1.2 OUTLINE OF OFFICIAL ARRANGEMENTS
2.2 VACCINE PRODUCTION AND RELATED ACTIVITIES
2.3 VETERINARY ADVISORY SERVICES
3.1 BACKGROUND TO THE RECOMMENDATIONS
3.7 VACCINE MARKETING, COMMERCIALIZATION AND LEGAL ASPECTS
3.8 TRAINING, BACK-UP SERVICES AND INFORMATION DISSEMINATION
ABH - Anaplasma, Babesia and Heartwater
CVL - Central Veterinary Laboratory
DANIDA - Danish International Development Agency
ECF - East Coast Fever
GLP - Good Laboratory Practices
GMP - Good Manufacturing Practices
IBAR - Inter-Bureau for Animal Resources
ILRI - International Livestock Research Institute
OAU - Organization of African Unity
SADC - Southern African Development Community
STRC - Scientific, Technical and Research Commission
TBD - Tick-borne Disease
TCP - Technical Cooperation Programme
UNDP - United Nations Development Programme
VPC - Vaccine Production Centre
The Government of Malawi has long recognized the severe constraint imposed upon the cattle industry by the presence of ticks and tick-borne diseases (TBD), in particular by East Coast Fever (ECF), the most important TBD in the country. The progress made in the 1970s by the FAO/UNDP project RAF/67/007 in developing a method for immunizing cattle against ECF, led the Government of Malawi to request assistance in order to exploit the new findings in Malawi. The assistance was received through project GCP/MLW/018/DEN "East Coast Fever Immunization", and projects TCP/MLW/4505 and TCP/MLW/6652 "Bridging Assistance to East Coast Fever Vaccine Production".
A routine diagnostic and research laboratory, the Central Veterinary Laboratory (CVL), was built in Lilongwe. This provided animal accommodation with handling facilities for animal disease research, and a 22 ha farm for producing cattle feed. Field immunization trials held at the CVL demonstrated the applicability of the infection-and-treatment method of vaccinating cattle against ECF in Malawi. The results confirmed similar observations from the United Republic of Tanzania, Kenya and Zambia, auguring well for a common regional approach in tackling ECF immunization.
The Organization of African Unity's Scientific, Technical and Research Commission (OAU/STRC) identified the CVL as a regional centre for eastern and central Africa, which was to focus on developing and introducing methods of immunizing cattle against TBD in the field using live vaccines. Towards the late 1980s, the Government of Malawi, through the Southern African Development Community (SADC), requested assistance from FAO in several areas of TBD control. The assistance was received through project GCP/RAF/247/NET "East Coast Fever Vaccine Production and Quality Control", with funding from the Government of the Netherlands, and through projects GCP/RAF/259/DEN and GCP/RAF/236/DEN on the production of Anaplasma, Babesia and Heartwater (ABH) vaccines for use in eastern Africa, funded by the Government of Denmark.
The above projects, based at the CVL, led to the construction of a purpose-built TBD Vaccine Production Centre (VPC). The new facility allowed production of better quality ECF vaccines by the incorporation of standardized vaccine quality control measures. The improvements also provided better facilities for training national and regional personnel in the various techniques essential in laboratory TBD diagnosis. The facility provided more efficient laboratory back-up service for other projects in the region, thus laying the foundation for the VPC to coordinate ticks and TBD control activities in the region.
Meetings held in Kampala, Uganda, under the joint auspices of FAO, OAU-IBAR, and ILRAD recommended the initiation of a pre-investment phase of a TBD control programme by immunization, with commercial cost-recovery of the vaccines and delivery in a sustainable manner. A follow-up Phase III of the TBD programme was planned to consolidate the advances made by Phase II, which came to an end in 1992. Phase III was to focus on the implementation of large-scale field immunization against the major TBDs affecting cattle in the region.
Projects GCP/RAF/291/DEN and GCP/RAF/293/BEL, which aimed to merge the activities of projects RAF/92/010, GCP/RAF/247/NET and GCP/RAF/259/DEN, were components of the FAO Coordinated Multi-Donor Programme for the Control of Ticks and TBD in Eastern, Central and Southern Africa. The principal donors were the Governments of Belgium, the Netherlands and Denmark.
