CN116600653A - Aquaculture feed - Google Patents

Abstract

The present invention relates to a feed premix comprising a high amount of eicosapentaenoic acid ("EPA") and docosahexaenoic acid ("DHA") as a feed premix for aquatic animals, preferably including fish and shrimp. More specifically, the present invention relates to a feed premix intended for ingestion by aquatic animals, said feed premix comprising a major amount of EPA and DHA derived from microbial sources, as well as vitamins a, K, D, C, E, cholesterol, phospholipids, arachidonic acid and small amounts of fish oil. The invention further relates to a method for producing a meat product, preferably an aquaculture meat product, by feeding animals, preferably fish, with a feed composition comprising DHA, preferably EPA and DHA, originating from a microbial source, and vitamin A, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, as well as small amounts of fish oil.

Description

Aquaculture feed

The present invention relates to a feed premix comprising a major amount of eicosapentaenoic acid ("EPA") and docosahexaenoic acid ("DHA") as well as vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid and a minor amount of fish oil; and to a final feed for farm animals, preferably for aquatic animals including fish and shrimp. More specifically, the present invention relates to a feed premix intended for ingestion by aquatic animals, said feed premix comprising a major amount of DHA, preferably EPA and DHA, as well as vitamins A, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids, arachidonic acid and a minor amount of fish oil. The invention further relates to a method for producing a meat product, preferably an aquaculture meat product, by feeding an animal, preferably a fish, with a feed composition comprising DHA, preferably EPA and DHA, and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, and a small amount of fish oil.

Aquaculture is a form of agriculture that involves the propagation, cultivation and sale of aquatic animals and plants in a controlled environment. Aquaculture is the fastest growing food production part of the world today. The world aquaculture industry produces about 6000 ten thousand tons of seafood each year, which is more than 700 million dollars worth. Today, farmed fish make up about 50% of the total fish consumption worldwide. This percentage is expected to increase due to the ever-decreasing fishing volume from fishing in marine and freshwater environments and the ever-increasing consumption of seafood (i.e., total and average). Today, the population of species in aquaculture production includes, for example: carp and other carp species; oyster; clams, clams and concha arcae; open shrimp (shramp) and prawn (prawn); salmon, trout and cucurbit; mussel; tilapia and other arrowheads; and scallops.

While some aquaculture species (e.g., tilapia) may be fed a full vegetarian diet, many other species are fed a meat diet. Typically, feeds for predatory fish include fish meal and fish oil derived from wild captured small ocean fish species (principally anchovies, horse mackerels, pollack, capelin, salmon (sandeel) and herring). These ocean fish are processed into fish meal and fish oil, and the end product is often a pellet or pellet feed, depending on the size of the fish. Other components of the aquaculture feed composition may include vegetable proteins, vitamins, minerals and pigments as desired.

Marine fish oils have traditionally been used as the sole dietary lipid source in commercial fish feeds because they are readily available, cost-competitive and such products contain abundant essential fatty acids. In addition, fish oils readily supply essential fatty acids required for normal growth, health, reproduction and bodily functions in 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; 22:6ω -3] is essential for fish growth and health and is often incorporated into commercial fish feeds via the addition of fish oil.

It is estimated that aquaculture feed compositions currently use about 87% of the global supply of fish oil as a lipid source. Since the annual fish oil production has not increased to over 150 ten thousand tons/year, the rapidly evolving aquaculture industry cannot continue to rely on limited marine ocean fish reserves as a fish oil supply. Thus, there is a great urgent need to find and achieve sustainable fish oil substitutes that can keep pace with the ever-increasing global demand for fish products.

U.S. Pat. No. 7,932,077 shows that recombinantly engineered yarrowia lipolytica (Yarrowia lipolytica) may be a useful additive for most animal feeds, including aquaculture feeds, as a means of providing the necessary omega-3 and/or omega-6 polyunsaturated fatty acids and based thereon unique protein: lipid: carbohydrate feed additives, as well as unique complex carbohydrate characteristics (comprising mannans: beta-glucan: chitin in a ratio of about 1:4:4.6).

U.S. patent application publication No. 2007/0226814 discloses a fish diet containing at least one biomass obtained from a fermenting microorganism, wherein the biomass contains at least 20% DHA relative to total fatty acid content. Preferred microorganisms for use as DHA source are organisms belonging to the genus different flagellum (Straminopiles).

However, an important factor in aquaculture is turnover rate. Turnover rate is determined by how fast the fish grow to a catchable size. It is highly affected by animal growth and thus growth reduction has a great economic impact on aqua farmers.

Fish oil substitutes with high concentrations of PUFA of at least 30% are highly advantageous in many respects. Formulations, which are usually liquid, are easier to incorporate into feeds. The aquatic animals can absorb higher amounts of nutrients while feeding the same amount of feed.

Unfortunately, the use of such highly concentrated alternatives has been problematic and may result in reduced growth of aquatic animals. This problem is currently addressed by diluting highly concentrated alternatives with low PUFA oils (e.g. rapeseed oil or soybean oil).

Thus, there remains a need in the aquaculture industry to find a composition of fish oil substitutes, preferably derived from microbial sources, enriched in PUFAs, preferably DHA, more preferably EPA and DHA, suitable for ingestion by aquatic animals to replace fish oil partially or completely, while maintaining the animal's performance and providing all necessary nutrients.

Disclosure of Invention

Surprisingly, we have found that the incorporation of oils (e.g. microbial oils or vegetable oils) having a high concentration of PUFAs (e.g. having a DHA content of at least 30%, preferably DHA and EPA) into animal feed does not result in reduced growth by the additional addition of additives (e.g. vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid), wherein at least one of the additives is preferably used in a higher dose than is normally used for animal feed.

In one aspect, the present invention relates to a feed premix intended for ingestion 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 consisting of vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid. In a preferred embodiment, the feed premix may be an oil.

