WO2022129592A1

WO2022129592A1 - Aquaculture feed

Info

Publication number: WO2022129592A1
Application number: PCT/EP2021/086625
Authority: WO
Inventors: David Christian Martin; Ester SANTIGOSA CULI
Original assignee: Dsm Ip Assets B.V.; Evonik Operations Gmbh
Priority date: 2020-12-17
Filing date: 2021-12-17
Publication date: 2022-06-23
Also published as: CN116600653A; CL2023001724A1; EP4262427A1

Abstract

The present invention relates to a feed premix feed premix comprising high amounts of eicosapentaenoic acid ("EPA") and docosahexaenoic acid ("DHA") as feed premix for animals, preferably aquatic animals including fish and shrimp. More particular, this invention relates to a feed premix intended for intake by aquatic animals comprising high amounts of EPA and DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid and low amounts of fish oil. The invention further concerns a method of producing a meat product, preferably anaquaculture meat product, by feeding an animal, preferably a fish, a feed composition, said method comprising the step of formulating a feed compositioncomprising DHA, preferably EPA and DHA, derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid and low amounts of fish oil.

Description

AQUACULTURE FEED

The present invention relates to a feed premix comprising high amounts of eicosapentaenoic acid ("EPA") and docosahexaenoic acid ("DHA") and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid and low amounts of fish oil and to a final feed for farm animals, preferably for aquatic animals including fish and shrimp. More particular, this invention relates to a feed premix intended for intake by aquatic animals comprising high amounts of DHA, preferably EPA and DHA, and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid and low amounts of fish oil. The invention further concerns a method of producing a meat product, preferably an aquaculture meat product, by feeding an animal, preferably a fish, a feed composition, said method comprising the step of formulating a feed composition comprising DHA, preferably EPA and DHA, and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid and low amounts of fish oil.

Aquaculture is a form of agriculture that involves the propagation, cultivation and marketing of aquatic animals and plants in a controlled environment. The aquaculture industry is currently the fastest growing food production sector in the world. World aquaculture produces approximately 60 million tons of seafood, which is worth more than $70 billion (US) annually. Today, farmed fish accounts for approximately 50% of all fish consumed globally. This percentage is expected to increase as a result of dwindling catches from capture fisheries in both marine and freshwater environments and increasing seafood consumption (i.e., total and per capita). Today, species groups in aquaculture production include, for example: carps and other cyprinids; oysters; clams, cockles and ark shells; shrimps and prawns; salmons, trouts and smelts; mussels; tilapias and other cichlids; and scallops.

While some aquacultured species (e.g., Tilapia) can be fed on an entirely vegetarian diet, many others species are fed a carnivorous diet. Typically, the feed for carnivorous fish comprises fishmeal and fish oil derived from wild caught species of small pelagic fish (predominantly anchovy, jack mackerel, blue whiting, capelin, sandeel and menhaden). These pelagic fish are processed into fishmeal and fish oil, with the final product often being either a pelleted or flaked feed, depending on the size of the fish. The other components of the aquaculture feed composition may include vegetable protein, vitamins, minerals and pigment as required.

Marine fish oils have traditionally been used as the sole dietary lipid source in commercial fish feed given their ready availability, competitive price and the abundance of essential fatty acids contained within this product. Additionally, fish oils readily supply essential fatty acids which are required for regular growth, health, reproduction and bodily functions within fish. More specifically, all vertebrate species, including fish, have a dietary requirement for both omega-6 and omega-3 polyunsaturated fatty acids ["PUFAs"]. Eicosapentaenoic acid ["EPA"; cis-5, 8,11 ,14,17-eicosapentaenoic acid; omega-3] and docosahexaenoic acid ["OHA"; cis- 4, 7, 10, 13, 16, 19-docosahexaenoic acid; 22:6 omega-3] are required for fish growth and health and are often incorporated into commercial fish feeds via addition of fish oils.

It is estimated that aquaculture feed compositions currently use about 87% of the global supply of fish oil as a lipid source. Since annual fish oil production has not increased beyond 1.5 million tons per year, the rapidly growing aquaculture industry cannot continue to rely on finite stocks of marine pelagic fish as a supply of fish oil. Thus, there is great urgency to find and implement sustainable alternatives to fish oil that can keep pace with the growing global demand for fish products.

U.S. Pat. No. 7,932,077 suggests recombinantly engineered Yarrowia lipolytica may be a useful addition to most animal feeds, including aquaculture feeds, as a means to provide necessary omega-3 and/or omega-6 PUFAs and based on its unique protein:lipid:carbohydrate feed additive , as well as unique complex carbohydrate profile (comprising an approximate 1 :4:4.6 ratio of mannan:beta-glucans:chitin).

U.S. Pat. Appl. Pub. No. 2007/0226814 discloses fish food containing at least one biomass obtained from fermenting microorganisms wherein the biomass contains at least 20% DHA relative to the total fatty acid content. Preferred microorganisms used as sources for DHA are organisms belonging to the genus Stramenopiles.

However, one important factor in aquaculture is the turnover rate. The turnover rate is determined by how fast the fish grow to a harvestable size. It is highly affected by the growth of the animals and therefore reduced growth has a large economic impact on aquaculture farmers.

Substitutes to fish oils, that have high concentrations of PUFAs of at least 30%, are highly advantageous in many ways. It is easier to incorporate the, often times liquid, formulation into the feed. The aquatic animal can take in a higher amount of nutrients, while ingesting the same amount of feed.

Unfortunately, the use of such highly concentrated substitutes has been an issue and can lead to reduced growth of the aquatic animals. This problem is currently solved by diluting the highly concentrated substitutes with oils low in PUFAs, such as rapeseed or soy oil. It therefore remains a need in aquaculture industry to find a composition of a fish oil substitute, preferably derived from microbial source, that is high in PUFAs, preferably DHA, more preferably EPA and DHA, suitable for intake by aquatic animals to replace fish oil partially or completely while maintaining the animal's performance and providing all necessary nutrients.

Summary of the invention

Surprisingly, we found that oils, such as microbial or vegetable oils, having high concentrations of PUFAs, such as a DHA content of at least 30%, preferably DHA and EPA, incorporated in animal feed does not result in a reduced growth by an extra addition of additives, such as vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid, wherein at least one of the additives is preferably used in higher dosages than normally used in animal feed.

In one aspect, the present invention relates to a feed premix intended for intake by aquatic animals, said feed premix comprising DHA, preferably EPA and DHA, at least 3, preferably 4, preferably 5, preferably 6, additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. In a preferred embodiment said feed premix may be an oil.

In a further embodiment, the present invention relates to animal feed, in particular an aquaculture feed comprising the feed premix in an amount between 1 and 10%, measured as a weight percent of the feed.

In a further embodiment, the present invention relates to animal feed, in particular an aquaculture feed comprising an oil containing high levels of DHA, preferably EPA and DHA and extra additions of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid and low amounts of fish oil.

In a further embodiment, the present invention relates to a method of producing a meat product, preferably an aquaculture meat product, by feeding an animal, preferably a fish, a feed composition, said method comprising the step of formulating a feed composition comprising DHA, preferably EPA and DHA, at least 5, preferably at least 6, additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid and low amounts of fish oil.

Detailed Description

Aquaculture is the practice of farming aquatic animals and plants. It involves cultivating an aquatic product (e.g., freshwater and saltwater animals) under controlled conditions. It involves growing and harvesting fish, shellfish, and aquatic plants in fresh, brackish or salt water.

Organisms grown in aquaculture may include fish and crustaceans. Crustaceans are, for example, lobsters, crabs, shrimp, prawns and crayfish. The farming of finfish is the most common form of aquaculture.

