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Sustainable, aff ordable and ethically acceptable

Sustainable, affordable and ethically acceptable

New protein sources for aquaculture feed

About 70 percent of fish and crustaceans produced in aquaculture are fed with protein-rich feed. If the aquaculture sector is to reach its growth targets by 2025, annual feed production must increase by approximately 38 million tonnes. This is not possible with today’s protein sources. The search for alternatives has become the central question in feed research on which the further development of aquaculture depends.

Algalimento

With their high protein and lipid content some microalgae species can potentially be used to replace plant and animal ingredients in fish feed. But production levels are tiny and the price correspondingly high. Shown, dried and powdered Tetraselmis produced by Algalimento, a company on the Canary Islands.

Global production of fishmeal is currently fluctuating around 4.5 million tonnes per year, of which 69 is used for fish feed. For fish oil, there is even less room for manoeuvre, as aquaculture already uses 75 of the annual production of around 0.9 million tonnes. Both resources are therefore scarce and correspondingly expensive. In order to meet the rising demand for protein sources for fish and shrimp feed, economical alternatives must be found that fulfil several criteria. By far the most important of these are nutritional properties, as the feed must reasonably meet the nutritional requirements of the animals and should be easily digestible and promote the growth of the fish. The fish must like the taste too, in order for them to willingly consume it, and it cannot negatively affect their physiology and health status. Protein alternatives should be available in large quantities, physically stable and easy to handle, so that they can be stored, processed and transported without problems. Using them for feed cannot have significant negative effects on the environment and must be ethically acceptable. This means that new raw materials and ingredients must not compete with food for humans or contribute to overgrazing, water scarcity and loss of natural biodiversity.

And because the feed infl uences the sensory and qualitative features of the fi sh and shrimp produced, the ingredients fi nally must give the products those health and taste advantages that consumers have rightly come to expect.

Th is small number of examples alone show how challenging and diffi cult the search is. Modern aquaculture feed is already an artifi cially balanced blend of the most varied ingredients, which only together optimally meet the nutritional requirements of the respective aquaculture species. Both plant-based protein components such as soy meal, corn gluten meal or rapeseed meal, as well as animal byproducts (e.g. meat, bone or poultry meal) are used as alternatives to fi sh meal in aquaculture feed. Th anks to new technologies, fungal and algal proteins, insect protein, olive pomace and even a variety of grasses are also being considered. Th e overarching goal of these eff orts remains to reduce the demand for fi shmeal and therefore the proportional demand for the wild catching of industrial fi sh for aquaculture feed. But using plant-based components in feed does not necessarily mean greater sustainability. Th e cultivation of soya beans, for example, has led to deforestation in some regions, which signifi cantly harms biodiversity and the climate.

It has already been demonstrated in feed experiments that the cultivation of carnivorous fi sh species such as salmon, trout or saltwater fi sh with feed that does not contain fi shmeal is possible in principle. However, across-the-board use of this scientifi cally blended feed in practice is not possible at the moment, because the cost is still too high and would make cultivation of the fi sh unprofi table. However, aquaculture feed is not just of interest to researchers and the reason for the foundation of numerous startups, it is also the business of many companies. In 2016, the feed specialist BioMar brought to market the fi rst commercial feed that included products made from microalgae, such as AlgaPrime DHA. By 2021, the quantity for salmon feed alone had risen to 1 million tonnes. Regardless of such isolated successes, the research costs and expenses for alternative sources of raw material for fi shmeal and fi sh oil remain extraordinarily high, however, particularly as plantbased raw materials often contain harmful ingredients called “antinutritional factors” (ANF) which are diffi cult or impossible for fi sh to digest and therefore harm their well-being and health. Th ey must be eliminated and deactivated by means of appropriate processes before use.

Microbial biomass and insect meal

An interesting approach to solving the protein problem is the use of microbial biomasses, such as those concentrated in microbial fl akes (biofl oc). Th ese proteinrich resources can be exploited as

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wet or dry meal (freeze-dried or oven-dried) and are consumed by many herbivorous and omnivorous fish species. Tests with tilapia hybrids show that with bioflocs, up to 50 percent of the protein in the feed can be provided without seriously restricting the growth of the fish. However the quantities available are still far too small to be of practical relevance for the time being. Irrespective of this, significant potential is attributed to microbial proteins or single cell proteins made up of bacteria, yeasts and microalgae for aquaculture feeds. These organisms have a high protein content of 45 to 65 and an amino acid profile that is comparable to fishmeal. The nutritional profile of yeasts and bacteria can also be controlled through the composition of the culture medium. The suitability and usability of these protein sources, must, however, be separately assessed at species level, because the absorption rates, digestibility and bioavailability of nutrients in the microbial biomasses can vary.

