Đề tài Development of an Improved Capability in support of N- Milestone 5

CSIRO-AAHL AUSAID DAH-VN CARD-072/04VIE Ministry of Agriculture & Rural Development 062/05VIE 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 5: Progress Report Date 31/12/2006 1 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_______________________________________________ 12 3 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 007611201/07/2006 – 31/12/2006 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 4 2. Project Abstract The projects 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 was to establish FMD diagnostics in the collaborating laboratories with a focus on the HCMC laboratory. All consumables and reagents were supplied to the laboratories in Vietnam for the second year of the project. AAHL staff prepared the protocols necessary for the training of laboratory scientists. Two scientists from the Regional Animal Health laboratory HCMC trained at AAHL in molecular techniques for the diagnosis of FMD and standardisation of the protocols and reagents to be used in the project for sequencing FMD isolates from Vietnam. The project carried out training at the HCMC and Hanoi laboratories in molecular technologies for diagnosis of FMD and reviewed ELISA results for the first round of sero- serveillance. An important part of the laboratory training was Quality Assurance (QA) in the laboratory to ensure tests were running according to a standard protocol and with Internal Quality Assurance (IQC) to allow AAHL staff to audit the results from the laboratory and give the appropriate data to allow trouble shooting. The QA records were reviewed during training visit to the laboratories. The Regional Animal Health Centre in Ho Chi Minh City also continued to train the field veterinarians on how to collect samples and how to collect information necessary for the history of the animals on the villages in the project. It was identified that data collected from the field was not suitable from the first round of sero-surveillance. The funding was supplied for the second year of field studies. Avian Influenza (AI) continues to delay the sero-surveillance in the field as field staff deal with the AI outbreak. . 5 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: 6 • Training of laboratory staff at AAHL. Two representatives from the HCMC laboratory were trained in molecular techniques for the detection of FMD genome and the standardisation of reagents and protocols needed to establish PCR in HCMC. Training was carried out under a quality system emphasizing the importance of Quality Assurance in the laboratory focusing on test maintenance, record keeping and data collection. • A consultant from AAHL completed training of HCMC-RAHC staff in molecular techniques and established the technology to be used to identify FMD isolates from the field in the HCMC laboratory. Again Quality Assurance, record keeping and data collection was emphasized. • 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, • Further training of field veterinarians was carried out with feedback of the results from first rounds of testing and experiences from the first round of sample collection to increase the quality of samples and data collected. • The consultant from AAHL also reviewed the transfer of ELISA technology to each laboratory and the implementation of a Quality Assurance system into the laboratory. The testing of sera in each laboratory was discussed with the Nth Vietnam sample collection needing to improve. Also training of RAHC-HCMC staff in FMD serology using cell culture to carry out the Virus Neutralisation Test (VNT) was part of training. • Sampling for next round of 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 • ELISA technology in use at collaborating laboratories in HCMC, Hanoi and Can Tho, for sero-typing of FMD tissue samples from the field. Sera from sero-surveillance tested at RAHC-HCMC and Hanoi laboratories. • Virus isolation in use at RAHC-HCMC. Molecular technology and serology by VNT using cell culture has started to be used. • Supply of consumables and further reagents for testing of samples by FMD ELISA, molecular technology and cell culture. • Project meeting held at AAHL with Director of DAH, Vietnam and Director of RAHO-HCMC to discuss project outcomes. Training in QA and biosafety given. Also held a meeting in Canberra with DAFF to discuss Australia’s preparedness for disease outbreaks. Project achieving aims for the DAH in establishing framework for control of FMD. Report from Vietnam: 30/06/31/12/ACTIVITIES from June 1st 2006 to Dec 31th 2006 7 1.Training No. Date & place Subject Person in charge Remark 1 July 2006 November 2006 -Expert from AAHL working in Viet Nam to review data from first round of sero-surveillance. Discuss improved data collection and records Expert from AAHL to complete molecular training and review FMD diagnostics in collaborating laboratories Chris Morrissy and Peter Durr Chris Morrissy Visit HCMC and Hanoi laboratories 2 23th. Oct. - 8th.Dec., 2006 AAHL, Australia -Sequencing and analysis of FMDV -Virus Inactivation by BEI to produce antigen for ELISA , practice on Blue Tongue virus and VSV - Chris Morrissy - Ian Pritchard - Tony Pye - Kim Newbery - Lynda Wright - Cathy Williams - 1 Participants from RAHC-HCMC 3 -13th. Nov. – 8th. Dec., 2006 - Analysis of FMDV sequencing data. -Virus Inactivation