OIE RL for West Nile Encephalitis Federica Monaco Italy

OIE Reference Laboratory Reports Activities in 2012

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

Address of laboratory Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise “G. Caporale” (IZSAM), Via Campo Boario, 64100 Teramo, ITALY

Tel.: (0039) 0861 33231

Fax: (0039) 0861 332251

e-mail address: [email protected]

website: www.izs.it

Fernando Arnolfo Name (including Title) of Head of General Director Laboratory (Responsible Official):

Federica Monaco Name(including Title and Position) of OIE Head of the diagnosis and surveillance of Reference Expert: the exotic viral diseases of animals laboratory Date of submission to the OIE January 25th, 2013

Instructions

This form should be used by an OIE Reference Laboratory to report activities that took place from January through December of the past year (2012), unless otherwise stated, and must be submitted by the end of January every year.

Only those activities that concern the disease (or topic) for which the laboratory is recognised by the OIE should be mentioned. The questionnaire structure follows the Terms of Reference (ToRs) for OIE Reference Laboratories, available at: http://www.oie.int/en/our-scientific-expertise/reference-laboratories/introduction/

Each ToR(blueitalicised text)has been placed as a heading covering the group of questions related to it.

Please note the reditalicisedtext is given as guidance and should be deleted from your report and substitute with your data. Examples are based onpast Annual Reports or have been invented.

The questionnaire represents a means of gathering information on activities carried out by OIE Reference Laboratories and making it available to OIE Member Countries and to the OIE Reference Laboratory network. This annual report will remain available for consultation on the OIE web site: (http://www.oie.int/en/our-scientific-expertise/reference-laboratories/annual-reports/):

ToR: To use, promote and disseminate diagnostic methods validated according to OIE Standards

Annual reports of OIE Reference Laboratories, 2012 1 OIE RL for « West Nile Encephalitis » – « Federica Monaco » – « Italy »

Test recommended by the OIE Total number of test performed last year

Indirect diagnostic tests Nationally Internationally

ELISA IgG 6139 tests 182 tests (Bolivia and Peru)

ELISA IgM 992 tests 194 tests (Bolivia, Peru and Croatia) Virus neutralization (microtitre format) 3514 tests 194 tests Plaque Reduction neutralization test 571 tests 194 tests (PRNT)

Direct diagnostic tests Nationally Internationally

Cell Culture (Vero) 49 tests - Real-time RT-PCRWNV lineage 1 76 tests - Real-time RT-PCRWNV lineage 2 58 tests - Real-time RT-PCRWNV lineages 1 and 2 7120 tests 46 tests (Tunisia) IF (Flavivirus Group) 49 tests - Genome sequencing 6 partial and 3 full genome - sequences Mosquitoes morphological identification 776 samples 154 samples (Tunisia)

ToR: To develop reference material in accordance with OIE requirements, and implement and promote the application of OIE Standards. Tostore and distribute to national laboratories biological reference products and any other reagents used in the diagnosis and control of the designated pathogens or disease.

2. Did your laboratory produce or store imported standard reference reagents officially recognised by the OIE or other international bodies?

X Yes No

3. Did your laboratory supply standard reference reagents to OIE Member Countries?

X Yes No

Name of Related Amount supplied Amount supplied recipient OIE Type of reagent Produced/ diagnostic nationally internationally Member available stored test (ml, mg) (ml, mg) Countries and of institutions Control positive Virus serum neutralizat ion Produced 1 ml - - (microtitre format) and PRNT Control negative Virus Produced 1 ml - - serum neutralizat

2 Annual reports of OIE Reference Laboratories, 2012 OIE RL for « West Nile Encephalitis » – « Federica Monaco » – « Italy »

Name of Related Amount supplied Amount supplied recipient OIE Type of reagent Produced/ diagnostic nationally internationally Member available stored test (ml, mg) (ml, mg) Countries and of institutions ion (microtitre format) and PRNT WNV monoclonal IF Produced 600 µl - - antibody Positive antigen for ELISA test ELISA IgG Produced 10 ml - - (Eg101 strain) Negative antigen ELISA IgG Produced 10 ml - - for ELISA test WNV strain Real-time Eg101 RT-PCR Italy (Italian Produced 200 ml - and Cell IIZZSS) Culture WNV strain B956 Real-time RT-PCR Italy (Italian Produced 60 ml - and Cell IIZZSS) Culture

