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OIE Reference Laboratory Reports Activities Activities in 2017 This report has been submitted : 2018-01-13 06:37:59 Name of disease (or topic) for which you are a designated OIE Reference Laboratory: Brucellosis (Brucella abortus, B. melitensis, B. suis) Address of laboratory: Istituto Zooprofilattico Sperimentale dell Abruzzo e del Molise, Via Campo Boario 64100 Teramo ITALY Tel.: +39 348 52 13 201 Fax: +39 0861 33 22 51 E-mail address: f.demassis@izs.it Website: www.izs.it Name (including Title) of Head of Laboratory (Responsible Official): Prof Mauro Mattioli, DVM, Professor, General Director Name (including Title and Position) of OIE Reference Expert: Dr Fabrizio De Massis, DVM Which of the following defines your laboratory? Check all that apply: Governmental Research OIE Reference Laboratory Reports Activities, 2017 1

ToR 1: To use, promote and disseminate diagnostic methods validated according to OIE Standards 1. Did your laboratory perform diagnostic tests for the specified disease/topic for purposes such as disease diagnosis, screening of animals for export, surveillance, etc.? (Not for quality control, proficiency testing or staff training) 2 OIE Reference Laboratory Reports Activities, 2017

Diagnostic Test Indicated in OIE Manual (/No) Total number of test performed last year Indirect diagnostic tests Nationally Internationally RBT 319,675 1,245 CFT 20,074 1,245 ELISA 28 0 I-ELISA (Bovine serum) 18 0 C-ELISA (Bovine / Ovine / Caprine sera) 7 1,243 C-ELISA (pig serum) 103 0 Milk ELISA 30 0 MIlk Ring Test 17 0 CFT - RB51 3030 0 SAT B. canis No 11 0 Direct diagnostic tests Nationally Internationally Bacterial isolation 1,066 0 Brucella identification and biotyping (human strains) 1 0 PCR 1,218 143 RT-PCR 0 143 PCR-RFLP (identification) 524 143 MLVA 22 34 MLST 65 143 WGS (SNP, cgmlst) 65 143 ToR 2: To develop reference material in accordance with OIE requirements, and implement and promote the application of OIE Standards. To store and distribute to national laboratories biological reference products and any other reagents used in the diagnosis and control of the designated pathogens OIE Reference Laboratory Reports Activities, 2017 3

or disease. 2. Did your laboratory produce or supply imported standard reference reagents officially recognised by the OIE? No 3. Did your laboratory supply standard reference reagents (non OIE-approved) and/or other diagnostic reagents to OIE Member Countries? 4 OIE Reference Laboratory Reports Activities, 2017

Type of reagent available Related diagnostic test Produced/ provide Amount supplied nationally (ml, mg) Amount supplied internationally (ml, mg) No. of recipient OIE Member Countries Region of recipients Rose Bengal Antigen RBT 168,200 135,040 80 1 Brucella RB51 Antigen CFT-RB51 2000 450 Milk Ring Test Antigen Milk Ring Test 150 20 0 1 Negative Buffalo Serum for Brucella abortus Negative control for all serological methods 20 1 Positive Bovine Serum for Brucella abortus CFT 740 Negative Standard Bovine Serum for Brucella abortus Negative control for all serological methods 404 OIE Reference Laboratory Reports Activities, 2017 5

National Standard Serum for Brucella abortus Positive control for all serological methods 114 Negative Bovine Milk for B. abortus MRT for control and ring trial 300 Positive Bovine Milk for B. abortus MRT for control and ring trial 1,000 300 Brucella spp. Strains 3 strains Brucella CFT Antigen CFT 500 127 4. Did your laboratory produce vaccines? No 5. Did your laboratory supply vaccines to OIE Member Countries? No ToR 3: To develop, standardise and validate, according to OIE Standards, new procedures for diagnosis and control of the designated pathogens or diseases 6. Did your laboratory develop new diagnostic methods validated according to OIE Standards for the designated pathogen or disease? 6 OIE Reference Laboratory Reports Activities, 2017

