Ministry of Agriculture & Rural Development Technical Report Development of an Improved Capability in support of National Bio-security for the Surveillance and Control of Foot & Mouth Disease in Cattle and Pigs Milestone National Reference Laboratory and Regional Laboratories operational and effective By Chris Morrissy Table of Contents Institute Information _ Project Abstract _ Executive Summary 4 Introduction & Background _ 5 National Reference Laboratory and Regional Laboratories operational and effective 5.1 5.2 Capacity Building _ 5.3 Implementation Highlights _ Publicity _ Conclusion _ Appendix _10 7.1 7.2 7.3 Appendix Standard coversheets, result sheets and IQC record keeping forms eg FMD 3ABC ELISA: _ 10 Appendix FMD Serotyping Results for Vietnam 2005 – 2009 _ 14 Appendix Sequencing information from project 2005 – 2009 _ 15 7.4 Appendix FMD serosurveilance results _ 24 Institute Information Project Name Vietnamese Institution Regional Animal Health Centre, Ho Chi Minh City (RAHO - ), South Vietnam Vietnamese Project Team Leader Dr Dong Manh Hoa Australian Organisation Australian Personnel Australian Animal Health Laboratory (AAHL), PMB 24, Geelong, 3213, Australia Mr Chris Morrissy Date commenced 01/06/2005 Completion date (original) 01/06/2008 Completion date (revised) Reporting period Contact Officer(s) In Australia: Team Leader Mr Chris Morrissy Diagnostic Virologist Supervisor Mammalian Virology Organisation Australian Animal Health Laboratory (AAHL), PMB 24, Geelong, 3213, Australia Name: Position: Telephone: Fax: +61 5227 5000 +61 5227 5555 Email: chris.morrissy@csiro.au In Australia: Administrative contact Mr Chris Morrissy Name: Patents Contracts Officer Position: Organisation Australian Animal Health Laboratory (AAHL), PMB 24, Geelong, 3213, Australia In Vietnam Telephone: Fax: Email: Dr Dong Manh Hoa Name: Director Position: Organisation Regional Animal Health Centre, Ho Chi Minh City (RAHO - ), South Vietnam +61 5227 5000 +61 5227 5555 christopher.morrissy@csiro.au Telephone: Fax: Email: + 84 8568220 + 84 8569050 rahchcmc@hcm.vnn.vn Project Abstract The project’s purpose was twofold - to develop capacity for FMD (and other disease) surveillance and diagnosis at both a laboratory and field level, and to investigate the serotypes of FMDV circulating in Vietnam and the reason for vaccine failures Regional laboratories were set up with the reagents and methods to allow a diagnostic capability for FMDV diagnosis and serology Control strategies for understanding of FMD epidemiology have been implemented through veterinary and laboratory training workshops The project has highlighted the importance of having a laboratory network to identify what is happening in the field and how to prevent and control disease outbreaks The pilot zones were established in provinces near the borders of Vietnam to study serotypes circulating in Vietnam and to determine their origin The number and quality of samples increased with each round of the project giving more data on the FMD situation in Vietnam Virus isolation and molecular studies can now be carried out on FMD samples from the field and molecular epidemiological studies of the FMDV isolates in these provinces has provided insights into the effectiveness of border control and origin of circulating FMDV Improved diagnostic capacity for FMD allows for the early detection and identification of disease enabling better control of disease and helps reduce loss of livestock and therefore increases productivity Executive Summary Over the life of the project there was improvement and advances in both activities in the field and laboratory The quality of both the diagnostic tests and the field data collected continued to improve throughout the project Throughout the project, consultants from AAHL worked with four diagnostic laboratories – RAHO – - HCMC, NCVD - Hanoi, RAHO – - Can Tho and RAHO – - Da Nang - to improve FMD diagnostics and monitor the progress of the project The RAHO - and NCVD laboratories now have virus isolation, virus neutralisation test, ELISA, PCR and sequencing techniques established for FMD diagnosis The project also worked with NAVETCO research laboratory to establish the capability for FMD serology by ELISA RAHO - is now well established in all these techniques and both RAHO - and NCVD laboratories are also applying the new technologies to other disease problems in Vietnam eg RAHO – have used virus isolation for Goat Pox, PRRS and CSF, NCVD used virus isolation to identify a new isolate of PRRS in Vietnam The Can Tho and Da Nang laboratories have the capability for FMD diagnosis for serotyping and post-vaccination serology by LP - ELISA In addition, RAHO - have learnt techniques in reagent production and have produced their own FMD antigen In developing such techniques they have also gained the ability to trouble-shoot the ELISAs and problems with growth of FMDV Staff from RAHO - and NCVD also visited AAHL during each year of the project for training and collaborative projects In regards to field activities, there were significant improvements in the quality and number of samples submitted to the laboratory for