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

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

In a further embodiment, the present invention relates to a method for producing a meat product, preferably an aquaculture meat product, by feeding animals, preferably fish, with a feed composition comprising the step of formulating the feed composition comprising DHA, preferably EPA and DHA, at least 5, preferably at least 6 additional ingredients selected from the group consisting of vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, and a small amount of fish oil.

Detailed Description

Aquaculture is a practice for growing aquatic animals and plants. It involves growing aquatic products (e.g., freshwater and saltwater animals) under controlled conditions. It relates to the growth and harvesting of fish, shellfish and aquatic plants in fresh, brackish or salty water.

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

It relates to the commercial cultivation of fish in tanks, ponds or marine pens, commonly used for food. The release of young fish into the field for recreational fishing or to supplement the natural number of species is commonly referred to as a fish hatchery. Of particular interest are salmon families such as sakura salmon (sakura kokumi (Oncorhynchus masou)), knoop salmon (large scale salmon (o.tshawythsch)), dog salmon (o.keta)), silver salmon (pacific silver salmon (o.kisutch)), smolt (o.gorbuscha)), red salmon (o.nerka)) and atlantic salmon (Salmo salar). Other finfish species of interest for aquaculture include, but are not limited to, various trout species, and white fish, such as tilapia (including species of the genus koilot (oreochrois), sarotheodon (sarotheodon) and tilapia), sea bass, catfish (catfish order), tuna (tuna of order), carp (Cyprinidae)), and codfish (Gadus)).

Aquaculture generally requires the preparation of aquaculture feed compositions to meet the dietary requirements of the cultivated animals. The dietary requirements vary from aquaculture species to aquaculture species, as does the dietary requirements of individual species at different stages of growth. Thus, a great deal of research was devoted to optimizing each aquaculture feed composition for each growth stage of the farmed organisms.

The aquaculture feed composition is composed of a micro component and a macro component. In general, all components used at levels greater than 1% are considered constant components. The feed ingredient used at a level of less than 1% is a micro component. They are premixed to achieve a uniform distribution of the micro-ingredients throughout the feed. Both the macro-and micro-ingredients are subdivided into components having nutritional and technical functions.

The components with technical functions improve the physical quality or appearance of the aquaculture feed composition.

Macrocomponents with nutritional functions provide the aquatic animals with the protein and energy required for growth and performance. For fish, the aquaculture feed composition should ideally provide for fish: 1) Fat, which acts as a source of fatty acids for energy (especially for heart and skeletal muscle); and 2) amino acids which serve as structural units of the protein. Fat also contributes to vitamin absorption; for example, vitamin a, vitamin K, vitamin D, vitamin E, vitamin K are fat soluble or can only be digested, absorbed and transported with fat. Carbohydrates, typically of vegetable origin (e.g. wheat, sunflower meal, corn gluten, soybean meal), are also often included in feed compositions, but are not an energy source for fish that is superior to protein or fat.

Fat is typically provided by incorporating fish meal (which contains small amounts of fish oil) and fish oil into an aquaculture feed composition. Extraction oils useful in aquaculture feed compositions include fish oils (e.g., from oily fish herring (menhaden), anchovy, herring (herring), capelin and cod liver) and vegetable oils (e.g., from soybean, rapeseed, sunflower and linseed). Generally, fish oil is the preferred oil because it contains long chain omega-3 polyunsaturated fatty acids [ "PUFA" ], EPA and DHA; in contrast, vegetable oils do not provide a source of EPA and/or DHA. These PUFAs are required for the growth and health of most aquaculture products. A typical aquaculture feed composition will comprise about 15-30% oil (e.g., fish, plants, etc.) measured as weight percent of the aquaculture feed composition.

The protein supplied in the aquaculture feed composition may be of vegetable or animal origin. For example, the animal-derived protein may be derived from marine animals (e.g., fish meal, fish oil, fish protein, krill meal, mussel meal, shrimp skin, squid meal, squid oil, etc.) or land animals (e.g., blood meal, egg meal, liver meal, meat and bone meal, silkworm, pupa meal, whey meal, etc.). The plant-derived proteins may include soybean meal, corn gluten meal, wheat gluten, cottonseed meal, rapeseed meal, sunflower meal, rice, and the like.

The technical functions of the macrocomponents may overlap, for example, wheat gluten may be used as a pelletization aid and have a relatively high nutritional value due to its protein content. Guar gum and wheat flour may also be mentioned.

Minor components include feed additives such as vitamins, trace minerals, feed antibiotics and other biological products. Minerals used at levels below 100mg/kg (100 ppm) are considered trace minerals or trace minerals.

The micro-components with nutrition function are biological products and trace minerals. They are involved in biological processes and are essential for good health and high performance. Additional vitamins may be mentioned, such as vitamin A, vitamin K3, vitamin D3, vitamin B1, vitamin B3, vitamin B6, vitamin B12, biotin, folic acid, pantothenic acid, nicotinic acid, choline chloride, inositol and para-aminobenzoic acid. Minerals such as calcium salt, cobalt salt, copper salt, iron salt, magnesium salt, phosphorus salt, potassium salt, selenium salt and zinc salt may be mentioned. Other components may include, but are not limited to, antioxidants, beta-glucans, bile salts, cholesterol, enzymes, monosodium glutamate, carotenoids, and the like.

The technical functions of the trace components are mainly related to granulation, detoxification, mildew prevention, oxidation resistance and the like.

In aquaculture, fish are typically fed at different dietary cycles as they grow. The weight of fish for different dietary periods may vary depending on the type of fish and/or aquaculture practices used.

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

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

In a further aspect, the present invention relates to a feed premix intended for ingestion 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 consisting of vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid. In a preferred embodiment, the feed premix may be an oil.

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

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 weight percent of the feed premix.

In 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%, at least 60%, preferably at least 65%, measured as weight percent of the feed additive.

In another aspect of the invention, the feed premix may be mixed with an oil selected from the group consisting of microbial oils and one or more vegetable oils to reduce the concentration of PUFAs and DHA, preferably EPA and DHA. In a more preferred embodiment, the vegetable oil is selected from the group consisting of: rapeseed oil and soybean oil.

