Classifications

• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
  • A23J1/04—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from fish or other sea animals
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J3/00—Working-up of proteins for foodstuffs
  • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
  • A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
  • A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K10/00—Animal feeding-stuffs
  • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
  • A23K10/14—Pretreatment of feeding-stuffs with enzymes
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K10/00—Animal feeding-stuffs
  • A23K10/20—Animal feeding-stuffs from material of animal origin
  • A23K10/22—Animal feeding-stuffs from material of animal origin from fish
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K10/00—Animal feeding-stuffs
  • A23K10/20—Animal feeding-stuffs from material of animal origin
  • A23K10/26—Animal feeding-stuffs from material of animal origin from waste material, e.g. feathers, bones or skin
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K50/00—Feeding-stuffs specially adapted for particular animals
  • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs

Definitions

• the present invention relates to a method for the treatment of aquatic animals, such as fish or prawns, in particular for the purpose of improving the bodyweight gain of the animals or for the purpose of reducing the conversion ratio of the feed used to feed the aquatic animals without reducing their bodyweight gain.
• the present invention further relates to feed compositions, methods for their production and uses thereof.
• Krill hydrolysates obtained by the methods described in WO 2010/030193 are one example of commercial protein additives available to feed manufacturers for the purpose of increasing feed intake and overall growth performance of farmed aquatic animals.
• the krill hydrolysates are obtained by contacting a disintegrated krill catch with an endoprotease (alkalase).
• the inventors have now surprisingly found that it is possible to provide an improved method fulfilling the above mentioned needs.
• said treatment comprises orally administering a feed to aquatic animals, wherein said feed comprises a hydrolyzed crustacean catch in an amount between 10 and 200 g/kg of dry weight of said feed, wherein said hydrolyzed crustacean catch is obtained by a method comprising providing a crustacean catch, disintegrating said crustacean catch to provide a disintegrated crustacean catch, contacting the disintegrated crustacean catch with at least one exogenous endoprotease and at least one exogenous exoprotease to provide the hydrolyzed crustacean catch, and wherein the aquatic animals are selected from anadromous fish, crustacean and mollusks.
• a method for the non-therapeutic treatment of aquatic animals wherein said treatment comprises orally administering a feed to aquatic animals, wherein said feed comprises a hydrolyzed crustacean catch in an amount between 10 and 200 g/kg of dry weight of said feed, wherein said hydrolyzed crustacean catch is obtained by a method comprising providing a crustacean catch, disintegrating said crustacean catch to provide a disintegrated crustacean catch, contacting the disintegrated crustacean catch with at least one exogenous endoprotease and at least one exogenous exoprotease to provide the hydrolyzed crustacean catch, and wherein the aquatic animals are selected from anadromous fish, crustacean and mollusks.
• the present invention further provides the use of the hydrolyzed crustacean catch, as detailed above, for reducing the conversion ratio of feed used to feed aquatic animals, without lowering their bodyweight gain, where said hydrolyzed crustacean catch, as detailed above, is orally administered to aquatic animals in an amount between 10 and 200 g/kg of dry weight of said feed, and wherein the aquatic animals are selected from anadromous fish, crustacean and mollusks.
• a method for preparing a coated pelletized feed for aquatic animals comprising providing a hydrolyzed crustacean catch, as detailed above, and depositing the hydrolyzed crustacean catch to at least one surface of a pelletized feed to form a coated pelletized feed for aquatic animals.
• the present invention further provides a feed composition
• a feed composition comprising: a) the hydrolyzed crustacean catch, as detailed above, b) one or more plant-based ingredients in a collective amount of at least 50 dry weight percent (dry wt.%), based on the total dry weight of the feed composition, and c) one or more additional ingredients selected from the group consisting of anti-caking agents, vitamins, minerals, various amino acids, free-flowing agents, animal feed flavors or the like.
• the feed composition is suitable as feed for aquatic animals, in particular for anadromous fish, crustacean and mollusks. Detailed description
• the present invention concerns a method for the non- therapeutic treatment of aquatic animals, which treatment comprises orally administering a feed to the aquatic animals, wherein said feed comprises a hydrolyzed crustacean catch in an amount between 10 and 200 g/kg of dry weight of said feed, wherein said hydrolyzed crustacean catch is obtained by a method comprising providing a crustacean catch, disintegrating said crustacean catch to provide a disintegrated crustacean catch, contacting the disintegrated crustacean catch with at least one exogenous endoprotease and at least one exogenous exoprotease to provide the hydrolyzed crustacean catch, wherein the aquatic animals are selected from anadromous fish, crustacean and mollusks.
