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A replacement of the sub-optimal live feeds used at hatcheries today with a new cryopreserved live diet for the improved and efficient production of juveniles in marine aquaculture

Periodic Reporting for period 1 - CryoPlankton (A replacement of the sub-optimal live feeds used at hatcheries today with a new cryopreserved live diet for the improved and efficient production of juveniles in marine aquaculture)

Reporting period: 2014-10-01 to 2015-03-31

Planktonic AS has succeeded in cryopreserving relatively large volumes of crustacean nauplii which can be used as a replacement of today's live feed (rotifers and Artemia nauplii) used at marine hatcheries. After thawing of the pouches, the crustacean nauplii is revived as live individuals, even after two year of storage. The revival rate is >90%, and relatively uniform for all volumes. Marine crustacean nauplii is regarded as the optimal feed for marine juveniles, and the new cryopreserved product can potentially revolutionize the marine aquaculture industry by solving the bottle-neck of the first feeding of fish larvae. A pilot market survey has resulted in very positive response from hatchery managers, and there is a high willingness to exchange today’s live feed used at hatcheries with our cryopreserved crustacean nauplii (hereafter called CryoProduct), as it is more user-friendly and most probably will give far better performances on the fish larvae at the hatchery.

The aim of the phase I study was to perform a feasibility study for the CryoProduct.

In the course of the project period for the SME instrument phase I, we have explored the following specific objectives for the feasibility study:

The specific objectives for the feasibility study in the Phase 1 project:
a. To establish co-operation with a distributor of fish feed in South Europe that will market and distribute the product in the Mediterranean, which is considered as a key market
b. Establish a co-operation with 1-3 producers of marine juveniles in the Mediterranean area that are willing to try out the new product
c. Perform a survey of the market potential for the product in the EU, including an assessment of willingness to pay for the product
d. Elucidate challenges regarded to logistics for distributing the product
e. Evaluate different technologies for up-scaling of the production of the cryopreserved product
f. Work out a cost accounting plan for the product
g. Accomplish a business plan


The objectives for the phase II project was originally detailed as:
1. To optimize pre-freezing conditions for target zooplankton and to optimize methods for cryopreservation of plankton samples with higher viability after thawing than today.
2. To scale-up the cooling and thawing methods for cryopreserving relatively large volumes of targeted zooplankton (250-1000 ml), while preserving the nutritional quality of zooplankton with regard to fatty acids, lipid classes, antioxidants, amino acids and proteins.
3. If needed, a detoxification protocol for the revived zooplankton prior to be offered as live feed for marine larvae will be developed.
4. To screen the cryopreserved organisms for possible harmful parasites, bacteria and viruses which might have negative influence on the fish in the breeding tanks and to assess the efficacy of possible eradication methods for them.
5. To develop and compare three ideal first feeding regimes in marine fish larvae cultivations: one regime consisting of fresh zooplankton in small scale cultivation, one consisting of cryopreserved zooplankton in small scale cultivation and one consisting of cryopreserved zooplankton in industrial scale cultivation. The regimes are determined through performance verification of the fish larvae using traditional analysis supported by molecular tools for the interpretation of the fish larva’s growth, stress-response, appetite and more.

In parallel to this project, Planktonic has produced 250 kg of the CryoProduct, and managed to produce large entities of pouches which is a suitable size for the hatcheries. Thus, objective number 1 in a phase II project is not of very high importance anymore, but still of interest since it would make the CryoProduct even more user-friendly if sizes of up to 1 kg could be produced. In addition, objective number 3 would be skipped in the phase II project, as we now have tested the CryoProduct on three different fish species without any negative consequenc
A producer of fish feed for juvenile fish species has been selected as one of the partners for the Phase II project, and was chosen because of their capacity as a future potential distributor in the South Europan market. The fish feed producer has a high competence in performing start-feeding trials of marine fish larvae, and the partner will be employed for doing large-scale trials at one or two marine hatcheries in the Mediterranean. In the phase II project, it is decided to invite two marine hatcheries as sub-contractors for testing the CryoProduct in large-scale, and the hatcheries are located in South Europe and Norway, respectively.

The cost of producing Artemia and rotifers ready to be used at the hatcheries display a high range, and in developed countries it is typically between 400 and 4000 €/kg for rotifers and 160 to 1150 for Artemia nauplii. The figures display the best case production cost versus the worst case production cost for rotifers and Artemia nauplii. In developing countries, it is often far cheaper to produce live feed, as the final costs are highly dependent of (1) cost of labour or (2) if yeast, live algae or algae paste is used as on-grow feed for the rotifers. Planktonic will aim to introduce the CryoProduct for prices that is competable with todays live feed, and we aim to enter volume markets as e.g. feed for sea bream and sea bass. Preliminary feed-back from hatchery managers indicate that the cost of the product for end-users are well within the willingness to pay for the CryoProduct as it most probably gives better performances of the fish larvae, as higher growth rates and reduced mortality.