It involves raising fish commercially in tanks, ponds, or ocean enclosures, usually for food. A facility that releases juvenile fish into the wild for recreational fishing or to supplement a species' natural numbers is generally referred to as a fish hatchery. Particularly of interest are fish of the salmonid group, for example, cherry salmon (Oncorhynchus rnasou), Chinook salmon (O. tshawytscha), chum salmon (O. keta), coho salmon (O. kisutch), pink salmon (O. gorbuscha), sockeye salmon (O. nerka) and Atlantic salmon (Salmo salar). Other finfish of interest for aquaculture include, but are not limited to, various trout, as well as whitefish such as tilapia (including various species of Oreochromis, Sarotherodon, and Tilapia), sea bass, catfish (order Siluri-formes), bigeye tuna (Thunnus obesus), carp (family Cyprinidae) and cod (Gadus).

Aquaculture typically requires a prepared aquaculture feed composition to meet dietary requirements of the cultured animals. Dietary requirements of different aquaculture species vary, as do the dietary requirements of a single species during different stages of growth. Thus, tremendous research is invested towards optimizing each aquaculture feed composition for each stage of growth of a cultured organism.

Aquaculture feed compositions are composed of micro and macro components. In general, all components, which are used at levels of more than 1 %, are considered as macro components. Feed ingredients used at levels of less than 1 % are micro components. They are premixed to achieve a homogeneous distribution of the micro components in the complete feed. Both macro and micro ingredients are subdivided into components with nutritional functions and technical functions.

Components with technical functions improve the physical quality of the aquaculture feed composition or its appearance.

Macro components with nutritional functions provide aquatic animals with protein and energy required for growth and performance. With respect to fish, the aquaculture feed composition should ideally provide the fish with: 1 ) fats, which serve as a source of fatty acids for energy (especially for heart and skeletal muscles); and, 2) amino acids, which serve as building blocks of proteins. Fats also assist in vitamin absorption; for example, vitamins A, K, D, E and K are fat-soluble or can only be digested, absorbed, and transported in conjunction with fats. Carbohydrates, typically of plant origin (e.g., wheat, sunflower meal, corn gluten, soybean meal), are also often included in the feed compositions, although carbohydrates are not a superior energy source for fish over protein or fat.

Fats are typically provided via incorporation of fish meals (which contain a minor amount of fish oil) and fish oils into the aquaculture feed compositions. Extracted oils that may be used in aquaculture feed compositions include fish oils (e.g., from the oily fish menhaden, anchovy, herring, capelin and cod liver), and vegetable oil (e.g., from soybeans, rapeseeds, sunflower seeds and flax seeds). Typically, fish oil is the preferred oil, because it contains the long chain omega-3 polyunsaturated fatty acids ["PUFAs"], EPA and DHA; in contrast, vegetable oils do not provide a source of EPA and/or DHA. These PUFAs are needed for growth and health of most aquaculture products. A typical aquaculture feed composition will comprise from about 15-30% of oil (e.g., fish, vegetable, etc.), measured as a weight percent of the aquaculture feed composition.

The protein supplied in aquaculture feed compositions can be of plant or animal origin. For example, protein of animal origin can be from marine animals (e.g., fish meal, fish oil, fish protein, krill meal, mussel meal, shrimp peel, squid meal, squid oil, etc.) or land animals (e.g., blood meal, egg powder, liver meal, meat meal, meat and bone meal, silkworm, pupae meal, whey powder, etc.). Protein of plant origin can include soybean meal, corn gluten meal, wheat gluten, cottonseed meal, canola meal, sunflower meal, rice and the like.

The technical functions of macro components can be overlapping as, for example, wheat gluten may be used as a pelleting aid and for its protein content, which has a relatively high nutritional value. There can also be mentioned guar gum and wheat flour.

Micro components include feed additives such as vitamins, trace minerals, feed antibiotics and other biologicals. Minerals used at levels of less than 100 mg/kg (100 ppm) are considered as micro minerals or trace minerals.

Micro components with nutritional functions are all biologicals and trace minerals. They are involved in biological processes and are needed for good health and high performance. There can be mentioned additional vitamins such as vitamins A, K3, D3, B1 , B3, B6, B12, biotin, folic acid, panthothenic acid, nicotinic acid, choline chloride, inositol and para-amino-benzoic acid. There can be mentioned minerals such as salts of calcium, cobalt, copper, iron, magnesium, phosphorus, potassium, selenium and zinc. Other components may include, but are not limited to, antioxidants, beta-glucans, bile salt, cholesterol, enzymes, monosodium glutamate, carotenoids, etc.

The technical functions of micro ingredients are mainly related to pelleting, detoxifying, mold prevention, antioxidation, etc.

In aquaculture, typically fish are fed in different dietary cycles as they grow. The weights of fish of different dietary cycles may vary depending on the type of fish and/or the aquaculture practice used.

In one aspect, the present invention relates to animal feed, in particular an aquaculture feed comprising an oil containing high levels of DHA, preferably EPA and DHA and extra additions of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid, preferably wherein the oil may be added after processing of the feed, e.g. into pellets.

The total concentration of fish oil in the feed is below 10%, preferably below 5%, preferably below 2.5%, preferably below 2%, preferably below 1.5%, preferably below 1 %, preferably below 0.5%, preferably below 0.1 %, measured as a weight percent of the feed.

In another aspect, the present invention relates to a feed premix intended for intake by aquatic animals, said feed premix comprising DHA, preferably EPA and DHA, at least 3, preferably 4, preferably 5, preferably 6, additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. In a preferred embodiment, said feed premix may be an oil.

The total concentration of DHA and EPA in the feed premix is at least 40%w/w, preferably at least 45%, preferably at least 50%, preferably at least 55%, preferably at least 58%, measured as a weight percent of the oil.

The total concentration of fish oil in the feed premix is below 50%, preferably below 25%, preferably below 20%, preferably below 15%, preferably below 10%, preferably below 5%, preferably below 1 %, measured as a weight percent of the feed premix.

In just another embodiment of the invention, the total unsaturated fatty acid concentration in the feed premix is at least 50%, preferably at least 55%, preferably at least 58% is at least 60%, preferably at least 65%, measured as a weight percent of the feed additive.

In another aspect of the invention, the feed premix can be mixed with an oil selected from the group consisting of, microbial oil and one or more vegetable oil(s) to reduce the concentrations of PUFAs, as well as DHA, preferably EPA and DHA. In a more preferred embodiment, the vegetable oil is selected from the group consisting of rape seed oil and soy oil.

In another embodiment of the invention, the feed premix may be an oil or a mixture of oils selected from the group consisting of fish oil, microbial oil and one or more vegetable oil(s) comprising at least 3, preferably 4, preferably 5, preferably 6, additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid.

The oil comprising DHA, preferably DHA and EPA, according to the present invention may be derived from a microbial source or a vegetable source. Preferably, the oil is an oil obtained from an algae, fungi or yeast. Preferred microbes are Thraustochytrids which are microorganisms of the order Thraustochytriales. Thraustochytrids include members of the genus Schizochytrium and Thraustochytrium and have been recognized as an alternative source of omega-3 fatty acids, including DHA and EPA. See U.S. Patent No. 5,130,242.

In a preferred embodiment the microorganism is a mutant strain of the species Schizochytrium. Schizochytrium strains are natural sources of PUFAs such as DHA and can be optimized by mutagenesis to be used as microbial source according to the present invention.

In a preferred embodiment of the invention, the concentration of EPA in the feed premix is at least 5%, preferably at least 6%, preferably at least 7%, preferably at least 8%, preferably at least 9%, preferably at least 10%, preferably at least 11 %, preferably at least 12 %, measured as a weight percent of the oil.