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

The oil comprising DHA, preferably DHA and EPA according to the present invention may be derived from microbial sources or from plant sources. Preferably, the oil is an oil obtained from algae, fungi or yeast. The preferred microorganism is Thraustochytriales (Thraustochytriales). Thraustochytriales include members of the genus Schizochytrium (Schizochytrium) and Thraustochytrium (Thraustochytrium), and have been considered as alternative sources 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 schizochytrium species. Schizochytrium strains are a natural source of PUFAs (e.g., DHA) and can be optimized by mutagenesis for use as a microbial source according to the 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 weight percent of 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 weight percent of 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 150ppm and 800ppm, preferably between 150ppm and 600ppm, preferably between 200ppm and 500ppm, preferably between 200ppm and 400ppm, preferably between 200ppm and 300ppm, preferably between 200ppm and 250 ppm.

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

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

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

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

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

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

In a preferred embodiment of the invention the concentration of vitamin K in the feed premix is between 10mg/kg and 20000mg/kg, preferably between 10mg/kg and 15000mg/kg, preferably between 10mg/kg and 10000mg/kg, preferably between 10mg/kg and 9000mg/kg, preferably between 10mg/kg and 8000mg/kg, preferably between 10mg/kg and 7000mg/kg, preferably between 10mg/kg and 6000mg/kg, preferably between 20mg/kg and 20000mg/kg, preferably between 30mg/kg and 20000 mg/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 1000IU/kg and 10000IU/kg, preferably between 1500IU/kg and 9500IU/kg, preferably between 1500IU/kg and 9000IU/kg, preferably between 1500IU/kg and 8500IU/kg, preferably between 1500IU/kg and 8000IU/kg, preferably between 1500IU/kg and 7500IU/kg, preferably between 1500IU/kg and 7000IU/kg, preferably between 1500IU/kg and 6500 IU/kg.

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

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

In a preferred embodiment of the invention, the concentration of cholesterol in the feed premix is at most 100000mg/kg, preferably at most 90000mg/kg, at most 80000mg/kg, preferably at most 75000 mg/kg.

In a preferred embodiment of the invention, the concentration of phospholipids in the feed premix is such that the final concentration of cholesterol in the feed, measured as weight percent of 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%, if added to 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 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 the final concentration of vitamin a in the feed, measured as weight percent of the feed, is at most 0.1%, preferably at most 0.08%, preferably at most 0.06%, preferably at most 0.04%, preferably at most 0.02%, if added to the feed.

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

In a further 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 vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, and further ingredients intended to be incorporated into an aquaculture feed (as defined above).

In another embodiment, the feed may comprise a total amount of EPA and DHA of at least about 0.8% as measured by weight percent of the feed. Such an amount (i.e. 0.8%) is generally suitable for supporting the growth of various animals grown in aquaculture, especially for the proper minimum concentration contained in the salmonids ration.

Most processes for making the aquaculture feed composition of the invention will start with microbial fermentation, wherein specific microorganisms are cultivated under conditions allowing the growth and production of a microbial oil comprising DHA, preferably EPA and DHA. At the appropriate time, microbial cells are harvested from the fermentation vessel. Such microbial biomass may be mechanically processed using various means, such as dewatering, drying, mechanical disruption, pelletization, and the like. The oil extracted from the biomass is then supplemented with appropriate amounts of vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid for use as ingredients in aquaculture feeds (preferably as a replacement for at least part of the fish oil used in standard aquaculture feed compositions). The aquaculture feed is then fed to the aquatic animals for a part of their lifetime such that DHA, preferably EPA and DHA, from the aquaculture feed is accumulated in the aquatic animals.

In another embodiment of the invention, an aquaculture feed according to the invention may be prepared as described in example 3. Water-soluble vitamins, such as vitamin C and vitamin E, may be added to the main raw materials for producing aquaculture feed (e.g. in the form of expanded feed). An amount of oil may be supplemented to reach the final concentration of DHA, preferably EPA and DHA, vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid in the feed as defined above, and the oil is applied to the feed surface by spraying the oil onto the surface of the feed or immersing the feed in the oil.

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

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 weight percent of the feed.

In a preferred embodiment of the invention, the concentration of DHA in the feed, measured as weight percent of 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%.

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

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

In a preferred embodiment of the invention, the concentration of vitamin E in the feed is between 250ppm and 800ppm, preferably between 250ppm and 700ppm, preferably between 250ppm and 600ppm, preferably between 300ppm and 550ppm, preferably between 300ppm and 400 ppm.

In a preferred embodiment of the invention, the concentration of vitamin a in the feed is between 1000IU/kg and 15000IU/kg, preferably between 1500IU/kg and 14500IU/kg, preferably between 1500IU/kg and 14000IU/kg, preferably between 2000IU/kg and 13500IU/kg, preferably between 2000IU/kg and 13000IU/kg, preferably between 2000IU/kg and 12000 IU/kg.

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

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

In a preferred embodiment of the invention, the concentration of cholesterol in the feed is at most 1000mg/kg, preferably at most 900mg/kg, at most 800mg/kg, preferably at most 750mg/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 weight percent of the feed.

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

In another aspect of the invention, the aquaculture feed is expanded feed pellets or compressed feed pellets. In a more preferred aspect, the aquaculture feed is a coated pellet. In a more preferred aspect, the feed additive is present in a coating of the feed.

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

In a further embodiment, the present invention relates to a method for producing a meat product, preferably an aquaculture meat product, by feeding animals, preferably fish, with a feed composition comprising the step of formulating a feed composition comprising DHA, preferably EPA and DHA, derived from a microbial source, as well as vitamin a, vitamin K, vitamin D, vitamin C, vitamin 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 a premix comprising DHA, preferably EPA and DHA, and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid derived from microbial sources, the amount of the premix in the feed being between 1% and 15%, preferably between 1% and 10%, preferably between 2% and 10%, preferably between 3% and 8%, preferably between 3% and 7%, preferably between 3% and 6%, measured as weight percent of the feed.