The Food Safety Authorities has been consulted, and the preliminary feed-back is that it will be possible and feasible to export the CryoProduct within EU. Special focus on harmful bacteria and parasites in the product has to be evaluated before the export will take place. Once approved by the Norwegian Food Authorities, it will also be approved in the EU.

We found that cryo-containers of volumes less than 200 liters (with liquid nitrogen) is the optimal size for storage, transport and distribution of the CryoProduct. Electrical Ultra-low freezers were also evaluated, but was found less reliable and more expensive than cryo-containers. Transport of biogenic material in open cryo-containers are subjected to safety rules, and a maximum size of cryo-container can be used, and also a special kind of vehicle and licence of the driver. The static holding time of a 175 litre cryo-container is 184 days at 20 degree celsius, but are severe reduced when lid are opened frequently. Once transported to the hatchery, it will probably be no need to replenish liquid nitrogen, as start feeding are performed in relatively short periods of up to 90 days for cold water species and 20-40 days for species as sea bream and sea bass.

From 2015 to 2018, a manual method for producing the CryoPlankton product will be used, whereas a more industrial scale production will be implemented from year 2019 and onwards. The bottle-neck for the production of the CryoPlankton is the capacity of controlled freezers, and probably, several such machines has to be purchased to increase the production of the CryoProduct to some hundred kg per day.

The CryoProduct is in size between rotifers and Artemia nauplii, and our product can exchange both types of live feed. The global market of Artemia cysts is estimated to 3000 tonnes per year. 80% of the Artemia cysts goes to shrimp production in developing countries. We assume that the fish larviculture requires roughly 500 tonnes of Artemia cysts per year, and it is estimated that the requirements of rotifers in fish larviculture constitute 50% of the Artemia sales. Thus, we estimate the global aquaculture market to 3150 tonnes of live feed, including rotifers and Artemia nauplii. The market is differentiated, and production cost of rotifers and Artemia nauplii for fish larviculture is mor
All members of Planktonic’s steering committee has agreed that the project shall continue, and it do not differ much from the business idea that were originally described in the application.
The new CryoProduct (live feed) is a potential ”game changer” and can become the de facto standard live start feed within marine fish larvae production globally. The new CryoProduct is cheap and simple to produce, and is targeted towards a large market with high willingness to pay. Planktonics CryoProduct is much less labor intensive and complicated to use compared to existing live feed concepts.

After completing the CryoPlankton project in SME instrument phase 1, we have got far more background data for our CryoProduct from production to logistics to the end-user. We have improved our understanding of the market we are about to enter with our CryoProduct, and we are prepared to apply for a SME instrument phase 2 project as a final tool for successfully launching our product within the EU and ROW market.
Start feeding of marine larvae is considered a bottle-neck for the success in marine aquaculture, and a live feed diet that match the nutritional demand of fish larvae is not commercially available today. Several experimental and industrial trials have documented that crustacean nauplii are optimal for marine larvae, and outcompetes today's feeding alternatives used at marine hatcheries. However, marine crustacean nauplii have until now not been commercially available in quantities needed for the marine larviculture industry. The new CryoProduct will most probably be highly demanded as it is a natural food item for fish larvae, and fish are evolutionary adapted to feed on crustacean nauplii.

Aquaculture has become a modern and dynamic industry in many European regions with a value amounting to € 3.2 billion that produces safe, high valuable and high quality products. The global developments and the strategic importance of aquaculture in terms of food security contribute to give this sector a promising future. Numbers of the FAO for the coming years support this: aggregated country productions from aquaculture are expected to grow at an average annual growth rate of 4.5 percent over the period 2010–2030.

The production potential of aquaculture is far from fully exploited in Europe as there are several constraints connected to the cultivation of several fish species. One innovation that is needed for a high production and sale of marine fish is the improvement of survival rate, and development during the larval stage of marine farmed fish species. Marine fish species are upcoming fish species and the production of seabream and seabass comprises ca 18 % of the total fish production in aquaculture and 95% of the total marin fish production in 2009 in Europe. Other species that have a high potential to succeed in the EU and worldwide with this innovation are e.g. sole (several species), Atlantic halibut, various species of wrasses (used as cleaner fish at salmon farms, tuna, groupers, meagre, breams (other than gilthead seabream)) and various species of humberjacks.

Currently, a complex feeding process during the larval phase of these species cannot overcome low survival rates and development of these species and therefore production and sales cannot be fully realized. This is even true for the production Gilthead seabream that is considered as successful in the EU. Furthermore, the production of juvenile fish larvae represents a high cost for the aquaculture industry. Maintaining and establishing cultures of live zooplankton feed is complex, and often will small modifications of e.g. the enrichment procedure result in high variability of the nutritional profile of the live feed.

Commercialisation of the CryoProduct will contribute to a higher and more efficient and new production of marine fish in aquaculture, which again will result in spin-off effects to other industries (e.g. our partner Sparos as a potential distrib
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