serosurveillance and serotyping for ELISA The latter allowed DAH to better understand and identify the circulating serotypes of FMD viruses in Vietnam in addition to gaining confidence in conducting large serosurveillance surveys There was also a significant improvement in the amount of data collected over each round of the project RAHO - now has an epidemiological unit and this project has assisted in developing the skills of the epidemiologists in serosurveillance, outbreak control and investigation of vaccine failure Information from the serosurveillance in the project was reviewed to give an indication on the success of vaccination and the prevalence of FMD infection This data has been presented at the OIE/SEAFMD regional meetings and to DAH The project was invited to a number of regional meetings to present data from Vietnam on control of FMD As a result of these outputs of the project, Vietnam has been used as an example for others in the region to show the type of information that can be collected on serosurveillance and outbreak investigation The project coordinated activities with the AusAID capacity building project to allow both projects to achieve their objectives, eg combined PCR and sequence training with sequencing FMD isolates along with AI, PRRS and CSF isolates from Vietnam Outbreaks of avian Influenza (AI) and PRRS had impact on both field and laboratory activities of this project with DAH staff being overloaded with work Introduction & Background Objectives of the project: To establish an effective laboratory network for the diagnosis and control of FMD by the provision of resources and training of staff in required methods and quality assurance To provide accurate data to explain failure of vaccination to control FMDV and to develop new effective vaccine application strategies Completing these objectives will improve the diagnostic capability of the veterinary laboratories in Vietnam and the training of DAH veterinarians in disease investigation and control This will strengthen the profile of DAH which will play a vital role in making Vietnam more economically competitive Improved animal health will lead to an increase in rural productivity though increased animal production and indirectly in increased crop production Healthy animals will enable small farmers to be more competitive in the local market Control of FMD and animal diseases in general will give poor farmers a more stable income stream and reduce their vulnerability to natural and economic problems Establishing a diagnostic network which extends from the North to South Vietnam, from the laboratory to the farm level, reinforced by training and education, will give Vietnam a working model on which to base disease control This will directly increase the competitiveness and productivity of the national agricultural system which includes the major areas of concern including the Mekong Delta and the Central Coast Specific Objectives for the Laboratories: • • • • • Laboratory training manuals incorporating diagnostic techniques Functional tests at RAHO – –HCMC and NCVD - Hanoi laboratories to allow it to operate as national reference laboratories Functional ELISA tests at RAHO – - Da Nang, RAHO – - Can Tho & NAVETCO laboratories to allow them to operate as regional laboratories Protocol manuals for submission and handling of samples and provision of feedback of results Laboratory Quality Control procedures documented and tested Internal Quality control results documented and reviewed for each laboratory Implementation Approach and Strategy The approach for technology transfer is well established at AAHL and has been successfully applied in previous projects in Vietnam, Thailand and Indonesia The project approach used was also thought to be the most appropriate for developing an understanding of FMD epidemiology in Vietnam The field studies and serosurveillance approaches were designed and planned in conjunction with DAH to provide the maximum necessary information to demonstrate the FMD situation in Vietnam and the effectiveness of FMD vaccines The diagnostic technologies that will be used in this approach are the standard diagnostic tests in use throughout the world to study FMD as directed by OIE AAHL has a lot of experience with field surveys for prevalence of antibodies, as in the ACIAR projects in Laos and Thailand on FMD The Philippines is another example where OIE standard diagnostic tests are being used to control and eradicate FMD National Reference Laboratories and Regional Laboratories operational and effective 5.1 Implementation Highlights The main achievements of the project were: • The following FMD diagnostics were established in the collaborating laboratories: RAHO - and NCVD laboratories have established virus isolation, virus neutralisation test, ELISA for antigen and antibody detection, PCR and sequencing RAHO – and RAHO – laboratories have the capability for FMD diagnosis for serotyping by ELISA (detection of antigen) from a FMD outbreak and serology by ELISA for post-vaccination surveillance • A quality system was implemented in each laboratory for FMD diagnostics which included