In another 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 weight percent of feed.

In another 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 an amount of 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 weight percent of feed.

In another 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 150ppm and 800ppm, preferably between 150ppm and 600ppm, preferably between 200ppm and 500ppm, preferably between 200ppm and 400ppm, preferably between 200ppm and 300ppm, preferably between 200ppm and 250 ppm.

In another 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 250ppm and 800ppm, preferably between 250ppm and 700ppm, preferably between 250ppm and 600ppm, preferably between 300ppm and 550ppm, preferably between 300ppm and 400 ppm.

In another 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 and DHA from a single microbial source in a total amount of at least about 0.8, measured as weight percent of feed. Such an amount (i.e. 0.8%) is generally suitable for supporting the growth of various animals grown in aquaculture, especially for the proper minimum concentration contained in the salmonids ration.

In another 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 expanded feed pellet or a compressed feed pellet.

In another 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 another 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 particles.

Embodiments of the present invention may be summarized as follows:

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

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

3. The feed premix of claim 1 or 2, wherein the oil is derived from a microbial or plant source.

4. A feed premix according to any one of claims 1 to 3, wherein

a. The concentration of vitamin C is such that the final concentration of vitamin C in the feed is between 200ppm and 600ppm, preferably between 200ppm and 500ppm, if added to the feed.

b. The concentration of vitamin E is such that if added to the feed the final concentration of vitamin E in the feed is between 250ppm and 700ppm, preferably between 300ppm and 600 ppm.

5. Feed premix according to any one of claims 1 to 4

a. The concentration of vitamin C is between 2000ppm and 60000ppm, preferably between 2000ppm and 50000 ppm.

b. The concentration of vitamin E is between 2500ppm and 70000ppm, preferably between 2500ppm and 60000 ppm.

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

7. 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 weight percent of the feed premix.

8. The feed premix of any one of claims 1-7, 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 10000IU/kg and 1500000IU/kg, preferably between 15000IU/kg and 1450000IU/kg, preferably between 15000IU/kg and 1400000IU/kg, preferably between 20000IU/kg and 1350000IU/kg, preferably between 20000IU/kg and 1300000IU/kg, preferably between 20000IU/kg and 1200000 IU/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 10mg/kg and 20000mg/kg, preferably between 10mg/kg and 15000mg/kg, preferably between 10mg/kg and 10000mg/kg, preferably between 10mg/kg and 9000mg/kg, preferably between 10mg/kg and 8000mg/kg, preferably between 10mg/kg and 7000mg/kg, preferably between 10mg/kg and 6000mg/kg, preferably between 20mg/kg and 20000mg/kg, preferably between 30mg/kg and 20000 mg/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 10000IU/kg and 1000000IU/kg, preferably between 15000IU/kg and 950000IU/kg, preferably between 15000IU/kg and 900000IU/kg, preferably between 15000IU/kg and 850000IU/kg, preferably between 15000IU/kg and 800000IU/kg, preferably between 15000IU/kg and 750000IU/kg, preferably between 15000IU/kg and 700000IU/kg, preferably between 15000IU/kg and 650000 IU/kg.

12. Feed premix according to any of claims 1 to 11, wherein the concentration of cholesterol in the feed premix is at most 100000mg/kg, preferably at most 90000mg/kg, at most 80000mg/kg, preferably at most 75000 mg/kg.

13. 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 weight percent of the feed premix.

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

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

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

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

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

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

20. The feed premix of claim 19, wherein the microorganism is a member of schizochytrium or thraustochytrium.

21. The feed premix of any one of claims 1-20, further comprising at least one additional component selected from the group consisting of: fat-soluble vitamins, water-soluble vitamins, trace minerals, carotenoids, polyunsaturated fatty acids, probiotics, prebiotics and macrominerals.

22. A feed intended for farm animals, the feed comprising an oil containing a high level of DHA and at least 5 additional ingredients selected from the group of vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, and wherein the concentration of DHA, measured as a weight percentage of the oil, is at least 30%.

23. The feed of claim 22, wherein the oil is derived from a microbial source or a plant source.

24. The feed of claim 22 or 23, wherein

a. The concentration of vitamin C is between 200ppm and 600ppm, preferably between 200ppm and 500 ppm.

b. The concentration of vitamin E is between 250ppm and 700ppm, preferably between 300ppm and 600 ppm.

25. The feed of any one of claims 22 to 24, wherein the oil comprises EPA, and wherein the concentration of EPA, measured as a weight percentage of the feed, is at least 0.1%, preferably at least 0.12%.

26. The feed of claim 25 comprising a total amount of EPA and DHA of at least 0.08%, measured as a weight percentage of the feed.

27. A feed intended for ingestion by farm animals, the feed comprising a feed premix according to any of claims 1 to 21.

28. The feed of claim 27, wherein the amount of feed premix of any one of claims 1 to 21 is such that the concentration of feed premix in the feed is between 1% and 10% measured as weight percent of the feed.

29. The feed according to any one of claims 22 to 28, wherein the total concentration of fish oil in the feed is less than 10%, preferably less than 5%, preferably less than 2.5%, preferably less than 2%, preferably less than 1.5%, preferably less than 1%, preferably less than 0.5%, preferably less than 0.1%, measured as weight percent of the feed.

30. The feed of any one of claims 22 to 29, wherein the feed does not comprise fish oil.

31. The feed according to any one of claims 22 to 30, wherein the concentration of vitamin a in the feed is between 1000IU/kg and 15000IU/kg, preferably between 1500IU/kg and 14500IU/kg, preferably between 1500IU/kg and 14000IU/kg, preferably between 2000IU/kg and 13500IU/kg, preferably between 2000IU/kg and 13000IU/kg, preferably between 2000IU/kg and 12000 IU/kg.

