (ILC) was organised in 2017 in order to assess both the methods and the results obtained by National Reference Laboratories (NRLs) in counting Nosema spp. spores by microscopy. Implementing their own routine conditions of analysis, the 23 participants were asked to perform an assay on a panel of ten positive and negative samples of crushed honey bee abdomens. They were asked to report results from a qualitative and quantitative standpoint. The assessment covered specificity, sensitivity, trueness and precision. Quantitative results were analysed in compliance with international standards NF ISO 13528 (2015) and NF ISO 5725-2 (1994). Three results showed a lack of precision and five a lack of trueness. However, overall results indicated a global specificity of 98% and a global sensitivity of 100%, thus demonstrating the advanced performance of the microscopic methods applied to Nosema spores by the NRLs. Therefore, the study concluded that using microscopy to detect and quantify spores of Nosema spp. was reliable and valid.

AGES Department for Apiculture and Bee Protection Spargelfeldstraß 3 191 1226 Vienna Austria

Finnish Food Safety Authority Evira Veterinary Bacteriology Research Department Neulaniementie 4 70210 Kuopio Finland

French Agency for Food Environmental and Occupational Health and Safety European Union Reference Laboratory for Bee Health Honey Bee Pathology Unit 105 route des Chappes CS 20111 06902 Sophia Antipolis France

Institute for Diagnosis and Animal Health NRL for Honey Bee Diseases and Other Useful Insects Dr N Staicovici street No 63 sector 5 050557 Bucharest Romania

Institute of Food Safety Animal Health and Environment BIOR Lejupes str 3 1076 Riga Latvia

Instituto Nacional Investigaçao Agraria e Veterinaria Rua Genaral Moraes Sarmento 1500 311 Lisbon Portugal

Istituto Zooprofilattico Sperimentale delle Venezie NRL for Beekeeping Viale dell'Università 10 35020 Legnaro PD Italy

Laboratorio Central de Veterinaria Ctra M 106 pK M 104 28110 Algete Spain

Laboratory for Honeybee Diseases APISlab University of Zagreb Faculty of Veterinary Medicine Department for Biology and Pathology of Fish and Bees Heinzelova 55 10000 Zagreb Croatia

Laboratory for Molecular Entomology and Bee Pathology Universiteit Gent Krijgslaan 281 S2 2de verdiep B 9000 Ghent Belgium

National Bee Unit FERA Sand Hutton YO41 ILZ York United Kingdom

National Food and Veterinary Risk Assessment Institute J Kairiukscio 10 LT 08409 Vilnius Lithuania

National Reference Laboratory for Bee Diseases Friedrich Loeffler Institut Südufer 10 17493 Greifswald Insel Riems Germany

National Veterinary Laboratory Abattoir street Albert Town MRS1123 Marsa Malta

National Veterinary Research Institute Department of Honey Bee Diseases 57 Partyzantow Avenue 24 100 Pulawy Poland

NFCSO Veterinary Diagnostic Directorate Molecular Biology Laboratory Tábornok utca 2 1143 Budapest Hungary

NRL Bee Health NDR VMI 15 Pencho Slaveykov Blvd 1606 Sofia Bulgaria

State Veterinary and Food Institute Janoskova 1611 58 026 01 Dolny Kubin Slovakia

State Veterinary Institute Olomouc NRL for Honey Bee Health Jakoubka ze Stribra 1 77900 Olomouc Czech Republic

Swedish University of Agricultural Sciences Department of Ecology Box 7044 750 07 Uppsala Sweden

Thessalonica Veterinary Center Laboratory of Bee Diseases 26th October str 80 54627 Thessalonica Greece

Veterinary and Food Laboratory Kreutzwaldi 30 51006 Tartu Estonia

Veterinary Faculty University of Ljubljana National Veterinary Institute Laboratory for Health Care of Bees Gerbičeva 60 1000 Ljubljana Slovenia

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$a An international inter-laboratory study on Nosema spp. spore detection and quantification through microscopic examination of crushed honey bee abdomens / $c V. Duquesne, C. Gastaldi, A. Del Cont, N. Cougoule, A. Bober, M. Brunain, G. Chioveanu, N. Demicoli, PD. Paulus, PF. Somalo, M. Filipova, E. Forsgren, A. Granato, K. Gurgulova, S. Heinikainen, A. Kärssin, I. Kinduriene, H. Köglberger, K. Oureilidis, Z. Ozolina, M. Pijacek, MP. Ocepek, MO. Schäfer, IT. Gajger, MJ. Valerio, M. Wakefield, S. Franco

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