• such method enables to reduce the conversion ratio of the feed used to feed these animals without reducing however the bodyweight gain, i.e. the average weight gain, or even enables to increase the bodyweight gain.
• conversion ratio of feed refers to a measure of an animal’s efficiency in converting feed mass increased body mass (e.g. muscle or roe mass for fish).
• conversion ratio of the feed is calculated by dividing the total weight of feed provided to the animals by the average daily bodyweight gain of the aquatic animals, all over a specified period.
• the inventors have now surprisingly found that when the disintegrated crustacean catch is further contacted with an exogenous exoprotease, an increase of the body weight gain or an increase of the body weight gain combined with a reduction of the feed conversion ratio can be obtained, as evidenced by the examples below. Moreover, small amounts of the hydrolyzed crustacean catch comprised between 10 and 200 g/kg of dry weight of the feed, can be used thereby resulting in a smaller cost for the supplementation of the feed with this feed additive.
• crustacean refers to any marine organism having an exoskeleton and being classified as part of the Crustacea subphylum.
• Non-limiting examples of crustaceans include krill, shrimp, lobsters, crab, water fleas, and/or barnacles.
• the crustacean used to provide the crustacean catch is krill, more preferably Antarctic krill ( Euphausia superba).
• crustacean catches can be provided fresh, or can have been previously frozen.
• the crustacean catch is a frozen crustacean catch.
• the crustacean catch is a fresh crustacean catch.
• the fresh crustacean catch is a crustacean catch caught within less than 24 hours, or less than 12 hours, or less than 2 hours, or less than 1 hour.
• the term “disintegrating” is intended to refer to mechanical breaking, granulation or fragmentation of the crustacean catch into smaller pieces or particles, with an average particle size of less than 25.0 millimeters (25.0 mm).
• Non-limiting examples of “disintegrated crustacean catch” are crustacean catch powders, fragments, particles, flakes, granules, grains or other components obtained by grinding a crustacean catch, with an average particle size of less than 25.0 mm.
• the particle sizes of the disintegrated crustacean catch can also be less than 20.0 mm, or less than 15.0 mm, or less than 10.0 mm, or less than 8.0 mm, or less than 7.0 mm, or less than 6.0 mm, or less than 5.0 mm.
• the average particle size can be measured by methods known in the art, for example, by particle size analyzers, methods such as measurement using light (light-scattering methods or turbidimetric methods), sedimentation methods (pipette analysis using an Andreassen pipette, sedimentation scales, photosedimentometers or sedimentation in a centrifugation force), pulse methods (Coulter counter), or sorting by means of gravitational or centrifugal force.
• particle size analyzers methods such as measurement using light (light-scattering methods or turbidimetric methods), sedimentation methods (pipette analysis using an Andreassen pipette, sedimentation scales, photosedimentometers or sedimentation in a centrifugation force), pulse methods (Coulter counter), or sorting by means of gravitational or centrifugal force.
• the disintegrated crustacean catch may be obtained by disintegrating the crustacean catch using any conventional means, such as using a knife shredder, a blender or a homogenizer.
• the temperature at which the disintegration process takes place may be around the ambient temperature of the water in which the crustacean catch was caught when the crustacean catch is a fresh crustacean catch.
• the crustacean catch is disintegrated at a temperature between -2.0 degrees Celsius (-2.0 °C) and +10.0 °C, preferably between 0.0 °C and +6.0 °C.
• the expression “hydrolyzed crustacean catch”, is intended to refer to a crustacean catch which has undergone hydrolysis.
• Hydrolysis is a chemical reaction which can be caused or mediated by a biological agent, such as a proteolytic enzyme (or protease), whereby the natural protein sequences become shorter (for example by breaking peptide bonds of the amino acid sequence primary structure) to form smaller peptides and/or free amino acids.
• the disintegrated crustacean catch needs to be hydrolyzed so that digestive enzymes, such as lipases and phospholipases, which are released from the crustacean catch upon death may be inactivated. If these digestive enzymes are not inactivated upon their release, there is a risk that they will hydrolyze the phospholipids and fatty acids within the crustacean catch.
• the disintegrated crustacean catch is contacted with a protease under certain conditions to form a hydrolyzed crustacean catch.
• the protease is generally chosen to specifically target the crustacean digestive enzymes while minimally damaging the other crustacean proteins.