In a preferred embodiment of the invention, the concentration of DHA in the feed premix is at least 30%, preferably at least 32%, preferably at least 35%, preferably at least 37%, preferably at least 38%, measured as a weight percent of the oil.

In a preferred embodiment of the invention, the concentration of vitamin C in the feed premix is such that if added to the feed the final concentration of vitamin C in the feed is between 150 and 800 ppm, preferably between 150 and 600 ppm, preferably between 200 and 600 ppm, preferably between 200 and 500ppm, preferably between 200 and 400ppm, preferably between 200 and 300ppm, preferably between 200 and 250ppm.

In a preferred embodiment of the invention, the concentration of vitamin E in the feed premix is such that if added to the feed the final concentration of vitamin E in the feed is between 250 and 800 ppm, preferably between 250 and 700 ppm, preferably between 250 and 600 ppm, preferably between 300 and 600 ppm, preferably between 300 and 550ppm, preferably between 300 and 400ppm.

In a preferred embodiment of the invention, the concentration of vitamin C in the feed premix is between 2000 and 60000 ppm, preferably 2000 and 50000 ppm, preferably 4000 and 50000 ppm, preferably 4000 and 30000 ppm, preferably 4000 and 25000 ppm.

In a preferred embodiment of the invention, the concentration of vitamin E in the feed premix is between 2500 and 70000 ppm, preferably between 25000 and 60000 ppm, preferably between 3000 and 35000 ppm, preferably between 3000 and 30000 ppm, preferably between 5000 and 30000 ppm.

In a preferred embodiment of the invention, the concentration of vitamin A in the feed premix is such that if added to the feed the final concentration of vitamin A in the feed is between 1000 and 15000 III per kg, preferably 1500 and 14500 IU per kg, preferably between 1500 and 14000 IU per kg, preferably between 2000 and 13500 IU per kg, preferably between 2000 and 13000 IU per kg, preferably between 2000 and 12000 IU per kg, preferably between 2000 and 12000 IU per kg.

In a preferred embodiment of the invention, the concentration of vitamin A in the feed premix is between 10000 and 1500000 IU per kg, preferably 15000 and 1450000 IU per kg, preferably between 15000 and 1400000 IU per kg, preferably between 20000 and 1350000 IU per kg, preferably between 20000 and 1300000 IU per kg, preferably between 20000 and 1200000 IU per kg, preferably between 20000 and 1200000 IU per kg.

In a preferred embodiment of the invention, the concentration of vitamin K in the feed premix is such that if added to the feed the final concentration of vitamin D in the feed is between 1 and 200 mg per kg, preferably between 1 and 150 mg per kg, preferably between 1 and 100 mg per kg, preferably between 1 and 90 mg per kg, preferably between 1 and 80 mg per kg, preferably between 1 and 70 mg per kg, preferably between 1 and 60 mg per kg, preferably between 2 and 200 mg per kg, preferably between 3 and 200 mg per kg.

In a preferred embodiment of the invention, the concentration of vitamin K in the feed premix is between 10 and 20000 mg per kg, preferably between 10 and 15000 mg per kg, preferably between 10 and 10000 mg per kg, preferably between 10 and 9000 mg per kg, preferably between 10 and 8000 mg per kg, preferably between 10 and 7000 mg per kg, preferably between 10 and 6000 mg per kg, preferably between 20 and 20000 mg per kg, preferably between 30 and 20000 mg per kg.

In a preferred embodiment of the invention, the concentration of vitamin D in the feed premix is such that if added to the feed the final concentration of vitamin D in the feed is between 1000 and 10000 III per kg, preferably between 1500 and 9500 IU per kg, preferably between 1500 and 9000 IU per kg, preferably between 1500 and 8500 IU per kg, preferably between 1500 and 8000 IU per kg, preferably between 1500 and 7500 IU per kg, preferably between 1500 and 7000 IU per kg, preferably between 1500 and 6500 IU per kg.

In a preferred embodiment of the invention, the concentration of vitamin D in the feed premix is between 10000 and 1000000 IU per kg, preferably between 15000 and 950000 IU per kg, preferably between 15000 and 900000 IU per kg, preferably between 15000 and 850000 IU per kg, preferably between 15000 and 800000 IU per kg, preferably between 15000 and 750000 IU per kg, preferably between 15000 and 700000 IU per kg, preferably between 15000 and 650000 IU per kg,

In a preferred embodiment of the invention, the concentration of cholesterol in the feed premix is such that if added to the feed the final concentration of cholesterol in the feed is up to 1000 mg per kg, preferably up to 900 mg per kg, up to 800 mg per kg, preferably up to 750 mg per kg.

In a preferred embodiment of the invention, the concentration of cholesterol in the feed premix is between up to 100000 mg per kg, preferably up to 90000 mg per kg, up to 80000 mg per kg, preferably up to 75000 mg per kg

In a preferred embodiment of the invention, the concentration of phospholipids in the feed premix is such that if added to the feed the final concentration of cholesterol in the feed is between 0.5 and 3 %, preferably between 0.5 and 2.5 %, preferably between 0.5 and 2 %, preferably between 0.5 and 1.5 %, preferably between 0.5 and 1.3 %, measured as a weight percent of the feed

In a preferred embodiment of the invention, the concentration of phospholipids in the feed premix is between 5 and 30 %, preferably between 5 and 25 %, preferably between 5 and 20 %, preferably between 5 and 15 %, preferably between 5 and 13 %, measured as a weight percent of the feed premix.

In a preferred embodiment of the invention, the concentration of arachidonic acid in the feed premix is such that if added to the feed the final concentration of vitamin A in the feed is up to 0.1 %, preferably up to 0.08%, preferably up to 0.06%, preferably up to 0.04%, preferably between up to 0.02%, measured as a weight percent of the feed.

In a preferred embodiment of the invention, the concentration of arachidonic acid in the feed premix is up to 1 %, preferably up to 0.8%, preferably up to 0.6%, preferably up to 0.4%, preferably between up to 0.2%, measured as a weight percent of the feed premix.

In another aspect, the present invention relates to a feed premix comprising DHA, preferably EPA and DHA, at least 3, preferably 4, preferably 5, preferably 6, additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid and further ingredients (as defined above) intended for incorporation into an aquaculture feed.

In another embodiment, the feed may comprise a total amount of EPA and DHA that is at least about 0.8%, measured as weight percent of the feed. This amount (i.e. , 0.8%) is typically an appropriate minimal concentration that is suitable to support the growth of a variety of animals grown in aquaculture, and particularly is suitable for inclusion in the diets of salmonid fish.

Most processes to make an aquaculture feed composition of the invention will begin with a microbial fermentation, wherein a particular microorganism is cultured under conditions that permit growth and production of microbial oils comprising DHA, preferably EPA and DHA. At an appropriate time, the microbial cells are harvested from the fermentation vessel. This microbial biomass may be mechanically processed using various means, such as dewatering, drying, mechanical disruption, pelletization, etc. Then, the oil extracted from the biomass is supplemented with the appropriate amounts of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid and used as an ingredient in an aquaculture feed (preferably as a substitute for at least a portion of the fish oil used in standard aquaculture feed compositions). The aquaculture feed is then fed to aquatic animals over a portion of their lifetime, such that DHA, preferably EPA and DHA, from the aquaculture feed accumulate in the aquatic animals.

In another embodiment of the invention, an aquaculture feed according to the invention can be made as described in example 3. Water-soluble vitamins, such as vitamins C and E, may be added to the main raw materials from which the aquaculture feed, such as in the form of extruded feed is produced. Oil may be supplemented with the appropriate amount to reach the final concentrations of DHA, preferably EPA and DHA, vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid in the feed as defined above and applied to the surface of the feed by spraying the oil onto the surface of the feed, or dipping the feed in the oil.