The substantive project: "Regional Pre-investment Project for the Commercial Production of Vaccines against Tick-borne Diseases" never became operational. Interim phases were operated first for 12 months in 1994, then for four, two and six months in 1995, for two six-month periods in 1996, and, finally, for six and three months in 1997. The Governments of Belgium, Denmark and the Netherlands agreed in principle to co-fund the Integrated Tick and TBD Control Programme in eastern, central and southern Africa. Owing to their concerns regarding the international technical assistance, considered top-heavy, and the legal aspects linked with the creation of a vaccine producing entity, the donors agreed with FAO to the interim phases in order to allow resolution of the issues before the project became fully operational. The Project Document was signed by FAO and the Government of Belgium on 3 December 1993. The implementing agency was the Ministry of Agriculture in each country, on behalf of the SADC. The total donor contribution to the project was $US 1 384 420 and $US 346 105 from the Governments of Denmark and Belgium, respectively. Project GCP/RAF/293/BEL, which started in January 1994, took over the funding of titrating activities from GCP/RAF/291/DEN when the latter came to an end on 30 June 1997. The Government of Malawi contributed $US 2 358 100 in kind. The project ended in October 1997.
The overall developmental objectives of the project were to:
- increase livestock production in sub-Saharan Africa, thus reducing the dependency of the local population on meat and milk imports and enhancing export potential;
- provide additional income for livestock owners by selling surpluses, thus contributing to an improved protein diet for the urban poor;
- reduce the overall regional level of potential environmental contamination with acaricides (including the level of residues in animal products); and
- reduce land degradation through improved livestock management systems.
The immediate objective of the project was to establish an operational, independent, financially responsible company capable of manufacturing vaccines and biological materials connected with the diagnosis and prevention of TBD.
The primary objective of the training undertaken at the VPC was to develop a cadre of national (and later regional) personnel equipped with the necessary techniques and skills in laboratory diagnostics, vaccine production, and immunization techniques for controlling the most significant TDBs of the region.
The mandate for VPC was to produce a vaccine against ECF (Theileria parva) in addition to three other major TBD blood vaccines for use against ABH. It was also to produce associated sero-diagnostic reagents. The major vaccine related activity was to assess ECF trivalent vaccine batch No. 0012, and produce the components for the trivalent vaccine batch No. 0013. Work continued through most of 1994. Towards the end of 1994, however, this activity was terminated; the production of sero-diagnostic reagents at the VPC continued.
Between January 1994 and September 1997, the following sero-diagnostic reagents were produced at the VPC:
4 727 slides of T. parva schizont antigen enough for 37 816 serological tests for ECF diagnosis using the IFAT test;
2 902 sides of B. bovis, enough for 23 216 tests for Babesia bovis infection;
5 398 slides of B. bigemina, enough for 43 184 tests for Babesia bigemina infection;
3 932 slides of A. centrale, enough for 31 456 tests for Anaplasma centrale infection;
1 661 slides of C. ruminantium, enough for 13 288 tests for the detection of Cowdria ruminatium infection (test in preliminary stages);
4 005 slides of T. mutans, enough for 32 040 tests for Theileria mutans infection.
In addition, three different batches of vaccine diluent were produced during the period, enough to constitute 24 140 doses of ECF vaccine.
The project assisted other projects in the region through activities of the veterinary advisory services and quality assurance sections of the VPC. The veterinary advisory services unit supervised TBD immunizations in Malawi. The staff at VPC assisted Government field staff whenever immunizations were planned and conducted. A total of 443 ECF immunizations was conducted between 1994 and 1997. Between 1990 and 1993, 1 320 and 1 272 vaccinations were carried out against anaplasmosis/babesiosis and heartwater, respectively.
The veterinary advisory services and quality assurance units of the VPC provided assistance in the various TBD sero-diagnostics to those projects or countries which were unable to conduct their own serology, or provided samples for reference purposes.
During the 1990-1997 period, the VPC received a total of $US 107 995 from the sale of ECF vaccine and related reagents, and $US 19 320 from the sale of ABH vaccines and related reagents.
