Fish feed or fish feed additive

Abstract

A fish feed additive or fish feed that includes guanadinoacetic acid, arginine, creatine, and / or phosphocreatine, and methods of making and using the fish feed additive or fish feed. When fed to fish residing in low oxygen conditions, the fish feed additive or fish feed can be effective to reduce or eliminate one or more symptoms of the low oxygen environment on the fish.

Description

FISH FEED OR FISH FEED ADDITIVECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 329,269, filed April 8, 2022, which is incorporated by reference herein in its entirety.BACKGROUND

[0002] Ocean deoxygenation is the reduction of the oxygen content of the oceans which can be caused by various factors such as anthropogenic emissions of carbon dioxide or eutrophication-driven excess production. It is manifest in the increasing number of coastal and estuarine hypoxic areas, or dead zones, and the expansion of oxygen minimum zones in the world’s oceans. The decrease in oxygen content of the oceans has been fairly rapid and poses a threat to all aerobic marine life, as well as to people who depend on marine life for nutrition or livelihood. During low O2 events fish typically experience decreased feeding, metabolic stress, a reduction in grow th, and a reduction in animal welfare. There is a need for fish feed additives and methods that reduce or eliminate the symptoms of residing in a low oxygen environment.SUMMARY OF THE INVENTION

[0003] In various aspects, the present invention provides a fish feed additive that includes guanadinoacetic acid.

[0004] In various aspects, the present invention provides a fish feed additive that includes guanadinoacetic acid that is 50 wt% to 99 wt% of the fish feed additive. The fish feed additive also includes arginine that is50 wt% of the fish feed additive.

[0005] In various aspects, the present invention provides a fish feed that includes a fish feed additive. The fish feed additive includes guanadinoacetic acid. The fish feed also includes a base feed component.

[0006] In various aspects, the present invention provides a fish feed that includes a fish feed additive. The fish feed additive is 50 wt% to 100 wt% guanadinoacetic acid. The fish feed additive is 0.01 wt% to 5 wt% of the fish feed. The fish feed also includes a base feed component.

[0007] In various aspects, the present invention provides a fish feed that includes guanadinoacetic acid. The guanadinoacetic acid is 0.01 wt% to 5% wt% of the fish feed. The fish feed also includes a base feed component.

[0008] In various aspects, the present invention provides a method of forming the fish feed described herein. The method includes adding the fish feed additive described herein to a base feed component to form the fish feed.

[0009] In various aspects, the present invention provides a method of feeding fish. The method includes feeding the fish the fish feed additive described herein or the fish feed described herein.

[0010] In various aspects, the present invention provides a method of feeding salmonid fish. The method includes feeding the salmonid fish a feed composition including a base feed and including a feed additive that is 0.01 wt% to 5 wt% of the fish feed. The feed additive is 50 wt% to 100 wt% guanadinoacetic acid. The salmonid fish reside in an environment having an oxygen saturation of 30% to 85%. The method is effective to improve to maintain overall fish health, growth rate, survival rate, rate of feed consumption, gill efficiency, rate of damaged gill cell repair, gill cell energy production, gill cell energy storage, gill processes including oxygen absorption, gill processes including ionic homeostasis, gill processes including osmoregulation, gill processes including pH control, blood anion gap, blood pH, blood sodium ion concentration, or a combination thereof, as compared to the same fish in the same environment and fed a diet free of the fish feed additive.

[0011] In various aspects, the present invention provides a fish feed additive for feeding to salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment. The fish feed additive includes arginine.

[0012] In various aspects, the present invention provides a fish feed for feeding to salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment. The fish feed includes a fish feed additive that includes arginine. The fish feed also includes a base feed component.

[0013] In various aspects, the present invention provides a method of feeding salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment. The method includes feeding the salmonids residing in the low oxygen environment a fish feed that includes arginine, a fish feed additive that includes arginine, or a combination thereof.

[0014] In various aspects, the present invention provides a fish feed additive for feeding to salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment. The fish feed additive includes creatine and / or phosphocreatine.

[0015] In various aspects, the present invention provides a fish feed for feeding to salmonids residing in a low oxygen environment or in preparation for residing in a low oxy gen environment. The fish feed includes a fish feed additive that includes creatine and / or phosphocreatine. The fish feed also includes a base feed component.

[0016] In various aspects, the present invention provides a method of feeding salmonids residing in a low oxygen environment or in preparation for residing in a low oxy gen environment. The method includes feeding the salmonids residing in the low oxygen environment a fish feed that includes creatine and / or phosphocreatine, a fish feed additive that includes creatine and / or phosphocreatine, or a combination thereof.

[0017] In various aspects, the fish feed additive of the present invention, the fish feed including the same, and / or methods of making or using the additive or fish feed, can have certain advantages over other fish feed additives and methods of using the same. For example, in various aspects, the fish feed additive of the present invention and / or the fish feed including the same can be effective to improve or maintain overall fish health, growth rate, survival rate, rate of feed consumption, gill efficiency, rate of damaged gill cell repair, gill cell energy production, gill cell energy storage, gill processes including oxygen absorption, gill processes including ionic homeostasis, gill processes including osmoregulation, gill processes including pH control, blood anion gap, blood pH, blood sodium ion concentration, fish productivity, reduce overall stress, reduce susceptibility to secondary illness, or a combination thereof, as compared to the same fish in the same environment and fed a diet free of the fish feed additive. In various aspects, the fish feed additive of the present invention and / or the fish feed including the same, when fed to a fish in a low oxygen environment, can be effective to reduce or eliminate one or more symptoms of the low oxygen environment on the fish. In various aspects, the fish feed additive of the present invention and / or the fish feed including the same may enable damaged gills to function more efficiently by supporting undamaged cells to maintain their gill function and prevent overall loss of fish condition while the damaged cells are being repaired. In various aspects, the fish feed additive of the present invention and / or the fish feed including the same may be used in reaction to or in preparation for sudden low oxygen events and / or gill damage, in reaction to known seasonal variations in oxygen concentration (e.g.,administration may begin prior to the onset of low' oxygen conditions), and / or for programmed handling or disease treatment.BRIEF DESCRIPTION OF THE FIGURES

[0018] The drawings illustrate generally, by way of example, but not by way of limitation, various aspects of the present invention.