No 7. Did your laboratory develop new vaccines according to OIE Standards for the designated pathogen or disease? No ToR 4: To provide diagnostic testing facilities, and, where appropriate, scientific and technical advice on disease control measures to OIE Member Countries 8. Did your laboratory carry out diagnostic testing for other OIE Member Countries? Name of OIE Member Country seeking assistance Date (month) No. samples received for provision of diagnostic support No. samples received for provision of confirmatory diagnoses NIGERIA January 1,243 sera 1,243 sera ZIMBABWE May 2,152 sera 2,152 sera 9. Did your laboratory provide expert advice in technical consultancies on the request of an OIE Member Country? Name of the OIE Member Country receiving a technical consultancy Purpose How the advice was provided TUNISIA Workshop on diseases of camelids Participation to the workshop as facilitator ZIMBABWE Epidemiological investigations by analysis of Zimbabwean sera Collaboration with the Central Veterinary Laboratory of Zimbabwe FAO/ IAEA Funded training to IZSAM for a colleague from Zimbabwe NIGERIA Epidemiological investigations by analysis of Nigerians sera Collaboration with the Central Veterinary Laboratory of Nigeria LEBANON OIE Twinning Project Final audit and final meeting with neighboring countries LEBANON workshop on HACCP system and audit procedures in dairy industry Participation to the workshop as facilitator OIE Reference Laboratory Reports Activities, 2017 7

ToR 5: To carry out and/or coordinate scientific and technical studies in collaboration with other laboratories, centres or organisations 10. Did your laboratory participate in international scientific studies in collaboration with OIE Member Countries other than the own? Title of the study Duration Purpose of the study Partners (Institutions) OIE Member Countries involved other than your country Epidemiological investigations by analysis of Nigerians strains and sera 1 year Epidemiological investigations by analysis of Nigerians sera Central Veterinary Laboratory Nigeria NIGERIA Epidemiological investigations by analysis of Zimbabwean sera 1 year Epidemiological investigations by analysis of Zimbabwean sera with RBT abd CFT Central veterinary Laboratory-Harare ZIMBABWE OIE Twinning between Ministry of Agriculture of Lebanon and Ministry of Health of Italy, through the Istituto Zooprofilattico Sperimentale dell Abruzzo e del Molise G. Caporale (IZSAM) as OIE Collaborating Centre for Veterinary Training, Epidemiology, Food Safety and Animal Welfare 3 years Twinning project Ministry of Agriculture of Lebanon Ministry of Health of Italy Istituto Zooprofilattico Sperimentale dell Abruzzo e del Molise G. Caporale LEBANON OIE Twinning contract between the OIE reference network of Istituti Zooprofilattici Sperimentali (IIZZSS) in Italy (Parent Collaborating Centre) and the Abu Dhabi Food Control Authority (ADFCA), the Animal Health Centre for Diagnostic and research in Abu Dhabi United Arab Emirates (Candidate Centre)forthe establishment and development of an OIE Collaborating Centre on camel diseases. 3 years Twinning project Abu Dhabi Food Control Authority ADFCA Italian IIZZSS Network UNITED ARAB EMIRATES 8 OIE Reference Laboratory Reports Activities, 2017

