Mandate and Activities of an OIE Reference Laboratory

Mandate and Activities of an OIE Reference Laboratory Nairobi Training 9-11 August 2011 Dr G Thobokwe BVI (OIE Sub Saharan Regional Reference Laboratory for FMD Diagnosis) gthobokwe@bvi.co.bw

ABOUT BVI 2

Introduction to BVI Established following the return of SAT viruses in N/Botswana 100% Shareholding to GoB Technical partnership with Merial signed May 1978 (10 year renewal) Mandate - research on viruses circulating in the region -make appropriate vaccines for safeguarding beef trade

Laboratory transfer A fully equipped laboratory module is sent to the Botswana Vaccine Institute for the production of FMD vaccine.

Human Resource Total complement of 102 Top Management 3 Senior Management= 8 Middle Management= 15 Tech support = 54 Support staff = 22 3 Merial expatriate to date 1 st Technical Manager(Late Mr. Dupasquier 07)accompanied by daughter with BVI staff 2006

New Process: Purification of antigens at Industrial scale

Diversification in Manufacturing and Sales 1978 1985 1992 FMD vaccine (AFTOVAX) Rinderpest vaccine (PESTOBOV 100)* Anthrax (CARBOVAX) Blackleg (ASYMPTOL) 1993 1994 CBPP (PERIBOV) 1998 Thermostable 2010 (PESTOBOV-50T)* Rinderpest Peste des Petits Ruminants PPR- VAC Purified FMD Vaccine (Aftovaxpur) and Antigen Banks *Product discontinued 2005 due to GREP recommendation 11

FMD diagnosis in Africa 1985 : Recognition by the O.I.E., the World Animal Health Organisation, as Regional Reference Laboratory for the Sub-Saharan Africa Provides the following; free laboratory diagnosis to SADC & SSA free technical assistance during field investigation and sample collection free packaging materials for submission Facilities used for training of SADC Vet officials of FMD Diagnosis (2 Zambians trained April 2008 for 3 weeks)

Diagnostic tests Detection of antibodies lpb ELISA and sbp ELISA 3ABC ELISA SNT Detection of virus Isolation on primary pig/lamb kidney cells Detection of viral antigen Sandwich ELISA Detection of viral genomic material PCR Sequencing

Relationship between outbreaks of FMDV type SAT 2 in Botswana, Namibia and Zambia in 2007 98 SAT2/ZAM/9/93* (AF367116)(buffalo) 100 SAT2/ZAM/10/93* (AF367117)(buffalo) 99 100 SAT2/ZAM/7/96* (AF367120)(buffalo) SAT2/ZAM/10/96* (AF367121)(buffalo) SAT2/RHO/1/48 (AJ251475) 70 100 SAT2/BOT/2/2007 SAT2/BOT/3/2007 SAT2/BOT/5/2007 SAT2/BOT/6/2007 SAT2/ZIM/10/2002 100 SAT2/ZIM/11/2002 SAT2/ZAM/1/2007 SAT2/ZAM/3/2007 96 100 100 SAT2/ZAM/2/2007 SAT2/NMB/1/2007 98 SAT2/NMB/4/2007 SAT2/BOT/24/2006 (buffalo) 99 SAT2/BOT/29/98* (AF367124)(buffalo) SAT2/BOT/31/98* (AF367125)(buffalo) SAT2/BOT/1/2005 SAT2/BOT/6/2005 SAT2/BOT/3/2005 SAT2/BOT/8/2005 100 SAT2/BOT/2/2005 SAT2/BOT/4/2005 SAT2/BOT/5/2005 71 SAT2/BOT/7/2005 0.02 Mid-point-rooted Neighbor-joining tree (Kimura 2-parameter model) constructed using MEGA 3.1 based on complete VP1 nt sequences.

