Chilean SRS salmon recombinant vaccine: High level biotechnology at the service of local economical development

Fundación Ciencia Para La Vida
Founded: 1997
Status: Private foundation
Headquarter: Santiago , Chile

Bios Chile
Founded: 1986
Status: Private company
Headquarter: Santiago , Chile

Fundación Chile
Founded: 1976
Status: Private foundation
Headquarter: Santiago , Chile

Novartis
Founded: 1996 (merger)
Status: Subsidiary
Headquarter: Basel , Switzerland

Chile is today the second largest salmon exporter in the world. However, every year, salmon farmers lose 200 million $ due to salmonoid rickettsial septicemia disease. Lead by Dr. Valenzuela, a group of private companies and Chilean foundations have developed a local solution to this problem by resorting to advanced biotechnology tools. Stemming from limited financial investments, a recombinant vaccine will open a 30 million $ market allowing for a 130 millions $ save in the Chilean salmon industry. This project was made possible thanks to the creation of synergies between the different key players and to significant technological transfer. It has proven the potential of biotechnology to bring profitably solution to industrial large scale problems.

Background

More that 200 million $ are lost every year in Chile ‘s salmon industry from P. salmonis infection.

Salmon has first been introduced in Chile in the early 90’s and within 15 years, it has become the second largest salmon exporter in the world accounting for a 1 700 million $ industry and more than 5% of Chile ‘s total exports. However, the apparition of salmonoid rickettsial septicemia (SRS) disease in 1998 caused by the bacteria Piscirickettsia salmonis , has caused important losses in this business central to Chilean economy. “We have calculated a 10% fish mortality and a resulting annual loss that should today be superior to 200 million $”, Dr. Gustavo Parada, Fundación Chile . Although bacteria can be found in other places, only in Chile is the disease responsible for such an important economical loss.

Contrary to most known bacteria, P. salmonis induces an intra-cellular infection rendering standard vaccine developments ineffective: available vaccines for this disease have not exceeded 30% efficiency. The risk associated with the development of a potentially costly vaccine for a regional market has put off major veterinary companies of proposing a solution to this problem.

A group of private companies and foundations led by Dr. Valenzuela decided to bring a biotechnology solution to the salmon industry.

For these reasons, a group of Chilean private companies and foundations leaded by Dr. Pablo Valenzuela decided in 1998 to develop a local solution to this problem. This project has involved a group of private companies as well as public and private foundations:

Fundación Ciencia para la Vida (FCV): Led by Pablo Valenzuela, this Chilean private non-profit research foundation was founded with the porpoise of stimulating the use of biotechnology in the productive sector.

BiosChile: This Chilean private company, also directed by Pablo Valenzuela, has a wide experience in genomics research and vaccine and diagnostics development.

Fundación Chile: This state foundation for the technological transfer in Chile was among other actions responsible for the initiative of introducing salmon culture in Chile .

AcquaHealth: A Canadian company specialised in fish vaccines formerly held by Cobequid.

Novartis Animal Health: One of the major veterinary companies, representing about 3% of the entire Novartis group.

These actors have put in common their financial and technical resources to obtain a product obeying the following characteristics:

Highly efficient vaccine: “Salmon is produced in intensive fish cultures which imply a low commercial margin market type. In order for this vaccine to present a practical solution to the problem and interest a major veterinary company, it should present over 70% efficiency ”, Fernando Flores, General Manager, Novartis Animal Health , Chile .

Low cost production: “The overall selling price of a salmon is inferior to ten dollars. Therefore, in order to be competitive, the production cost of a vaccine should be on the order of ten cents or lower ”, Gustavo Parada, Sea Resources Manager, Fundación Chile .

Finally, the leaders of the project were decided to address this problem by using to state of the art research: “We believe that the use of biotechnology to solve important economical problems will represent a boost on biotechnology development in Chile ”, Pablo Valenzuela, President, Fundación Ciencia para la Vida and BiosChile .

From science to an industrial activity

A recombinant vaccine was developed based on three bacterial antigens and membranes polysaccharides.

Due to the specificities of the P. Salmonis bacteria, standard vaccine approaches based on attenuated or dead bacteria have low efficiency. For that reason, a recombinant vaccine was developed. The genes coding for antigen proteins that induce an efficient immunity response by the fish organism are introduced in the DNA sequence of a E. Coli bacteria that will produce a large quantity of these proteins. When the produced proteins exceed 20% of the bacteria’s volume, their membranes are broken down and the resulting, almost non-purified, product is obtained. This product consists of a mixture of the three antigen proteins and the membrane polysaccharide which are then added to an oil substrate. Through injection of this solution, salmon fish can be immunized to P. Salmonis infection.

Finally Novartis has decided to increase the added value of this product by adding to the final substrate the antigens for other salmon fish diseases. This way Novartis has created a vaccine capable of immunizing salmon simultaneously to different infections; a multivalent vaccine. Two products will finally be commercialised: Forte 3 that combines SRS vaccine with Infectious Pancreatic Necrosis and Vibrio ordalii , and Forte 4 with the addition of the Aeromuna antigen.

The combination of resources of the different partners through the different development stages led to a final multivalent vaccine with over 80% efficiency.

The development of the SRS vaccine was performed in several different stages:

Sequencing and gene identification: First the genome of the P. Salmonis bacteria was sequenced to identify the genes that could be used for antigen generation. In order to save development costs, raw sequencing was outsourced to as specialized company. Based in comparisons with other known bacterial genes, 16 potential antigens were identified in this phase

Antigen identification: The different antigens were tested in order to select the most promising combination. 3 antigens were selected to compose the most promising vaccine for this disease.