Following the OAU/FAO/ILRAD recommendation that a pre-investment phase be initiated for tick-borne control by immunization, projects GCP/RAF/236/DEN and GCP/RAF/299/NET together implemented the structural adjustments to the premises and redeployment of staff to meet requirements for Good Manufacturing Practices (GMP) and Good Laboratory Practices (GLP). The restructuring involved the merger of the activities of ECF and ABH projects under a single management. Under this management three departments were created as follows; Vaccine Production, Quality Assurance and Product Improvement, and Veterinary Advisory Services. Alterations to the buildings and reassignment of staff to the new departments were completed on 28 March 1994.
Following FAO's contractual arrangement with the International Livestock Research Institute (ILRI), the production process of the interim batch of the vaccine commenced early in 1996 and was completed in December 1996. The modifications introduced by ILRI incorporated the three different constituent stocks of the trivalent vaccine into one representative composite stabilate. The other change involved storing the stabilate in plastic straws instead of tubes. Titration procedures took place at the Lilongwe VPC and terminated in October 1997.
2.5.1.1 Reference vaccine stocks
The seed stocks used to produce the interim batch were Theileria parva Muguga, Theileria parva Kiambu 5, and Theileria parva Serengeti. When these are transformed, they form the constituent stocks for the trivalent ECF vaccine (Muguga cocktail). The seed material was transported from the VPC to ILRI early in 1996, and the preliminary infectivity testing and characterization of the seed material was completed on 3 May 1996. These characterization studies were made to confirm that the materials at ILRI were of the same profile as those procured at the VPC.
The procedure for stabilate production, which involves infecting cattle and ticks with the appropriate seed stock, was carried out between May and August 1996. Infected ticks from each of the three stocks were then processed, and an assessment was made of the T. parva infection rates in ticks before preparing reference stabilates for each of the vaccine seed stocks. The reference stabilates, Kiambu 5, Muguga, and Serengeti, were prepared on 10, 12 and 17 September 1996, respectively.
2.5.1.2 Production of the composite stabilate
To produce a composite stabilate of the ECF trivalent vaccine, it is important that the resultant mixture of stabilate contain an equal number of Theileria parva sporozoites from the three components of the vaccine. To ensure this, batches of ticks infected with reference parasite stocks of Muguga, Kiambu 5, and Serengeti were mixed in proportions ensuring an equal number of infected tick salivary gland acini from each of the component reference stocks. The composite stabilate was prepared from this pooled mixture of ticks.
The infectivity assessment for each stock was made separately, so that the predetermined proportions were used in constituting the pooled mixture of ticks. From the pooled mixture, a five-litre ECF vaccine stabilate was prepared on 25 September 1996. Since the stabilate represented all three parasite stocks used in the production of the trivalent vaccine, it was designated "FAO 1 composite stabilate". This five-litre stabilate was dispensed into 10 000 plastic straws and stored in liquid nitrogen. Each straw contained a 0.5 ml inoculum of the composite vaccine. Storage in a single container instead of three separate ones for each of the constituent stabilates, saved storage space and eliminated the necessity for repeated resuscitation cycles before using the stabilate.
The various vaccine production processes, the seed material characterization, the production and characterization of reference stabilates, the production of the FAO 1 composite stabilates, and the preliminary determination of an immunizing dose were completed in January 1996.
The production process of the interim batch of vaccine fulfilled the recommended standards proposed by the Standards Committee representing the FAO, OAU, and ILRI.
The primary goal of the titration exercise was to derive, through experimentation in cattle, a dilution of the FAO 1 stabilate that would be safe and efficacious for use in field immunizations against ECF. The titration investigation was carried out at the VPC, which offers excellent facilities for experimental work of this kind. The investigation spanned a period of six months between March and September 1997 and was carried out in groups of matched Friesian steers from the same farm.
The objective of experimental activity was to evaluate the performance of various stabilate dilutions, following a series of immunization and challenge experiments, in order to select the best performing stabilate dilution. Very close clinical and parasitological monitoring of immunized cattle was necessary to gather sufficiently detailed data for the vaccine evaluation.