4. Did your laboratory produce diagnostic reagents other than the OIE-approved standard reference reagents? X Yes No

5. Did your laboratory produce vaccines?

Yes X No

6. Did your laboratory supply vaccines to OIE Member Countries?

Yes X No

ToR: To develop, standardise and validate, according to OIE Standards, new procedures for diagnosis and control of the designated pathogens or diseases 7. Did your laboratory develop new diagnostic methods validated according to OIE Standards for the designated pathogen or disease? X Yes No

8. Did your laboratory develop new vaccines according to OIE Standards for the designated pathogen or disease? Yes X No

Name of the new test or diagnostic Description and References (Publication, website, etc.) method or vaccine developed

Virus neutralisation test in microtiter format OIE Manual chapter on West Nile Fever

ToR: To provide diagnostic testing facilities, and, where appropriate, scientific and technical advice on disease control measures to OIE Member Countries

9. Did your laboratory carry out diagnostic testing for other OIE Member Countries?

X Yes No

Name of OIE Date No. samples received for No. samples received for Member Country (dd/mm) provision of diagnostic support provision of confirmatory seeking assistance (i.e. from surveillance campaign) diagnoses Namibia 20/01 2 samples -

Peru 06/06 - 116 samples

Bolivia 20/09 - 66 samples

Croatia 18/06 - 12 samples

Tunisia 10/09 46 samples -

10. Did your laboratory provide expert advice in technical consultancies on the request of an OIE Member Country?

X Yes No

Name of the OIE Member Country How the advice was Purpose receiving a technical consultancy provided Centro de Diagnóstico de Sanidad Interpretation of serological results Remote assistance Animal SENASA, Perù collected during the surveillance activities Etlik Veterinary Control Central To provide support to the Turkish Institute Twinning project Research Institute , Ankara, Turkey to enhance capacity and improve scientific capabilities in relation to diagnosis and surveillance of West Nile Disease Croatian veterinary Institute, Split, Support to interpret the diagnostic results Remote and in loco Croatia and to draw a national surveillance plan assistance

ToR: To carry out and/or coordinate scientific and technical studies in collaboration with other laboratories, centres or organisations 11. Did your laboratory participate in international scientific studies in collaboration with OIE Member Countries other than the own? X Yes No

OIE Member Countries Title of the study Duration Purpose of the study Partners (Institutions) involved other than your country Origin and evolution 3 years Assessment of the Institut de la Recherche Tunisia of recent vector- role played by the Vétérinaire Morocco borne virus most common Biopharma Croatia

incursions in the species of migrating Croatian Veterinary Mediterranean Basin birds in the new Institute recent WNV and USUV incursions; Study on the prevalence and possible genetic evolution of USUV and WNV isolates circulating in Italy and in Mediterranean Countries correlated to their pathogenicity; Development of diagnostic techniques to identify and characterize the different strains of Flaviviruses. Transnational 3 years Development and Etlik Central Veterinary Turchia network for early validation of a Control and Research Morocco detection of network for the Institute; Tunisia emerging animal collection and the Biopharma; diseases in the analysis of data Institut de la Recherche Mediterranean Basin generated from Vétérinaire; surveillance systems; National Center for Analysis of Zoosanitary Vigilance. environmental variables related to vector/disease association; Evaluation of inactivated and recombinant WNV vaccines used in Morocco.

ToR: To collect, process, analyse, publish and disseminate epizootiological data relevant to the designated pathogens or diseases

12. Did your Laboratory collect epizootiological data relevant to international disease control? X Yes No

13. Did your laboratory disseminate epizootiological data that had been processed and analysed?

X Yes No

14. 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

b) International conferences:...... 7 c) National conferences:...... 2 d) Other:...... internet

 For publications see annex: “List of publications”

ToR: To provide scientific and technical training for personnel from OIE Member Countries

To recommend the prescribed and alternative tests or vaccines as OIE Standards

15. Did your laboratory provide scientific and technical training to laboratory personnel from other OIE Member Countries?

X Yes No

a) Technical visits:...... b) Seminars:...... c) Hands-on training courses:...... d) Internships (>1 month):......