• proteolytic enzyme refers to an enzyme that catalyses the cleavage of peptide bonds within a polypeptide or protein, the internal peptide bonds of a polypeptide or protein.
• endoprotease refers to an enzyme that catalyses the cleavage of peptide bonds within a polypeptide or protein, the internal peptide bonds of a polypeptide or protein.
• exoprotease refers to a protease that can remove the terminal amino acid(s) of a peptide or protein by cleaving peptide bonds.
• a terminal amino acid is an amino acid that is the N-terminal or C-terminal amino acid of a protein or peptide.
• suitable exogenous endoproteases and exogenous exoproteases include serine proteases, cysteine proteases, aspartic proteases, metalloproteases, threonine proteases, glutamic proteases and asparagine proteases.
• the at least one exogenous endoprotease and the at least one exogenous exoprotease are each independently selected from serine, cysteine, aspartic and metalloproteases.
• the at least one exogenous endoprotease and the at least one exogenous exoprotease are independently selected form serine proteases and metalloproteases.
• the at least one exogenous endoprotease is a metalloprotease and the at least one exogenous exoprotease is a serine protease.
• the at least one exogenous endoprotease and the at least one exogenous exoprotease according to the method of the present invention can also be each independently selected from acid proteases, neutral proteases and basic (or alkaline) proteases.
• Acid proteases are proteases that exhibit maximum activity and stability in acidic conditions (pH 2.0 - 5.0) and are typically inactive at pH values above 6.0. Acidic proteases generally have a low isoelectric point and have a low basic amino acid content.
• Neutral proteases are typically active in a narrow pH rage (pH 5.0 - 8.0), whereas alkaline proteases are characterized by their high activity at alkaline pH, typically above pH 9.0.
• the at least one exogenous endoprotease is a neutral endoprotease or an alkaline endoprotease.
• the at least one exogenous endoprotease is a neutral endoprotease.
• the at least one exogenous exoprotease is an alkaline or a neutral exoprotease.
• the at least one exogenous exoprotease is a neutral exoprotease.
• the at least one exogenous endoprotease is a neutral endoprotease and the at least one exogenous exoprotease is a neutral exoprotease.
• the at least one exogenous endoprotease and the at least one exogenous exoprotease according to the present invention can further be derived from any organism, including plants, animals, bacteria, viruses or fungi.
• the at least one exogenous endoprotease and the at least one exogenous exoprotease are obtained from bacteria of the genus Bacillus. Suitable exogenous endoproteases and the exogenous exoproteases are obtained from the bacteria Bacillus Licheniformis.
• the at least one endoprotease and the at least one exoprotease are exogenous proteases. This implies that the hydrolyzed crustacean catch results from the hydrolysis of a crustacean catch by additional proteases that are not naturally present in the crustacean catch and that are added to the catch in order to hydrolyze proteins.
• the disintegrated crustacean catch will be contacted with an amount of at least 0.1 g/kg, or at least 0.2 g/kg, or at least 0.5 g/kg, or at least 1.0 g/kg, or at least 2.0 g/kg of the at least one exogenous endoprotease, relative to the total weight of said disintegrated crustacean catch.
• the disintegrated crustacean catch will be contacted with an amount of at least 0.01 g/kg, or at least 0.02 g/kg, or at least 0.05 g/kg, or at least 0.1 g/kg, or at least 0.2 g/kg of the at least one exogenous exoprotease, relative to the total weight of said disintegrated crustacean catch.
• the disintegrated crustacean catch will be contacted with an amount of at least 0.1 g/kg of the at least one exogenous endoprotease and an amount of at least 0.01 g/kg of the at least one exogenous exoprotease, or least 0.2 g/kg of the at least one endoprotease and an amount of at least 0.02 g/kg of the at least one exogenous exoprotease, or least 0.5 g/kg of the at least one exogenous endoprotease and an amount of at least 0.05 g/kg of the at least one exogenous exoprotease, or least 1 .0 g/kg of the at least one exogenous endoprotease and an amount of at least 0.1 g/kg of the at least one exogenous exoprotease, or least 2.0 g/kg of the at least one exogenous endoprotease and an amount of at least 0.2 g/kg of the at least
• the amounts of the at least one exogenous endoprotease and the at least one exogenous exoproteases refer either to the amount of the exogenous (endo-/exo-)protease, i.e. the exogenous (endo-/exo-)protease, when only one exogenous (endo-/exo-)protease is added to the disintegrated crustacean catch, or to the sum of the amounts of exogenous (endo-/exo- )proteases, when more than one exogenous (endo-/exo-)protease is added to the disintegrated crustacean catch.