2.5.2.1 The titration procedure and the results obtained
The six-month investigation was planned as a three-stage titration procedure, with each of the three stages comprising an immunization and a challenge experiment. Thus, the whole titration involved six separate experiments, each lasting approximately 28 days (up to 35 days with the immunization investigations). The titration involved inoculating groups of cattle with varying doses of the FAO 1 stabilate, against a standardized treatment with a long-acting oxytetracycline in the typical version of infection-and-treatment immunization. The optimal immunizing dose was derived at the end of the investigation through observations of the performance of various stabilate dilutions in the different vaccine treatments. The clinical and parasitological observations in various treatment groups were compared, using a standardized format of Theileria parva reaction classification, in order to determine the best vaccine treatment.
Friesian males aged 6-9 months, from a farm in southern Malawi which was free of ECF and other TBD infections were used. The choice of Bos taurus cattle fulfilled the recommendations made by the FAO/ILRI/OAU Standards Committee.
2.5.2.2 Titration results and recommendations
Although the investigation was executed in three separate stages and comprised six experiments, the various experimental activities were interlinked. Thus the results of the first stage of the titration formed the basis for planning the second stage, and the results of the second stage were the basis for designing the third stage of titration.
The results of the first-stage titration identified the broad range of immunizing material for the FAO 1 stabilate as between 1:60 and 1:256; the second stage narrowed down the optimal immunizing dose to between 1:60 and 1:100, while the third stage predicted the immunizing dose as 1:80 and investigated both end-points.
The results indicated the possibility that all three test dilutions could be used to immunize cattle. However, where large numbers of cattle were involved, strong immunization reactions could be expected in a percentage of animals inoculated with the 1:60 dose, and a small percentage of poor vaccine-take could be expected in the 1:100 dilution vaccination. The 1:80 dose occupied the middle ground and was therefore thought to be the safest.
A wealth of information and data is available from the various consultancy reports regarding the best management structure for the VPC and how to guarantee the production of quality vaccines for the region. Considerable time was spent in 1994 examining various aspects and functions of the VPC. These included: personnel needs, the management structure, vaccine production, improvements in quality-vaccine distribution and marketing, and the VPC's envisaged role in backstopping other projects. In-depth consultancies were carried out on vaccine production, commercialization, and distribution. Additional studies were undertaken on legal aspects pertaining to the transformation of the VPC into a corporate body to be entrusted with the commercial production of vaccines.
With the VPC compliance to the general guidelines for vaccine production, enhanced confidence and an increase in the demand for vaccines from the VPC can be expected. An increase in demand would, in its turn, ensure long-term sustainability. It is recommended that all the proposals pertaining to the VPC be compiled into a single set of recommendations in order to initiate the implementation process.
It is recommended that the VPC be divided into three functional departments, covering vaccine production, quality assurance and improvement, and veterinary advisory services. Following a period of VPC inactivity, revitalization and revamping of the various sections should be undertaken as soon as possible.
It is recommended that appropriately qualified and experienced personnel be deployed in the various sections of the VPC in order to meet the proposed standards and requirements set by the Vaccine Standards Committee for the production of quality vaccines, effectively backstop field immunization programmes in the region, and perform appropriate applied research. While preference should be given to recruiting laboratory technicians nationally, the higher scientific and managerial posts should be filled by experts in TBD and theileriosis research.
It is recommended that the major functions of the VPC be spelt out unambiguously, and that the VPC be equipped and funded adequately in order to perform its mandated functions appropriately. The functions should be inclusive of vaccine production and titration; marketing; production of related biologicals; storing of vaccine stocks; field immunization back-up; training; information dissemination; and research and development of the vaccines produced.
It is recommended that some of these functions take place in collaboration with other organizations, research institutes, private concerns and companies.
The standards for producing live TBD vaccines were compiled by the OAU/FAO/ILRAD Standards Committee and circulated to all interested parties. They were based largely upon existing documents from vaccine producing laboratories in Australia, Kenya, Malawi (VPC), and South Africa. The standards considered all pertinent factors involved in the production process including buildings and grounds; manufacturing facilities; seed stock and stabilate production procedures; quality control; donor animals and ticks; vaccine storage; and the finished products, among others.
It is recommended that vaccines at the VPC be produced in general compliance with the code of Good Manufacturing Practices established in these standards.