99 99 99 77 SAT2/BOT/6/2008 Semboyo, Khundu, Maun 78 SAT2/BOT/8/2008 Semboyo, Thaba, Maun 71 SAT2/BOT/10/2008 Katuba-Senboyo, Khundu, Maun SAT2/BOT/9/2008 Semboyo, Thaba, Maun SAT2/BOT/5/2007 } SAT2/BOT/2/2007 Habu, Maun Veterinary District 99 SAT2/BOT/3/2007 SAT2/BOT/7/2008 Moporota, Naana, Maun 96 SAT2/BOT/11/2008 Moporota, Nkene, Maun SAT2/BOT/6/2007 Nxame Crush, Habu, Maun SAT2/ZIM/10/2002 99 SAT2/ZIM/11/2002 99 SAT2/BOT/1/2005 SAT2/BOT/2/2005 SAT2/BOT/P3/98 (buffalo 29) (AF367124) SAT2/BOT/P3/98 (buffalo 31) (AF367125) SAT2/BOT/24/2006 (buffalo) Muchenje, Kasane 99 SAT2/BOT/14/2008 SAT2/BOT/15/2008 Satau Crush, Khundu SAT2/ZAM/1/2007 88 SAT2/ZAM/3/2007 } Kazungula, Southern Province SAT2/ZAM/2/2007 93 SAT2/NMB/1/2007 } Caprivi Strip SAT2/NMB/4/2007 SAT2/ZAM/8/2008 SAT2/NMB/2/2008 86 SAT2/NMB/1/2008 Eastern Caprivi SAT2/NMB/3/2008 SAT2/NMB/4/2008

Situation ctd Kenya SA1 Sept 09 to date Kenya O Sept 09 to date Kenya SAT 2 Sept 09 to date Zam, O Nov 2010 Namibia SAT1 Apr 2010 Malawi, SAT2, 2007-2009 Zambia SAT 1 2004,2006, 2008 Namibia SAT 2, 2007-2009 Zambia, SAT 2 2007-2009 Botswana SAT 2, 2008 Botswana zone 1, SAT 1-2006, SAT 2-2005 and 2008 Botswana zone 2, SAT2, 2007-2011? Zim SAT 2 May 2010 Bots SAT2 Aug 2010 Botswana SAT 2-2002(zone 6), SAT 1-2003(zone 6), SAT 2-2006(zone 7) RSA SAT1,2 Aug 2010

Reference Laboratories and Collaborating Centres Additional Reference Centres Regional/National Reference Centres Pool 3 O, A, Asia 1 Pool 1 O, A, Asia 1 Pool 7 O, A Pool 5 O, A, SAT 1, 2 Pool 2 O, A, Asia 1 Pool 4 A, O, SAT 1, 2, 3 Pool 6 SAT 1, 2, 3 FMD - Free Free. Virus present in game parks Intermediate, sporadic Endemic Free with vaccination Countries with multiples zones Source: Annual OIE/FAO FMD Reference Laboratory Network Report, 2007

FAO Progressive control pathway - risk reduction approach not a top down prescribed approach: but each MS encouraged to develop national risk reduction strategies that are supportive to the regional effort 5 Officially free without vaccination No circulation / containment zones only 4 Officially free with vaccination No circulation / containment zones only 3 Approaching freedom Outbreaks < once / year 2 Critical points addressed incidence 1 Critical risk points identified, strategy being developed Stages 0-3 = infected countries/zones 0 Risk not controlled Continuous FMDV circulation

Africa Roadmap progression to 2020 after Nairobi and Algiers Workshops 2009 2020

Mandate of OIE Ref Labs 22

Summary of OIE Reference Laboratories and collaborating Centres in 2009

Distribution of OIE Ref Labs

Ref Labs mandate Reference Laboratories of the OIE shall have as their principal mandate: to function as a centre of expertise and standardization for a designated disease(s) or topics; to store and distribute to national laboratories biological reference products and any other reagents used in the diagnosis and control of the designated disease(s) or topics; to develop new procedures for diagnosis and control of the designated disease(s) or topics; to gather, process, analyze and disseminate epizootiological data relevant to their specialty; to place expert consultants at the disposal of the OIE

They may also contribute to: provision of scientific and technical training for personnel from OIE Member Countries and Territories; provision of diagnostic testing facilities to OIE Member Countries and Territories. In the case of results that are confirmed positive for diseases that are reportable to OIE, the Reference Laboratory should immediately inform the OIE Delegate of the Member Country or Territory from which the samples originated as well as the OIEHeadquarters; organization of scientific meetings on behalf of the OIE; coordination of scientific and technical studies in collaboration with other laboratories or organizations; publication and dissemination of any information in their sphere of competence that may be useful to OIE Members.