standardised methods, IQC and better record keeping Standardised methods introduced for ELISAs, cell culture, PCR, sequencing and data collection ( eg ELISA methods attached to this email with this report ) Standard coversheets, result sheets and IQC record keeping forms ( eg ELISA forms appendix ) • Increased collaboration between the laboratories since the inception of the project • Improvement in the quality and number of samples submitted to the laboratory for serotyping by ELISA ( summary of serotyping results appendix ) This gave DAH a better understanding of the circulating serotypes of FMD viruses in Vietnam The improvement in sample collection also allowed for virus isolation from field samples that had not been possible prior to the project and in turn enabled sequence data to be obtained from FMD virus isolates • Sequencing, genotyping and analysis of approximately 100 Vietnamese FMD isolates collected from 2006 onwards The isolates were sequenced and analysed at AAHL by a scientists from AAHL and RAHO – The sequence data was then sent to WRL for confirmation and comparison to other FMD isolates This information was shared with SEAFMD in support of the regional project to control FMD • Sequencing/Genotyping 2006/2008 Serotype O: topotypes Cathay (33%), ME-SA (PanAsia) (6%) & SEA (Myanmar 98) (41%) Pigs: mainly Cathay & SEA Cattle: mainly SEA Serotype A: Thailand/Malaysia 97 Vaccine used for FMD Serotype A needed to change from A 22 to A Malaysia 97 (also antigen for serology) Serotype Asia 1: Jiangsu-China-2005 (isolated from the Nth & Centre) and Myanmar 98 (isolated from the Centre) Confirmed sources of Asia virus into Vietnam • The analysis of FMD isolate sequence data from Vietnam was used to compare sequences to FMD sequences from around the world and is shown in appendix • Improvement in the quality and number of serum samples submitted to the laboratory for serology by ELISA This gave DAH a better understanding of the animals exposed to FMD and the vaccination coverage in Vietnam Two rounds of serosurvellance each year with first round collected prior to vaccination and the second round collected after vaccination The data form serosurvellance showed vaccine coverage and number of animals exposed to FMD (eg summary table example attached to email) Recommendations for improved serosurveillance can be found • A number of sera samples were retested to compare antibody titres for samples from the provinces to allow identification by serology of the circulating isolate The results indicated that the combined use of the non-structural 3ABC ELISA and the structural LP-ELISA could be used to identify the serotype of FMD that had been circulating in the field (eg appendix ) • Throughout the project AAHL consultants visited the laboratories to: Establish FMD Elisas and standardised techniques Validate an Elisa using antigen produced at RAHO – using Vietnam isolates The production of FMD Elisa antigen allows the laboratories to be more selfsufficient and is the start of the laboratories capability to produce its own reagants Establish and review cell culture and virus isolation for growth of FMD isolates from the field Cell culture is important to grow FMD virus to allow further analysis of FMD field isolates by PCR and sequencing Cell culture has also been used for serology (VNT) and for isolation of other disease agents, such as goat pox, CSF and PRRS Review molecular techniques and establish a workflow for sample processing and testing to ensure quality results Evaluate quality assurance records and data collection to ensure test records were being maintained and results were being interpreted correctly Provided advice on field information required and produced data collection form Analyse field and laboratory results to provide epidemiological input Supply consumables and reagents for testing of samples by FMD Elisa, molecular technology and cell culture Train staff in biosafety techniques and assist with their implementation Assess quality assurance procedures and provide input to quality assurance manuals Assess accurately of tests by Proficiency Testing Samples supplied to laboratories to test results produced were accurate 5.2 Capacity Building The project has provided training and technology transfer of FMD diagnostics to each laboratory involved in the project Reagents and standard methods were supplied to each laboratory giving them the diagnostic capability for FMDV diagnosis Serology and serotyping (detection of antigen) by Elisa is now being practiced at all of the laboratories and RAHO – and NCDV have also established virus isolation, cell culture, virus neutralisation for serology, molecular and sequencing techniques Training and education of field veterinarians in sample and data collection showed an impact with an increase in quality and numbers of samples collected and submitted to the laboratory These skills will be vital in implementation of the National FMD Control Program 5.3 Publicity The CARD AusAID project received publicity in Vietnam, Australia and internationally through the training programs and also through the achievements in understanding