For reasons of economy of scale, T. Parva stabilate for titration should be produced in bulk, preferably in five-litre volumes.
It is recommended that the three-step procedure used in titrating the FAO 1 stabilate be adopted as a standard procedure for all future vaccine titrations. The advantage of this procedure is that it is cheap but able to generate a great deal of statistically-valid data.
It is also recommended that these titration procedures be carried out in groups of matched Bos taurus cattle from the same source.
The business consultant proposed commercialization of the VPC's activities by transforming the VPC into a corporate body as a limited company, incorporating the laws of Malawi. The VPC management would comprise a Chief Executive Officer and a Vaccine Production Administrator, supported by technical heads of the three departments. Potentially huge markets for the vaccine were thought to exist in Tanzania, Uganda, Kenya, and Zambia but market penetration and distribution management plans should be drawn up and implemented.
The legal consultant recommended that the commercialization of the VPC take place within an established legal framework in order to safeguard government interests and those of other interested parties.
It is therefore recommended that the privatization of VPC be initiated only after an interim period in which the services of the VPC are treated as being primarily of public good. Further commercialization of the VPC should take place when the regional market for these vaccines has been adequately tested.
The VPC should back up the various national field delivery vaccine programmes in the region, and assist in particular with investigations of possible vaccine breakdowns or breakthroughs. It should also conduct training workshops for national and regional personnel to acquire the necessary skills and techniques used in vaccine production, laboratory diagnosis, and immunizing and monitoring vaccinated cattle. Information on the status of vaccine production and vaccine performance should be disseminated to the various recipient countries and end users on a regular basis using the appropriate tools and media.
It is strongly recommended that the VPC personnel participate in applied research as part of the vaccine improvement endeavours, particularly focusing on other potential vaccine strains from the region. These investigations should take place with the collaboration of other institutions.
Specifically, it is recommended that a validation cross-immunity trial be undertaken in order to assess the level of protection provided by the Boleni vaccine from Zimbabwe to cattle vaccinated using other major Theileria parva stocks from the region. This trial should be undertaken as a matter of priority since the Boleni vaccine is currently being used to immunize cattle without the usual oxytetracycline blockage. This makes the vaccination procedure very cheap.
PROJECT STAFF
Dates of Service
Name Function Starting Date Concluding Date
International
F.L. Musisi Chief Technical Adviser 1 Jan. 1995 18 Jan. 1997
J.J. Mutugi Chief Techncial Adviser 19 Jan. 1997 30 Sept. 1997
J.C. Quiroga Veterinary Advisory Services 1 Jan. 1994 28 Feb. 1995
L.M. Njuguna Senior Laboratory Technician 1 Jan. 1994 31 Dec. 1994
17 June 1997 28 June 1997
S.E. Moyo Administrative Assistant 1 Jan. 1994 31 Jan. 1995
J.C. Zenengeya Secretary 1 Jan. 1997 30 June 1997
E. Makawa Secretary 28 April 1997 30 Sept. 1997
National
P.T. Kafuwa Senior Technical Officer 1988
H.W. Sakala Senior Technical Assistant 1990
K.E. Chamambala Technical Assistant 1986
M.A.O. Pangani Technical Assistant 1988
L.R. Kasiya Technical Assistant 1989
L.D. Chombo Technical Assistant 1989