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

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

34. The feed according to any one of claims 22 to 33, wherein the concentration of cholesterol in the feed is at most 1000mg/kg, preferably at most 900mg/kg, at most 800mg/kg, preferably at most 750mg/kg.

35. The feed according to any one 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 weight percent of the feed.

36. The feed according to any one of claims 22 to 35, wherein the concentration of arachidonic acid in the feed is at most 0.1%, preferably at most 0.08%, preferably at most 0.06%, preferably at most 0.04%, preferably at most 0.02%, measured as weight percent of the feed.

37. The feed of any one of claims 22 to 36, further comprising one or more ingredients selected from the group consisting of: fish meal, krill meal, soy concentrate, corn gluten, wheat gluten, pea protein, wheat flour, fish oil, vitamins, mineral premixes, addition of synthetic phosphorus and combinations thereof.

38. The feed of any one of claims 22 to 37, wherein the aquaculture feed is expanded feed pellets or compressed feed pellets.

39. The feed of any one of claims 22 to 37, wherein the feed pellet is a coated feed pellet.

40. The feed of any one of claims 22 to 39, wherein the feed premix of any one of claims 1 to 21 is present in a coating of the coated feed particles.

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

42. The use according to claim 39, wherein the oil is derived from microbial or plant sources.

43. The use according to claim 41 or 42, wherein

a. The concentration of vitamin C is between 200ppm and 600ppm, preferably between 200ppm and 500 ppm.

b. The concentration of vitamin E is between 250ppm and 700ppm, preferably between 300ppm and 600 ppm.

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

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

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

47. The use according to any one of claims 41 to 46, wherein the feed comprises a total amount of EPA and DHA of at least 0.08%, measured as a weight percentage of the feed.

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

49. The use of any one of claims 41 to 48, wherein the feed does not comprise fish oil.

50. The use according to any one of claims 41 to 49, wherein the concentration of vitamin a in the feed is between 1000IU/kg and 15000IU/kg, preferably between 1500IU/kg and 14500IU/kg, preferably between 1500IU/kg and 14000IU/kg, preferably between 2000IU/kg and 13500IU/kg, preferably between 2000IU/kg and 13000IU/kg, preferably between 2000IU/kg and 12000 IU/kg.

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

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

53. The use according to any one of claims 41 to 52, wherein the concentration of cholesterol in the feed is at most 1000mg/kg, preferably at most 900mg/kg, at most 800mg/kg, preferably at most 750mg/kg.

54. Use according to any one 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 weight percent of the feed.

55. The use according to any one of claims 41 to 54, wherein the concentration of arachidonic acid (ARA) in the feed is at most 0.1%, preferably at most 0.08%, preferably at most 0.06%, preferably at most 0.04%, preferably at most 0.02%, measured as a weight percentage of the feed.

56. The use of any one of claims 41 to 55, wherein the feed further comprises one or more ingredients selected from the group consisting of: fish meal, krill meal, soy concentrate, corn gluten, wheat gluten, pea protein, wheat flour, fish oil, vitamins, mineral premixes, addition of synthetic phosphorus, and combinations thereof.

57. The use according to any one of claims 41 to 56, wherein the feed is a puffed feed pellet or a compressed feed pellet.

58. The use of any one of claims 41 to 57, wherein the feed pellet is a coated feed pellet.

59. The use according to any one of claims 41 to 58, wherein a feed premix according to any one of claims 1 to 9 is present in the coating of the coated feed particles.

Definition of the definition

The invention comprises the following steps: 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 specification herein, and should not be construed as limited to any particular embodiment or aspect.

Farm animals: the term "farm animal" or "animal" refers to an animal raised on a farm that is primarily used for production purposes, such as for producing meat, milk, eggs, or hair. Examples of farm animals include aquatic animals, cattle, pigs, sheep, goats, poultry, such as turkeys, chickens or ducks.

Aquatic animals: the term "aquatic animals" refers to the class of crustaceans, including but not limited to, prawns and prawns, as well as fish, including but not limited to, croaker, megalobrama, cheilus grahami, weever, carangid, daceae (bocachico), bream, kohlrabi, megacyprinus carpio, carp, catfish , louse, red salmon (char), salmon, cobia, cod, perch, croaker, barracuda (drum), eel, goby, goldfish, spiny poda, grouper, body of parapleus, halibut, java, wild dace, garter, loach, mackerel the fish feed comprises the following materials of eye-shading fish, silver perch, mud fish, mullet, paco, green belly fish (pearspot), silver salmon, sea perch, shuttle, pomfret, group bream, salmon, filial heterobranchia catfish, larch, sea bass, sea bream, flash fish (shiner), perk, snakehead (sleep), snakehead, porgy, saw cover fish (snook), sole, blue fish, sturgeon, tipper, fragrant fish, dead sea bream, crown fish (terror), tilapia, trout, tuna, turbot, white trout, glass weever and white salmon.

Feed premix: the term feed premix according to instant invention refers to a formulation intended to be ingested by farm animals, preferably aquatic animals, comprising DHA, preferably EPA and DHA, derived from a single microbial source, as well as vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid as active ingredients. The incorporation of the feed additives as exemplified above into animal feed, such as fish feed, is actually carried out using concentrates or premixes. Premix means a preferably homogeneous mixture of one or more minor components with the diluent and/or carrier. The premix is used to promote uniform dispersion of the minor ingredients in the larger mixture. The premix according to the invention may be added to the feed ingredient as a solid (e.g. as a water-soluble powder) or as a liquid. In a preferred embodiment of the invention, the feed premix may be an oil. The oil may be added after feed processing (e.g., processing into pellets).

Feed or aquaculture feed: the term "feed" or "aquaculture feed" or "feed composition" refers to any compound, preparation or mixture suitable or intended for ingestion by farm animals, particularly aquatic animals. Animal feeds for aquatic animals typically contain high protein and energy concentrations such as fish meal, molasses, oligosaccharide concentrates, vitamins, minerals, enzymes, direct fed microorganisms, amino acids and/or other feed ingredients (e.g. as a premix). Aquaculture feed refers to a manufactured or artificial ration (i.e., a compound feed) that is most commonly produced in the form of flakes or pellets for supplementing or replacing natural feed. Typically, the aquaculture feed may be in the form of flakes or pellets, such as expanded pellets.