• the disintegrated crustacean catch can be contacted with the at least one exogenous endoprotease and the at least one exogenous exoprotease at a pH ranging from 5.0 to 9.0, preferably, ranging from 7.0 to 8.0.
• the disintegrated crustacean catch can be contacted with the at least one exogenous endoprotease and the at least one exogenous exoprotease at a temperature ranging from 30.0 °C to 80.0 °C, or between 40.0 °C and 70.0 °C, or between 50.0 °C and 60.0 °C.
• the disintegrated crustacean catch can be contacted either sequentially or simultaneously with the at least one exogenous endoprotease and the at least one exogenous exoprotease. Therefore, according to certain embodiments of the method according to the present invention, the disintegrated crustacean catch is contacted simultaneously with the at least one exogenous endoprotease and the at least one exogenous exoprotease.
• the disintegrated crustacean catch are contacted with the at least one exogenous endoprotease and the at least one exogenous exoprotease for a duration ranging from 1 minute to 24 hours, or from 5 minutes to 3 hours, or from 10 minutes to 2 hours.
• the disintegrated crustacean catch is contacted sequentially with the at least one exogenous endoprotease and the at least one exogenous exoprotease.
• the disintegrated crustacean catch is first contacted with the at least one exogenous endoprotease, and subsequently with the at least one exogenous exoprotease.
• the inventors have found that the disintegrated crustacean catch could be advantageously contacted with the at least one exogenous exoprotease for a longer time period than with the at least one exogenous endoprotease.
• the duration of the contacting step with the at least one exoprotease can range from 30 minutes to 24 hours, or from 1 hour to 3 hours, while the duration of the contacting step with the at least one endoprotease can be reduced to less than 2 hours, or less than 1 hour, or less than 30 minutes, or less than 15 minutes, or less than 10 minutes or even less than 5 minutes.
• the term “sequentially” is intended to refer to two separate steps occurring either directly after one another, or to a second step occurring substantially later than a first step.
• the contacting of the disintegrated crustacean catch with the at least one exogenous exoprotease can take place at any point in time after the contacting with the at least one exogenous endoprotease.
• the hydrolysis may be stopped by inactivating the proteases. This may advantageously be achieved by raising the pH and/or the temperature.
• the inventors have surprisingly found that by performing the hydrolysis in the presence of at least one exogenous endoprotease and at least one exogenous exoprotease, a hydrolyzed crustacean catch having improved properties could be obtained. It was observed that the hydrolyzed crustacean catch according to the method of the present invention, as detailed above, resulted in increased bodyweight gain as well as specific growth rate (SGR) as compared to conventional hydrolysates when orally administered to aquatic animals, wherein the aquatic animals are selected from anadromous fish, crustacean and mollusks.
• SGR specific growth rate
• the hydrolyzed crustacean catch comprises a modified distribution of total amino acids, free amino acids, as well as a different distribution of molecular weight peptide size, as compared to conventional hydrolyzed crustacean catches.
• the hydrolyzed crustacean catch comprises a higher proportion of small molecular weight peptides ( ⁇ 200 Da).
• distribution of many amino acids (total and free amino acids) is altered in the hydrolyzed crustacean catch, as detailed above, as compared with conventional hydrolyzed crustacean catches. The combination of these factors appears to have a beneficial effect on the overall performance of aquatic animals to which a feed comprising the hydrolyzed crustacean catch according to the method of the present invention, as detailed above, is orally administered.
• the method according to the present invention comprises administration of a feed which is in the form of a pelletized feed.
• Non-limiting examples of suitable pelletized feeds include feeds in the form of particulate chunks or pieces formed by either a press or extrusion process.
• the pellets can vary in size and/or shape, depending on the process or the equipment.
• fish pelleted feeds have a cylindrical shape.
• the size of the pelleted feed to be used can be determined by the skilled person based on the size of the aquatic animal.
• the pelletized feed is a coated pelletized feed comprising a core, and at least one coating layer. More preferably, at least part of the hydrolyzed crustacean catch according to the present invention is comprised in said coating layer. More preferably, the hydrolyzed crustacean catch is substantially only present in the coating layer, and this in an amount between 10 and 200 g/kg of dry weight of the feed.