by BEI to produce antigen for ELISA , practice on Blue Tongue virus and VSV - Chris Morrissy - Ian Pritchard - Tony Pye - Kim Newbery - Lynda Wright - Cathy Williams - 1 participant from RAHC-HCMC 4 13th. -18th. Nov, 2006 In Australia -Review the project - Chris Morrissy - Dr.Bui Quang Anh (Director General of DAH of Viet Nam) - Dr.Dong Manh Hoa ( Director of RAHC-HCMC, main Viet Nam partner) 2. Field activities 2.1.1. Visiting 8 provinces in the project (Middle and Southern ) , Oct. 2006. • Transfer the equipments and documents ( form, questionnaire) for sample collection. • Transfer money for sample collection. • Feed back the results from second 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) 8 2.1.2. Visiting 2 provinces in the project ( North ) , Oct. 2006. • Transfer the equipments and documents (form, questionnaire) for sample collection. • Transfer money for sample collection. • Feed back the results from second 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 third 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 ), total 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 : 103 tissue samples from the North , 2.3.2 RAHC-HCMC : 189 tissue samples from the South . 3. Laboratory Activities 3.1 National Vaterinary Diagnostic Centre- Ha Noi ( NVDC-Ha Noi): 3.1.1 Testing on tissue samples for FMDV typing: 103 samples, all of them are serotype O 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 Testing on sera for FMDV antibodies detection : 480 sera, test finished 3.2.2 Sending 480 sera to RAHC-HCMC for retest : 480 sera 3.3 Regional Animal Health Center Can Tho (RAHC-Can Tho) 3.3.1 Testing on sera for FMDV antibodies detection : 480 sera, test finished 3.2.2 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: 189 samples, 01 of them is serotype Asia 1, 179 samples are serotype O and 09 samples are negative Remark: test covers 3 serotype: serotype A, O and Asia 1 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 Isolate FMDV by BHK cell line from 32 selected samples, 01 serotype Asia1 and 31 serotype O. 3.4.4 Application of SNT protocol in testing phase : Adapt 5 FMDV on BHK cell line to produce reference virus for using in SNT for • Serotype O, PanAsia ( ME) topotype : 01 • Serotype O, South EastAsia ( SEA) topotype : 01 • Serotype O, Cathay topotype : 01 • Serotype A : 01 • Serotype Asia 1: 01 9 • Titrate these viruses Setup SNT using the serum from vaccine trial on cattle as reference serum and to the vaccine which was selected for applying in FMD National Control program since 2006. 3.4.5 RT-PCR application Extract RNA from FMDV isolates • Serotype O: 74 (52 from the South and 22 from the North) • Serotype A: 9 • Serotype Asia 1: 4 Make cDNA from these RNA Send cDNA and amplicons to AAHL for sequencing The number of FMDV cDNA sequenced at AAHL: • Serotype Asia 1: 4 • Serotype A: 9 • Serotype O: 55 (52 from the South and 03 from the North) 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, knowledge on the selection of the right vaccine and improving the efficacy of vaccination 5.3 Capacity Building Training and education of field veterinarians in disease prevention, disease investigation and sample collection has continued with lessons learnt from the first round of sero-surveillance and sample collection. This training has already shown an impact with an increase in quality of sample collection and number of samples collected and submitted to the laboratory. The project has provided training and technology transfer of FMD diagnostics to each laboratory involved in the project. Reagents and standard methods have been supplied to each laboratory giving them the diagnostic capability for FMDV diagnosis and serology using ELISA technology, this technology is now being practised at each laboratory. The RAHC- HCMC has virus isolation and cell culture techniques established and has begun to use molecular technology and serology by VNT using cell culture. RAHC-HCMC now has capacity to carry out RT-PCR for FMD for detection of FMD antigen and for sequence analysis. 5.4 Publicity The CARD AusAID project have received publicity through the training programs and also through the achievements so far in the project in understanding FMD in Vietnam. FMD is a 10 disease on importance in Vietnam and this has put our project into the lime light. The project has been publicised through a press release in Australia and articles in news letters including the SEAFMD newsletter and on the internet. The results from the project have been presented at OIE/SEAFMD meetings during the project. 5.5 Project Management Implementation of the project is on schedule. AAHL has kept the project on target and has supported or requests from the Vietnamese partners. Training has been carried out in Australia and Vietnam with both partners benefiting from the information that is being generated. Implementation in Vietnam is also continuing to go well. Trainees have been supplied in a timely manner, from DAH. The field work has been organised well with data and samples being collected from farms in each pilot zone. Training and the transfer of information to other regional laboratories in Vietnam is also occurring, RAHC-HCMC is supplying support to these laboratories. 6. Report on Cross-Cutting Issues 6.1 Environment No direct environmental impacts associated with project activities have been identified. On a broader