In another embodiment, the invention relates to a feed, wherein the concentration of the supplemented feed premix, comprising DHA, preferably EPA and DHA, at least 3, preferably 4, preferably 5, preferably 6, additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid, in the feed is between 1 and 15%, preferably between 1 and 10%, preferably between 2 and 10%, preferably between 3 and 10%, Preferably between 3 and 8%, preferably between 3 and 7%, preferably between 3 and 6%, measured as a weight percent of the feed.

In a preferred embodiment of the invention, the concentration of EPA in the feed is at least 0.05%, preferably at least 0.06%, preferably at least 0.07%, preferably at least 0.08%, preferably at least 0.09%, preferably at least 0.1 %, preferably at least 0.11%, preferably at least 0.12 %, preferably at least 0.15 %, preferably at least 0.2 %, preferably at least 0.25 %, preferably at least 0.3 % measured as a weight percent of the feed.

In a preferred embodiment of the invention, the concentration of DHA in the feed is at least 0.3%, preferably at least 0.32%, preferably at least 0.35%, preferably at least 0.37%, preferably at least 0.38%, preferably at least 0.4%, preferably at least 0.45%, preferably at least 0.5%, preferably at least 0.55%, preferably at least 0.6%, measured as a weight percent of the feed.

In a preferred embodiment of the invention, the total amount of EPA and DHA that is at least 0.05 %, preferably 0.06 %, preferably 0.07 %, 0.08%, measured as a weight percent of the feed.

In a preferred embodiment of the invention, the concentration of vitamin C in the feed is between 150 and 800 ppm, preferably between 150 and 600 ppm, preferably between 200 and 600 ppm, preferably between 200 and 500ppm, preferably between 200 and 400ppm, preferably between 200 and 300ppm, preferably between 200 and 250ppm.

In a preferred embodiment of the invention, the concentration of vitamin E in the feed is between 250 and 800 ppm, preferably between 250 and 700 ppm, preferably between 250 and 600 ppm, preferably between 300 and 600 ppm, preferably between 300 and 550ppm, preferably between 300 and 400ppm. In a preferred embodiment of the invention, the concentration of vitamin A in the feed is between 1000 and 15000 III per kg, preferably 1500 and 14500 IU per kg, preferably between 1500 and 14000 IU per kg, preferably between 2000 and 13500 IU per kg, preferably between 2000 and 13000 IU per kg, preferably between 2000 and 12000 IU per kg, preferably between 2000 and 12000 IU per kg,

In a preferred embodiment of the invention, the concentration of vitamin K in the feed is between 1 and 200 mg per kg, preferably between 1 and 150 mg per kg, preferably between 1 and 100 mg per kg, preferably between 1 and 90 mg per kg, preferably between 1 and 80 mg per kg, preferably between 1 and 70 mg per kg, preferably between 1 and 60 mg per kg, preferably between 2 and 200 mg per kg, preferably between 3 and 200 mg per kg.

In a preferred embodiment of the invention, the concentration of vitamin D in the feed is between between 1000 and 10000 IU per kg, preferably between 1500 and 9500 IU per kg, preferably between 1500 and 9000 IU per kg, preferably between 1500 and 8500 IU per kg, preferably between 1500 and 8000 IU per kg, preferably between 1500 and 7500 IU per kg, preferably between 1500 and 7000 IU per kg, preferably between 1500 and 6500 IU per kg,

In a preferred embodiment of the invention, the concentration of cholesterol in the feed is up to 1000 mg per kg, preferably up to 900 mg per kg, up to 800 mg per kg, preferably up to 750 mg per kg.

In a preferred embodiment of the invention, the concentration of Phospholipids in the feed is between 0.5 and 3 %, preferably between 0.5 and 2.5 %, preferably between 0.5 and 2 %, preferably between 0.5 and 1.5 %, preferably between 0.5 and 1.3 %, measured as a weight percent of the feed.

In a preferred embodiment of the invention, the concentration of arachidonic acid in the feed is up to 0.1 %, preferably up to 0.08%, preferably up to 0.06%, preferably up to 0.04%, preferably between up to 0.02%, measured as a weight percent of the feed.

In another aspect of the invention, the aquaculture feed is an extruded feed pellet or a pressed feed pellet. In a more preferred aspect, the aquaculture feed is a coated pellet. In a more preferred aspect, the feed additive is present in the coating of the feed.

In a further embodiment, the present invention relates to the use of an animal feed comprising a microbial oil with high PUFA concentrations for replacing all or part of fish oil in the feed in combination with high levels of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid for maintaining animal growth compared to the use of fish oil alone as additional PUFA source.

In a further embodiment, the present invention relates to a method of producing a meat product, preferably an aquaculture meat product, by feeding an animal, preferably a fish, a feed composition, said method comprising the step of formulating a feed composition comprising DHA, preferably EPA and DHA, derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid.

In a preferred embodiment of the invention, the method of producing a meat product, preferably an aquaculture meat product, comprises the step of formulating a feed composition comprising the premix comprising DHA, preferably EPA and DHA, derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid, in the feed in an amount between 1 and 15%, preferably between 1 and 10%, preferably between 2 and 10%, preferably between 3 and 10%, Preferably between 3 and 8%, preferably between 3 and 7%, preferably between 3 and 6%, measured as a weight percent of the feed.

In a further embodiment of the invention, the method of producing a meat product, preferably an aquaculture meat product, comprises the step of formulating a feed composition comprising DHA in an amount of at least 0.3%, preferably at least 0.32%, preferably at least 0.35%, preferably at least 0.37%, preferably at least 0.38%, preferably at least 0.4%, preferably at least 0.45%, preferably at least 0.5%, preferably at least 0.55%, preferably at least 0.6%, measured as a weight percent of the feed.

In a further embodiment of the invention, the method of producing a meat product, preferably an aquaculture meat product, comprises the step of formulating a feed composition comprising EPA in amount of least 0.05%, preferably at least 0.06%, preferably at least 0.07%, preferably at least 0.08%, preferably at least 0.09%, preferably at least 0.1 %, preferably at least 0.11 %, preferably at least 0.12 %, preferably at least 0.15 %, preferably at least 0.2 %, preferably at least 0.25 %, preferably at least 0.3 % measured as a weight percent of the feed.

In a further embodiment of the invention, the method of producing a meat product, preferably an aquaculture meat product, comprises the step of formulating a feed composition comprising vitamin C in an amount between 150 and 800 ppm, preferably between 150 and 600 ppm, preferably between 200 and 600 ppm, preferably between 200 and 500ppm, preferably between 200 and 400ppm, preferably between 200 and 300ppm, preferably between 200 and 250ppm. In a further embodiment of the invention, the method of producing a meat product, preferably an aquaculture meat product, comprises the step of formulating a feed composition comprising vitamin E in an amount between 250 and 800 ppm, preferably between 250 and 700 ppm, preferably between 250 and 600 ppm, preferably between 300 and 600 ppm, preferably between 300 and 550ppm, preferably between 300 and 400ppm.

In a further embodiment of the invention, the method of producing a meat product, preferably an aquaculture meat product, comprises the step of formulating a feed composition comprising a total amount of EPA and DHA derived from a single microbial source that is at least about 0.8%, measured as weight percent of the feed. This amount (i.e. , 0.8%) is typically an appropriate minimal concentration that is suitable to support the growth of a variety of animals grown in aquaculture, and particularly is suitable for inclusion in the diets of salmonid fish.