• the inventors have indeed surprisingly found that when the hydrolyzed crustacean catch is present in the coating layer, improved effects in terms of bodyweight gain and feed conversion ratio could be overserved. Without being bound by theory, it seems that when the hydrolyzed crustacean catch is present in the coating layer, it will dissolve more efficiently in water, thereby increasing the overall attractiveness of the feed. As mentioned above, the inventors have found that the upper limit for the amount of hydrolyzed crustacean catch comprised in the feed should be at most 200 g/kg of dry weight of said feed.
• the feed comprises the hydrolyzed crustacean catch in an amount of less than 180, preferably less than 150, more preferably less than 120, even more preferably less than 100, most preferably less than 80 g/kg dry weight of said feed.
• the amount of hydrolyzed crustacean catch, as detailed above, in the feed is at least 12, preferably at least 15, more preferably at least 20 g/kg dry weight of said feed.
• the feed comprises the hydrolyzed crustacean catch in an amount between 12 and 180, preferably between 15 and 150, more preferably between 20 and 100 g/kg dry weight of said feed.
• aquatic animal refers to any animal, either vertebrate or invertebrate, which lives in water for the most or all of its lifetime.
• the method according to the present invention is applicable for the treatment of aquatic animals selected from anadromous fish, crustacean and mollusks.
• the aquatic animal is a crustacean.
• Non-limiting examples of crustaceans include prawns, shrimp, lobsters, crabs, water fleas, and/or barnacles.
• the aquatic animal is a prawn or a shrimp.
• farmed species suitable for the method according to the present invention include whiteleg shrimp (also known as Pacific white shrimp or king prawn, Litopenaeus vannamei, formerly known as Penaeus vannamei ), giant tiger prawn (also known as Asian tiger shrimp or black tiger shrimp, Penaeus monodon), Indian white prawn (also known as Indian prawn, Fenneropenaeus indicus, formerly known as Penaeus indicus) and giant river prawn (also known as giant freshwater prawn, Macrobrachium rosenbergii).
• the method according to the present invention is particularly suitable for the treatment of farmed shrimp or prawn as these animals are typically grown under high density conditions which induce high stress and reduced appetite. It is therefore essential to use feed ingredients that can trigger feed intake and initiate body weight gain in order to obtain efficient growth.
• the aquatic animal is an anadromous fish.
• Anadromous fish are those that spawn in freshwater, migrate to the ocean to forage and mature, and return to freshwater to spawn and begin the cycle again.
• the aquatic animal is an anadromous carnivorous fish or an anadromous omnivorous fish.
• Non-limiting examples of anadromous carnivorous fish or anadromous omnivorous fish include pink salmon (Oncorhynchus gorbuscha), chum salmon (Oncorhynchus keta), coho salmon (Oncorhynchus kisutch), masu salmon (Oncorhynchus masou), rainbow trout (Oncorhynchus mykiss), sockeye salmon (Oncorhynchus nerka), Atlantic salmon (Salmo salar), sea trout (Salmo trutta morpha trutta), striped bass (Morone saxatilis), and sturgeon (Acipenseridae spp.).
• the method according to the present invention is particularly suitable for the treatment of juvenile anadromous fish which have been transferred from a freshwater environment to a marine environment for farming purposes.
• This is indeed a critical stage in the farming process, where most of the body weight gain of the animals will occur. It is also a period where the fish will typically be under substantial stress.
• juvenile fish such as salmon smolt
• normal aquaculture growing conditions include substantial density in the enclosure. It is therefore essential to use feed ingredients that can trigger feed intake and initiate body weight gain to obtain efficient growth.
• the aquatic animal is an anadromous carnivorous fish from the Salmonidae family (salmonids), most preferably an anadromous salmonid selected from pink salmon (Oncorhynchus gorbuscha), chum salmon (Oncorhynchus keta), coho salmon (Oncorhynchus kisutch), masu salmon (Oncorhynchus masou), rainbow trout (Oncorhynchus mykiss), sockeye salmon (Oncorhynchus nerka), Atlantic salmon (Salmo salar) and sea trout (Salmo trutta morpha trutta).
• Salmonidae family Salmonidae family
• the aquatic animal is an anadromous salmonid from the genus Salmo, even more preferably a salmonid from the species Salmo salar (Atlantic salmon).
• Salmonids are among the most important fish species used in aquaculture worldwide, particularly in Europe. Globally, Salmonid aquaculture production grew over ten-fold between 1982 and 2007, reaching a 10.7 billion USD globally.
