Đề tài Development of an Improved Capability in supporte in Cattle and Pigs - Milestone 9

Ministry of Agriculture & Rural Development Progress 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 9: 5th 6 monthly Report Date 01/07/2007 to 31/12/12/2007 Table of Contents 1. Institute Information ____________________________________________ 3 2. Project Abstract _______________________________________________ 4 3. Executive Summary ____________________________________________ 4 4. Introduction & Background ______________________________________ 5 5. Progress to Date _______________________________________________ 6 5.1 Implementation Highlights ___________________________________ 6 5.2 Smallholder Benefits ________________________________________ 7 5.3 Capacity Building___________________________________________ 8 5.4 Publicity __________________________________________________ 8 5.5 Project Management ________________________________________ 8 6. Report on Cross-Cutting Issues __________________________________ 8 6.1 Environment_______________________________________________ 8 6.2 Gender and Social Issues ____________________________________ 8 7. Implementation & Sustainability Issues ____________________________ 9 7.1 Issues and Constraints ______________________________________ 9 7.2 Options ___________________________________________________ 9 7.3 Sustainability ______________________________________________ 9 8. Next Critical Steps ____________________________________________ 10 9. Conclusion __________________________________________________ 10 10. Statuatory Declaration ___________________ Error! Bookmark not defined.5 11. Project progress against proposed objectives, outputs, activities and inputs___________________________________________________ 16 1. Institute Information Project Name Vietnamese Institution Regional Animal Health Centre, Ho Chi Minh City (RAHC-HCMC), South Vietnam. Vietnamese Project Team Leader Dr. Dong Manh Hoa Australian Organisation Australian Animal Health Laboratory (AAHL), PMB 24, Geelong, 3213, Australia Australian Personnel Mr Chris Morrissy Date commenced 01/06/2005 Completion date (original) 01/06/2008 Completion date (revised) Reporting period 01/07/2007 – 31/12/2007 Contact Officer(s) In Australia: Team Leader Name: Mr Chris Morrissy Telephone: +61 3 5227 5000 Position: Diagnostic Virologist Supervisor Mammalian Virology Fax: +61 3 5227 5555 Organisation Australian Animal Health Laboratory (AAHL), PMB 24, Geelong, 3213, Australia Email: chris.morrissy@csiro.au In Australia: Administrative contact Name: Mr Chris Morrissy Telephone: +61 3 5227 5000 Position: Patents Contracts Officer Fax: +61 3 5227 5555 Organisation Australian Animal Health Laboratory (AAHL), PMB 24, Geelong, 3213, Australia Email: chris.morrissy@csiro.au In Vietnam Name: Dr. Dong Manh Hoa Telephone: + 84 8 8568220 Position: Director Fax: + 84 8 8569050 Organisation Regional Animal Health Centre, Ho Chi Minh City (RAHC-HCMC), South Vietnam. Email: rahchcmc@hcm.vnn.vn 2. Project Abstract The project’s purpose is to determine why there are vaccine failures, and to investigate what serotypes of FMDV are circulating in Vietnam so that the correct vaccines can be used for these serotypes. Regional laboratories are setup 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 and laboratory testing for both virus typing and sero- surveillance have begun. 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 are established in provinces near the borders of Vietnam to study serotypes circulating in Vietnam and to determine their origin. The quality and collection of samples have increased giving more data on the FMD situation in Vietnam. Molecular epidemiological studies of the FMDV isolates in these provinces will provide insights to the effectiveness of border control and the origin of FMDV circulating in Vietnam each year. Virus isolation and molecular studies have started to be carried out on FMD samples from the field. A diagnostic capacity for FMD would allow the early detection and identification of disease enabling better control of disease and help reduce loss of livestock and so productivity. 3. Executive Summary The main focus of this reporting period is the sequencing, genotyping and analysis of 50 Vietnamese FMD isolates collected in 2006-07 and the analysis of the data from sero-surveillance. The isolates were sequenced and analysed at AAHL by a scientist from RAHO-HCMC and AAHL scientists. The sequence data was then sent to WRL for confirmation and comparison to other FMD isolates. This information is given to SEAFMD as part of the regional project to control FMD. The from the sero-surveillance collected to date in the project was reviewed to give a 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 (see copy of PowerPoint presentation attached). There has been improvement in the quality and number of samples submitted to the laboratory for serotyping by ELISA allowing DAH to better understand and identify the circulating serotypes of FMD viruses in Vietnam. The project epidemiological and sero-survellance programs are now operational with the capability to determine the serotypes circulating in Vietnam and the effectiveness of the FMD vaccination campaign. The establishment FMD diagnostics in the collaborating laboratories is continuing with the HCMC laboratory having FMD diagnostics established for virus