[0019] FIG. 1 illustrates modelled weight gain of fish subjected to various feed / 02 conditions, in accordance with various aspects.

[0020] FIG. 2 illustrates the anion gap of blood of fish subjected to various feed / 02 conditions, in accordance with various aspects.

[0021] FIG. 3 illustrates blood pH of fish subjected to various feed / 02 conditions, in accordance with various aspects.

[0022] FIG. 4 illustrates blood sodium ion concentration of fish subjected to various feed / 02 conditions, in accordance with various aspects.DETAILED DESCRIPTION OF THE INVENTION

[0023] Reference will now be made in detail to certain aspects of the disclosed subject matter, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter.

[0024] Throughout this document, values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. For example, a range of “about 0.1% to about 5%” or “about 0.1% to 5%” should be interpreted to include not just about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.

[0025] In this document, the tenns “a,” “an,” or “the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to anonexclusive “or” unless otherwise indicated. The statement “at least one of A and B” or “at least one of A or B” has the same meaning as “A, B, or A and B.” In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting: information that is relevant to a section heading may occur within or outside of that particular section.

[0026] In the methods described herein, the acts can be carried out in a specific order as recited herein. Alternatively, in any aspect(s) disclosed herein, specific acts may be carried out in any order without departing from the principles of the invention, except when a temporal or operational sequence is explicitly recited. Furthermore, specified acts can be carried out concurrently unless explicit claim language recites that they be carried out separately or the plain meaning of the claims would require it. For example, a claimed act of doing X and a claimed act of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process.

[0027] The term “about” as used herein can allow for a degree of variability in a value or range, for example, within 10%, within 5%, or within 1% of a stated value or of a stated limit of a range, and includes the exact stated value or range.

[0028] The term “substantially” as used herein refers to a majority of, or mostly, as in at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or 100%.

[0029] The term “substantially free of’ as used herein can mean having none or having a trivial amount of, such that the amount of material present does not affect the material properties of the composition including the material, such that about 0 wt% to about 5 wt% of the composition is the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4, 3.5, 3, 2.5, 2, 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0. 1, 0.01, or about 0.001 wt% or less, or about 0 wt%.Fish feed additive.

[0030] The present invention provides a fish feed additive that includes guanadinoacetic acid. The fish feed additive can be suitable for feeding to fish that reside in any aqueous environment, such as an environment having normal oxygen levels (e g., having an oxygen saturation of >95% to <100%), or such as an environment having low oxygen levels. Low oxygen levels can include water conditions having an oxygen saturation of 10% to 95%, 30% to 85%, 50% to 70%, or less than or equal to 95% and greater than or equal to 10%, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90%. An environment having normal oxygen levels can have an oxygen concentration of 6.5 mg / L or more, or 5.5 mg / L or more. An environment having low oxygen levels can have an oxygen concentration of 0. 1 mg / L to 6.5 mg / L, or 0. 1 mg / L to 5.5 mg / L, or 0.1 mg / L to 4 mg / L, or less than or equal to 6.5 mg / L and greater than or equal to 0.1 mg / L, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, or 6 mg / L.

[0031] The fish feed additive can be suitable for feeding to any suitable type of fish. For example, the fish can be a carp, anchovy, tuna, salmon, tilapia, Alaska pollack, catfish, cod, or a combination thereof. The fish can be a salmonid. The salmonid can be any suitable type of salmonid, such as Atlantic salmon, Chinook salmon, Chum salmon, Coho salmon, Masu salmon, Pink salmon, Sockeye salmon, Australian salmon, Danube salmon, Hawaiian salmon, Indian salmon, or a combination thereof. The fish feed additive be for feeding to a salmonids in a low oxygen environment.

[0032] When fed to fish, the fish feed additive can have a beneficial effect on the fish. The beneficial effect can occur when the fish feed additive is fed to fish in normal oxygen conditions or under low oxygen conditions. When fed to fish, such as fish residing in a normal oxygen environment or under a low oxygen environment, the fish feed additive can be effective to improve to maintain overall fish health, growth rate, survival rate, rate of feed consumption, gill efficiency, rate of damaged gill cell repair, gill cell energy production, gill cell energy storage, gill processes including oxygen absorption, gill processes including ionic homeostasis, gill processes including osmoregulation, gill processes including pH control, blood anion gap, blood pH, blood sodium ion concentration, or a combination thereof, as compared to the same fish in the same environment and fed a diet free of the fish feed additive or fed a diet including less of the fish feed additive. When fed to fish residing in a low oxygen environment or in preparation for residing in a low oxygen environment, the fishfeed additive can be effective to reduce or eliminate one or more symptoms of the low oxygen environment on the fish.

[0033] The fish feed additive can be fed to the fish as any suitable proportion of the diet of the fish. For example, the fish feed additive can be for feeding to the fish as 0.01 wt% to 5 wt% of the diet of the fish, 0.05 v % to 0.5 v .%, 0. 1 wt% to 0.2 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% of the diet of the fish.