• Anadromous salmonids are usually farmed in two stages. First the salmonids are hatched from eggs and raised on land in freshwater tanks. After 12 to 18 months, the smolts are then transferred to floating sea cages or net pens, typically anchored in sheltered bays or fjords along a coast. After transfer, the smolt are fed (pelleted) feed for another 12 to 24 months, before they develop into adult stage and are considered ready to be harvested.
• Smoltification is a complex developmental transformation involving morphological, biochemical, physiological and behavioral changes that preadapt young parr for life in high salinity water (Hogasen, HR (1998). Physiological changes associated with the diadromous migration of salmonids. Canadian Special Publication of Fisheries and Aquatic Sciences 127, 128 p.). Light and temperature regimes can be manipulated artificially to induce early smoltification.
• the morphological changes include a slimmer body form and alterations in body coloration (darkened fins, dark back, white belly and silver sides) that help to conceal with the pelagic environment. Atlantic salmon smolts originally weigh 50-80 grams and can be grown up to 1 kg before being transferred to seawater.
• the term “smolt” refers to anadromous salmonids which have undergone smoltification but have not yet reached full adult size (adult stage).
• the term “smolt” thus encompasses fish which a grown in seawater up to the moment they reach their full adult size, typically 12-24 months on farms, sometimes referred to as “post-smolts”.
• the aquatic animal is a smolt, most preferably an Atlantic salmon ( Salmo salar) smolt.
• Another aspect of the present invention is a feed composition for aquatic animals, in particular the aquatic animals, as detailed above, comprising the hydrolyzed crustacean catch as detailed above.
• the feed composition which is suitable as a feed for aquatic animals, in particular the aquatic animals, as detailed above, comprises: a) the hydrolyzed crustacean catch, as detailed above, in an amount of between 10 and 200 g/kg of dry weight of said feed composition; b) one or more plant-based food ingredients in a collective amount of at least 50 dry weight percent (dry wt.%), based on the dry weight of the feed composition; and c) one or more additional ingredients comprising anti-caking agents, vitamins, minerals, various amino acids, free-flowing agents, animal feed flavors or the like.
• the plant-based food ingredients may be derived from any plant source.
• suitable plant sources include soy, rapeseed, wheat, corn, lupine, pea, sunflower, rice and the like.
• the feed composition does not contain any added ingredients or food contaminant that are poisons or toxins, e.g. substances that have an inherent property and in amounts to induce death or illness in animals, including the aquatic animals.
• the feed composition is in a solid form.
• the feed composition is in the form of a pellet.
• the hydrolyzed crustacean catch can be distributed within and throughout the pellets.
• the pellet is a coated pellet, comprising a core and at least one coating layer.
• At least part of the hydrolyzed crustacean catch is comprised in the coating layer. More preferably, the core of the coated pellet comprises said one or more plant-based ingredients, whereas the at least one coating layer comprises said hydrolyzed crustacean catch.
• the upper limit for the amount of hydrolyzed crustacean catch comprised in the feed composition should be at most 200 g/kg of dry weight of said feed.
• the feed composition comprises the hydrolyzed crustacean catch in an amount of less than 180, preferably less than 150, more preferably less than 120, even more preferably less than 100, most preferably less than 80 g/kg dry weight of said feed.
• the amount of hydrolyzed crustacean catch, as detailed above, in the feed composition is at least 12, preferably at least 15, more preferably at least 20 g/kg dry weight of said feed.
• the feed composition comprises the hydrolyzed crustacean catch in an amount between 12 and 180, preferably between 15 and 150, more preferably between 20 and 100 g/kg dry weight of said feed.
• the methods to manufacture a coated pelletized feed is also an aspect of the present invention.
• coated pelletized feed suitable for aquatic animals, in particular the aquatic animals, as detailed above, can be prepared by a variety of methods known in the art.
• the method for preparing the coated pelletized feed comprises: a) providing a hydrolyzed crustacean catch, as detailed above; and b) depositing said hydrolyzed crustacean catch to at least one surface of a pelletized feed to form a coated pelletized feed for aquatic animals.
• a total of 1440 fertilized Atlantic salmon ( Salmo salar) eggs were hatched and raised in freshwater aquaria for 12 months. All fish were fed a commercial freshwater diet prior to the start of the trial. The fish were then starved for one day and randomly distributed over 18 fiberglass tanks (1 x 1 m 2 ) comprising each 80 animals and assigned to one out of the six test diets in triplicate. The fish were fed in excess appetite by automatic feeders at intervals of 38 seconds duration, every 32 minutes, which amounts to 48 meals per day on average, for a feeding period of 42 days. Unconsumed feed was collected daily, dried (40 °C, 24 h) and weighed to record the daily feed consumption. Recovery tests were performed for each test diet in order to correct for variable loss of dry matter.