FMD in Vietnam FMD is a disease of importance in Vietnam and the region and this put this project into the lime light The project has been publicised through a press releases in Australia and articles in newsletters including the SEAFMD newsletter and on the internet The results from the project have been presented at: o o o o OIE/SEAFMD meetings (eg presentation attached to email) EU FMD 2008 (eg presentation attached to email) WAVLD 2007, 2009 Lower and Upper Mekong Working Group meetings in the region With the laboratory and epidemiological capacity now available in the collaborating laboratories, particularly RAHO – 6, there is now the potential for a smaller, more focused study on vaccination failure This would be best limited to a smaller number of provinces in southern Vietnam, with a study protocol aimed specifically at investigating vaccination effectiveness As the collaborating laboratories are now implementing a range of FMD diagnostic techniques it is considered that these abilities will be sustained and transferred to other laboratories Conclusion The project achieved its objectives by helping to improve the FMD and general diagnostic capacity of the network of veterinary laboratories The RAHO – and Hanoi laboratories have provided and are continuing to provide support and training to other laboratories in the network The project has highlighted the need for training of field veterinarians in the collection of data and how to ensure the correct information is obtained from the farmer The staff at RAHO – will continue to deal directly with the veterinarians in the field to ensure the correct information is collected in future surveys The establishment of cell culture has allowed the growth of FMD isolates from the field that could not be previously identified at RAHO - and at NCVD The increased FMD diagnostic capacity, combination of ELISA, PCR and cell culture for detection of FMD, has increased the number of FMD outbreaks identified in Vietnam The use of cell culture for growth of FMDV has resulted in the availability of higher quality samples for sequencing The introduction of molecular techniques at RAHO – has allowed sequencing results to be obtained quickly and has allowed informed decisions to be made about serotypes of FMD circulating in Vietnam and the correct vaccine to use in each region to control FMD AAHL, DAH and WRL have cooperated on analysis of the sequence data from Vietnam isolates Cell culture has also allowed the production of ELISA antigen which is a key reagent in FMD diagnostics RAHO – is now producing own reagent which it has transferred to NCVD, RAHO – and RAHO – This project continues to be important for Vietnam with the repeated cyclical outbreaks of FMD The Vietnam Government and RAHO - have invited AAHL to give advice on FMD control and have used the project as a model for the implementation of their control plan Despite the overwhelming requirement for control and surveillance of AI and more recently PRRS, the Vietnam Government is committed to the control of FMD This is evidenced by the currently active National FMD Control Program The project highlighted the need to have coordination between the laboratory and the field staff in the control of FMD and in understanding the serosurveillance data post-vaccination and post-exposure to FMD from the laboratory results In the laboratory it is important to have the correct reagents to match the circulating field virus ( also highlighting the importance of improved FMDV identification ) and in the field the field data to allow the interpretation of the laboratory results The whole veterinary network is important in the control of disease, there needs to be strong link between the field and laboratory to ensure both the actions in the field and the results in the laboratory are correct This project set the standard for what was required for a laboratory network to function and the lessons learnt were used in handling HPAI and PRRS outbreaks in Vietnam and the projects and investigations that followed Appendix Standard coversheets, result sheets and IQC record keeping forms eg FMD 3ABC ELISA: 9.11 3ABC C-ELISA Test Coversheet FMD 3ABC COMPETITION ELISA SAN: SAMPLE IDENTIFICATION: TEST (3ABC C-ELISA): SUPERVISOR: DATE PLATE COATED: OPERATOR: DATE OF TEST: OPERATOR: WASHING METHOD: PLATE SHAKER: INCUBATOR: PIPETTERS: NUMBER OF PLATES: 3ABC ANTIGEN DILUTION: DETECTING ANTIBODY BATCH: DETECTING ANTIBODY BATCH: SERUM CONTROLS & SAMPLE DILUTIONS: COATING BUFFER DATE: BLOCKING BUFFER DATE: CONJUGATE BATCH: CONJUGATE DILUTION: SUBSTRATE INCUBATION TIME: Date Reported Signed 9.12 Result page/Plate Format ELISA PLATE FORMAT S1 S1 S9 S9 S17 S17 S25 S25 S33 10 S33 11 12 A C++ C++ B S2 S2 S10 S10 S18 S18 S26 S26 S34 S34 C++ C++ C S3 S3 S11 S11 S19 S19 S27 S27 S35 S35 C+ C+ D S4 S4 S12 S12 S20 S20 S28 S28 S36 S36 C+ C+ E S5 S5 S13 S13 S21 S21 S29 S29 S37 S37 C- C- F S6 S6 S14 S14 S22 S22 S30 S30 S38 S38 G S7 S7 S15 S15 S23 S23 S31 S31 S39 S39 S16 S16 S24 S24 S32 S32 S40 S40 OD MA X OD MA X CC OD MA X OD MA X CC H S8 S8 9.13 Quality Control Sheet C++ Date C+ C- OD Max Comments Antigen Serotype Expected