This also applies to insect meals, which would also be a promising protein alternative due to their nutritional profile. Insects do not compete with human food sources, have short life cycles and can grow on a variety of substrates. They are therefore receiving increasing attention in many countries. The European Union approved processed animal protein from insects (i.e. insect meal) for use in fish feed from July 2017 in Regulation (EU) 2017/893. International research is currently concentrating on multiple insect species that are of interest for feed.

The main focus, apart from the yellow mealworm (Tenebrio molitor), the common housefly (Musca domestica), tropical house cricket (Gryllodes sigillatus) and Turkestan cockroach (Blatta lateralis) as well as the domestic silk moth (Bombyx mori), is mainly on the black soldier fly (Hermetia illucens), the larvae of which come relatively close to the raw protein content of fishmeal. However, the biochemical composition, amino acid and fatty acid profile of the larvae vary significantly depending on the stage of development and feed substrate, making their exploitation in feed difficult and costly. If the larvae grow on food waste, for example, which would be very advantageous from a hygiene perspective, they put on a lot of fat, which however contains hardly any omega-3 and more omega-6 fatty acids instead. Since lipids are prone to oxidative decomposition, larvae meal such as this usually needs to have the fat removed, which means additional costs. Generally, the lipid quality of the insect larvae can be manipulated through the substrate used. In order to increase the EPA and DHA content, for example, the larvae can be fed on fish waste, but this can also be used directly as raw material for fishmeal, which is more costeffective. The nutritional value and composition of the larvae also depend very significantly on their stage of development. The closer they are to pupation, the greater their chitin content, which influences the proportion of fibre and thus digestibility.

Do insect meals improve the immune system?

Properly prepared, dried and defatted, and appropriately dosed, larvae meal of the black soldier fly can replace a portion of the protein in fish feed. For cultivation of young sea trout, a test was done with feed that contained 20 percent insect larvae meal, and no difference in growth was detected compared to the standard feed containing fishmeal. This is not surprising, since flying insects are also part of the food spectrum of numerous wild salmonids in natural waters. It is more unusual that larvae meal is also suitable for the cultivation of shrimp. For experiments with Pacific white shrimp (Litopenaeus vannamei), 20 of the fishmeal was replaced with de-fatted insect meal without the growth of the crustaceans being negatively affected. The researchers suspect that the insect larvae meal improves the immune system and intestinal microbiome of the shrimp. They detected an increased resistance to AHPND (acute hepatopancreatic necrosis disease). This only works if the proportion of larvae meal in the shrimp feed is not too high, however, as replacing 30 or more of the fishmeal results in degenerative effects. Some of the intestinal cells died due to programmed cell death (apoptosis), which triggered further damage.

Insect larvae meals would be suitable in principle, but their use at a large scale is currently not very feasible. For one, the production quantities are nowhere near large enough to consistently supply the aquaculture industry. According to cautious estimates, barely more than 30,000 tonnes is currently being produced globally on an annual basis. This may change soon however, as significant sums are being invested in insect cultivation in some countries. The Israeli startup Entoprotech alone is investing around 30 million US dollars in its soldier fly project “Waste2Protein”. This means there is a real chance that the second obstacle to widespread use, the relatively high price for insect meal, may be overcome, since although this alternative protein source is not exactly cheap, if the promised increase in production actually occurs, insect meal could potentially compete with fishmeal on price within a few years.

AlgaPrime from Corbion

Feed specialist BioMar brought to market the first commercial feed that included products made from microalgae, such as AlgaPrime DHA (pictured).

Feed proteins made from leaf and grass biomass

Plant-based biomasses harvested from leaves and grasses are raw materials that are available on a huge scale and attractively priced, but are technically demanding to process. The few studies carried out to date on Leucaena and cassava leaf meal as well as with leaf protein concentrate from rye grass and alfalfa

have shown that plant leaves can be used as dietetic protein sources in fish feed. The results of the tested diets are still contradictory in parts, however this can be explained by differences in the protein content and amino acid profiles, their digestibility, their taste or the ANF mentioned above. Recent experiments with protein from the drumstick tree (Moringa oleifera), a fastgrowing, drought-resistant plant from the Indian subcontinent, have achieved promising results with tilapia, grass carp, rainbow trout, shrimp and other species. Moringa meal has a high nutritional value, is rich in fibres, proteins, vitamins, minerals and lipids and also has remarkable pharmacological qualities. It is claimed that these plants, among other characteristics, have antiinflammatory and anticarcinogenic, antidiabetic, antioxidative, antimycotic and antibacterial properties. However, further studies are necessary before they can be used in aquaculture in order to determine the appropriate dosing.