• the fish were exposed to continuous light (12:12h) throughout the trial and kept at ambient water temperature of average 12 °C for the duration of the trial. All tanks were supplied with running seawater at a flow of 24 L/min. Oxygen content of the outlet water from each tank ranged from 83% to 95% saturation throughout the trail. The total fish biomass was recorded at the start and end of the trail. The total biomass and number of fish per tank were used to calculate the fish body weight.
• composition of the diets was designed according to meet energy and nutrient requirements of salmon feed.
• the composition of the basal formulation of these diets is shown in Table 1.
• Table 1. Ingredient composition of the basal formulation.
• the basal formulation was used to produce the seven experimental diets.
• the diets as described in Table 1 , were supplemented with fish meal, wheat, fish oil and rapeseed oil on top of the basal formulation.
• the krill hydrolysate diets contained 25 g/kg or 50 g/kg of different types of krill hydrolysates.
• the amount of fish meal, wheat, fish oil, rapeseed oil and water was adjuster to maintain a constant overall nutritional composition (iso-caloric, constant protein and lipid levels) throughout each of the diets, as described in Table 3.
• the weight percentage of each of the components is relative to the total weight of the test diets. All diets were produced as pelleted feed in a commercial feed mill.
• the characteristics of the krill hydrolysates 1 to 3 are summarized in Table 2.
• the krill hydrolysates were deposited as coating layer to the feed pellets.
• the krill hydrolysates KH1, KH2 and KH3 were obtained as follows. 500 kg of frozen krill was shredded using a knife cutter to a particle size comprised between 3 and 6 mm. The temperature of the krill was comprises between 1 and 2 °C. A mixture of 500 kg of fresh water and protease was added and incubated at 55 °C.
• the shells were separated from the main process line using a decanter.
• the hydrolyzed krill in the main process line was then pasteurized to ensure hygienic quality and to inactivate the proteases. Pasteurization was performed at 90 °C for 10 minutes.
• the krill hydrolysates were subsequently dried using a sedicanter to mechanically separate the excess water, followed by low temperature vacuum drying.
• Each of the three krill hydrolysates were added to the basal formulation in combination with other additives, so as to maintain a stable nutritional balance throughout the test diet compositions, as detailed above.
• Example 2 Effect of krill hydrolysate supplementation on daily feed intake.
• Table 5 demonstrates that supplementation with krill hydrolysates increases feed intake relative to the control diet. Most importantly, it was observed that supplementation with the either 2.5 wt.% or 5.0 wt.% of the krill hydrolysates according to the invention (Examples 4 - 5, Ex 4 - 5) results in an increase in feed intake as compared to conventional krill hydrolysates (Comparative Examples 1 - 3, CEx 1 - 3) after both 15 and 42 days in smolts.
• Table 6 demonstrates that supplementation with krill hydrolysates increases final body weight and daily specific growth rate (SGR) relative to the control diet. Most importantly, it was observed that supplementation with the either 2.5 wt.% or 5.0 wt.% of the krill hydrolysates according to the invention
• Examples 4 - 5 results in an increase in final body weight as well as in daily specific growth rate (SGR) as compared to conventional krill hydrolysates (Comparative Examples 1 - 3) after both 15 and 42 days in smolts.
• SGR daily specific growth rate
• Examples 4 and 5 Based on the results of Table 5 and Table 6, it can be concluded that the dietary treatments prepared according to the invention (Examples 4 and 5) perform best.
• the dietary preparations comprise the krill hydrolysate KH4 at concentrations of 2.5 wt.% (Example 4) or 5.0 wt.% (Example 5)
• smolt salmons perform better in terms of final growth and growth rate. Also, the feed intake response was found to be increased.
• Penaeus vannamei are imported as postlarvae and reared into a water recirculation system containing artificial seawater at a salinity of 20 g L 1 . They are raised on an artificial diet and after weaned onto a pelleted feed. Both feed are distributed 12 times a day. Water temperature is kept constant at 27°C by means of an automatic heating system. A complete biological/mechanical filter and regular water changes keep the total ammonia at ⁇ 0.05 mg L ⁇ 1 and nitrites at ⁇ 0.8 mg L ⁇ 1 .