ToR 6: To collect, process, analyse, publish and disseminate epizootiological data relevant to the designated pathogens or diseases 11. Did your Laboratory collect epizootiological data relevant to international disease control? 12. Did your laboratory disseminate epizootiological data that had been processed and analysed? 13. What method of dissemination of information is most often used by your laboratory? (Indicate in the appropriate box the number by category) a) Articles published in peer-reviewed journals: 5 1. Barbara Bonfini, Giuseppina Chiarenza, Valentina Paci, Flavio Sacchini, Romolo Salini, Gesualdo Vesco, Sara Villari, Katiuscia Zilli, Manuela Tittarelli. Cross-reactivity in serological tests for brucellosis: comparison of immune response of Escherichia coli O157:H7 and Yersinia enterocolitica O:9 vs Brucella spp. (2017) (Accettato e in stampa su Veterinaria Italiana). 2. Savini L., Candeloro L., Conte A., De Massis F., Giovannini A., 2017. Development of a forecasting model for brucellosis spreading in the Italian cattle trade network aimed to prioritise the field interventions. PLoS ONE 12(6): e0177313 https://doi.org/10.1371/journal.pone.0177313 3. Di Sabatino D., Garofolo G., Di Provvido A., Zilli K., Foschi G., Di Giannatale E., Ciuffetelli M., De Massis F., 2017. Brucella suis biovar 2 MLST ST16 in wild boars from Abruzzi region. Introduction from ern? Infection, Genetics and Evolution, 55: 63-67. doi: 10.1016/j.meegid.2017.08.031 4. Garofolo G, Di Giannatale E, Platone I, Zilli K, Sacchini L, Abass A, Ancora M, Cammà C, Di Donato G, De Massis F, Calistri P, Drees KP, Foster JT. Origins and global context of Brucella abortus in Italy. 2017 BMC Microbiol. 2017 Feb 2;17(1):28. doi: 10.1186/s12866-017-0939-0. 5. Ndengu M., Matope G., de Garine-Wichatitsky, Tivipasi M., Scacchia M., Bonfini B., Pfukenyi D., 2017. Seroprevalence of brucellosis in cattle and selected wildlife species at selected livestock/wildlife interface areas of the Gonarezhou national Park, Zimbabwe. Preventive Veterinary Medicine 146 (2017), 158-166. doi:10.1016/j.prevetmed.2017.08.004 b) International conferences: 2 1 F. De Massis, K. Zilli, M. Ancora, G. Di Donato, L. Sacchini, E. Di Giannatale. Update about typing of Brucella field strains isolated from livestock and wildlife populations in Italy, 2007-2015 Epidemics6, Sixth International Conference on Infectious Diseases Dynamics, 29th November 1st December Sitges, Spain. 2 Paola Roncada, Cristian Piras, Isabella Alloggio, Viviana Greco, Tiziana Di Febo, Ivanka Krasteva, Andrea Urbani, Giuliano Garofolo, Manuela Tittarelli, Luigi Bonizzi, Alessio Soggiu. Proteomics to improve serodiagnosis of Brucellosis. Proceeding of XII Annual Conference of Italian Proteomics Association. June 12th-15th 2017. Pag.43 ISBN978-88-7959-961-0 c) National conferences: 1 1 Garofolo G., Munoz P. M., Blasco J. M., Jay M., Mick V., Adone R., Di Giannatale E., Zilli K., Sacchini L., de Miguel M. J., De Massis F., Scacchia M., Tittarelli M, 2017. Origins and spread of brucellosis in wildlife and domestic animals in. Networking: tool for an excellent Research. Atti del IV Convegno Nazionale sulla Ricerca in Sanità Pubblica Veterinaria Roma, 6 aprile 2017. Veterinaria Italiana, Collana di Monografie, Monografia n. 26, P78. ISSN 0505-401x. d) Other: (Provide website address or link to appropriate information) 1 Collective, 2017. Demande d expertise sur l évaluation approfondie et réactualisée de mesures de maîtrise du foyer de brucellose chez les bouquetins du Bargy Saisine «2016-SA-0229 brucellose des bouquetins du Bargy» - AVIS de l Agence nationale de sécurité sanitaire de l alimentation, de l environnement et du travail relatif à «l évaluation approfondie et réactualisée de mesures de maîtrise du foyer de brucellose chez les bouquetins du Bargy» https://www.anses.fr/fr/system/files/saba2016sa0229.pdf OIE Reference Laboratory Reports Activities, 2017 9