[0034] Guanadinoacetic acid can be any suitable proportion of the fish feed additive. For example, the guanadinoacetic acid can be 1 wt% to 100 wt% of the fish feed additive, 10 wt% to 100 wt%, 50 wt% to 100 wt%, 90 wt% to 100 wt%, or less than or equal to 100 wt% and greater than or equal to 1 wt%, 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 94, 96, 98, or 99 wt% of the fish feed additive.

[0035] The fish feed additive can further include one or more secondary components. The one or more secondary components can form any suitable proportion of the fish feed additive, such as 0 wt% of the fish feed additive, or such as 0 wt% to 1 wt% of the fish feed additive, or 1 wt% to 99 wt% of the fish feed additive, 1 wt% to 90 wt%, 1 wt% to 50 wt%, 1 wt% to 20 wt%, 0. 1 wt% to 3 wt%, or less than or equal to 90 wt% and greater than or equal to 0.1 wt%, 1, 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, or 85 wt% of the fish feed additive. The fish feed additive can have any suitable weight ratio of the guanadinoacetic acid to the one or more secondary components, such as 99: 1 to 1 :99, 99: 1 to 1 : 1 , or less than or equal to 99: 1 and greater than or equal to 1:99, 1:95, 1:90, 1:85, 1:80, 1:75, 1:70, 1:65, 1:60, 1 :55, 1 :50, 1 :45, 1 :40, 1 :35, 1 :30, 1:25, 1:20, 1: 15, 1: 10, 1:5, 1 :4, 1:3, 1:2, 1 : 1, 2: 1, 3:1, 4: 1, 5: 1, 10: 1, 15: 1, 20: 1, 25: 1, 30: 1, 35: 1, 40: 1, 45: 1, 50: 1, 55:1, 60:1, 65:1, 70: 1, 75: 1, 80: 1, 85: 1, 90: 1, or 95:1. The one or more secondary components can include creatine, phosphocreatine, arginine, or a combination thereof. The one or more secondary components can include arginine. In various aspects, the fish feed additive can include guanadinoacetic acid and arginine, and can be substantially free of other materials. In various aspects, the fish feed additive can include guanadinoacetic acid and a secondary component that is arginine, creatine, and / or phosphocreatine, and can be substantially free of other materials.Fish feed.

[0036] The present invention provides a fish feed that includes guanadinoacetic acid. The fish feed can be suitable for feeding to fish that reside in any environment, such as an environment having normal oxygen levels, or such as an environment having low oxygen levels. The fish feed can include the fish feed additive described herein including guanadinoacetic acid, and a base feed component.

[0037] The fish feed additive can be any suitable proportion of the fish feed. For example, the fish feed additive can be 0.01 wt% to 5 wt% of the fish feed, 0.05 wt% to 0.5 wt%, 0. 1 wt% to 0.2 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% of the fish feed.

[0038] The base feed component can be any suitable base feed. The base feed can be an aquafeed, such as a manufactured or artificial diet to supplement or to replace natural feeds in the aquaculture industry (e.g., the fish or salmonid industry ). These prepared feeds are most commonly produced in flake, pellet, or tablet form. The base feed is composed of several ingredients in various proportions complementing each other to form a nutntionally complete diet for the fish.

[0039] The base feed can include one or more proteins, carbohydrates, fats, fibers, vitamins, minerals, phosphorus sources, or a combination thereof. The base feed can include wheat, wheat flour, soybean meal, fish meal, fish oil, poultry oil, vegetable oil (e g., from soybeans, rapeseeds, sunflower seeds, or flax seeds), cottonseed meal, peanut meal, canola meal, distillers grain with solubles, legume meal, binder attractant, additives, toxin absorbers, or a combination thereof. Additives can include enzymes, growth promoters, heath additives, or a combination thereof. Health additives can include immunostimulants, probiotics, vaccines, or a combination thereof.

[0040] The base feed component can be any suitable proportion of the fish feed. For example, the base feed component can be 95 wt% to 99.99 wt% of the fish feed, 99.5 wt% to 99.95 wt%, or less than or equal to 99.99 wt% and greater than or equal to 95 wt%, 95.5, 96, 96.5, 97, 97.5, 98, 98.5, 99, 99.2, 99.4, 99.6, 99.8, or 99.9 wt% of the fish feed.

[0041] The guanadinoacetic acid of the fish feed additive can be any suitable proportion of the fish feed, such as 0.01 wt% to 5 wt% of the fish feed, 0.05 wt% to 0.5 wt% of the fish feed, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02,0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% of the fish feed.

[0042] If the fish feed additive contains arginine, the arginine from the fish feed additive can form any suitable proportion of the fish feed. For example, arginine in the fish feed additive can be 0.001 wt% to 4 wt% of the fish feed, or 0.01 wt% to 2 wt% of the fish feed, or less than or equal to 4 wt% and greater than or equal to 0.001 wt%, 0.005, 0.01, 0.05, 0.1, 0.2, 0.4, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, 3, 3.2, 3.4, 3.6, or 3.8 wt%.

[0043] If the fish feed additive contains creatine and / or phosphocreatine, the arginine from the fish feed additive can form any suitable proportion of the fish feed. For example, creatine and / or phosphocreatine in the fish feed additive can be 0.001 wt% to 4 wt% of the fish feed, or 0.01 wt% to 2 wt% of the fish feed, or less than or equal to 4 wt% and greater than or equal to 0.001 wt%, 0.005, 0.01, 0.05, 0.1, 0.2, 0.4, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, 3, 3.2, 3.4, 3.6, or 3.8 wt%.