• a total of 980 shrimp weighing 0.5-2 grams are used to randomly compose 14 groups of 70 individuals. Each group is housed in a feeding unit. Each experimental diet is randomly assigned to 3 tanks. During the feeding trial period, the groups of shrimp receive the respective diets at the predetermined percentages of their initial mean body weight (MBW). This is adjusted daily according to the expected growth, and observed mortality and feed consumption per group. The total weight of the groups is measured at the start and at the end of the trials. A subsampling is performed every two weeks and growth curves are adjusted accordingly.
• MMW mean body weight
• the number of shrimp is reduced to 30 shrimp per tank for performing the palatability trial.
• Feeding tanks are kept in full recirculation except during the periods of when palatability feed samples will be collected. At this time, tanks are placed in individual circulation to avoid exchange of dietary attractants in between treatments.
• feed portions are added to the respective tanks. Thirty minutes after feeding, the leftover portions are recovered and dried. Weights of each feed leftover portions are measured and percentage of feed intake per group is calculated.
• shrimp faeces are collected suing a faeces precipitation system coupled to the shrimp feeding tank.
• the daily samples are stored at -20°C until at least the minimal wet weight of the faeces has been collected from each replicate tank.
• the frozen feaces are free-dried before analysis.
• the analysis is performed for protein (Kjeldahl) and T1O2 based on UV-spectroscopy.
• the outcome in each experimental group is compared with the control group and with every other experimental group using a two-way ANOVA statistical analysis followed by a Turkey’s multiple comparisons test.
• the composition of the diets are designed to meet energy and nutrient requirements of shrimp feed.
• a basal formulation are used to produce five experimental diets. Four of the diets are supplemented with 40 or 80g/kg of different types of krill hydrolysates.
• the composition of the basal formulation is adjusted to maintain a constant overall nutrition composition (iso-caloric, constant protein and lipid levels) throughout each of the diets. The weight percentage of each of the components is relative to the total weight of the test diets. All diets were produced as pelleted feed in a commercial feed mill.
• 4.00 wt.% of krill hydrolysate is added through topcoating in Examples 6 and 7 (Ex 6 and 7). In Examples 7 and 8 and Comparative Example 9 (Ex 7, 8 and CEx 9), 4.00 wt.% of krill hydrolysate is added by inclusion. Titanium oxide is added as a tracer for the digestibility analysis.
• Example 5 Effect of krill hydrolysate supplementation on growth performance Supplementation with krill hydrolysates increases feed intake, relative to the control diet. Most importantly, supplementation the krill hydrolysates according to the invention (Examples 6 - 8) results in an increase in feed intake as compared to conventional krill hydrolysates (Comparative Example 9). Applying the krill hydrolysate as a top-coating (Example 6) also increases feed intake as compared to the same amount of krill hydrolysate added by inclusion into the diet (Example 8).
• Example 6 Effect of krill hydrolysate supplementation on body weight and daily specific growth rate Supplementation with krill hydrolysates increases final body weight and daily specific growth rate (SGR) relative to the control diet. Most importantly, supplementation with krill hydrolysates according to the invention (Examples 6 - 8) results in an increase in final body weight as well as in daily specific growth rate (SGR) as compared to conventional krill hydrolysates (Comparative Example 9) in shrimp. Applying the krill hydrolysate as a top-coating (Example 6) also increases final body weight as well as in daily specific growth rate (SGR) as compared to the same amount of krill hydrolysate added by inclusion into the diet (Example 8).
• Example 7 Effect of krill hydrolysate supplementation on feed palatability
• Supplementation with krill hydrolysates increases feed palatability relative to the control diet. Most importantly, supplementation with krill hydrolysates according to the invention (Examples 6 - 8) results in an increase in feed palatability as compared to conventional krill hydrolysates (Comparative Example 9) in shrimp. Applying the krill hydrolysate as a top-coating (Example 6) also increases feed palatability as compared to the same amount of krill hydrolysate added by inclusion into the diet (Example 8).
• Supplementation with krill hydrolysates increases feed digestibility relative to the control diet. Most importantly, supplementation with krill hydrolysates according to the invention (Examples 6 - 8) results in an increase in feed digestibility as compared to conventional krill hydrolysates (Comparative Example 9) in shrimp. Applying the krill hydrolysate as a top-coating (Example 6) also increases feed digestibility as compared to the same amount of krill hydrolysate added by inclusion into the diet (Example 8).

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