Expected Expected Inhibition Inhibition Inhibition 80 – 100% 50 – 80% < 30% 0.8 – 1.5 SAN Staff 9.14 Summary FMD – 3ABC C-ELISA 10 11 12 13 14 15 16 17 18 19 20 Coat plates with 50µl/well 3ABC(D) baculovirus-expressed antigen (crude soluble protein) diluted 1:2000 in Carbonate Bicarbonate Coating Buffer Incubate shaking for hour at 37OC (Can be left at 4OC overnight) Wash plates times with PBS Block plate with 100uL/well Blocking buffer Incubate shaking for 30minutes at 37OC Wash plates times with PBS Add 40uL Blocking buffer to each well and then add 10uL sera and mix up and down with pipette ODmax and Background (Bkg) wells get 50uL Blocking buffer only Incubate shaking for 30 minutes at 37OC DO NOT WASH PLATE Dilute chicken egg antibody pool 1:100 in Blocking buffer and add 50uL to all wells, directly on top of test sera Bkg wells get 50uL buffer only Incubate shaking for 30 minutes at 37OC During this incubation prepare the conjugate (minimum of 30 minutes prior to use.) Wash plate times with PBS Add 50µl/well conjugate in Incubate shaking for 1hour at 37OC Prepare substrate (DO NOT ACTIVATE) Wash plates times with PBS Activate Substrate and add 100µl of Substrate to each well Incubate at room temp 10 minutes (NOT shaking) Stop with 100µl Stopping Solution Read on plate reader for inhibition with 450nm Appendix FMD Serotyping Results for Vietnam 2005 - 2009 Year Total Samples (NCVD & RAHO – 6) Samples Positive for Serotype O Samples Positive for Serotype A Samples Positive for Serotype Asia Negative / Unsuitable 2005 251 207 17 13 12/2 216 * RAHO - 180 17 11 6/2 803 663 128/4 506 467 33/4 224 177 7/0 123 115 7/0 154 86 59 2006 2007 2008 *poor quality sample was a problem in north 22 14 0 8/0 2/0 2009 Appendix 3: Examples of sequence information form the project Sequencing Results for FMDV Serotype O 2008 – 09 48 FMDV/Type O/08-6415-E2 65 FMDV/Type O/08-6415 E3 FMDV/Type O/08-6924 E4 100 FMDV/Type O/08-5515-12 99 71 FMDV/Type O/08-5515-14 49 O/UKG/35/2001|AJ539141| ME-SA 56 O/India/53/79|AF292107| ME-SA O/O1 Manisa/87|AY593823| ME-SA 70 67 O/India/R2/75|AF204276| ME-SA O/MYA/7/98|DQ164925| SEA 56 94 O/CAM/3/98|AJ294910| SEA O/SUD/2/86|DQ165075| EA3 O/KEN/83/79|AJ303511| EA1 99 68 94 O/UGA/5/96|AJ296327| EA1 73 O/GHA/5/93|AJ303488| WA 100 O/CIV/8/99|AJ303485| WA O/MAL/1/98|DQ165074| EA2 O/UGA/3/2002|DQ165077| EA2 100 63 O/KEN/5/2002|DQ165073| EA2 O/ISA/9/74|AJ303502| ISA-1 92 100 O/ISA/8/83|AJ303503| ISA-1 O/ISA/1/62|AJ303500| ISA-1 O/ISA/1/74|AJ303501| ISA-2 100 61 O/JAV/5/72|AJ303509| ISA-2 O/Corrientes/Arg/06|DQ834727| EURO-SA 70 O/O3 Venezuela/51|AJ004645| EURO-SA 91 O/O2Brescia/ITL/47|M55287| EURO-SA 58 39 O/O1BFS 1860/UK/67|AY593815| EURO-SA O/HKN/21/70|AJ294911| Cathay O/HKN/6/83|AJ294919| Cathay 100 O/PHI/7/96|AJ294926| Cathay 100 O/Yunlin/TAW/97|AF308157| Cathay 84 44 FMDV/Type O/08-4856 100 FMDV/Type O/08-4923-4 64 FMDV/Type O/08-4923-3 100 FMDV/Type O/09-1045-KH2 FMDV/Type O/09-1045 KH3 100 FMDV/Type O/09-1045 KH5 FMDV/Type O/09-1045 KH6 0.01 Report on 34 FMDV O VP1 sequences from Vietnam received from AAHL Software: MEGA No of Taxa : 173 Data File : n:\evd\meg\db\fmdv\o\VITaahl1.meg Data Title : Vietnam Data Type : Nucleotide (Coding) Analysis : Phylogeny reconstruction Tree Inference : ============================== ->Method : Neighbor-Joining ->Phylogeny Test and options : Bootstrap (500 replicates; seed=64238) Include Sites : ============================== ->Gaps/Missing Data : Pairwise Deletion ->Codon Positions : 1st+2nd+3rd+Noncoding Substitution Model : ============================== ->Model : Nucleotide: Maximum Composite Likelihood ->Substitutions to Include : d: Transitions + Transversions ->Pattern among Lineages : Same (Homogeneous) ->Rates among sites : Uniform rates No of Sites : 639 No Of Bootstrap Reps = 500 Only bootstrap vales of 70% and above are shown 8 9 104A A HL 0738K TPHCM 84A A HL 0748BG2 BaRiaV ungTau 86A A HL 0745 DongNai 107A A HL 0758T TPHCM 106A A HL 0751QM TPHCM 87A A HL 0750 PhuY en 85A A HL 0748PH1 BaRiaV ungTau 103A A HL 06467 Soc tr ang 90A A HL 0779DH1 LongA n 82A A HL 0708 LamDong 89A A HL 0762 DongNai 112A A HL 06466 TPHCM O/V IT/7/2006 O/V IT/5/2006 O/V IT/4/2006 O/V IT/6/2006 O/TA I/15/05* ( TRRL) O/TA I/14/05* (TRRL) 0 O/TA I/3/2003 ( DQ164981) O/MA Y /1/2005 O/MA Y /2/2005 0 O/V IT/6/2005 0 O/MY A /4/2004 O/MY A /5/04* (TRRL) O/TA I/21/05* (TRRL) O/MA Y /9/2007 O/MA Y /7/2007 9 O/MA Y /4/2007 O/MA Y /5/2007 9 O/MA Y /6/2007 66A A HL ThanhHoa O/MA Y /5/2006 0 O/TA I/8/2004 O/TA I/37/04* ( TRRL) O/MY A /3/04* ( TRRL) O/MY A /4/04* ( TRRL) O/TA I/5/99 O/MY A /2/2000 (DQ164927) O/V IT/4/2005 O/TA I/36/04* (TRRL) 0 O/MY A /1/04* ( TRRL) O/MY A /2/04* ( TRRL) O/TA I/189/87* ( TRRL) O/TA I/2/2000 ( DQ164979) 0 O/MA Y /1/2002 ( DQ164923) O/TA I/44- 2/02R2B3* ( TRRL) O/TA I/8/99 (DQ164977) O/BUR/2/89 0 9 O/V IT/2/97 O/V IT/7/97 O/CA M/12/94 O/CA M/3/98 O/CA M/6/99 0 O/CA M/1/98 O/CA M/2/98 O1/Manis a/TUR/69 ( A J251477) O/LA O/17/2003 ( DQ164912) 0 O/LA O/2/2006 O/LA O/19/2003 ( DQ164913) O/LA O/20/2003 O/MA Y /1/2006 O/MA Y /2/2006 O/MA Y /7/2005 O/MA Y /3/2005 0 O/MA Y /4/2005 O/MA Y /6/2005 O/MA Y /3/2006 O/MA Y /9/2005 9 O/LA