Type of technical training Country of origin of the expert(s) No. participants from the provided (a, b,c or d) provided with training corresponding country

c Thailand 1 c Tunisia 1

ToR: To maintain a system of quality assurance, biosafety and biosecurity relevant for the pathogen and the disease concerned

16. Does your laboratory have a Quality Management System certified according to an International Standard?

X Yes No

Quality management system adopted

ISO/IEC 17025

17. Is your laboratory accredited by an international accreditation body?

X Yes No

Test for which your laboratory is accredited Accreditation body ELISA IgG ILAC (Accredia) Plaque Reduction neutralization test (PRNT) ILAC (Accredia) Virus neutralization (microtitre format) ILAC (Accredia)

Real-time RT-PCR WNV lineage 1 ILAC (Accredia)

18. 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 2012, Chapter 1.1.3 or Manual of Diagnostic Tests for Aquatic Animals 2012, Chapter 1.1.1)

X Yes No

ToR: To organise and participate in scientific meetings on behalf of the OIE

19. Did your laboratory organise scientific meetings on behalf of the OIE? X Yes No

National/ Title of event Co- Date Locatio No. International organiser (mm/yy) n Participants

International Kick-Off Meetings: “Origin and - 04/12 Teram 22 meeting evolution of recent vector-borne o, Italy virus incursions in the Mediterranean Basin” and “Transnational network for early detection of emerging animal diseases in the Mediterranean Basin”

20. Did your laboratory participate in scientific meetings on behalf of the OIE? X Yes* No

*The table content refers to the participation in international meetings

Title of event Date Location Role (speaker, presenting Title of the work (mm/yy) poster, short presented communications)

Chronic Wasting 06/12 Seoul, South Speaker Current West Nile Disease Disease Korea Status and Research 6th Annual Meeting 06/12 Brighton, UK Presenting poster West Nile Disease in EPIZONE “Viruses on Sardinian wild birds the move” 6th Annual Meeting 06/12 Brighton, UK Presenting poster West Nile and Usutu virus EPIZONE “Viruses on vectors in Italy the move” 15th International 06/12 Bangkok, Presenting poster Importance of dogs as Congress of Infectious Thailand sentinels of West Nile virus Diseases activity in urban and suburban areas 15th International 06/12 Bangkok, Presenting poster West Nile Disease (WND) in Congress of Infectious Thailand Sicily Diseases Endemic and Emerging 06/12 Istanbul, Presenting poster West Nile Disease in the Infectious Diseases of Turkey Mediterranean Basin: Priority in the Middle activities and results of East and North Africa: collaboration between the Research Istituto G. CaporaleTeramo Opportunities and (Italy) and Middle East and

Biosafety in a North African countries Changing Environment Epidemiological and 10/12 Zagreb, Croatia Speaker West Nile virus in Italy, five clinical aspects of years of epidemic West Nile virus infection in Croatia and neighbouring countries Arbo-Zoonet final 09/12 Orvieto, Italy Presenting poster An integrated information meeting, Joint system for the management Conference on of West Nile Disease Emerging and Re- surveillance in Italy. emerging epidemics affecting global health Arbo-Zoonet final 09/12 Orvieto, Italy Presenting poster Analysis of surveillance meeting, Joint systems in place in European Conference on Mediterranean countries for Emerging and Re- West Nile virus (WNV) and emerging epidemics Rift Valley fever (RVF) affecting global health

ToR: 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 21. Did your laboratory exchange information with other OIE Reference Laboratories designated for the same pathogen or disease? X Yes No

22. 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? X Yes No

Purpose of the proficiency tests: Role of your Reference Participating OIE (validation of a diagnostic protocol: Laboratory No. Ref. Labs/ specify the test; quality control of (organiser/participant) participants organising OIE Ref. vaccines: specify the vaccine type, etc.) Lab. Validation of the virus neutralisation test in Co-organiser 2 APHIS, USDA OIE Ref. microtiter format by using PRNT as gold Lab.For WN(co- standard organizer) Validation single stage conventional, nested, Co-organiser 2 APHIS, USDA OIE Ref. and real time PCR assays by side-by-side Lab. For WN (co- performance comparison organizer)

23. 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?