isolation, virus neutralisation test, ELISA, PCR and sequencing. The NCVD-Hanoi laboratory has diagnostics for virus isolation, ELISA and PCR established and is implementing sequencing capability. The Can Tho and Da Nang laboratory have the capability for FMD diagnosis for serotyping and serology by ELISA. One consultant worked with the RAHO-HCMC laboratory to compare and establish the FMD ELISAs using antigen produced at HCMC using Vietnam isolates. The ELISA test was standardised using Vietnam antigen and reference serum from AAHL and Vietnam reference sera collected during the sero-surveillance. The ELISA was validated and the Vietnam antigen is able to be used for serology and as a control in the serotyping ELISA. AAHL project leader visited HCMC and Hanoi to review FMD diagnostics and QA and analyse the data collect in the project from serosurveillance and disease investigations. Quality of diagnostic tests continues to improve and data collected as improved in most of the provinces carrying out sero- surveillance. Both the HCMC and Hanoi laboratories are applying the new technologies to other disease problems in Vietnam eg Hanoi used virus isolation to isolate new isolate of PRRS in Vietnam. The project was invited to number of regional meetings to present data from Vietnam on control of FMD. The project has coordinated activities with the AusAID capacity building project to allow both projects to achieve its objectives, eg combined PCR and sequence training with sequencing FMD isolates along with AI, PRRS and CSF isolates from Vietnam. Avian Influenza (AI) continues to delay the sero-surveillance in the field as field staff deal with the AI outbreak. 4. Introduction & Background Objectives of the project: 1. 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. 2. 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. Implementation Approach and Strategy The project approach is thought to be the most appropriate for developing an understanding of FMD epidemiology in Vietnam. The approach for technology transfer is well established at AAHL and has been successfully applied in previous projects in Vietnam, Thailand and Indonesia. The field studies and epidemiology and sero-surveillance approaches have been 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. 5. Progress to Date 5.1 Implementation Highlights Main achievements for this reporting period were: • Sequencing, genotyping and analysis of 50 Vietnamese FMD isolates collected in 2006-07 and the analysis of the data from sero-surveillance. The isolates were sequenced and analysed at AAHL by a scientist from RAHO-HCMC and AAHL scientists. The sequence data was then sent to WRL for confirmation and comparison to other FMD isolates. This information was shared with SEAFMD as part of the regional project to control FMD. • A consultant from AAHL (project leader) reviewed data from the field in the HCMC laboratory and outlined the improvements required in data collection. The data from the sero-surveillance collected to date in the project was reviewed to give a 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 (see copy of PowerPoint presentation attached). 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. Plans were made to further investigate the difference between vaccine coverage in each of the provinces and to determine why vaccination of pigs was giving a poor result. • There has been improvement in the quality and number of samples submitted to the laboratory for serotyping by ELISA allowing DAH to better understand and identify the circulating serotypes of FMD viruses in Vietnam. The improvement in sample collection has allowed 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. The project epidemiological and sero-survellance programs are now operational with the capability to determine the serotypes circulating in Vietnam and the effectiveness of the FMD vaccination campaign. • The establishment FMD diagnostics in the collaborating laboratories is continuing with the HCMC laboratory having FMD diagnostics established for virus isolation, virus neutralisation test, ELISA, PCR and sequencing. The NCVD-Hanoi laboratory has diagnostics for virus isolation, ELISA and PCR established and is implementing sequencing capability. The Can Tho and Da Nang laboratory have the capability for FMD diagnosis for serotyping and serology by ELISA. The laboratories have worked closer together and collaborated more since the project began. • One AAHL consultant worked with the RAHO-HCMC laboratory to compare and establish the FMD ELISAs using antigen produced at HCMC using Vietnam isolates. The ELISA test was standardised using Vietnam antigen and reference serum from AAHL and Vietnam reference sera collected during the sero-surveillance. The ELISA was validated and the Vietnam antigen is able to be used for serology and as a control in the serotyping ELISA. The production of FMD ELISA antigen allows the laboratories to be more self sufficient and is the start of the laboratories capability to produce its own reagents. • TTraining of field veterinarians is on going to increase the quality of sample and data collection to ensure issues of vaccine failure can be addressed. This training has continued throughout the project to ensure field veterinarians will understand the importance of data collection to the final disease diagnosis. • AAHL consultant reviewed 