Role in reporting

OIE and its network of expertise (Reference Laboratories and Collaborating Centres) must: improve early warning in the event of outbreaks of animal disease, including zoonoses and the knowledge of the animal health situation in the World, with regard to OIE listeddisease and to emerging and reemerging disease that are not listed

Notification of Animal Diseases including Zoonoses The General Assembly Decision of 2004 determined that OIE Reference Laboratories must immediately communicate positive findings of a reportable disease to the veterinary authority of the respective Member Country and to the OIE Prior to publishing these results and if the biological sample is provided by a country other than that in which the RL is located OIE needs the agreement by the respective Member Country and a precise identification of the origin ofthe samples Requirements for confirmations prevent a premature or erroneous report from a laboratory A Delegate who does not share information about the possible occurrence of a disease has no grounds for objection if the OIE informs other Members (Art. 4 and 9 of the Organic Statutes) The OIE may also report unofficial (but reliable) information of global health concern

Situation ctd Kenya SA1 Sept 09 to date Kenya O Sept 09 to date Kenya SAT 2 Sept 09 to date Zam, O Nov 2010 Namibia SAT1 Apr 2010 Malawi, SAT2, 2007-2009 Zambia SAT 1 2004,2006, 2008 Namibia SAT 2, 2007-2009 Zambia, SAT 2 2007-2009 Botswana SAT 2, 2008 Botswana zone 1, SAT 1-2006, SAT 2-2005 and 2008 Botswana zone 2, SAT2, 2007-2011? Zim SAT 2 May 2010 Bots SAT2 Aug 2010 Botswana SAT 2-2002(zone 6), SAT 1-2003(zone 6), SAT 2-2006(zone 7) RSA SAT1,2 Aug 2010

Annual reporting to OIE eg 2007 Summary of general activities related to the disease 1. Test(s) in use/or available for the specified disease at your laboratory Test For Specificity Total ELISA Antibody SAT1/SAT2/SAT3/O/ 591 A ELISA Antigen SAT1/SAT2/SAT3/O/ 29 A VN Antibody SAT1/SAT2/SAT3/O/ 79 A CFT50% Antigen SAT1/SAT2/SAT3/O/ 29 A RT-PCR Molecular SAT1/SAT2/SAT3/O/ 4 A Nucleotide Molecular SAT1/SAT2/SAT3/O/ 4 Sequencing A BHK-21 cell culture Virus isolation / 32

2. Production and distribution of reagents Rabbit and bovine hyper immune sera Guinea-pig hyper immune sera Negative bovine sera SAT1, SAT2, SAT3, O and A FMDV for VNT Inactivated antigens for ELISA OIE Regional Reference Laboratory for the Sub-Saharan continent o o o o Negative bovine sera 2 x 300 ml Bovine positive control sera SAT1/SAT2/SAT3 300 ml of each Rabbit antisera SAT1/SAT2/SAT3 100 ml of each ELISA FMD antigen SAT1/SAT2/SAT3 200 ml of each Botswana National Veteinary Laboratory o o o o Negative bovine sera 500 ml Bovine positive control sera SAT1/SAT2/SAT3 50 ml of each Rabbit antisera SAT1/SAT2/SAT3 100 ml each ELISA FMD antigen SAT1/SAT2/SAT3 250 ml each

International harmonisation and standardisation of methods for diagnostic testing or the production and testing of vaccines Annual EU-FMD profficiency testing Since 2008; reference labs vaccine matching tests harmonisation Post vaccination sero-monitoring SADC ELISA Profficiency testing distribution of samples

Results --- r1 values by VNT r1 values for sample 4 A TUR 24/07 Lab Code BVS 610 637 645 679 6104 Pool 1 0.7 0.59 0.36 0.59 0.56 0.77 4 0.35 0.4 0.18 0.11 0.11 0.16 5 0.14 0.07 0.06 0.08 0.03 0.18 6 0.51 0.51 0.8 0.47 0.5 0.56 7 0.47 0.47 0.85 0.32 0.70 0.45

70 60 50 40 30 C a s e s vaccinated not vaccinated calves vaccinated calves not vaccinated Total Cases Primo vaccination Booster vaccination 20 10 0

Antibody titres progression - Bots 2.5 2.0 1.5 1.0 Sat 1 Sat 2 Sat 3 0.5 0.0 D0 D14 D28 D42 D58 D80 D92 D120

SUMMARY OIE Reference Laboratories and Collaborating Centers have mandates to assist the OIE in developing international standards. The OIE is very dependent on and appreciative of the expertise at the OIE Reference Laboratories and Collaborating Centers to accomplish its mission of improving animal health worldwide

Asante sana No NSP = maker vaccines High quality = aqueous and oil based vaccines Reduced dose volumes & increased margins Minimal side effects Antigen shelf life increased from 2 to 5 years Increased international Acceptance = Antigen banks for non vaccinating countries Increased throughput to BMC