X Yes No

Name(s) of relevant OIE Title of the project or contact Scope Reference Laboratories Revision of the OIE Manual Updating and revising the protocols of APHIS, USDA OIE Ref. Lab. For chapter on West Nile Fever old procedures and adding new WN diagnostic procedures

ToR: To organise inter-laboratory proficiency testing with laboratories other than OIE Reference Laboratories for the same pathogens and diseases to ensure equivalence of results.

24. Did your laboratory organise or participate in inter-laboratory proficiency tests with laboratories other than OIE Reference Laboratories for the same disease?

X Yes No

No. participating Participating OIE Member Purpose for inter-laboratory test comparisons1 laboratories Countries

Harmonising diagnostic tests 2 Italy (Sardinia region)

ToR: To place expert consultants at the disposal of the OIE 25. Did your laboratory place expert consultants at the disposal of the OIE? X Yes No

Kind of consultancy Location Subject (facultative ) Harmonization of the diagnostic tests included Teramo Revision of chapter for WNF in the West Nile Fever chapter of the OIE Manual ad hoc Group Paris Validation of diagnostic tests

Annex : List of publications

List of publications: 1. Savini G., Capelli G., Monaco F., Polci A., Russo F., Di Gennaro A., Marini V., Teodori L., Montarsi F., Pinoni C., Pisciella M., Terregino C., Marangon S., Capua I., Lelli R. 2012. Evidence of West Nile virus lineage 2 circulation in Northern Italy. Vet Microbiol. doi:10.1016/j.vetmic.2012.02.018. 2. Mulatti P., Capelli G., Lorenzetto M., Terregino C., Monaco F., Bonfanti L., Marangon S. Surveillance of West Nile Virus in north-eastern Italy, 2011: equine IgM and mosquitoes to early detect virus circulation. Zoonoses and Public Health. 2012 Sep 13. doi: 10.1111/zph.12013. 3. Lelli R, Calistri P, Bruno R, Monaco F, Savini G, Di Sabatino D, Corsi I, Pascucci I. West Nile transmission in resident birds in Italy. Transbound Emerg Dis. 2012 Oct; 59 (5):421-8. doi: 10.1111/j.1865-1682.2011.01287.x. Epub 2011 Dec 28. 4. Barbić L., Listeš E., Katić S., Stevanović V., Madić J., Starešina V., Šeparović S., Di Gennaro A., Savini. G. Spreading of West Nile virus infection in Croatia. Vet Microbiol. 2012 Oct 12;159(3-4):504-8. doi: 10.1016/j.vetmic.2012.04.038. Epub 2012 May 9. 5. Di Francesco G., Savini G., Maggi A., Cavaliere N., D’Angelo A. R., Marruchella G. 2012. Equine meningo-encephalitis caused by Halicephalobus gingivalis: a case report observed during West Nile disease surveillance activities. Veterinaria Italiana 48: 431-443.

International conferences: 1. Goffredo M. “Training on Identification of Culicoides and Mosquitoes at Species Level and Molecular Typing of Culicoides Species”, TAIEX 46424 Ankara, Turkey, 17 - 18 April 2012.

1 See Interlaboratory test comparisons in: Laboratory Proficiency Testing at: www.oie.int/en/our-scientific-expertise/reference-laboratories/proficiency-testing see point 1.3