In a further embodiment of the invention, the method of producing a meat product, preferably an aquaculture meat product, comprises the step of formulating a feed composition, wherein the feed is an extruded feed pellet or a pressed feed pellet.

In a further embodiment of the invention, the method of producing a meat product, preferably an aquaculture meat product, comprises the step of formulating a feed composition, wherein the feed pellet is a coated feed pellet.

In a further embodiment of the invention, the method of producing a meat product, preferably an aquaculture meat product, comprises the step of formulating a feed composition, wherein the feed premix is present in the coating of the coated feed pellet.

Embodiments of the invention can be summarized as follows:

1 . A feed premix intended for intake by a farm animal comprising an oil containing high levels of DHA and at least 3 additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid, and wherein the concentration of DHA is at least 30 % w/w measured as a weight percent of the oil.

2. The feed premix according to claim 1 comprising at least 4, preferably at least 5 additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid.

3. The feed premix according to claim 1 or 2, wherein the oil is derived from a microbial source or vegetable source.

4. The feed premix according to any of claims 1 to 3, wherein a. The concentration of vitamin C is such that if added to the feed the final concentration in the feed is between 200 and 600 ppm, preferably between 200 and 500ppm. b. The concentration of Vitamin E is such that if added to the feed the final concentration in the feed is between 250 and 700 ppm, preferably between 300 and 600 ppm.

5. The feed premix according to any of claims 1 to 4 a. The concentration vitamin C is between 2000 and 60000 ppm, preferably between 2000 and 50000ppm. b. The concentration of Vitamin E is between 2500 and 70000 ppm, preferably between 2500 and 60000 ppm.

6. The feed premix according to any of claims 1 to 5, wherein the oil comprises EPA and wherein the concentration of EPA is at least 10%, preferably at least 12 %, measured as a weight percent of the oil.

7. The feed premix according to any of claims 1 to 6, wherein the total concentration of fish oil in the feed premix is below 50%, preferably below 25%, preferably below 20%, preferably below 15%, preferably below 10%, preferably below 5%, preferably below 1 %, preferably below 0.1 %, measured as a weight percent of the feed premix.

8. The feed premix according to any of claims 1 to 7, wherein the feed premix does not comprise fish oil.

9. The feed premix according to any of claims 1 to 8, wherein the concentration of vitamin A in the feed premix is between 10000 and 1500000 III per kg, preferably 15000 and 1450000 IU per kg, preferably between 15000 and 1400000 IU per kg, preferably between 20000 and 1350000 IU per kg, preferably between 20000 and 1300000 IU per kg, preferably between 20000 and 1200000 IU per kg, preferably between 20000 and 1200000 IU per kg.

10. The feed premix according to any of claims 1 to 9, wherein the concentration of vitamin K in the feed premix is between 10 and 20000 mg per kg, preferably between 10 and 15000 mg per kg, preferably between 10 and 10000 mg per kg, preferably between 10 and 9000 mg per kg, preferably between 10 and 8000 mg per kg, preferably between 10 and 7000 mg per kg, preferably between 10 and 6000 mg per kg, preferably between 20 and 20000 mg per kg, preferably between 30 and 20000 mg per kg.

11 .The feed premix according to any of claims 1 to 10, wherein the concentration of vitamin D in the feed premix is between 10000 and 1000000 IU per kg, preferably between 15000 and 950000 III per kg, preferably between 15000 and 900000 IU per kg, preferably between 15000 and 850000 IU per kg, preferably between 15000 and 800000 IU per kg, preferably between 15000 and 750000 IU per kg, preferably between 15000 and 700000 IU per kg, preferably between 15000 and 650000 IU per kg.

12. The feed premix according to any of claims 1 to 11 , wherein the concentration of cholesterol in the feed premix is between up to 100000 mg per kg, preferably up to 90000 mg per kg, up to 80000 mg per kg, preferably up to 75000 mg per kg.

13. The feed premix according to any of claims 1 to 12, wherein the concentration of phospholipids in the feed premix is between 5 and 30 %, preferably between 5 and 25 %, preferably between 5 and 20 %, preferably between 5 and 15 %, preferably between 5 and 13 %, measured as a weight percent of the feed premix.

14. The feed premix according to any of claims 1 to 13, wherein the concentration of arachidonic acid (ARA) in the feed premix is up to 1 %, preferably up to 0.8%, preferably up to 0.6%, preferably up to 0.4%, preferably between up to 0.2%, measured as a weight percent of the feed premix.

15. The feed premix according to any of claims 1 to 14, wherein the feed premix is an oil.

16. The feed premix of any of claims 1 to 15, wherein the oil comprises EPA and wherein the total concentration of DHA and EPA is at least 50%w/w, preferably at least 55%, preferably at least 58%, measured as a weight percent of the oil.

17. The feed premix of claim 16, wherein the oil comprises DHA, EPA and ARA.

18. The feed premix of any of claims 1 to 17, wherein the total unsaturated fatty acid concentration is at least 60%, preferably at least 65%, measured as a weight percent of the feed premix.

19. The feed premix of any of claims 1 to 18, wherein the oil is derived from a microbial source and wherein said microorganism is an algae, fungi or yeast.

20. The feed premix of claim 19, wherein the microorganism is a member of the genus Schizochytrium or Thraustochytrium.

21 .The feed premix according to any of claims 1 to 20, further comprising at least one additional component selected from the group consisting of fat-soluble vitamins, water soluble vitamins, trace minerals, carotenoids, polyunsatured fatty acids, probiotics, prebiotics and macro minerals. A feed intended for intake by a farm animal comprising an oil containing high levels of DHA and at least 5 additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phoshpholipids and arachidonic acid, and wherein the concentration of DHA is at least 30 % measured as a weight percent of the oil. The feed according to claim 22, wherein the oil is derived from a microbial source or vegetable source. The feed according to claim 22 or 23, wherein a. The concentration vitamin C is between 200 and 600 ppm, preferably between 200 and 500ppm. b. The concentration of Vitamin E between 250 and 700 ppm, preferably between 300 and 600 ppm. The feed according to any of claims 22 to 24, wherein the oil comprises EPA and wherein the concentration of EPA is at least 0.1 %, preferably at least 0.12 %, measured as a weight percent of the feed. Feed according to claim 25, comprising a total amount of EPA and DHA that is at least 0.08%, measured as a weight percent of the feed. A feed intended for intake by a farm animal comprising the feed premix according to any of claims 1 to 21 . Feed according to claim 27, wherein the amount of the feed premix according to any of claims 1 to 21 is such that the concentration in the feed is between 1 and 10%, measured as a weight percent of the feed. Feed according to any of claims 22 to 28, wherein the total concentration of fish oil in the feed is below 10%, preferably below 5%, preferably below 2.5%, preferably below 2%, preferably below 1.5%, preferably below 1 %, preferably below 0.5%, preferably below 0.1 %, measured as a weight percent of the feed. Feed according to any of claims claim 22 to 29, wherein the feed does not comprise fish oil. Feed according to any of claims 22 to 30, wherein the concentration of vitamin A in the feed is between 1000 and 15000 III per kg, preferably 1500 and 14500 IU per kg, preferably between 1500 and 14000 IU per kg, preferably between 2000 and 13500 IU per kg, preferably between 2000 and 13000 IU per kg, preferably between 2000 and 12000 III per kg, preferably between 2000 and 12000 IU per kg.

32. Feed according to any of claims 22 to 31 , wherein the concentration of vitamin K in the feed is between 1 and 200 mg per kg, preferably between 1 and 150 mg per kg, preferably between 1 and 100 mg per kg, preferably between 1 and 90 mg per kg, preferably between 1 and 80 mg per kg, preferably between 1 and 70 mg per kg, preferably between 1 and 60 mg per kg, preferably between 2 and 200 mg per kg, preferably between 3 and 200 mg per kg.