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 and for isolation of other disease agents. ELISA and molecular techniques were also reviewed along with Quality Assurance record keeping and data collection to ensure test records were being maintainedand results were being interpreted correctly. . • Sampling for next round sera collection completed for each province for epidemiological studies and sero-surveillance. The collection of tissue samples from outbreaks continued to increase giving valuable information on the serotypes circulating in Vietnam • Virus isolation and cell culture in the process established at the Hanoi laboratory. • The supply of consumables and further reagents for testing of samples by FMD ELISA, molecular technology and cell culture were supplied. • Funding for serosurveillance was supplied to the RAHO-HCMC for the final collection of samples. Report from Vietnam: ACTIVITIES from June. 1st. 2007 to Dec. 31th. 2007 1. Training No. Date & place Subject Person in charge Remark 1 Sept. 2007 August – Sept 2007 - Expert from AAHL working in Viet Nam Expert from AAHL working in Viet Nam Lynda Wright Chris Morrissy - Visit 4 laboratories , checking the activities of these laboratories; delivery FMDV antigen which was produced by RAHC-HCMC and other reagents for FMD LP- ELISA to RAHC-Can Tho, Da Nang and NCVD-Ha Noi - Visit Hnaoi & HCMC to review serosurvalence data and application of FMD diagnostics 2. Field activities 2.1.1. Visit to 8 provinces in the project (Middle and South Vietnam), Oct. 2007. • Transfer the equipments and documents ( form, questionnaire) for sample collection • Transfer money for sample collection • Feed back the results from fourth testing round and share the experience on sample collection with the field veterinarian staff to increase the quality of collected samples • Staff in charge : Dr. Hoa, Dr. Vu , Dr. Ha ( RAHC-HCMC) - Dr. Quan , Dr.Quang (RAHC-Da Nang ) – Dr. Thanh , Dr. Dung ( RAHC-Can Tho) 2.1.2. Visiting 2 provinces in the project (North Vietnam), Oct. 2007. • Transfer the equipments and documents (form, questionnaire) for sample collection • Transfer money for sample collection • Feed back the results from fourth testing round and share the experience on sample collection with the field veterinarian staff to increase the quality of collected samples • Staff in charge : Dr. Cam , Dr. Tung ( NVDC- Ha Noi) 2.2. Collect sera samples from 10 provinces for the fifth round of surveillance , 120 cattle sera and 120 pig sera from each province. 2.2.1 NVDC-Ha Noi : 02 provinces (Quang Ninh and Lang Son) , total 480 sera samples. 2.2.2 RAHC-Da Nang : 02 provinces (Kontum and Quang Nam) , total 480 sera samples. 2.2.3 RAHC-Can Tho : 02 provinces (An Giang, Kien Giang ), toatl 480 sera. 2.2.4 RAHC-HCMC : 04 provinces ( Binh Phuoc, Tay Ninh, Long An, Dong Thap), total 960 sera 2.3 Collect tissue samples for FMDV typing 2.3.1 NVDC-Ha Noi: 19 tissue samples from the North , 2.3.2 RAHC-HCMC: 00 tissue samples from the South (No FMD outbreak in the South since May ) 3. Laboratory Activities 3.1 National Vaterinary Diagnostic Center- Ha Noi ( NVDC-Ha Noi): 3.1.1 Testing on tissue samples for FMDV typing : 19 samples, 12 of them are serotype O and 7 are serotype Asia 1 3.1.2 Testing on sera for FMDV antibodies detection : 480 sera , test finished 3.2 Regional Animal Health Center Da Nang (RAHC-Da Nang) 3.2.1 Sending 480 sera to RAHC-HCMC for retest : 480 sera 3.3 Regional Animal Health Center Can Tho (RAHC-Can Tho) 3.2.1 Sending 480 sera to RAHC-HCMC for retest : 480 sera 3.4 Regional Animal Health Center HCMC(RAHC-HCMC) 3.4.1 Testing on tissue samples for FMDV typing : 00 samples (no FMD outbreak in the South since May 2007) 3.4.2 Testing on sera for FMDV antibodies detection: 1920 sera ( 960 sera in the region and retest 960 sera from RAHC-Can Tho and Da Nang ). Test finished 3.4.3 Production FMDV for Virus Neutralization Test (VNT) Passage FMD viruses 3-4 time on BHK cell culture to produce stock virus for use in VNT Batches of FMDV for VNT: • Serotype O : • ME topotype : 02 • SEA topotype: 02 • Cathay topotype : 02 • Serotype A : 02 • Serotype Asia 1: 02 Titrate VNT virus stocks and keep at -80oC ready for using in VNT. 3.4.4 Production FMDV ELISA antigen Produce a second batch of FMDV antigens • Serotype O : 50 ml • Serotype A : 50ml • Serotype Asia 1: 50ml Test this antigen with reference sera panel from AAHL Deliver this antigen and other reagents for FMDV LP- ELISA to RAHC- Can Tho, Da Nang and NCVD (10ml Serotype O and 10ml Serotype A antigen for each laboratory) for testing the repeatability of the ELISA test while using this antigen in another laboratory. 3.4.5 Application RT-PCR • Extract RNA from isolated FMDV • Serotype O: 34 • Serotype A: 1 • Make cDNA from these RNA . • Sending cDNA and amplicons to AAHL for sequencing ( all of them are good for sequencing at AAHL) Report by: Ngo Thanh Long Note: Further detail on activities for reporting period in log frame. 5.2 Smallholder Benefits All pig and cattle and producers are potential beneficiaries. Those that take up the advice and use vaccine according to recommendations will benefit financially through reduced losses due to death and disease in their cattle and pigs. The benefits of a more profitable farming operation flow to all family members. Farmers and district veterinarians will have improved knowledge and skill in disease prevention,