Dry meals made from Kikuyu grass (Cenchrus clandestinus) have already been successfully tested as a protein replacement in diets for tilapia rendalli. Kikuyu grass has a relatively high protein content and a good amino acid profile. It comes from the east African highlands but has spread worldwide, is easy to cultivate, still largely unexploited and is correspondingly cheap. An initial feed test with 20 Kikuyu grass meal shows that it is highly usable. However, for high admixtures around 30, the growth of the fish decreased, which the researchers attributed to the increased fibre content as well as the lack of important amino acids, above all methionine and lysine. This deficit could however be slightly alleviated by adding methionine as provided by the company Evonik. The Danish aquafeed manufacturer Aller Aqua, which is testing the usability of 7,000 tonnes of an organic protein concentrate made from grass from BioRefine in trout feed, demonstrates that feeding fish with grass protein is not just a fad among eccentric researchers. According to Aller Aqua, the protein concentrate has a similar nutritional profile to soya bean meal and it can also be produced regionally and in a climate and environmentefficient way.

Olive pomace and micro and macroalgae

Due to their high concentration of valuable lipids, water-soluble carbohydrates and polyphenols, for which antimicrobial, antioxidative and antimycotic properties are claimed, the byproducts of olive oil manufacturing are also being considered as potential raw materials for fish feed. Byproducts such as olive pomace occur in considerable quantities when pressing olive oil and can, for example, be used as an alternative to wheat bran. This raw material has already been successfully tested in feed tests on the African Clarias catfish and tilapia.

However, the greatest hopes of the feed industry are currently resting on the production of micro and macroalgae, especially as they contribute not just to the solution of the nutrition problems in aquaculture, but also to the improvement of marine health and climate protection (algae bind carbon dioxide and reduce the quantities of phosphorus and nitrogen in the seas.) Macroalgae forests also act as a nursery and hiding place for many marine animals, thereby promoting biodiversity underwater. The European Commission, along with the European Climate, Environment and Infrastructure Executive Agency (CINEA) and a consortium of several interest groups in the area of algae cultures, founded the European platform “EU4Algae”, which aims to advance development in this area.

Microalgae have the potential to replace fishmeal and fish oil in aquaculture feeds. With raw protein contents of up to 65 and lipid contents of up to 40, their nutritional profile is very comparable to terrestrial plant and animal sources. However, the current global production of microalgae (autotrophs and heteretrophs), at an estimated 50,000 tonnes per year, only covers 0.7 of the actual protein demand of the feed industry, which is much too little to substitute for fishmeal protein in fish feed. In addition, with prices between 10 and 30 US dollars per kg, microalgae cost significantly more than soya meal, which is currently at around 0.60 USD per kg. It can therefore hardly be expected that microalgae will become a viable alternative protein source for fish feed. However interest from the industry is significant and even growing, since it was recently discovered that the photoautotrophic marine diatom Phaeocystis pouchetii not only contains high concentrations of the omega-3 essential fatty acids EPA and DHA and vitamins, but can also deter sea lice when added to salmon feed. The results of feed experiments show that the number of lice parasites on salmon that were fed with feed containing diatoms was significantly lower. The scientists suspect that this effect was caused by oxylipin, a fatty acid-based compound containing oxygen. However, microalgae are also increasing in importance for aquaculture as a source of minerals. For example, the microalga Nannochloropsis oceanica is rich in selenium, which is indispensable for the healthy development of the fish and the functioning of their immune system. Selenium deficiencies can inhibit growth, increase mortality and hinder immune response. Algae selenium also has high bioavailability, i.e. it is present in a form that can easily be absorbed by fish intestine. Because there are still many open questions regarding the potential of microalgae, the EU Horizon 2020 fund is supporting the corresponding GAIN project research.

Although the potential of macroalgae is often overestimated (the proportion of raw protein is very variable and ranges from less than 1 to 48 of the dry weight of the biomass), it could also play a certain role in fish nutrition in aquaculture, because its proteins have just as many or more essential amino acids as agricultural crops and fishmeal. If the high content of complex polysaccharides (fibre), which negatively affects the digestibility of the algal protein and the nutritional value, can eventually successfully be reduced, macroalgae could also be considered for aquaculture feed. All the more so since they contain bioactive compounds that are linked to higher stress resistance and improved immune function for fish. It appears that macroalgae in feed stimulate the appetite and thereby indirectly contribute to better growth of fish and crustaceans. Finally, however, none of the alternative protein sources can currently completely replace fishmeal. A high degree of flexibility is being demanded of the feed industry regarding the composition of its aquaculture feed, especially since raw material prices are volatile and are likely to rise further. mk

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