33. Feed according to any of claims 22 to 32, wherein the concentration of vitamin D in the feed is between 1000 and 10000 IU per kg, preferably between 1500 and 9500 IU per kg, preferably between 1500 and 9000 IU per kg, preferably between 1500 and 8500 IU per kg, preferably between 1500 and 8000 IU per kg, preferably between 1500 and 7500 IU per kg, preferably between 1500 and 7000 IU per kg, preferably between 1500 and 6500 IU per kg,

34. Feed according to any of claims 22 to 33, wherein the concentration of cholesterol in the feed is up to 1000 mg per kg, preferably up to 900 mg per kg, up to 800 mg per kg, preferably up to 750 mg per kg.

35. Feed according to any of claims 22 to 34, wherein the concentration of Phospholipids in the feed is between 0.5 and 3 %, preferably between 0.5 and 2.5 %, preferably between 0.5 and 2 %, preferably between 0.5 and 1.5 %, preferably between 0.5 and 1 .3 %, measured as a weight percent of the feed.

36. Feed according to any of claims 22 to 35, wherein the concentration of arachidonic acid in the feed is up to 0.1 %, preferably up to 0.08%, preferably up to 0.06%, preferably up to 0.04%, preferably between up to 0.02%, measured as a weight percent of the feed.

37. Feed according to any of claims 22 to 36, further comprising one or more ingredients selected from the group consisting of fish meal, krill meal, soya concentrate, corn gluten, wheat gluten, pea protein, wheat flour, fish oil, a vitamin, mineral premix, mineral premix plus synthetic phosphorus and combinations thereof.

38. Feed according to any of claims 22 to 37, wherein the aquaculture feed is an extruded feed pellet or a pressed feed pellet.

39. Feed according to any of claims 22 to 37, wherein the feed pellet is a coated feed pellet. 40. Feed according to any of claims 22 to 39, wherein the feed premix according to any of claims 1 to 21 is present in the coating of the coated feed pellet.

41. Use of an animal feed comprising an oil containing high levels of DHA for replacing all or part of fish oil in the feed in combination with at least 5 additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid, wherein the concentration of DHA is at least 0.30 %, preferably at least 0.35%, measured as a weight percent of the feed.

42. The use according to claim 39, wherein the oil is derived from a microbial source or vegetable source.

43. Use according to claim 41 or 42, wherein a. The concentration of Vitamin C is between 200 and 600 ppm, preferably between 200 and 500ppm. b. The concentration of Vitamin E between 250 and 700 ppm, preferably between 300 and 600 ppm.

44. Use according to any of claims 41 to 43, wherein the oil comprises EPA and wherein the concentration of EPA is at least 0.1 %, preferably at least 0.12 %, measured as a weight percent of the feed.

45. Use of an animal feed to any of claims 41 to 44 comprising a feed premix according to any of claims 1 to 21 in such an amount that the concentration of the premix in the feed is between 1 and 10%, measured as a weight percent of the feed.

46. The use according to any of claims 41 to 45 for maintaining animal growth compared to the use of fish oil alone as additional PUFA source.

47. Use according to any of claims 41 to 46, wherein the feed comprises a total amount of EPA and DHA that is at least 0.08%, measured as a weight percent of the feed.

48. Use according to any of claims 41 to 47, wherein the total concentration of fish oil in the feed is below 10%, preferably below 5%, preferably below 2.5%, preferably below 2%, preferably below 1.5%, preferably below 1 %, preferably below 0.5%, preferably below 0.1 %, measured as a weight percent of the feed.

49. Use according to any of claims 41 to 48, wherein the feed does not comprise fish oil.

50. Use according to any of claims 41 to 49, wherein the concentration of vitamin A in the feed is between 1000 and 15000 IU per kg, preferably 1500 and 14500 IU per kg, preferably between 1500 and 14000 IU per kg, preferably between 2000 and 13500 IU per kg, preferably between 2000 and 13000 IU per kg, preferably between 2000 and 12000 III per kg, preferably between 2000 and 12000 IU per kg.

51 . Use according to any of claims 41 to 50, wherein the concentration of vitamin K in the feed is between 1 and 200 mg per kg, preferably between 1 and 150 mg per kg, preferably between 1 and 100 mg per kg, preferably between 1 and 90 mg per kg, preferably between 1 and 80 mg per kg, preferably between 1 and 70 mg per kg, preferably between 1 and 60 mg per kg, preferably between 2 and 200 mg per kg, preferably between 3 and 200 mg per kg.

52. Use according to any of claims 41 to 51 , wherein the concentration of vitamin D in the feed is between 1000 and 10000 IU per kg, preferably between 1500 and 9500 IU per kg, preferably between 1500 and 9000 IU per kg, preferably between 1500 and 8500 IU per kg, preferably between 1500 and 8000 IU per kg, preferably between 1500 and 7500 IU per kg, preferably between 1500 and 7000 IU per kg, preferably between 1500 and 6500 IU per kg.

53. Use according to any of claims 41 to 52, wherein the concentration of cholesterol in the feed is up to 1000 mg per kg, preferably up to 900 mg per kg, up to 800 mg per kg, preferably up to 750 mg per kg.

54. Use according to any of claims 41 to 53, wherein the concentration of Phospholipids in the feed is between 0.5 and 3 %, preferably between 0.5 and 2.5 %, preferably between 0.5 and 2 %, preferably between 0.5 and 1.5 %, preferably between 0.5 and 1 .3 %, measured as a weight percent of the feed.

55. Use according to any of claims 41 to 54, wherein the concentration of arachidonic acid (ARA) in the feed is up to 0.1 %, preferably up to 0.08%, preferably up to 0.06%, preferably up to 0.04%, preferably between up to 0.02%, measured as a weight percent of the feed.

56. Use according to any of claims 41 to 55, wherein the feed further comprises one or more ingredients selected from the group consisting of fish meal, krill meal, soya concentrate, corn gluten, wheat gluten, pea protein, wheat flour, fish oil, a vitamin, mineral premix, mineral premix plus synthetic phosphorus and combinations thereof.

57. Use according to any of claims 41 to 56, wherein the feed is an extruded feed pellet or a pressed feed pellet.

58. Use according to any of claims 41 to 57, wherein the feed pellet is a coated feed pellet.

59. Use according to any of claims 41 to 58, wherein the feed premix according to any of claims 1 to 9 is present in the coating of the coated feed pellet. Definitions

Invention: As used herein the term "invention" or "present invention" is intended to refer to all aspects and embodiments of the invention as described in the claims and Ospecification herein and should not be read so as to be limited to any particular embodiment or aspect.

Farm animals: The term "farm animal" or "animal" refers to animals bred on farms mainly for production purposes, for example for the production of meat, milk, eggs or wool. Examples of farm animals include aquatic animals, cattle, pigs, sheep, goat, poultry, such as turkey, chickens or ducks.

Aquatic Animal: The term “aquatic animal” refers to crustaceans including but not limited to shrimps and prawns and fish including but not limited to amberjack, arapaima, barb, bass, bluefish, bocachico, bream, bullhead, cachama, carp, catfish, catla, chanos, char, cichlid, cobia, cod, crappie, dorada, drum, eel, goby, goldfish, gourami, grouper, guapote, halibut, java, labeo, lai, loach, mackerel, milkfish, mojarra, mudfish, mullet, paco, pearlspot, pejerrey, perch, pike, pompano, roach, salmon, sampa, sauger, sea bass, seabream, shiner, sleeper, snakehead, snapper, snook, sole, spinefoot, sturgeon, sunfish, sweetfish, tench, terror, tilapia, trout, tuna, turbot, vendace, walleye and whitefish.