[0044] In various aspects, the fish feed additive is vacuum coated on the base feed component. For example, the vacuum coating can be performed by subjecting the base feed component to a vacuum, adding the fish feed additive thereto (optionally dispersed in a carrier liquid, such as an oil), and releasing the vacuum to cause the fish feed additive to be drawn within the pores of the base feed component. The oil can be any suitable oil, such as fish oil, poultry oil, or a combination thereof. In various aspects, the fish feed additive is combined with a base feed component and the combined mixture is extruded to form the fish feed.

[0045] The fish feed can have any suitable physical form that is palatable to fish. For example, the fish feed can have the form of a flake, tablet, pellet, or a combination thereof.Method of making fish feed.

[0046] In various aspects, the present invention provides a method of forming the fish feed described herein that includes a fish feed additive and a base feed component. The method can be any suitable method that forms the fish feed described herein. For example, the method can include combining the fish feed additive and the base feed component together to form the fish feed. The method can include extruding a mixture of the fish feed additive and the base feed component to form the fish feed. In some aspects, the method includes vacuum coating the fish feed additive onto and into the base feed component. For example, the vacuum coating can be performed by subjecting the base feed component to a vacuum, adding the fish feed additive thereto (optionally dispersed in a carrier liquid, such as an oil), and releasing the vacuum to cause the fish feed additive to be drawn within the pores of the base feed component.Method of feeding fish,

[0047] In various aspects, the present invention provides a method of feeding fish. The method includes feeding the fish the fish feed additive described herein and / or the fish feed described herein. The method can include feeding the fish while the fish reside in water having any suitable oxygen concentration, such as an environment having normal oxygen levels (e.g., having an oxygen saturation of >95% to <100%), or such as an environment having low oxygen levels. Low oxygen levels can include water conditions having an oxygen saturation of 10% to 95%, 30% to 85%, 50% to 70%, or less than or equal to 95% and greater than or equal to 10%, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90%. An environment having normal oxygen levels can have an oxygen concentration of 6.5 mg / L or more, or 5.5 mg / L or more. An environment having low oxygen levels can have an oxygen concentration of 0.1 mg / L to 6.5 mg / L, or 0.1 mg / L to 5.5 mg / L, or 0.1 mg / L to 4 mg / L, or less than or equal to 6.5 mg / L and greater than or equal to 0.1 mg / L, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, or 6 mg / L. The method can include feeding the fish in preparation for the fish to reside in water having such an oxygen concentration, such as 1-7 days ahead of time, 1-4 weeks ahead of time, or 1-6 months ahead of time.

[0048] During performance of the method, the fish feed additive can form any suitable proportion of the diet of the fish. For example, the fish feed additive can be 0.01 wt% to 5 wt% of a diet of the fish, or 0.05 wt% to 0.5 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16,0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% ofthe diet of the fish.

[0049] The fish can be any suitable fish. The fish can be a carp, anchovy, tuna, salmon, tilapia, Alaska pollack, catfish, cod, or a combination thereof. The fish can be a salmonid. The salmonid can be any suitable type of salmonid, such as Atlantic salmon, Chinook salmon, Chum salmon, Coho salmon, Masu salmon, Pink salmon, Sockeye salmon, Australian salmon, Danube salmon, Hawaiian salmon, Indian salmon, or a combination thereof

[0050] The method can have a beneficial effect on the fish. The beneficial effect can occur when the fish feed additive is fed to fish in normal oxygen conditions or under low oxygen conditions. For fish residing in a normal oxygen environment or under a low oxygen environment, the method can be effective to improve to maintain overall fish health, growth rate, survival rate, rate of feed consumption, gill efficiency, rate of damaged gill cell repair, gill cell energy production, gill cell energy storage, gill processes including oxygen absorption, gill processes including ionic homeostasis, gill processes including osmoregulation, gill processes including pH control, blood anion gap, blood pH, blood sodium ion concentration, or a combination thereof, as compared to the same fish in the same environment and fed a diet free of the fish feed additive or fed a diet including less of the fish feed additive. For fish residing in a low oxygen environment or in preparation for residing in a low oxygen environment, the method can be effective to reduce or eliminate one or more symptoms of the low oxygen environment on the fish.Fish feed additive including arginine, fish feed including same, and method of feeding fish the fish feed additive and / or the fish feed.

[0051] In various aspects, the present invention provides a fish feed additive for feeding to fish such as salmonids. The fish can reside in an aqueous environment having normal oxygen levels or such as an aqueous environment having low oxygen levels. Low oxygen levels can include water conditions having an oxygen saturation of 10% to 95%, 30% to 85%, 50% to 70%, or less than or equal to 95% and greater than or equal to 10%, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90%. An environment having normal oxygen levels can have an oxygen concentration of 6.5 mg / L or more, or 5.5 mg / L or more. An environment having low oxygen levels can have an oxygen concentration of 0. 1 mg / L to 6.5 mg / L, or 0.1 mg / L to 5.5 mg / L, or 0.1 mg / L to 4 mg / L, or less than or equal to 6.5 mg / L and greater than or equal to 0.1 mg / L, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, or 6 mg / L. The fish feed additive can include arginine. For example, arginine can be any suitable proportion of the fish feed additive, such as 1 wt% to 100 wt% of the fish feed additive, 1 wt% to 90 wt%, 1 wt% to 50 wt%, 1 wt% to 20 wt%, or less than or equal to 90 wt% and greater than or equal to 1 wt%, 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 94, 96, 97, or 98 wt% of the fish feed additive. The fish feed additive can be for feeding to fish at any suitable concentration of their diet, such wherein the fish feed additive is 0.01 wt% to 5 wt% of the diet of the fish, 0.05 wt% to 0.5 wt%, 0. 1 wt% to 0.2 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1 , 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% of the diet of the fish.