Polyunsaturated fatty acids [ "PUFA" ]: the terms polyunsaturated fatty acid [ "PUFA" ] the terms "polyunsaturated fatty acid" and "PUFA" include not only the free fatty acid form but also other forms, such as Triacylglycerols (TAGs) in Phospholipid (PL) form and other forms of esterified forms. Additional details regarding the differences between "saturated fatty acids" and "unsaturated fatty acids", "monounsaturated fatty acids" and "polyunsaturated fatty acids" [ "PUFA" ], and "omega-6 fatty acids" [ "00-6" or "n-6" ] and "omega-3 fatty acids" [ "00-3" or "n-3" ] are provided in U.S. Pat. No. 7,238,482.

Eicosapentaenoic acid [ EPA ]: the term "eicosapentaenoic acid" [ "EPA" ] is the common name of cis-5, 8,11,14, 17-eicosapentaenoic acid. Such fatty acids are 20:5 omega-3 fatty acids. Unless specifically stated otherwise, the term EPA as used in this disclosure will refer to an acid or acid derivative (e.g., glyceride, ester, phospholipid, amide, lactone, salt, etc.).

Docosahexaenoic acid [ DHA ]: the term "docosahexaenoic acid" [ "DHA" ] is the common name of cis-4, 7,10,13,16, 19-docosahexaenoic acid. Such fatty acids are 22:6 omega-3 fatty acids. Unless specifically stated otherwise, the term DHA as used in this disclosure will refer to an acid or derivative of an acid (e.g., glyceride, ester, phospholipid, amide, lactone, salt, etc.).

Vitamin C: for the purposes of the present invention, vitamin C, for example, is provided as a product, for example under the trademarkStay-35 as a supplement to animal feed to contain DHA from microbial sources and vitamins A, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and peanutsThe tetraenoic acid is suitably administered to the animal in the form of a feed premix or in the form of a feed comprising DHA from a microbial source and vitamin A, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid. The feed may be supplemented by: vitamin C is blended into conventional feed or a premix of feed components and vitamin C is first prepared and then mixed with the other feed components. The feed may be any feed. The term feed as used herein includes both solid feed and liquid feed.

Vitamin E: for the purposes of the present invention, vitamin E, for example, is provided as, for example, a trademarkE50 SD-available commercial preparations are suitably administered to animals as supplements to animal feed in the form of feed premixes comprising DHA from microbial sources and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, or in the form of feeds comprising DHA from microbial sources and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid. The feed may be supplemented by: vitamin E is blended into conventional feed or a premix of feed components and vitamin E is first prepared and then mixed with the other feed components. The feed may be any feed. The term feed as used herein includes both solid feed and liquid feed.

Vitamin a: for the purposes of the present invention, vitamin A, for example, is provided as, for example, a trademarkA1000 is available as a supplement to animal feed in the form of a feed premix comprising DHA from microbial sources and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, or in the form of a feed premix comprising DHA from microbial sources and vitamin a, vitamin CThe feed forms of vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid are suitably administered to animals. The feed may be supplemented by: the vitamin a is blended into a conventional feed or a premix of the feed components and vitamin a is first prepared and then mixed with the other feed components. The feed may be any feed. The term feed as used herein includes both solid feed and liquid feed.

Vitamin K: for the purposes of the present invention, vitamin K is vitamin K3, for example as a vitamin K which is for example under the trademarkK3 MNB is suitably administered to an animal as a supplement to an animal feed in the form of a feed premix comprising DHA from a microbial source and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, or in the form of a feed comprising DHA from a microbial source and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid. The feed may be supplemented by: vitamin K is blended into conventional feed or a premix of feed components and vitamin K is first prepared and then mixed with other feed components. The feed may be any feed. The term feed as used herein includes both solid feed and liquid feed.

Vitamin D: for the purposes of the present invention, vitamin D is vitamin D3, for example, as a vitamin D, for example under the trademarkD3-500 is available as a supplement to animal feed in the form of a feed premix comprising DHA from microbial origin and vitamin A, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, or in the form of a feed premix comprising DHA from microbial origin and vitamin A, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and flowersThe feed form of tetraenoic acid is suitably administered to the animal. The feed may be supplemented by: the vitamin D is blended into a conventional feed or a premix of the feed components and vitamin D is first prepared and then mixed with the other feed components. The feed may be any feed. The term feed as used herein includes both solid feed and liquid feed.

Cholesterol: for the purposes of the present invention, cholesterol is suitably administered to animals as a supplement to animal feed in the form of a feed premix comprising DHA from microbial origin and vitamin A, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, or in the form of a feed comprising DHA from microbial origin and vitamin A, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid. The feed may be supplemented by: the cholesterol is blended into conventional feed or a premix of the feed components and cholesterol is first prepared and then mixed with the other feed components. The feed may be any feed. The term feed as used herein includes both solid feed and liquid feed.

Arachidonic acid: for the purposes of the present invention, arachidonic acid (ARA), for example, as a commercial preparation available, for example, under the trademark ARASCOTM, is suitably administered to animals as a supplement to animal feed, in the form of a feed premix comprising DHA from microbial sources and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, or in the form of a feed comprising DHA from microbial sources and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid. The feed may be supplemented by: the arachidonic acid is admixed to conventional feed or a premix of the feed components and arachidonic acid is first prepared and then mixed with the other feed components. The feed may be any feed. The term feed as used herein includes both solid feed and liquid feed.

