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Cryopreservation of marine planktonic crustacean nauplii for innovative and cost-effective live feed diet in fish juvenile aquaculture

Periodic Reporting for period 2 - CryoPlankton2 (Cryopreservation of marine planktonic crustacean nauplii for innovative and cost-effective live feed diet in fish juvenile aquaculture)

Reporting period: 2017-02-01 to 2018-04-30

While the global production of farmed seafood grows, European production has stagnated; a central reason is lack of low-cost, high-quality live-feed for the many commercially valuable fish species whose complex larvae-state feeding needs are not fulfilled by current feed options. If the European aquaculture industry is to grow, it is thus of critical importance to bring new live feeds appropriate to complex feeding needs to market. The live feeds that exist on the market today, is rotifers and Artemia. Rotifers and Artemia are sub-optimal as prey items for marine larvae in aquaculture, and because of that, the aquaculture industry struggles with low growth rates and high mortalities in the early phase when producing fish juveniles. The natural prey items, which marine larvae are evolutionary adapted to feed on, is marine zooplankton, and especially small crustaceans as copepods.

Our SME, Planktonic AS, has succeeded in cryopreserving marine crustacean plankton in user-friendly pellets, and to revive them as live individuals after thawing. The ease-of-use CryoPlankton meets the nutritional requirements of fish larvae.

Aquaculture is a major industry in the EU, worth €3.5bn and responsible for more than 88,000 jobs within some 14,000 businesses. If the CryoPlankton product meet the expectations, it will most probably be a major contribution to realize the production potential of marine fish in aquaculture in the EU. This will result in many new jobs, and primarily in the Mediterranean region. Many high-valued marine species cannot be cultivated when offered low quality prey items as rotifers and Artemia. Fish species that have complex live feed needs in the larval stage are potential the new aquaculture species in the EU, and examples are high priced fish species as amberjack, groupers and more. All these species require a live feed meeting their nutritional needs as the CryoPlankton. To have a self-supporting supply of seafoods to the EU, it is important to strengthen the aquaculture industry in the region, and the CryoPlankton will be a major contributor to meet this goal.

The objectives of the CryoPlankton2 project is to upscale the CryoPlankton production process, and associated technologies, towards industrial production volumes. We are about to demonstrate the performance and commercial potential of the CryoPlankton and its process under operating production and end-user conditions. The work in the project involves also development and upscaling of both our upstream and downstream supply chain to ensure rapid market uptake and to meet expected market demand. We are doing continuous market surveillance and analysis covering the EU and worldwide marine aquaculture feed markets to determine optimal market entry and distribution, and we are implementing a detailed innovation management strategy covering a plan for stakeholder engagement, project result exploitation and dissemination.
During the project period, we have made effort on optimizing the production protocols of the CryoPlankton for improving the vitality of crustacean nauplii after thawing, and to even out variabilities in quality between batches. Upscaling of the production is a challenge, and most of the work in the project have been allocated to this part. Feeding protocols have been optimized for several species of fish, including sea bream, sea bass, ballan wrasse, lump fish and sole.
A bio-security evaluation has been performed, with a screening of microorganisms necessary for the registration of the CryoProduct. Efficient logistic systems, identification of end-users needs and a reliable commercialisation plan for the best possible market introduction have been evaluated.

At the end of the project, after 27 months, we have achieved to increase our technology level from TRL7 to TRL9. We have succeeded to handle raw material consisting of planktonic crustacean nauplii from harvesting site to production plant in an effcient way. To transport a live product with negligible changes in product quality is a challenge, but we have found thresholds on temperature and densities of the product when transported. Also, a maximum transport longevity without any change in quality have been identified. We have made progress on the cryopreservation technology, and the quality of the CryoPlankton is close to natural.

Results on sea bass display 75% higher growth rate, almost 50% higher survival, and a far more stress-resistant fish when offered CryoPlankton compared to the fish that have been feeding on rotifers/artemia. CryoPlankton can be fed to the sea bass larvae as the first prey item, and replaces rotifers, AF artemia and EG artemia.
Similar results was achieved on sea bream as for sea bass, with about 50% higher growth rate, and a higher survival.
For both species sea bream/sea bass, an unexpected result on Vibrio bacterium was observed, as no fish larvae were infected by this pathogenic bacteria. The absence of Vibrio when fed CryoPlankton was demonstrated in all performed trials on sea bass and sea bream. Vibrio is the pathogen that is causing the most problem in larviculture in South Europe.

Many trials have been done on species as lump fish, ballan wrasse, Solea senegalensis, all displaying high performances when fed the CryoPlankton.

The growth of ballan wrasse larvae display a doubling in size compared to the control fish. The growth is maintained, and after 3 months, the CryoPlankton fish is still about the double size of the control fish. For lump fish and sole, about 50% higher growth can be expected when replacing CryoPlankton with conventional live feeds.
We have documented that performances of the fish larvae are far better when offered the CryoPlankton versus the normal diet used at marine hatcheries. Growth have been demonstrated to be more than 100% higher when offered CryoPlankton compared to the fish that solely have been feeding on other live feeds or formulated dry diets. For lump fish, the mortality is 4-6 times higher when the larvae is offered formulated dry diets compared to the CryoPlankton. Bacterial supply to the fish tanks have turned out to be far less when using the CryoPlankton compared to when for instance formulated micro-diets are used. At one of the lump fish hathceries which are using the CryoPlankton on a permanent basis, the hatchery manager could allocate a person that previously were cleaning fish tanks as the sole work-function, to other important duties at the hatchery. Fish health in aquaculture is very important, and the industry is struggling with many fish diseases. Thus, a new diet that contributes to far less pathogens are requested by the industry. Absence of Vibrio bacterium when using CryoPlankton is definitely a progress beyond the state of the art, and will have major impact of production of sea bass and sea bream in South Europe.

Eight end-users have started to use our product on a permanent basis. With the good results these end-users obtain on their fish juveniles, we experience much attention, and inquires on the CryoPlankton product. Future aquaculture can be done far more rational, and the profitability can increase significantly. Employment are expected to increase when production of juvenile fish can be produced optimally with a diet that meets the fish nutrititional requirements, and especially in areas as in the Mediterranean, where marine aquaculture is a relatively large industry.
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