O/3/2006 O/LA O/4/2006 O/LA O/31/2003 ( DQ164918) O/LA O/28/2003 ( DQ164916) O/LA O/23/2003 (DQ164915) O/TA I/20/04R2* ( TRRL) O/V IT/1/2004 ( DQ165032) O/LA O/21/2003 (DQ164914) O/LA O/5/2006 O/LA O/30/2003 ( DQ164917) O/LA O/12/2003 (DQ164910) 0 O/LA O/11/2003 ( DQ164909) O/LA O/13/2003 ( DQ164911) O/LA O/3/2003 ( DQ164908) 98A A HL 06452T TienGiang 0 101A A HL 06459CT1 LongA n 9 O/CA M/4/2006 O/V IT/17/2005 O/V IT/14/2002 ( DQ165026) O/V IT/1/2003 ( DQ165030) 0 0 O/CA M/2/2000 ( A J318828) O/CA M/4/2000 (A J318829) O/TA I/2/2003 (DQ164980) 9 O/V IT/8/2004 9 O/V IT/7/2005 O/V IT/7/2004 9 O/V IT/16/2002 (DQ165027) O/V IT/12/2002 ( DQ165024) O/V IT/3/2005 O/TA I/1/2000 O/V IT/6/2002 ( DQ165020) O/V IT/20/2002 ( DQ165029) O/V IT/7/2002 O/V IT/10/2002 ( DQ165023) O/TA W/2/99 O/SKR/1/2000 ( A J318854) O/JPN/2000 ( A B050978) O/SKR/2/2002 O/TA I/9/99 ( DQ164978) O/HKN/6/83 109A A HL 06482T Soc Tr ang 113A A HL 06482 Soc Tr ang 99A A HL 06453V Soc Tr ang 100A A HL 06457H CaMau 114A A HL 06443 CanTho 110A A HL 06476 115A A HL 06457 CaMau 102A A HL 06463 V inhLong 105A A HL 0739 CanTho 92A A HL 0757 V inhLong 88A A HL 0753 CanTho 111A A HL 0716 75 CanTho 116A A HL 0715C V inhLong O/V IT/3/2006 0 O/V IT/1/2006 O/V IT/2/2006 83A A HL 0716 CanTho 95A A HL 06430 HauGiang 96A A HL 06434B HauGiang 9 93A A HL 0779CD1 LongA n 97A A HL 06443T Bac Lieu 9 O/MA Y /8/2005 O/V IT/9/2005 9 O/V IT/1/2005 0 O/PHI/21/2003 (DQ164953) O/PHI/23/2003 ( DQ164954) O/TA I/54- 1/05B1* ( TRRL) O/TA I/54- 2/05B1* ( TRRL) O/TA I/53-2/05* ( TRRL) O/TA I/64-1/05* ( TRRL) 0 O/TA I/64- 2/05* (TRRL) O/TA I/35/05B2* ( TRRL) 0 108A A HL 0735 DongThap O/V IT/2/2004 ( DQ165033) O/V IT/3/2004 ( DQ165034) O/V IT/11/2005 9 O/TA W/81/97 0 O/TA W/4/99 O/TA W/83/97 O/HKN/16/96 O/PHI/5/95 ( DQ164946) O/PHI/7/96 O/V IT/3/97 O/V IT/13/2002 ( DQ165025) O/HKN/7/96 O/HKN/20/96 9 O/HKN/12/2005 O/HKN/15/2005 O/HKN/14/2005 O/HKN/9/2005 O/HKN/10/2005 9 O/HKN/17/2005 O/HKN/18/2005 O/HKN/19/2005 0 O/HKN/20/2005 O/HKN/25/2005 O/HKN/24/2005 O/HKN/3/2006 4 O/HKN/22/2005 O/HKN/23/2005 O/HKN/1/2006 O/HKN/5/2006 O/HKN/2/2006 O/HKN/4/2006 O1/BFS 1860/UK/67 (J02185) 9 9 N.J Knowles & J Wadsworth, March 2008 Page of 0.02 SE A M E -SA CAT H AY E URO -SA SEA topotype 8 9 9 1 0 0.02 104A A HL 0738K TPHCM 84A A HL 0748BG2 BaRiaV ungTau 86A A HL 0745 DongNai 107A A HL 0758T TPHCM 106A A HL 0751QM TPHCM 87A A HL 0750 PhuY en 85A A HL 0748PH1 BaRiaV ungTau 103A A HL 06467 Soc trang 90A A HL 0779DH1 LongA n 82A A HL 0708 LamDong 89A A HL 0762 DongNai 112A A HL 06466 TPHCM O/V IT/7/2006 O/V IT/5/2006 O/V IT/4/2006 O/V IT/6/2006 O/TA I/14/05* (TRRL) 0 O/TA I/3/2003 (DQ164981) O/MA Y /1/2005 O/MA Y /2/2005 9 O/V IT/6/2005 0 O/MY A /4/2004 O/MY A /5/04* (TRRL) O/TA I/21/05* (TRRL) O/MA Y /9/2007 O/MA Y /7/2007 9 O/MA Y /4/2007 O/MA Y /5/2007 O/MA Y /6/2007 66A A HL ThanhHoa 67A A HL LangSon O/MA Y /5/2006 0 O/TA I/8/2004 O/TA I/37/04* (TRRL) O/MY A /3/04* (TRRL) O/MY A /4/04* (TRRL) O/TA I/5/99 O/MY A /2/2000 (DQ164927) O/V IT/4/2005 O/TA I/36/04* (TRRL) 0 O/MY A /1/04* (TRRL) O/MY A /2/04* (TRRL) 8 O/TA I/189/87* (TRRL) O/TA I/2/2000 (DQ164979) 0 O/MA Y /1/2002 (DQ164923) O/TA I/44-2/02R2B3* (TRRL) O/TA I/8/99 (DQ164977) O/BUR/2/89 9 O/V IT/2/97 O/V IT/7/97 O/CA M/12/94 O/CA M/3/98 O/CA M/6/99 0 O/CA M/1/98 O/CA M/2/98 O/TA I/15/05* (TRRL) ME-SA topotype O1/Manis a/TUR/69 (A J251477) O/LA O/17/2003 (DQ164912) O/LA O/2/2006 O/LA O/19/2003 (DQ164913) O/LA O/20/2003 O/MA Y /1/2006 O/MA Y /2/2006 O/MA Y /7/2005 O/MA Y /3/2005 0 O/MA Y /4/2005 O/MA Y /6/2005 O/MA Y /3/2006 O/MA Y /9/2005 9 O/LA O/3/2006 O/LA O/4/2006 O/LA O/31/2003 (DQ164918) O/LA O/28/2003 (DQ164916) O/LA O/23/2003 (DQ164915) O/TA I/20/04R2* (TRRL) O/V IT/1/2004 (DQ165032) O/LA O/21/2003 (DQ164914) O/LA O/5/2006 O/LA O/30/2003 (DQ164917) O/LA O/12/2003 (DQ164910) 0 O/LA O/11/2003 (DQ164909) O/LA O/13/2003 (DQ164911) O/LA O/3/2003 (DQ164908) 98A A HL 06452T TienGiang 0 101A A HL 06459CT1 LongA n 9 O/CA M/4/2006 O/V IT/17/2005 O/V IT/14/2002 (DQ165026) O/V IT/1/2003 (DQ165030) O/CA M/2/2000 (A J318828) O/CA M/4/2000 (A J318829) O/TA I/2/2003 (DQ164980) 9 O/V IT/8/2004 9 O/V IT/7/2005 O/V IT/7/2004 9 O/V IT/16/2002 (DQ165027) O/V IT/12/2002 (DQ165024) O/V IT/3/2005 O/TA I/1/2000 O/V IT/6/2002 (DQ165020) O/V IT/20/2002 (DQ165029) O/V IT/7/2002 O/V IT/10/2002 (DQ165023) O/TA W/2/99 O/SKR/1/2000 (A J318854) O/JPN/2000 (A B050978) O/SKR/2/2002 O/TA I/9/99 (DQ164978) 0 9 8 1 0 0.01 Page of CATHAY topotype O/HKN/6/83 9 9 0 7 9 0 0.02 Page of 109A A HL 06482T Soc Tr ang 113A A HL 06482 Soc Tr ang 99A A HL 06453V Soc Trang 100A A HL 06457H CaMau 114A A HL 06443 CanTho 110A A HL 06476 115A A HL 06457 CaMau 102A A HL 06463 V inhLong 105A A HL 0739 CanTho 92A A HL 0757 V inhLong 88A A HL 0753 CanTho 111A A HL 0716 75 CanTho 116A A HL 0715C V inhLong O/V IT/3/2006 O/V IT/1/2006 O/V IT/2/2006 83A A HL 0716 CanTho 95A A HL 06430 HauGiang 96A A HL 06434B HauGiang 9 93A A HL 0779CD1 LongA n 97A A HL 06443T Bac Lieu O/MA Y /8/2005 O/V IT/9/2005 9 O/V IT/1/2005 0 O/PHI/21/2003 (DQ164953) O/PHI/23/2003 (DQ164954) O/TA I/54-1/05B1* ( TRRL) O/TA I/54- 2/05B1* (TRRL) O/TA I/53- 2/05* ( TRRL) O/TA I/64-1/05* ( TRRL) 0 