R.G. Mwangonde Technical Assistant 1989
R. Dalikeni Driver
Consultants
L.M. Njuguna Senior Laboratory Technician 17 Sept. 1997 28 June 1997
P.E. Hermans Veterinary Consultant 10 Jan. 1994 2 Feb. 1994
P. Van der Stichele Communications Consultant 1 March 1994 30 March 1994
R. Dagliesh TBD Consultant 13 Feb. 1994 2 March 1994
P.J. Nelson Business Dev. Consultant 7 Feb. 1994 2 March 1994
25 April 1994 31 May 1994
J. Ntambirweki Legal Consultant 13 Feb. 1994 16 April 1994
P.D. Kemp Acaricides Consultant 17 April 1994 30 April 1994
S.P. Kamwendo National Project Director 1 Jan. 1994 31 Dec. 1994
R.C. De Rooij LIvestock Consultant 1 Oct. 1994 16 Nov. 1994
FELLOWSHIPS AND WORKSHOPS
A2.1 FELLOWSHIPS
Participants Study Place Date
T. Kasaluka BTEC Higher National Britain Aug. 1993 - Aug. 1995
Diploma, Applied Biology
R. Mwangonde Veterinary laboratory Harare, Zimbabwe 1994
P.T. Kafuwa Sero-diagnostic techniques Nairobi, Kenya Oct. 1994 - Nov. 1994
A2.2 WORKSHOPS
Technical Protozoology techniques VPC, Malawi 14-18 March 1994
assistants
2 veterinarians; Laboratory diagnosis for VPC, Malawi 12-30 Sept. 1994
6 lab. technicians TBDs
42 field ECF immunization Arusha, 26-28 July 1994
veterinarians techniques Tanga,
Tanzania
23 veterinarians; ECF immunization Mwanza 26-28 July 1994
2 livestock techniques
officers
MAJOR ITEMS OF EQUIPMENT PROVIDED
Cost
Quantity Item ($ US)
1 Auto dispenser 2 846
1 Autoclave, large 5 302
1 Autoclave, portable 826
1 Balance, electronic 1 594
2 Cabinet, drying 3 558
2 Centrifuge 2 790
1 Centrifuge, M/300 6 204
5 Incubator, cooled 8 227
1 Deioniser 382
1 Distilling apparatus 438
1 Drill, electric 306
1 Freezer 680
1 Hood, laminar 4 840
1 Hood, laminar 2 520
1 Hood, laminar 4 599
3 Stirrer, magnetic 495
2 Mixer, maxi 192
1 Microscope, D/View 8 000
2 Microscope, stereo 6 000
1 Mixer, Omni 4 395
2 Cooker, pressure 321
1 Printer, Rejafix 4 664
1 Refrigerator 474
1 Revco 7 002
1 Revco 4 037
1 Shaker, Spiramix 818
2 Balance, trip 2 952
3 Pump, Vacuum 509
1 Waterbath 234
1 pH meter 799
1 Balance, electronic 2 745
3 Centrifuge 4 381
3 Cabinets, filing 783
Cost
Quantity Item ($ US)
1 Freezer 538
1 Freezer 404
1 Freezer 840
1 Freezer 404
1 Incubator 4 173
1 Incubator, cooled 1 645
1 Incubator, large 933
1 Hood, laminar 1 763
1 Stirrer, magnetic, large 365
1 Stirrer, magnetic, small 165
5 Refrigerator 2 372
1 Sealer 252
2 Water bath 842
2 Computer, IBM 4 465
2 Computer, laptop 7 650
1 Computer, Pentium 4 145
2 Cabinet, dry ice 760
1 Generator, diesel 22 883
1 Storage tank, LN, 55 l 9 082
1 Storage tank, LN, 30 l 1 765
1 Storage tank, LN, 35 l 16 446
5 Transporter, LT 8 594
2 Computer, Epsom 1 550
1 Printer, Laser Jet 995
1 Switchboard, telephone 6 493
1 Balance, haematocrit 763
1 Centrifuge, standard 2 027
2 Cabinet, dry ice 380
2 Cabinet, filing 522
1 Freezer 404
1 Freezer 404
DOCUMENTS PREPARED DURING THE PROJECT
Commercial rationale for a production and distribution component of a multi-donor programme for tick and tick-borne diseases control in Eastern, Central and Southern Africa. P.J. Nelson.
Commercial rationale for vaccine production and distribution. P.J. Nelson.
Technical review of the production and quality control of tick-borne disease vaccines in Vaccine Production Unit, Malawi. R.J. Dalgliesh.
Legal aspects of establishing a vaccine production centre as a legal entity. J. Ntambirweki.
Structure of the veterinary services and the status of the privatization effort in selected African countries. P.E. Hermans.
The production of East Coast Fever vaccines within the framework of the FAO Coordinated Multi-Donor Programme for Integrated Tick and Tick-borne Disease Control in Eastern, Central and Southern Africa. R.C. De Rooij, T. Lang.