Phospholipids for the purposes of the present invention, phospholipids are suitably administered to animals, for example, as a commercial formulation available, for example, under the trademark ARASCOTM, as a supplement to animal feed, in the form of a feed premix comprising DHA from a microbial source and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, or in the form of a feed comprising DHA from a microbial source and vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid. The feed may be supplemented by: the phospholipids are blended into conventional feeds or a premix of the feed components and phospholipids is first prepared and then mixed with the other feed components. The feed may be any feed. The term feed as used herein includes both solid feed and liquid feed.

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

Vegetable oil: "vegetable oil" refers to any edible oil obtained from a plant. Vegetable oils are typically extracted from seeds or grains of plants. The term "triacylglycerols" [ "TAG" ] refers to neutral lipids composed of three fatty acyl residues esterified to glycerol molecules.

Microbial oil: the term "microbial oil" refers to an oil that has been separated from cellular material (e.g., microorganisms that are synthetic oils). Microbial oils can be obtained by a variety of methods, the simplest of which involves separate physical means. For example, mechanical disruption using various press configurations (e.g., screw press, piston, bead mill, etc.) can separate the oil from the cellular material. Alternatively, the extraction may be via treatment with various organic solvents (e.g., hexane), via enzymatic extraction, via osmotic shock, via ultrasonic extraction, via supercritical fluid extraction (e.g., CO ₂ Extraction), oil extraction via saponification and via a combination of these methods. The extracted oil may be further purified or concentrated. In a preferred embodiment, the microbial oil is the source Oil from schizochytrium species. ATCC PTA-10208, e.g. under the trademarkOr->Commercial oil products are available.

Examples

Example 1

The amounts of EPA (in percent of total fatty acids [ "% TFA" ]) and DHA provided in typical fish oils vary, as do the ratios of EPA to DHA. Typical values are summarized in table 6, which also shows the fatty acid composition profile of the microbial oil used in the feed additive of the present invention.

Table 1 fatty acid feed additives for various fish oils compared to the present invention

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Example 2: preparation of pressed fish feed

The main raw materials are ground and mixed. The micro ingredients and/or feed premix are then added to the mixer and the homogeneous mixture is adjusted by adding water and steam to the material in the preconditioner. This starts the cooking process in the starch fraction (adhesive component). The material is fed into a granulator. The material is forced through the die of the granulator and the ribbons are broken into pellets outside the die. Low in moisture content and does not require a dry feed.

Additional oil is then sprayed onto the surface of the pellets, but since the pellets are quite dense, the total lipid content is rarely over 24%. The added oil may be fish oil, microbial/algal or vegetable oil, such as rapeseed oil or soybean oil, or a mixture of oils. After oiling, the pellets were cooled in a cooler and bagged. The final compacted fish feed contains 10ppm to 5000ppm of the composition as described in the invention.

Example 3: method for preparing puffed fish feed

The main raw materials are ground and mixed. The micro ingredients and/or feed premix are added to the mixer. The homogeneous mixture is conditioned by adding water and steam to the material in a preconditioner. Additional oil may also be added to the material at this stage. This starts the cooking process in the starch fraction (adhesive component). The material was fed into an extruder. The extruder may be of the single screw or twin screw type. The materials are further mixed due to the rotational movement of the materials in the extruder. Additional oil, water and steam may be added to the feed in the extruder. At the end of the extruder, the temperature of the mass was above 100 ℃, and the pressure was above ambient. Forcing the material through openings in the die plate of the extruder. As a result of the decrease in temperature and pressure, some of the moisture will immediately evaporate (flash) and the extruded material becomes porous. The ribbons were cut into pellets with a rotating knife. The moisture content is quite high (18-28%) and thus the pellets are immediately dried in a dryer to a moisture content of about 10%.

After the dryer, more oil may be added to the feed by spraying the oil onto the surface of the feed or immersing the feed in the oil. It is advantageous to add the oil to the feed in a closed container with a gas pressure below ambient pressure (vacuum coating) so that the porous feed pellets will absorb more oil. Feeds containing more than 40% lipid can be produced in this way. After the coating machine, the feed is cooled and bagged. As mentioned above, the oil may be added at several places in the process and may be fish oil, micro-organisms/algae or vegetable oil, such as rapeseed oil or soybean oil, or a mixture of oils.

Fish require proteins, fats, minerals and vitamins to grow and remain healthy. Daily ration of carnivorous fish is particularly important. Initially in the farming of carnivorous fish, whole or ground fish is used to meet the nutritional needs of farmed fish. Ground fish mixed with various types of dry raw materials (e.g. fish meal and starch) are known as soft or semi-moist feed. As agriculture becomes industrialized, soft feed or semi-moist feed is replaced by pressed dry feed. This itself is gradually replaced by expanded dry feed.

Expanded feeds are almost universally used today for the cultivation of a variety of fish species (e.g. salmonids, cod, sea bass and sea bream of various types).

The main protein source in dry fish feed is fish meal of different quality. Other animal protein sources are also used in 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, such as chicken meal. These are generally cheaper than fish meal and fish oil. However, in some geographical areas, the use of such raw materials in the production of feeds for animals and fish for the production of food has been banned.

It is also known to use vegetable proteins such as wheat gluten, maize (corn) gluten, soy protein, lupin meal, pea meal, soy meal, canola meal, sunflower seed meal and rice flour.

Example 4:

aquaculture feed can be produced according to the formulation described in table 3.

TABLE 2 aquaculture ration

Example 5:

aquaculture feed may be produced according to the formulation described in table 4.

TABLE 3 aquaculture ration

Composition of the components	[％]
		Fish meal	25.00
Wheat	12.80
		Wheat gluten	10.00
Soybean protein concentrate	18:00
		Rapeseed meal	10.00
Mineral mixture	1.00
		Vitamin mixture	1.30
Monocalcium phosphate	1.00
		L-lysine	0.30
DL-methylthioammoniaAcid(s)	0.20
		Vitamin C (rovimin STAY-C35)	200ppm
Vitamin E (ROVIMIX E50 SD)	400ppm
		Vitamin A	1.8ppm

Composition of the components	[％]
		Vitamin K	3ppm
Vitamin D	0.04ppm
		Cholesterol	250ppm
Phospholipid	1.5
		Arachidonic acid	0.01
Rapeseed oil	10.40
		Fish oil	0.00
Microbial oil	10.00

Example 6:

aquaculture feed may be produced according to the formulation described in table 5.