Production optimization and efficiency testing: At this stage, it was found that reducing the purification stages and therefore leading to the presence of membrane polysaccharides in the final product increased the antigen activity of the vaccine. This will also results in lower production costs. Fish were infected with P. Salmonis and the survival rate was compared between immunized and non immunized specimens. Global efficiency for this vaccine reached 90%.

Multivalent efficiency testing: This stage involved the development of the multivalent vaccines and testing its efficiency in laboratory and natural conditions. The survival rate of specimens immunized with the multivalent vaccines was superior to 80%. At this point Novartis decided to go up for full-scale vaccine production.

Large scale production: Large scale production for the commercialisation of the vaccine is currently being performed in Novartis industrial facilities in New England , Canada .

Overall, the full project took about 7 years from the initial research phases up to full scale production and cost less than 5 million $ for all the development phases. Full scale commercialisation for the SRS multivalent vaccine is scheduled for September 2006.

Fig. 1: The project development track for the SRS vaccine.

SRS vaccine impacts

• Besides a 30 million $ market for Novartis, a 130 million $ net gain for salmon fish farmers and important financial counterparts for the actors of this project, this success has also impacted on the potential of Chilean biotechnology.

The success of the SRS project will impact in all actors involved in this project:

• Novartis: The SRS multivalent vaccine to be commercialised will address a population of salmon, Salmon Salas and Salmon Rain-bow, for which no effective vaccine exists today. “From our experience, we expect to cover 80% of the fish population within 5 years which corresponds to an annual overall turnover of 30 million $”, Fernando Flores, General Manager, Novartis Animal Health Chile.

• Salmon farmers: Taking into account the efficiency of the vaccine, its cost and the fish farming cost, net gains for salmon farmers could be superior to 130 million $, that is over 7% gain per salmon fish which is quite significant in a mass export industry like salmon.

• BioChile, FCV and Fundación Chile : Alongside this direct gain for Novartis and fish farmers, the actors in this project were also entitled to financial counterparts for the commercialisation of this product : “For the licensing deal, royalties were also negotiated with Novartis. Depending on the market potential of the product, these royalties for these cases are usually in the order of 10%”, Gustavo Parada, Sea Resources Manager, Fundación Chile . The success of this project has also obvious impacts on the credibility of the actors involved, either to obtain funds for new researches or to negotiate early licensing deals with industrial companies in other projects.

• Chilean biotechnology: Finally, the impacts on this sector could not be neglected. This project has shown the potential of biotechnology “made in Chile ” to directly solve industrial problems that affect Chilean economy and may help boost investments in this sector.

View of the future

• A new project for increasing the sustainability of the Chilean salmon industry may be the next biotechnological success for the key player of the SRS vaccine project.

Motivated by the success of this experience and the acquired know-how through this project, both on the technical aspects of vaccine development and on the coordination of different resources in different structures, FCV and Fundación Chile have already began researches for the development of new vaccines that still affect the Chilean salmon fish population such as Streptococcus phocae .

One of the most promising projects in development by FCV for the salmon industry is the development of genetically modified lupine, a leguminous plant whose seeds have been used from ancient times for fish feeding. “In order to farm 1kg of salmon, 2kg of fish are needed as feed. This is not sustainable environmentally or economically. We are developing modified lupine seeds that also express the fundamental amino acids for fish culture and that could help solve this problem in the salmon industry”, Pablo Valenzuela, President, Fundación Ciencia para la Vida and BiosChile.

Finally, new products are also being developed that could help other exporting industries in Chile such as copper mining and forestry.

Lessons and key success factors

• This case illustrates the viability of biotechnology projects, even with low available resources, that can be put to use to aid in large scale industrial problems.

Two main lessons can be taken from the analysis of this case:

• Viability of biotechnology research with limited financial resources: “This project shows that making good use of available human resources and education makes it possible to perform very successful biotechnology research, with direct social impacts even when financial resources are scarce like in Chile”, Vivian Wilhem, Associate researcher, Fundación Ciencia para la Vida.

• Potential of biotech to solve industrial problems: “With this success we have demonstrated that biotechnology can give effective solutions and increase value of economical activities in the Chilean context”, Pablo Valenzuela, President, Fundación Ciencia para la Vida and BiosChile.

• Taking advantage of the skills of all the partners and of past experiences to create positive synergies while assuring a successful technological transfer were key factors that explain the success of this project.

Three main factors can explain the success of the SRS vaccine project:

• Partnership synergy: “None of the individual actors of this project had the necessary skills and means to succeed in such a project. However, thanks to BiosChile’ genomic competences, Fundación para la Vida ‘s scientific skills, Fundación Chile’s fish testing facilities and know-how and Novartis’ production and commercialisation means, we were able to be successful and develop a product that will greatly profit both its developers and the salmon industry in Chile”, Pablo Valenzuela, President, Fundación Ciencia para la Vida and BiosChile

• Technological transfer: Although Chile is a country with little technological background, effective technological transfer from the USA and Canada made possible to develop, mostly with Chilean means, a biotech vaccine of international level. “Thanks to the help we got from Novartis in Canada, today we have the facilities and the know-how to perform almost all experimental stages on our Chilean facilities”, Erwin Landskron, Experimental Unit Manager, Novartis Animal Health Chile

• Experienced team: “The long time biotechnology development experience of Pablo Valenzuela and that of Fundación Chile in the salmon industry were key factors to lead the project and to make all key decisions during the research phases”, Gustavo Parada, Sea Resources Manager, Fundación Chile.