[0052] The present invention provides a fish feed for feeding to fish such as salmonids, such as for feeding to fish in a low oxygen environment. The fish feed includes the fish feed additive that includes arginine. The fish feed also includes a base feed component. The fish feed can include any suitable amount of arginine, such as arginine at a concentration of 0.01 wt% to 5 wt% of fish feed, 0.05 wt% to 0.5 wt%, 0. 1 wt% to 0.2 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% of the fish feed

[0053] In various aspects, the present invention provides a method of feeding fish such as salmonids, such wherein the fish reside in a low oxygen environment. The method includes feeding the salmonids residing in the low oxygen environment the fish feed of thepresent invention that includes arginine, or the fish feed additive of the present invention that includes arginine, or a combination thereof. The method can provide arginine at any suitable concentration in the diet of the fish, such as at a concentration of 0.01 wt% to 5 wt% of the diet of the fish, 0.05 wt% to 0.5 wt%, 0.1 wt% to 0.2 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% of the diet of the fish.Fish feed additive including creatine and / or phosphocreatine, fish feed including same, and method of feeding fish the fish feed additive and / or the fish feed.

[0054] In various aspects, the present invention provides a fish feed additive for feeding to fish such as salmonids. The fish can reside in an aqueous environment having normal oxygen levels or such as an aqueous environment having low oxygen levels. Low oxygen levels can include water conditions having an oxygen saturation of 10% to 95%, 30% to 85%, 50% to 70%, or less than or equal to 95% and greater than or equal to 10%, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90%. An environment having normal oxygen levels can have an oxygen concentration of 6.5 mg / L or more, or 5.5 mg / L or more. An environment having low oxygen levels can have an oxygen concentration of 0. 1 mg / L to 6.5 mg / L, or 0.1 mg / L to 5.5 mg / L, or 0.1 mg / L to 4 mg / L, or less than or equal to 6.5 mg / L and greater than or equal to 0.1 mg / L, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, or 6 mg / L. The fish feed additive can include creatine and / or phosphocreatine. For example, creatine and / or phosphocreatine can be any suitable proportion of the fish feed additive, such as 1 wt% to 100 wt% of the fish feed additive, 1 wt% to 90 wt%, 1 wt% to 50 wt%, 1 wt% to 20 wt%, or less than or equal to 90 wt% and greater than or equal to 1 wt%, 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 94, 96, 97, or 98 wt% of the fish feed additive. The fish feed additive can be for feeding to fish at any suitable concentration of their diet, such wherein the fish feed additive is 0.01 wt% to 5 wt% of the diet of the fish, 0.05 wt% to 0.5 wt%, 0. 1 wt% to 0.2 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% of the diet of the fish.

[0055] The present invention provides a fish feed for feeding to fish such as salmonids, such as for feeding to fish in a low oxygen environment. The fish feed includes the fish feed additive that includes creatine and / or phosphocreatine. The fish feed alsoincludes a base feed component. The fish feed can include any suitable amount of creatine and / or phosphocreatine, such as creatine and / or phosphocreatine at a concentration of 0.01 wt% to 5 wt% of fish feed, 0.05 wt% to 0.5 wt%, 0.1 wt% to 0.2 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 v .% of the fish feed.In various aspects, the present invention provides a method of feeding fish such as salmonids, such wherein the fish reside in a low oxygen environment. The method includes feeding the salmonids residing in the low oxygen environment the fish feed of the present invention that includes creatine and / or phosphocreatine, or the fish feed additive of the present invention that includes creatine and / or phosphocreatine, or a combination thereof. The method can provide creatine and / or phosphocreatine at any suitable concentration in the diet of the fish, such as at a concentration of 0.01 wt% to 5 wt% of the diet of the fish, 0.05 wt% to 0.5 wt%, 0. 1 wt% to 0.2 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% of the diet of the fish.

[0056] In various aspects, the present invention provides a method of feeding fish such as salmonids, such wherein the fish reside in a low oxygen environment. The method includes feeding the salmonids residing in the low oxygen environment the fish feed of the present invention that includes creatine and / or phosphocreatine, or the fish feed additive of the present invention that includes creatine and / or phosphocreatine, or a combination thereof. The method can provide creatine and / or phosphocreatine at any suitable concentration in the diet of the fish, such as at a concentration of 0.01 wt% to 5 wt% of the diet of the fish, 0.05 wt% to 0.5 wt%, 0.1 wt% to 0.2 wt%, or less than or equal to 5 wt% and greater than or equal to 0.01 wt%, 0.02, 0.04, 0.06, 0.08, 0.10, 0.12, 0.14, 0.15, 0.16, 0.18, 0.20, 0.25, 0.3, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.5, 3, 3.5, 4, or 4.5 wt% of the diet of the fish.Examples

[0057] Various aspects of the present invention can be better understood by reference to the following Examples which are offered by way of illustration. The present invention is not limited to the Examples given herein.Methods.