2. Savini G., Monaco F., Briguglio P., Pinto S., Pinoni C., Polci A., Di Gennaro A., Pais M., Teodori L., Bruno R., Ruiu A., Lelli R. West Nile disease in Sardinian wild birds. 6th Annual Meeting EPIZONE Brighton, UK, 12 - 14 June 2012. 3. Fares F., Savini G., Goffredo M., Cipriani M., Di Gennaro A., Marini V., Santilli A., Ippoliti C., Monaco F., Lelli R. West Nile and Usutu virus vectors in Italy. 6th Annual Meeting EPIZONE, "Viruses on the move", Brighton, UK, 12 - 14 June 2012. 4. Guercio A., Savini G., Monaco F., Calistri P., Bruno, S. Di Bella, P. Di Marco, V. Di Marco, A. Torina, G. Purpari, R. Lelli. West Nile Disease (WND) in Sicily. 15th International Congress of Infectious Diseases, Bangkok, Thailand, 13 - 16 June 2012. 5. Purpari G., Savini G., Monaco F., Calistri P., Di Gennaro A., Cannella V ., Vitale F., Mira F., Di Bella C., Guercio A., Lelli R. Importance of dogs as sentinels of West Nile virus activity in urban and suburban areas. 15th International Congress of Infectious Diseases, Bangkok, Thailand, 13 - 16 June 2012. 6. Savini G., Monaco F., El Harrak M., Sghaier S., Erturk A., Kennerman E., Calistri P., Lelli R. West Nile Disease in the Mediterranean Basin: activities and results of collaboration between the Istituto G. Caporale Teramo (Italy) and Middle East and North African countries. “Endemic and Emerging Infectious Diseases of Priority in the Middle East and North Africa: Research Opportunities and Biosafety in a Changing Environment”. Istanbul, Turkey, 18 - 21 June 2012. 7. Lelli R. Current West Nile Disease Status and Research. “Chronic Wasting Disease”. Seoul, South Korea, 18 - 23 June 2012. 8. Cito F., Narcisi V., Danzetta M.L., Iannetti S., Di Sabatino D., Bruno R., Carvelli A., Atzeni M., Sauro F., Calistri P. Analysis of surveillance systems in place in European Mediterranean countries for West Nile virus (WNV) and Rift Valley fever (RVF). Arbo-Zoonet final meeting, Joint Conference on Emerging and Re-emerging epidemics affecting global health. Orvieto, Italia, 19 - 21 September 2012. 9. Calistri P., Iannetti S., Danzetta M.L., Narcisi V, Cito F., Di Sabatino D., Bruno R., Sauro F., Atzeni M., Carvelli A., Giovannini A. The Components of “One World – One Health” Approach. Arbo-Zoonet final meeting, Joint Conference on Emerging and Re-emerging epidemics affecting global health. Orvieto, Italia, 19 - 21 September 2012. 10. Danzetta M.L., Iannetti S., Di Sabatino D., Bruno R., Cioci D., Cito F., Narcisi V., Primavera A. , Carvelli A., Atzeni M., Sauro F., Calistri P. An integrated information system for the management of West Nile Disease surveillance in Italy. Arbo-Zoonet final meeting, Joint Conference on Emerging and Re- emerging epidemics affecting global health. Orvieto, Italia, 19 - 21 September 2012. 11. Savini G. West Nile virus in Italy, five years of epidemic. Symposium “Epidemiological and clinical aspects of West Nile virus infection in Croatia and neighbouring countries” Zagreb, Croatia, 25 October 2012 (oral presentation). National conferences: 1. Marruchella G., Di Francesco G., Vita S., Portanti O., Marini V., d'Angelo A.R., Savini G Infezione da virus west nile: protocolli immunoistochimici a confronto nel modello murino. XIV Congresso Nazionale della S.I.Di.L.V. Sorrento, 24 - 26 October 2012. 2. Savini G., Puggioni G., Di Gennaro A., Rossi R., Di Francesco G., Rocchigiani A., Polci A., Marini V., Pinoni C., Arru D., Rolesu S., Lorusso A., Monaco F. Ceppi del virus della West Nile appartenenti al lineage 2 isolati in uccelli selvatici in Sardegna. XIV Congresso Nazionale della S.I.Di.L.V. Sorrento, 24 - 26 October 2012 (oral presentation). 3. Di Francesco G., Savini G., Maggi A., Cavaliere N., D’Angelo A. R., Marruchella G. Meningo-encefalite equina da Halicephalobus gingivalis: contributo casistico nell’ambito del piano di sorveglianza West Nile disease. IX Congresso Nazionale dell’Associazione Nazionale di Patologia Veterinaria, Perugia, 25-26 May 2012. Other communications The public web site (www.izs.it) disseminating information and data on West Nile is kept continuously updated in order to have:

 the latest on the Italian and European Regulations issued by the Italian Ministry of Health;  the current (2012) and past (2008, 2009, 2010 and 2011) epidemiological situations in Italy;  maps on entomological and serological surveillance activities;  the current epidemiological situations in the Mediterranean Basin;  scientific documents on-line.

Annual reports of OIE Reference Laboratories, 2012 11