TABLE 4 aquaculture ration

Composition of the components	[％]
		Fish meal	10.00
Wheat	10.00
		Wheat gluten	16.00
Soybean protein concentrate	16.00
		Soybean meal	14.8
Rapeseed meal	9.00
		Choline chloride 60%	0.2
Mineral mixture	1.00
		Vitamin mixture	1.00
Guar gum	0.2
		Monocalcium phosphate	1.00
L-lysine	0.50
		DL-methionine	0.30
Yttrium	0.0002
		Vitamin C (rovimin STAY-C35)	200ppm

Composition of the components	[％]
		Vitamin E (ROVIMIX E50 SD)	250ppm
Vitamin A	1.8ppm
		Vitamin K	3ppm
Vitamin D	0.04ppm
		Cholesterol	250ppm
Phospholipid	1.5
		Arachidonic acid	0.01
Rapeseed oil	16.7
		Fish oil	0
Microbial oil	2.5

Example 7:

aquaculture feed may be produced according to the formulation described in table 6.

TABLE 5 aquaculture ration

Composition of the components	[％]
		Fish meal	10.00
Wheat	10.00
		Wheat gluten	16.00
Soybean protein concentrate	16.00
		Soybean meal	14.8
Rapeseed meal	9.00
		Choline chloride 60%	0.2
Mineral mixture	1.00
		Vitamin mixture	1.00
Guar gum	0.2
		Monocalcium phosphate	1.00
L-lysine	0.50
		DL-methionine	0.30

Composition of the components	[％]
		Yttrium	0.0002
Vitamin C (rovimin STAY-C35)	200ppm
		Vitamin E (ROVIMIX E50 SD)	250ppm
Vitamin A	0.9ppm
		Vitamin K	3ppm
Vitamin D	0.01ppm
		Cholesterol	250ppm
Phospholipid	0.5
		Arachidonic acid	0.01
Rapeseed oil	14.2
		Fish oil	0
Microbial oil	5

Example 8:

aquaculture feed may be produced according to the formulation described in table 7.

TABLE 6 aquaculture ration

Composition of the components	[％]
		Fish meal	10.00
Wheat	10.00
		Wheat gluten	16.00
Soybean protein concentrate	16.00
		Soybean meal	14.8
Rapeseed meal	9.00
		Choline chloride 60%	0.2
Mineral mixture	1.00
		Vitamin mixture	1.00
Guar gum	0.2
		Monocalcium phosphate	1.00

Composition of the components	[％]
		L-lysine	0.50
DL-methionine	0.30
		Yttrium	0.0002
Vitamin C (rovimin STAY-C35)	200ppm
		Vitamin E (ROVIMIX E50 SD)	250ppm
Vitamin A	0.9ppm
		Vitamin K	3ppm
Vitamin D	0.01ppm
		Cholesterol	250ppm
Phospholipid	1
		Arachidonic acid	0.01
Rapeseed oil	9.2
		Fish oil	0
Microbial oil	10

Example 9:

aquaculture feed may be produced according to the formulation described in table 9.

TABLE 7 aquaculture ration

Composition of the components	[％]
		Fish meal	25.00
Wheat	12.90
		Wheat gluten	11.00
Soybean protein concentrate	18:00
		Rapeseed meal	8.7
Mineral mixture	1.00
		Vitamin mixture	1.00
Monocalcium phosphate	1.00
		L-lysine	0.50

Composition of the components	[％]
		DL-methionine	0.30
Vitamin C (rovimin STAY-C35)	500ppm
		Vitamin E (ROVIMIX E50 SD)	600ppm
Vitamin A	1.8ppm
		Vitamin K	3ppm
Vitamin D	0.04ppm
		Cholesterol	150ppm
Phospholipid	1.2
		Arachidonic acid	0.01
Rapeseed oil	15.80
		Fish oil	0.00
Microbial oil	4.50

Example 10:

aquaculture feed may be produced according to the formulation described in table 10.

TABLE 8 aquaculture ration

Composition of the components	[％]
		Fish meal	25.00
Wheat	12.90
		Wheat gluten	11.00
Soybean protein concentrate	18:00
		Rapeseed meal	8.7
Mineral mixture	1.00
		Vitamin mixture	1.00
Monocalcium phosphate	1.00
		L-lysine	0.50
DL-methionine	0.30
		Vitamin C (rovimin STAY-C35)	500ppm

Composition of the components	[％]
		Vitamin E (ROVIMIX E50 SD)	600ppm
Vitamin A	0.9ppm
		Vitamin K	3ppm
Vitamin D	0.01ppm
		Cholesterol	500ppm
Phospholipid	0.93
		Arachidonic acid	0.01
Rapeseed oil	14.30
		Fish oil	0.00
Microbial oil	6.00

Claims (15)

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

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

3. The feed premix of claim 1 or 2, wherein the oil is derived from a microbial or plant source.

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

5. Feed premix according to any one 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 weight percent of the feed premix.

6. A feed intended for farm animals, the feed comprising an oil containing a high level of DHA and at least 5 additional ingredients selected from the group of vitamin a, vitamin K, vitamin D, vitamin C, vitamin E, cholesterol, phospholipids and arachidonic acid, and wherein the concentration of DHA, measured as a weight percentage of the oil, is at least 30%.

7. The feed of claim 6, wherein the oil is derived from a microbial source or a plant source.

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

9. A feed intended for ingestion by farm animals, the feed comprising a feed premix according to any of claims 1 to 5.

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

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

12. The use according to claim 11, wherein the oil is derived from microbial or plant sources.

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

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

15. The use according to any one of claims 11 to 14, wherein the feed comprises EPA and DHA in a total amount of at least 0.08% as measured by weight percent of the feed.