[0058] The trial was conducted at Cargill Innovation center, Colaco (CIC) with fish having been produced on site. Atlantic salmon (Salmo salar) with an approximate weight of 90 g were distributed into 9 tanks of 500 L filtered sea water (32 ppm NaCl, 11 °C, 24 h light). 142 fish were allocated per tank, to produce a starting biomass of 11 kg / m3. Fish were acclimatized for 28 days with the Saturation of O2 being reduced gradually (100% O2 sat. - 60% O2 sat.) as well as flow reduced (10.5 L / min-7.2 L / min) for 6 tanks. The remaining 3 tanks were maintained at 100% O2 sat, together with a 10.5 L / min flow rate. All tanks during the acclimatization period were fed a standard Cargill commercial diet (Micro G 100- caliber 100 (4mm)) using an SFR calculated with the EGI electronic resource (EWOS Growth indicator).

[0059] Post acclimatization, all fish per tank were anesthetized (1.5 mL benzocaine / 10 L) and weight and length recorded. 10 fish per tank were removed for lethal sampling (4.5 mL Benzocaine / 10 L). Two 0.5 mL blood samples were removed per fish for blood parameter analysis. The first sample was analyzed in-situ (i-STAT, Abbott, Abbott park, IL, USA) and second sent for external biochemical analysis (VeHiCe, Puerto Montt, Chile). The second gill arch, from the right-hand side of the fish was removed and stored (10% buffered formalin) for later histological analysis, and the left second gill arch was removed and stored (rapid freezing -80 °C with liquid N2) for later qPCR analysis.

[0060] All tanks then began a period of “experimental feeding” with 3 of the 9 tanks being fed a commercial Cargill diet (Micro GlOO-caliber 100 (4mm)), with the inclusion of Creamino® (Guanadinoacetic acid) (Alzchem, Trosberz, Germany), at 0. 15% w / w (added to dry Micro G100 pellet via oil (Fish) top vacuum coating - Forberg Vacuum coater, Norway). The remaining tanks were fed standard Micro G100 diet -caliber 100 (4 mm). All tanks were fed according to an SFR calculated using the EGI electronic resource. After 28 days, weight and length of all fish was recorded, with 10 fish being removed per tank for lethal sampling, in the manner described previously.

[0061] Environmental and feeding conditions were maintained for a further 28 days, after which an identical sampling was conducted, with all fish then being euthanized (4.5 mL Benzocaine / 10 L), and end of trial declared.Results: Productivity data.

[0062] Results showed that compared to standard Cargill Micro G feed at 60% O2 sat., the addition of CreAmino® at 0.15% w / w did not cause an improvement or reduction infish grow th or condition when measured both at 28 days and 56 days after beginning the experimental feeding period. As discussed below, the use of CreAmino alone did cause an improvement in certain blood parameters that may be beneficial for fish survival and wellbeing at low O2 conditions. Preliminary results using the same base feed but with 3 wt% total arginine in the feed instead of CreAmino caused a reduction in modelled growth, but also caused an improvement in certain blood parameters that may be beneficial for fish survival and well-being at low O2 conditions.Results: Blood parameters.

[0063] Blood parameter analysis showed that compared to fish fed standard Cargill Micro G feed at 60% O2 sat. and 100% O2 sat., the addition of CreAmino® at 0.15% w / w caused a pattern of blood biochemistry measurements that were similar to those of fish subject to 100% O2 sat. and fed standard Cargill Micro G feed, and dissimilar to fish fed standard Cargill Micro G feed exposed to an environment of 60% O2. This was particularly noted in anion gap, pH, and electrolyte balance measurements, all of which are known to be controlled by gills. Differences in these parameters were seen at both 28 days post beginning experimental feeding with other parameters being seen at 56 days after beginning experimental feeding.

[0064] FIG. 1 illustrates modelled weight gain of fish subjected to various feed / Ch conditions, in accordance with various aspects. FIG. 2 illustrates the anion gap of blood of fish subjected to various feed / Ch conditions, in accordance with various aspects. FIG. 3 illustrates blood pH of fish subjected to various feed / Ch conditions, in accordance with various aspects. FIG. 4 illustrates blood sodium ion concentration of fish subjected to various feed / 02 conditions, in accordance with various aspects.

[0065] The terms and expressions that have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features show n and described or portions thereof, but it is recognized that various modifications are possible within the scope of the aspects of the present invention. Thus, it should be understood that although the present invention has been specifically disclosed by specific aspects and optional features, modification and variation of the concepts herein disclosed may be resorted to by those of ordinary skill in the art, and that such modifications and variations are considered to be within the scope of aspects of the present invention.Exemplary Aspects.

[0066] The following exemplary aspects are provided, the numbering of which is not to be construed as designating levels of importance:

[0067] Aspect 1 provides a fish feed additive comprising: guanadinoacetic acid.

[0068] Aspect 2 provides the fish feed additive of Aspect 1, wherein the fish feed additive is for feeding to fish residing in a low oxygen environment or in preparation for residing in a low oxygen environment.

[0069] Aspect 3 provides the fish feed additive of Aspect 2, wherein the fish residing in the low oxygen environment comprise salmonid fish.

[0070] Aspect 4 provides the fish feed additive of any one of Aspects 2-3, wherein the low oxygen environment has an oxygen saturation of 10% to 95%.

[0071] Aspect 5 provides the fish feed additive of any one of Aspects 2-4, wherein the low oxygen environment has an oxygen saturation of 30% to 85%.

[0072] Aspect 6 provides the fish feed additive of any one of Aspects 2-5, wherein the low oxygen environment has an oxygen saturation of 50% to 70%.

[0073] Aspect 7 provides the fish feed additive of any one of Aspects 1-6, wherein when fed to fish, the fish feed additive is effective to improve to maintain overall fish health, growth rate, survival rate, rate of feed consumption, gill efficiency, rate of damaged gill cell repair, gill cell energy production, gill cell energy storage, gill processes including oxygen absorption, gill processes including ionic homeostasis, gill processes including osmoregulation, gill processes including pH control, blood anion gap, blood pH, blood sodium ion concentration, or a combination thereof, as compared to the same fish in the same environment and fed a diet free of the fish feed additive.