ToR 7: To provide scientific and technical training for personnel from OIE Member Countries To recommend the prescribed and alternative tests or vaccines as OIE Standards 14. Did your laboratory provide scientific and technical training to laboratory personnel from other OIE Member Countries? a) Technical visits: 2 b) Seminars: 3 c) Hands-on training courses: 0 d) Internships (>1 month): 0 Type of technical training provided (a, b, c or d) Country of origin of the expert(s) provided with training No. participants from the corresponding country a Zimbabwe 1 a Zimbabwe 2 b Algeria, Angola, Botswana, Libya, Mauritania, Morocco, Namibia, South, Tanzania, Tunisia, Zambia and Zimbabwe 87 b Etiopia 25 b Italy 60 b United Arab Emirates 60 ToR 8: To maintain a system of quality assurance, biosafety and biosecurity relevant for the pathogen and the disease concerned 15. Does your laboratory have a Quality Management System certified according to an International Standard? Quality management system adopted ISO 17025 Certificate scan (PDF, JPG, PNG format) Certificato Accreditamento 2017.pdf 16. Is your laboratory accredited by an international accreditation body? 10 OIE Reference Laboratory Reports Activities, 2017

Test for which your laboratory is accredited Complement Fixation test (CFT) RBT i-elisa (milk and serum) FPA CFT - RB51 Brucella spp. Isolation Brucella spp. Isolation from food matrix Identification and typing c-elisa Accreditation body 17. Does your laboratory maintain a biorisk management system for the pathogen and the disease concerned? (See Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, Chapter 1.1.4) ToR 9: To organise and participate in scientific meetings on behalf of the OIE 18. Did your laboratory organise scientific meetings on behalf of the OIE? National/ International Title of event Co-organiser Date (mm/yy) Location No. Participants National National workshops on Brucellosis diagnosis and epidemiology Ministry of Health, Italy 10/17 IZSAM, Teramo-Italy 60 International IZSAM in : the strengthening of research capacity through the consolidation of a collaborative network March 2017 IZSAM, Teramo-Italy 86 International Strengthening ERFAN through the support of OIE Reference Laboratories/CollaboratingCentres in the SADC Region University of Pretoria, South October 2017 Faculty of Veterinary Science, Pretoria 30 19. Did your laboratory participate in scientific meetings on behalf of the OIE? OIE Reference Laboratory Reports Activities, 2017 11

Title of event Date (mm/yy) Location Role (speaker, presenting poster, short communications) Title of the work presented Workshop on diseases of camelids 12/17 Tunisia Expert and facilitator Eléments nécessaires pour mettre un réseau de surveillance de santé animale pour les camelins Consultant Meeting on Advanced Diagnostic Technologies for Detection and Differentiation of Animal and Zoonotic Diseases 04/17 Vienna Speaker The use of Whole Genome Sequencing and Multiple Locus Variablenumber Tandem Repeat Analysis in the Understanding of the Epidemiology of Brucellosis Transboundary Animal Diseases Diagnosis: Early Detection and Characterization Focus on the differential diagnosis of major transboundary and zoonotic animal diseases. Zoonotic diseases Brucella 10/2017 Ethiopia Speaker Epidemics6, Sixth International Conference on Infectious Diseases Dynamics 10/17 Sitges, Barcelona, Spain presenting poster Update about typing of Brucella field strains isolated from livestock and wildlife populations in Italy, 2007-2015 18th International Symposium of The World Association Of Veterinary Laboratory Diagnosticians (WALVD) 06/17 Sorrento, Italy presenting poster The population structure of brucella ovis: insights from italian genomes. XII Annual Conference of Italian Proteomics Association 06/17 Lecce, Italy presenting poster Proteomics to improve serodiagnosis of Brucellosis IV Convegno Nazionale sulla Ricerca in Sanità Pubblica Veterinaria 04/17 Roma, Italy speaker Origins and spread of brucellosis in wildlife and domestic animals in ToR 10: To establish and maintain a network with other OIE Reference Laboratories designated for the same pathogen or disease and organise regular inter-laboratory proficiency testing to ensure comparability of results 12 OIE Reference Laboratory Reports Activities, 2017