Feed Premix: The term feed premix according to the invention refers to a formulation comprising DHA, preferably EPA and DHA, derived from a single microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid as active ingredient intended for intake by a farm animal, preferably an aquatic animal. The incorporation of the feed additive as exemplified herein above to animal feeds, for example fish feeds, is in practice carried out using a concentrate or a premix. A premix designates a preferably uniform mixture of one or more micro-ingredients with diluent and/or carrier. Premixes are used to facilitate uniform dispersion of micro-ingredients in a larger mix. A premix according to the invention can be added to feed ingredients as solids (for example as water soluble powder) or liquids. In a preferred embodiment of the invention, the feed premix may be an oil. Said oil may be added after processing of the feed, e.g. into pellets.

Feed or Aquaculture feed: The term “Feed” or “Aquaculture feed” or “feed composition” refers to any compound, preparation, or mixture suitable for, or intended for intake by farm animals, preferably aquatic animals. An animal feed for aquatic animals typically comprises high protein and energy concentrations, such as fish meal, molasses, oligosaccharide concentrates as well as vitamins, minerals, enzymes, direct fed microbial, amino acids and/or other feed ingredients (such as in a premix). Aquaculture feed refers to a manufactured or artificial diet (i.e. , formulated feed) to supplement or to replace natural feed, which is most commonly produced in form of flakes or pellets. Typically, an aquaculture feed may be in the form of flakes or pellets, for example extruded pellets.

Polyunsaturated fatty acids ["PUFAs"]: The term polyunsaturated fatty acids ["PLIFAs"] The term "polyunsaturated fatty acid" and "PUFA" include not only the free fatty acid form, but also other forms, such as triacylglycerols (TAG) in the form of, phospholipid (PL) and other forms of esterified forms. Additional details concerning the differentiation between "saturated fatty acids" versus "unsaturated fatty acids", "monounsaturated fatty acids" versus "polyunsaturated fatty acids" ["PUFAs"], and "omega-6 fatty acids" ["00-6" or "n-6"] versus "omega-3 fatty acids" ["00-3" or "n-3"] are provided in U.S. Patent 7,238,482.

Eicosapentaenoic acid [EPA]: The term "Eicosapentaenoic acid" ["EPA"] is the common name for eis-5, 8, 11 ,14, 17-eicosapentaenoic acid. This fatty acid is a 20:5 omega-3 fatty acid. The term EPA as used in the present disclosure will refer to the acid or derivatives of the acid (e.g., glycerides, esters, phospholipids, amides, lactones, salts or the like) unless specifically mentioned otherwise.

Docosahexaenoic acid [DHA]: The term "Docosahexaenoic acid" ["DHA"] is the common name for eis-4, 7, 10, 13, 16, 19-docosahexaenoic acid. This fatty acid is a 22:6 omega-3 fatty acid. The term DHA as used in the present disclosure will refer to the acid or derivatives of the acid (e.g., glycerides, esters, phospholipids, amides, lactones, salts or the like) unless specifically mentioned otherwise.

Vitamin C: For the purposes of the invention, vitamin C, e.g., as a commercial formulation such as available under the Trademark ROVIMIX® Stay-C® 35, is suitably administered to the animal as supplement to animal feed in the form of a feed premix comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid or in the form of a feed comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. Feed may be supplemented by admixing vitamin C to regular feed or by first preparing a premix of a feed component and vitamin C and subsequent mixing the premix with other feed components. The feed can be any feed. The term feed as used herein comprises both solid and liquid feed.

Vitamin E: For the purposes of the invention, vitamin E, e.g., as a commercial formulation such as available under the Trademark ROVIMIX® E50 SD, is suitably administered to the animal as supplement to animal feed in the form of a feed premix comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid or in the form of a feed comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. Feed may be supplemented by admixing vitamin E to regular feed or by first preparing a premix of a feed component and vitamin E and subsequent mixing the premix with other feed components. The feed can be any feed. The term feed as used herein comprises both solid and liquid feed.

Vitamin A: For the purposes of the invention, vitamin A, e.g., as a commercial formulation such as available under the Trademark ROVIMIX® A 1000, is suitably administered to the animal as supplement to animal feed in the form of a feed premix comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid or in the form of a feed comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. Feed may be supplemented by admixing vitamin A to regular feed or by first preparing a premix of a feed component and vitamin A and subsequent mixing the premix with other feed components. The feed can be any feed. The term feed as used herein comprises both solid and liquid feed.

Vitamin K: For the purposes of the invention, vitamin K is vitamin K3, e.g., as a commercial formulation such as available under the Trademark ROVIMIX® K3 MNB, is suitably administered to the animal as supplement to animal feed in the form of a feed premix comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid or in the form of a feed comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. Feed may be supplemented by admixing vitamin Kto regular feed or by first preparing a premix of a feed component and vitamin K and subsequent mixing the premix with other feed components. The feed can be any feed. The term feed as used herein comprises both solid and liquid feed.

Vitamin D: For the purposes of the invention, vitamin D is vitamin D3, e.g., as a commercial formulation such as available under the Trademark ROVIMIX® D3-500, is suitably administered to the animal as supplement to animal feed in the form of a feed premix comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid or in the form of a feed comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. Feed may be supplemented by admixing vitamin D to regular feed or by first preparing a premix of a feed component and vitamin D and subsequent mixing the premix with other feed components. The feed can be any feed. The term feed as used herein comprises both solid and liquid feed. Cholesterol: For the purposes of the invention, cholesterol, is suitably administered to the animal as supplement to animal feed in the form of a feed premix comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid or in the form of a feed comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. Feed may be supplemented by admixing cholesterol to regular feed or by first preparing a premix of a feed component and cholesterol and subsequent mixing the premix with other feed components. The feed can be any feed. The term feed as used herein comprises both solid and liquid feed.

Arachidonic acid: For the purposes of the invention, arachidonic acid (ARA), e.g., as a commercial formulation such as available under the Trademark ARASCO™, is suitably administered to the animal as supplement to animal feed in the form of a feed premix comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid or in the form of a feed comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. Feed may be supplemented by admixing arachidonic acid to regular feed or by first preparing a premix of a feed component and arachidonic acid and subsequent mixing the premix with other feed components. The feed can be any feed. The term feed as used herein comprises both solid and liquid feed.

Phospholipids: For the purposes of the invention, phospholipids, e.g., as a commercial formulation such as available under the Trademark ARASCO™, is suitably administered to the animal as supplement to animal feed in the form of a feed premix comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid or in the form of a feed comprising DHA derived from a microbial source and vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid. Feed may be supplemented by admixing phospholipids to regular feed or by first preparing a premix of a feed component and phospholipids and subsequent mixing the premix with other feed components. The feed can be any feed. The term feed as used herein comprises both solid and liquid feed.

Fish oil: The term "Fish oil" refers to oil derived from the tissues of an oily fish. Examples of oily fish include, but are not limited to: menhaden, anchovy, herring, capelin, cod and the like. Fish oil is a typical component of feed used in aquaculture.

Vegetable oil: "Vegetable oil" refers to any edible oil obtained from a plant. Typically plant oil is extracted from seed or grain of a plant. The term "triacylglycerols" ["TAGs"] refers to neutral lipids composed of three fatty acyl residues esterified to a glycerol molecule.