[0074] Aspect 8 provides the fish feed additive of any one of Aspects 1-7, wherein when fed to fish residing in a low oxygen environment or in preparation for residing in a low oxygen environment, the fish feed additive is effective to reduce or eliminate one or more symptoms of the low oxygen environment on the fish.

[0075] Aspect 9 provides the fish feed additive of any one of Aspects 1-8, wherein when fed to fish residing in a low oxygen environment or in preparation for residing in a low oxy gen environment, the fish feed additive is effective to maintain or improve overall fish health, growth rate, survival rate, rate of feed consumption, gill efficiency, rate of damagedgill cell repair, gill cell energy production, gill cell energy storage, gill processes including oxygen absorption, gill processes including ionic homeostasis, gill processes including osmoregulation, gill processes including pH control, blood anion gap, blood pH, blood sodium ion concentration, or a combination thereof, as compared to the same fish in the same environment and fed a diet free of the fish feed additive.

[0076] Aspect 10 provides the fish feed additive of any one of Aspects 1-9, wherein the fish feed additive is for feeding to carp, anchovy , tuna, salmon, tilapia, Alaska pollack, catfish, cod, or a combination thereof

[0077] Aspect 11 provides the fish feed additive of any one of Aspects 1-10, wherein the fish feed additive is for feeding to salmonid fish.

[0078] Aspect 12 provides the fish feed additive of any one of Aspects 1-11, wherein the fish feed additive is for feeding to a salmonid fish comprising Atlantic salmon, Chinook salmon, Chum salmon, Coho salmon, Masu salmon, Pink salmon, Sockeye salmon, Australian salmon, Danube salmon, Hawaiian salmon, Indian salmon, or a combination thereof.

[0079] Aspect 13 provides the fish feed additive of any one of Aspects 1-12, wherein the fish feed additive is for feeding to fish as 0.01 wt% to 5 wt% of a diet of the fish.

[0080] Aspect 14 provides the fish feed additive of any one of Aspects 1-13, wherein the fish feed additive is for feeding to fish as 0.05 wt% to 0.5 wt% of a diet of the fish.

[0081] Aspect 15 provides the fish feed additive of any one of Aspects 1-14, wherein the fish feed additive is for feeding to fish as 0. 1 wt% to 0.2 wt% of a diet of the fish.

[0082] Aspect 16 provides the fish feed additive of any one of Aspects 1-15, wherein the guanadinoacetic acid is 1 wt% to 100 wt% of the fish feed additive.

[0083] Aspect 17 provides the fish feed additive of any one of Aspects 1-16, wherein the guanadinoacetic acid is 90 wt% to 100 wt% of the fish feed additive.

[0084] Aspect 18 provides the fish feed additive of any one of Aspects 1-17, further comprising one or more secondary components.

[0085] Aspect 19 provides the fish feed additive of Aspect 18, wherein the one or more secondary components are 1 wt% to 90 wt% of the fish feed additive.

[0086] Aspect 20 provides the fish feed additive of any one of Aspects 18-19, wherein the one or more secondary components are 1 wt% to 50 wt% of the fish feed additive.

[0087] Aspect 21 provides the fish feed additive of any one of Aspects 18-20, wherein the one or more secondary components are 1 wt% to 20 wt% of the fish feed additive.

[0088] Aspect 22 provides the fish feed additive of any one of Aspects 18-21, wherein the fish feed additive has a weight ratio of the guanadinoacetic acid to the one or more secondary components of 99: 1 to 1:99.

[0089] Aspect 23 provides the fish feed additive of any one of Aspects 18-22, wherein the fish feed additive has a weight ratio of the guanadinoacetic acid to the one or more secondary components of 99: 1 to 1: 1.

[0090] Aspect 24 provides the fish feed additive of any one of Aspects 18-23, wherein the secondary component comprises creatine, phosphocreatine, arginine, or a combination thereof.

[0091] Aspect 25 provides the fish feed additive of any one of Aspects 18-24, wherein the secondary component comprises arginine.

[0092] Aspect 26 provides the fish feed additive of Aspect 25, wherein the fish feed additive has a weight ratio of the guanadinoacetic acid to arginine of 99: 1 to 1 :99.

[0093] Aspect 27 provides the fish feed additive of any one of Aspects 25-26, wherein the fish feed additive has a weight ratio of the guanadinoacetic acid to arginine of 99:1 to 1 : 1.

[0094] Aspect 28 provides a fish feed additive comprising: guanadinoacetic acid that is 50 wt% to 99 wt% of the fish feed additive; and arginine that is 1 wt% to 50 wt% of the fish feed additive.

[0095] Aspect 29 provides a fish feed comprising: the fish feed additive of any one of Aspects 1-28; and a base feed component.

[0096] Aspect 30 provides the fish feed of Aspect 29, wherein the fish feed additive is 0.01 wt% to 5 wt% of the fish feed.

[0097] Aspect 31 provides the fish feed of any one of Aspects 29-30, wherein the fish feed additive is 0.05 wt% to 0.5 wt% of the fish feed.

[0098] Aspect 32 provides the fish feed of any one of Aspects 29-31, wherein the fish feed additive is 0. 1 wt% to 0.2 wt% of the fish feed.

[0099] Aspect 33 provides the fish feed of any one of Aspects 29-32, wherein the base feed component is 95 wt% to 99.99 wt% of the fish feed.