20. Did your laboratory exchange information with other OIE Reference Laboratories designated for the same pathogen or disease? 21. Was your laboratory involved in maintaining a network with OIE Reference Laboratories designated for the same pathogen or disease by organising or participating in proficiency tests? Purpose of the proficiency tests: 1 Role of your Reference Laboratory (organiser/ participant) No. participants Participating OIE Ref. Labs/ organising OIE Ref. Lab. 2017 ILPT Brucella bacteriological and molecular identification methods, organised by EU RL ANSES Participant 27 EU Countries ANSES France EU RL for Brucellosis EU Bovine Brucellosis Serum Proficiency Test 2016 organised by EU RL ANSES (feed-back results and follow-up received on 2017 Participant 27 EU Countries ANSES France EU RL for Brucellosis 1 validation of a diagnostic protocol: specify the test; quality control of vaccines: specify the vaccine type, etc. 22. Did your laboratory collaborate with other OIE Reference Laboratories for the same disease on scientific research projects for the diagnosis or control of the pathogen of interest? Title of the project or contract Scope Name(s) of relevant OIE Reference Laboratories ARIMNET (BrucMedNet) Improvement of epidemiological and serological tools for diagnosis and control of Brucellosis in the Mediterranean region. IZSAM Scientific coordinator (OIE RL and NRL for Brucellosis)-Italy, Portugal, Greece, Tunisia, Egypt, Spallanzani Institute, Milan- Italy and IZS Sicily-Italy ANIHWA BRUEpidia Brucellosis in wildlife and domestic animals: development of a direct diagnostic method and assessment of genetic diversity of Brucella melitensis and Brucella suis biovar 2 strains in the EU IZSAM (OIE RL and NRL for Brucellosis)- Italy, Spain, France (OIE and EU RL for Brucellosis) ANIHWA (EpiRisk) Integrated epidemiological models for risk based surveillance approaches. IZSAM Scientific coordinator (OIE RL and NRL for Brucellosis)- Italy, Spain (IRTA; Barcellona), The Netherlands (CVI, part of Wageningen UR now called WBVR (Wageningen BioVeterinary Research), Israel (Kimron Veterinary Institute) OIE Reference Laboratory Reports Activities, 2017 13

ToR 11: To organise inter-laboratory proficiency testing with laboratories other than OIE Reference Laboratories for the same pathogens and diseases to ensure equivalence of results 23. Did your laboratory organise or participate in inter-laboratory proficiency tests with laboratories other than OIE Reference Laboratories for the same disease? Note: See Interlaboratory test comparisons in: Laboratory Proficiency Testing at: http://www.oie.int/en/our-scientific-expertise/reference-laboratories/proficiency-testing see point 1.3 Purpose for inter-laboratory test comparisons 1 No. participating laboratories Region(s) of participating OIE Member Countries 11th Italian Brucellosis microbiological ring trial for culture and sensitivity from various matrix 19 18th Brucellosis Italian Proficiency Ring-Trial serum. Laboratory capability to conduct specific diagnostic tests 74 5th Italian Brucellosis microbiological ring trial for culture and sensitivity from food matrix 8 1st International Brucella MLVA ring trial 5 ToR 12: To place expert consultants at the disposal of the OIE 24. Did your laboratory place expert consultants at the disposal of the OIE? 14 OIE Reference Laboratory Reports Activities, 2017

Kind of consultancy Location Subject (facultative) Expert on disease surveillance systems and facilitator in the framework of a workshop on diseases of camelids Tunisia Participation to the workshop as expert identified by the OIE 25. Additional comments regarding your report: - Organised by EFSA; Participation to the Animal Health and Welfare Network Task Force on Zoonosis Data Collection; - Organised by EFSA; Participation to the Working group on Zoonosis SINZOO; - Organised by EU; Chairmanship and participation to the Task force on monitoring animal disease eradication programmes in Member States Brucellosis subgroup. OIE Reference Laboratory Reports Activities, 2017 15