O/TA I/64-2/05* ( TRRL) O/TA I/35/05B2* (TRRL) 108A A HL 0735 DongThap O/V IT/2/2004 ( DQ165033) O/V IT/3/2004 ( DQ165034) O/V IT/11/2005 9 O/TA W/81/97 O/TA W/4/99 O/TA W/83/97 O/HKN/16/96 O/PHI/5/95 ( DQ164946) O/PHI/7/96 O/V IT/3/97 O/V IT/13/2002 (DQ165025) O/HKN/7/96 O/HKN/20/96 O/HKN/12/2005 O/HKN/15/2005 O/HKN/14/2005 O/HKN/9/2005 O/HKN/10/2005 9 O/HKN/17/2005 O/HKN/18/2005 O/HKN/19/2005 O/HKN/20/2005 O/HKN/25/2005 O/HKN/24/2005 O/HKN/3/2006 O/HKN/22/2005 O/HKN/23/2005 O/HKN/1/2006 O/HKN/5/2006 O/HKN/2/2006 O/HKN/4/2006 No of Taxa : 154 Data File : n:\evd\meg\db\fmdv\o\VIT2006B.MEG Data Title : Vietname - AAHL Data Type : Nucleotide (Coding) Analysis : Phylogeny reconstruction Tree Inference : ============================== Method : Neighbor-Joining Phylogeny Test and options : Bootstrap (1000 replicates; seed=64238) Include Sites : ============================== Gaps/Missing Data : Pairwise Deletion Codon Positions : 1st+2nd+3rd+Noncoding Substitution Model : ============================== Model : Nucleotide: Kimura 2-parameter Substitutions to Include : d: Transitions + Transversions Pattern among Lineages : Same (Homogeneous) Rates among sites : Uniform rates No of Sites : 639 No Of Bootstrap Reps = 1000 9 * Not a WRLFMD ref no IVRI, Indian Veterinary Research Institute (Mukteswar) TRRL, Thailand Regional Reference Laboratory, Pak Chong N.J Knowles, 12 Feb 2007 ME-SA SEA 9 EURO-SA CATHAY 0 O / IR N / / 0 O / IR N / / 0 O / IR N / / 0 O / IR N / / 0 O / IR N / / 0 O /P A K /9 /2 0 O /P A K /1 /2 0 O /N E P /2 /2 0 O /N E P /5 /2 0 ( D Q 6 ) O /B H U /1 /2 0 ( D Q ) O /B H U /4 /2 0 O /B H U /4 /2 0 ( D Q ) O /B H U /4 /2 0 ( D Q ) O /B H U /2 /2 0 ( D Q ) O /B H U /2 /2 0 ( D Q 4 ) O /B H U /3 /2 0 ( D Q 5 ) O /B H U /3 /2 0 ( D Q ) O /B H U /4 /2 0 ( D Q ) O /P A K /9 /2 0 O /P A K /1 /2 0 O /P A K /6 /2 0 O /P A K /4 /2 0 O /P A K /1 /2 0 9 O /J O R /5 /2 0 O /J O R /6 /2 0 O / IR N / / 0 O / IR N / / 0 O / IR N / / 0 O / IR N / / 0 9 O /M A Y /6 /2 0 ( D Q 5 ) O /M A Y /7 /2 0 O /M A Y /9 /2 0 0 O /M A Y /7 /2 0 O /M A Y /3 /2 0 0 O /M A Y /4 /2 0 O /M A Y /3 /2 0 O /M A Y /6 /2 0 O /M A Y /1 /2 0 O /M A Y /2 /2 0 O / IN D / / * ( IV R I) O / IN D / / * ( IV R I) 0 O / IN D / 5 / * ( IV R I) O /L A O /1 /2 0 ( D Q 1 ) O /L A O /1 /2 0 ( D Q 9 ) 0 O /L A O /5 /2 0 O /L A O /2 /2 0 ( D Q ) O /L A O /2 /2 0 ( D Q ) O /L A O /2 /2 0 ( D Q ) O / V IT / / 0 ( D Q ) O /L A O /3 /2 0 O /L A O /4 /2 0 9 0 O / V IT / / 0 V IT / - * ( A A H L ) O / V IT / / 0 O / T A I/ / 0 ( D Q ) O /J P N /2 0 ( K a n n o ) O / T A I/ / 9 ( D Q ) O /S K R /2 /2 0 O /S K R /1 /2 0 ( A J 8 ) O /T A W /2 /9 O /M A Y /2 /2 0 ( A J 8 ) O / T A I/ / 0 O / V IT / / 0 O / V IT / / 0 0 O / V IT / / 0 O /L A O /1 /2 0 ( D Q ) O /L A O /2 /2 0 0 O / M a n is a / T U R / ( A J 7 ) O /L A O /4 /9 ( D Q ) 9 O / T A I/ / 9 ( D Q 7 ) O /M A Y /1 /2 0 ( D Q ) 0 O / T A I/ 4 - / R B * ( T R R L ) 9 O /M Y A /1 /8 ( D Q ) O / T A I/ / * ( T R R L ) 9 O / T A I/ / 0 0 O / T A I/ / 0 O /M A Y /5 /2 0 O /M Y A /3 /0 * ( T R R L ) O /M Y A /4 /0 * ( T R R L ) O /M Y A /1 /9 O /M Y A /5 /9 ( D Q ) O /M Y A /2 /2 0 ( D Q ) O /M A Y /2 /2 0 O /M A Y /3 /2 0 ( D Q ) 0 O / T A I/ / 0 O / V IT / / 0 O /M A Y /5 /2 0 ( D Q ) 9 O /M A Y /6 /2 0 ( D Q 2 ) O /L A O /4 /2 0 ( D Q ) O /L A O /2 /2 0 O /M Y A /7 /2 0 ( D Q ) O / T A I/ / 0 9 O /M A Y /1 /2 0 O /M A Y /2 /2 0 O / T A I/ / 0 ( D Q ) O / V IT / / 0 V IT / - * ( A A H L ) O / V IT / / 0 O / V IT / / 0 0 O / V IT / / 0 V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / 2 - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) O / V IT / / 0 V IT / - * ( A A H L ) O /B F S /U K /6 O /H K N /6 /8 O / P H I/ / 0 ( D Q ) O / P H I/ / 0 O / P H I/ / 0 ( D Q ) O /T A W /4 /9 O / P H I/ / ( D Q ) O / P H I/ / O /H K N /1 /2 0 O /H K N /1 /2 0 0 O / T A I/ / 0 O / V IT / / 0 ( D Q 3 ) 0 V IT / - * ( A A H L ) O / V IT / 1 / 0 0 V IT / 3 - * ( A A H L ) O / V IT / / 0 O / V IT / / 0 9 V IT / - * ( A A H L ) 0 O /M A Y /8 /2 0 V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / 1 - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) V IT / - * ( A A H L ) O / V IT / / 0 V IT / - * ( A A H L ) 0 A/VIT/10/2005 A/VIT/44-05* (AAHL) A/VIT/43-05* (AAHL) A/VIT/8/2005 78 A/VIT/13/2005 98 A/VIT/45-05* (AAHL) A/TAI/45/04* (TRRL) 95 A/TAI/48-2/04R2* (TRRL) A/TAI/66/04R1* (TRRL) A/TAI/3-3/05* (TRRL) 75 A/TAI/1/05R2B2* (TRRL) A/TAI/4/05R1* (TRRL) A/TAI/6/05* (TRRL) A/TAI/3/2005 A/VIT/10/2004 A/VIT/6/2004 95 97 A/VIT/42-04* (AAHL) A/TAI/9/2004 A/VIT/4/2004 70 A/LAO/36/2003 A/LAO/1/2006 A/MAY/4/2003 A/TAI/8/04R2* (TRRL) A/VIT/12/2004 A/TAI/28-04* (TRRL) A/TAI/11/2003 70 A/TAI/12/2003 A/TAI/6/2004 A/TAI/11/04* (TRRL) A/TAI/2/04R2* (TRRL) A/TAI/10/2003 72 A/TAI/4/2003 A/TAI/5/2003 A/TAI/4/04R2* (TRRL) A/TAI/7/2003 A/TAI/6-3/04* (TRRL) A/MAY/1/2003 77 A/MAY/3/2003 A/TAI/8/2003 A/TAI/3507/47R2* (TRRL) A/VIT/11/2004 A/VIT/5/2004 94 99 A/VIT/9/2004 A/TAI/11/2005 A/VIT/18/2005 A/VIT/14/2005 100 A/VIT/46-05* (AAHL) A/TAI/7/2002 A/MAY/2/2002 100 85 A/TAI/3/2002 A/TAI/3/2001 A/TAI/2/2002 A/TAI/1/2001 A/TAI/2/97 (EF208778) A/TAI/6/99 A/TAI/2/98 A/TAI/1/99 A/TAI/4/2005 A/TAI/7/2005 100 