Microbial oil: The term "microbial oil" refers to oil that has been separated from cellular materials, such as the microorganism in which the oil was synthesized. Microbial oils are obtained through a wide variety of methods, the simplest of which involves physical means alone. For example, mechanical crushing using various press configurations (e.g., screw, expeller, piston, bead beaters, etc.) can separate oil from cellular materials. Alternatively, oil extraction can occur via treatment with various organic solvents (e.g., hexane), via enzymatic extraction, via osmotic shock, via ultrasonic extraction, via supercritical fluid extraction (e.g., CO2 extraction), via saponification and via combinations of these methods. An extracted oil may be further purified or concentrated. In a preferred embodiment, the microbial oil is an oil derived from a species of Schizochytrium sp. ATCC PTA-10208, as for example the commercial oil product available under the Trademarks OvegaGold® or Veramaris®.

Examples

Example 1

The amount of EPA (as a percent of total fatty acids ["% TFAs"]) and DHA provided in typical fish oils varies, as does the ratio of EPA to DHA. Typical values are summarized in Table 6, which also shows a fatty acid composition profile of the microbial oil used in the feed additive of the present invention.

Table 1 Fatty acid feed additive of various fish oil compared to the present invention

Example 2: Preparation of Pressed Fish Feed

The main raw materials are ground and mixed. Microingredients and/or a feed premix are then added to the mixer and the homogenous mix is conditioned by adding water and steam to the mass in a preconditioner. This starts a cooking process in the starch fraction (the binding component). The mass is fed into a pellet mill. The mass is forced through the mill's die and the strings are broken into pellets on the outside of the die. The moisture content is low and drying of the feed is not necessary.

Additional oil is then sprayed onto the surface of pellets, but as the pellets are rather compact, the total lipid content rarely exceeds 24 %. The added oil may be fish oil, microbial/algal or vegetable oils, for example rape seed oil or soy oil, or a mixture of oils. After oil coating, the pellets are cooled in a cooler and bagged. The final pressed fish feed contains 10 to 5000 ppm of the composition as described in the invention.

Example 3: Method for Preparation of Extruded Fish Feed The main raw materials are ground and mixed. Micro ingredients and/or a feed premix are added to the mixer. The homogenous mix is conditioned by adding water and steam to the mass in a preconditioner. Additional oil may also be added to the mass at this stage. This starts a cooking process in the starch fraction (the binding component). The mass is fed into an extruder. The extruder may be of the single screw or the twin-screw type. Due to the rotational movement of the mass in the extruder, the mass is further mixed. Additional oil, water and steam may be added to the mass in the extruder. At the end of the extruder, the mass has a temperature above 100 °C and a pressure above ambient pressure. The mass is forced through the openings in the extruder's die plate. Due to the relief in temperature and pressure, some of the moisture will evaporate immediately (flash off) and the extruded mass becomes porous. The strings are cut into pellets by a rotating knife. The water content is rather high (18-28 %) and the pellets are therefore immediately dried to approximately 10 % water content in a dryer.

After the dryer, more oil may be added to the feed by spraying oil onto the surface of the feed, or by dipping the feed in oil. It is advantageous to add the oil to the feed in a closed vessel where the air pressure is below ambient (vacuum coating) so that the porous feed pellets absorb more oil. Feed containing more than 40 % lipid may be produced this way. After the coater, the feed is cooled and bagged. Oil may be added at several places in the process as explained above, and may be fish oil, microbial/algal or vegetable oils, by example rape seed oil or soy oil, or a mixture of oils.

Fish need protein, fat, minerals and vitamins in order to grow and to be in good health. The diet of carnivorous fish is particularly important. Originally in the farming of carnivorous fish, whole fish or ground fish were used to meet the nutritional requirements of the farmed fish. Ground fish mixed with dry raw materials of various kinds, such as fish meal and starch, was termed soft or semi-moist feed. As farming became industrialized, soft or semi-moist feed was replaced by pressed dry feed. This was itself gradually replaced by extruded dry feed.

Today, extruded feed is nearly universal in the farming of a number of fish species such as various types of salmonid, cod, sea bass and sea bream.

The dominant protein source in dry feed for fish has been fish meal of different qualities. Other animal protein sources are also used for dry fish feed. Thus, it is known to use blood meal, bone meal, feather meal and other types of meal produced from other slaughterhouse waste, for example chicken meal. These are typically cheaper than fish meal and fish oil. However, in some geographic regions, there has been a prohibition against using such raw materials in the production of feeds for food-producing animals and fish. It is also known to use vegetable protein such as wheat gluten, maize (corn) gluten, soya protein, lupin meal, pea meal, bean meal, rape meal, sunflower meal and rice flour.

Example 4: Aquaculture feed can be produced according to the formulation described in table 3.

Table 2 Aquaculture diet

Example 5: Aquaculture feed can be produced according to the formulation described in table 4. Table 3 Aquaculture diet

Example 6:

Aquaculture feed can be produced according to the formulation described in table 5. Table 4 Aquaculture diet

Example 7:

Aquaculture feed can be produced according to the formulation described in table 6.

Table 5 Aquaculture diet

Example 8:

Aquaculture feed can be produced according to the formulation described in table 7.

Table 6 Aquaculture diet

Example 9:

Aquaculture feed can be produced according to the formulation described in table 9.

Table 7 Aquaculture diet

Example 10:

Aquaculture feed can be produced according to the formulation described in table 10.

Table 8 Aquaculture diet

Claims

35 CLAIMS

1. A feed premix intended for intake by a farm animal comprising an oil containing high levels of DHA and at least 3 additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid, and wherein the concentration of DHA is at least 30 % w/w measured as a weight percent of the oil.

4. The feed premix according to any of claims 1 to 3, wherein the oil comprises EPA and wherein the concentration of EPA is at least 10%, preferably at least 12 %, measured as a weight percent of the oil.

5. The feed premix according to any of claims 1 to 4, wherein the total concentration of fish oil in the feed premix is below 50%, preferably below 25%, preferably below 20%, preferably below 15%, preferably below 10%, preferably below 5%, preferably below 1 %, preferably below 0.1 %, measured as a weight percent of the feed premix.

6. A feed intended for intake by a farm animal comprising an oil containing high levels of DHA and at least 5 additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phoshpholipids and arachidonic acid, and wherein the concentration of DHA is at least 30 % measured as a weight percent of the oil.

7. The feed according to claim 6, wherein the oil is derived from a microbial source or vegetable source.

8. The feed according to any of claims 6 or 7, wherein the oil comprises EPA and wherein the concentration of EPA is at least 0.1 %, preferably at least 0.12 %, measured as a weight percent of the feed.

9. A feed intended for intake by a farm animal comprising the feed premix according to any of claims 1 to 5.

10. Feed according to any of claims 6 to 9, wherein the total concentration of fish oil in the feed is below 10%, preferably below 5%, preferably below 2.5%, 36 preferably below 2%, preferably below 1.5%, preferably below 1 %, preferably below 0.5%, preferably below 0.1 %, measured as a weight percent of the feed.

11. Use of an animal feed comprising an oil containing high levels of DHA for replacing all or part of fish oil in the feed in combination with at least 5 additional ingredients selected from the group of vitamins A, K, D, C, E, cholesterol, phospholipids and arachidonic acid, wherein the concentration of DHA is at least 0.30 %, preferably at least 0.35%, measured as a weight percent of the feed.

12. The use according to claim 11 , wherein the oil is derived from a microbial source or vegetable source.

13. Use according to any of claims 11 or 12, wherein the oil comprises EPA and wherein the concentration of EPA is at least 0.1 %, preferably at least 0.12 %, measured as a weight percent of the feed.

14. The use according to any of claims 11 to 13 for maintaining animal growth compared to the use of fish oil alone as additional PUFA source.

15. Use according to any of claims 11 to 14, wherein the feed comprises a total amount of EPA and DHA that is at least 0.08%, measured as a weight percent of the feed.