A/MAY/5/2005 A/TAI/2/2005 A/TAI/14-1/04* (TRRL) A/TAI/118/87* (EF208777) A/IND/68/2001* (AF390659) A/IND/7/82 (1980) 86 No of Taxa : 93 Data File : n:\evd\meg\db\fmdv\a\VIT2005B.MEG Data Title : Vietnam A (AAHL) Data Type : Nucleotide (Coding) Analysis : Phylogeny reconstruction Tree Inference : ============================== Method : Neighbor-Joining Phylogeny Test and options : Bootstrap (1000 replicates; seed=64843) Include Sites : ============================== Gaps/Missing Data : Pairwise Deletion Codon Positions : 1st+2nd+3rd+Noncoding Substitution Model : ============================== Model : Nucleotide: Kimura 2-parameter Substitutions to Include : d: Transitions + Transversions Pattern among Lineages : Same (Homogeneous) Rates among sites : Uniform rates No of Sites : 642 N.J Bootstrap 12 = 1000 No Of Knowles,RepsFeb 2007 * Not a WRLFMD ref no IVRI, Indian Veterinary Research Institute (Mukteswar) TRRL, Thailand Regional Reference Laboratory, Pak Chong N.J Knowles, 12 Feb 2007 99 99 75 71 99 75 100 A/IND/17/77* (AF204108) A22/IRQ/24/64 (AJ251474) A/IRN/87 A/SAU/41/91 A/IRN/22/99 (EF208772) A/IRN/1/96 (EF208771) A/SAU/23/86 86 A/BHU/41/2002 100 A/BHU/7/2003 A/BHU/27/2003 100 78 ASIA A/IND/16/2000* (AF390610) A/IND/78/2000* (AF390664) A/IND/50/2000* (AF390653) A/IND/24/2001* (AF390624) A/IND/38/2000* (AF390643) A/IND/173/2000* (AF390616) A/IND/84/2000* (AF390668) A/IND/126/2000* (AF390599) A/IND/80/2000* (AF390665) 99 100 A15/Bangkok/TAI/60 (AY593755) A/EGY/1/2006 (EF208757) A/K35/80* A/K5/80* A10/HOL/42 (M20715) A24/Cruzeiro/BRA/55 (K03340) 99 AFRICA 87 EURO-SA 97 89 100 86 0.02 A/PHI/1/77 A/PHI/10/75 A/PHI/1/76 Asia Sequence sp147 Da Nang sp148 North 3kimron iso61.seq isrl3-63.seq IND 63-72.seq YNBS-58.seq pak iso3.seq Jiangsu-China-2005.seq spl47 spl48 AFG 1-2001 BHU 27-2002 HNK-CHA-05.seq Leb83.seq leb-89 iso89.seq 0.1 Appendix Results from Serosurveilance in 2007 from all provinces in the Project The table shows the ELISA results for each province with the number of animals positive for each serotype after vaccination and the number of animals positive to nonstructural antibodies (3ABC ELISA positives) and hence exposed to FMDV Province BinhPhuoc Vaccinated O A Asia 3ABC Cow 101 80 79 56 68 Pig DongThap Species 120 15 0 Cow 109 43 87 57 Pig TayNinh 0 0 Cow 60 Pig LongAn 0 0 Cow 60 42 59 59 13 Pig KienGiang 0 0 Cow 120 32 30 23 40 Pig 0 56 37 44 120 0 0 Cow 67 29 40 17 0 0 Cow 120 95 119 115 Pig 0 0 Cow *1 1 0 Pig KonTum 79 Pig QuangNam 120 Pig AnGiang 117 Cow 0 0 Vaccinated with O, A Asia vaccine Vaccinated in Feb 2007 Samples Collected April 10 2007 Cow Pig QuangNinh LangSon Note: Below are tables showing identification of FMDV circulating in Dong Thap by serology Serosurveillance data used to show serotypes of FMD circulating in Dong Thap in 2007 The Data shows that both serotype A & O may be circulating in 2007 Field isolates send to the laboratory for serotyping have only identified serotype O Serology suggests both O & A Sample B09 B10 B15 B16 B17 B19 B20 B21 B22 B23 B26 B27 B29 B31 B32 B33 B34 B35 B44 B46 Field Sample number Serotype O Titer final PI 1280 52 320 58 80 53 80 65 80 57 640 65 2560 56 320 55 1280 54 1280 53 320 58 80 58 80 63 80 57 80 56 80 61 80 57 1280 56 80 60 320 56 Elisa Result PI Type O LP ELISA RESULT 2007 Serotype A Serotype Asia Titer final PI Titer final PI 320 57 80 57 160 56 40 52 1280 54 80 53 1280 54 80 54 320 53 40 56 2560 56 80 67 2560 67 80 74 320 61 40 65 320 56 80 57 320 57 80 57 2560 65 320 57 320 56 80 52 320 64 80 56 640 58 80 53 640 65 160 53 320 72 160 53 640 67 320 52 2660 63 320 54 80 57 40 52 320 65 80 63 Elisa Result PI Titre Type A Elisa Result PI Titre Asia Titre B-242 68 160 70 160 38 neg B-250 55 160 68 160 73 80 B-253 74 320 70 320 71 160 B-254 70 320 71 320 70 160 B-257 67 160 60 320 29 neg B-260 65 160 53 640 69 80 B-263 70 160 62 640 65 160 B-264 59 160 57 320 41 63 160 75 640 75 160 All 3ABC Positive Outbreak O 2006/07 & A 2005 Can Serology be used to identify circulating FMDV? Serum from Dong Thap 2007 neg B-265 Serum from Dong Thap 2007 Vaccinated in 2007 with O, A Asia vaccine B-269 72 160 77 1280 77 320 14A-B12 67 160 55 320 76 80 14A-B14 72 160 70 320 69 80 14A-B24 74 320 59 640 73 80 14A-B25 70 320 57 640 70 80 14A-B47 65 80 64 160 65 80 14A-B48 73 160 59 320 72 80 14A-B49 65 160 57 640 54 160 14A-B87 56 160 51 640 63 80 14A-B88 80 160 51 640 71 80 14A-B89 71 160 68 640 74 80 Vaccinated in 2007 with O, A Asia vaccine All 3ABC Negative Outbreak O 2006/07 & A 2005 Can Serology be used to identify circulating FMDV? ... provided and are continuing to provide support and training to other laboratories in the network The project has highlighted the need for training of field veterinarians in the collection of data and. .. diagnosis and control of FMD by the provision of resources and training of staff in required methods and quality assurance To provide accurate data to explain failure of vaccination to control FMDV and. .. has assisted in developing the skills of the epidemiologists in serosurveillance, outbreak control and investigation of vaccine failure Information from the serosurveillance in the project was