[0100] Aspect 34 provides the fish feed of any one of Aspects 29-33, wherein the base feed component is 99.5 wt% to 99.95 wt% of the fish feed.

[0101] Aspect 35 provides the fish feed of any one of Aspects 29-34, wherein the guanadinoacetic acid of the fish feed additive is 0.01 wt% to 5 wt% of the fish feed.

[0102] Aspect 36 provides the fish feed of any one of Aspects 29-35, wherein the guanadinoacetic acid of the fish feed additive is 0.05 wt% to 0.5 wt% of the fish feed.

[0103] Aspect 37 provides the fish feed of any one of Aspects 29-36, wherein arginine from the fish feed additive is 0.001 wt% to 4 wt% of the fish feed.

[0104] Aspect 38 provides the fish feed of any one of Aspects 29-37, wherein arginine from the fish feed additive is 0.01 wt% to 2 wt% of the fish feed.

[0105] Aspect 39 provides the fish feed of any one of Aspects 29-38, wherein the fish feed additive is vacuum coated on the base feed component, or wherein a mixture of the fish feed additive and the base feed component are extruded.

[0106] Aspect 40 provides the fish feed of any one of Aspects 29-39, wherein a mixture of the fish feed additive and a carrier liquid is vacuum coated on the base feed component.

[0107] Aspect 41 provides the fish feed of Aspect 40, wherein the carrier liquid comprises an oil.

[0108] Aspect 42 provides the fish feed of any one of Aspects 29-41, wherein the base feed component comprises proteins, carbohydrates, fat, fiber, vitamins, minerals, a phosphorus source, or a combination thereof.

[0109] Aspect 43 provides the fish feed of any one of Aspects 29-42, wherein the fish feed has the form of a flake, tablet, pellet, or a combination thereof.

[0110] Aspect 43.5 provides a fish feed comprising: guanadinoacetic acid that is 0.01 wt% to 5 wt% of the fish feed; and a base feed component.

[0111] Aspect 44 provides a fish feed comprising: a fish feed additive that is 50 wt% to 100 wt% guanadinoacetic acid, wherein the fish feed additive is 0.01 wt% to 5 wt% of the fish feed; and a base feed component

[0112] Aspect 45 provides a method of forming the fish feed of any one of Aspects 29-44, the method comprising:adding the fish feed additive of any one of Aspects 1-28 to the base feed component to form the fish feed.

[0113] Aspect 46 provides the method of Aspect 45, wherein adding the fish feed additive to the base feed component comprises vacuum coating an oil composition comprising the fish feed additive onto the base feed component, or wherein adding the fish feed additive to the base feed component comprises forming a mixture of the fish feed additive and the base feed component and the method further comprises extruding the mixture to form the fish feed.

[0114] Aspect 47 provides a method of feeding fish, the method comprising: feeding the fish the fish feed additive of any one of Aspects 1-28 and / or the fish feed of any one of Aspects 29-44.

[0115] Aspect 48 provides the method of Aspect 47, wherein the fish reside in an environment having an oxygen saturation of 10% to 95%.

[0116] Aspect 49 provides the method of any one of Aspects 47-48, wherein the fish reside in an environment having an oxygen saturation of 30% to 85%.

[0117] Aspect 50 provides the method of any one of Aspects 47-49, wherein the fish reside in an environment having an oxygen saturation of 50% to 70%.

[0118] Aspect 51 provides the method of any one of Aspects 47-50, wherein the fish feed additive is 0.01 wt% to 5 wt% of a diet of the fish.

[0119] Aspect 52 provides the method of any one of Aspects 47-51, wherein the fish feed additive is 0.05 wt% to 0.5 wt% of a diet of the fish.

[0120] Aspect 53 provides the method of any one of Aspects 47-52, wherein the fish are carp, anchovy, tuna, salmon, tilapia, Alaska pollack, catfish, cod, or a combination thereof.

[0121] Aspect 54 provides the method of any one of Aspects 47-53, wherein the fish are salmonids.

[0122] Aspect 55 provides the method of any one of Aspects 47-54, wherein the method is effective to reduce or eliminate one or more symptoms of the low oxygen environment on the fish.

[0123] Aspect 56 provides the method of any one of Aspects 47-55, wherein the method is effective to improve to maintain overall fish health, growth rate, survival rate, rate of feed consumption, gill efficiency, rate of damaged gill cell repair, gill cell energy production, gill cell energy' storage, gill processes including oxygen absorption, gill processesincluding ionic homeostasis, gill processes including osmoregulation, gill processes including pH control, blood anion gap, blood pH, blood sodium ion concentration, or a combination thereof, as compared to the same fish in the same environment and fed a diet free of the fish feed additive.

[0124] Aspect 57 provides a method of feeding salmonid fish, the method comprising: feeding the salmonid fish a feed composition comprising a base feed and comprising a feed additive that is 0.01 wt% to 5 wt% of the fish feed, wherein the feed additive is 50 wt% to 100 wt% guanadinoacetic acid; wherein the salmonid fish reside in an environment having an oxygen saturation of 30% to 85%, and wherein the method is effective to improve to maintain overall fish health, growth rate, survival rate, rate of feed consumption, gill efficiency, rate of damaged gill cell repair, gill cell energy production, gill cell energy storage, gill processes including oxygen absorption, gill processes including ionic homeostasis, gill processes including osmoregulation, gill processes including pH control, blood anion gap, blood pH, blood sodium ion concentration, or a combination thereof, as compared to the same fish in the same environment and fed a diet free of the fish feed additive.

[0125] Aspect 58 provides a fish feed additive for feeding to salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment, the fish feed additive comprising arginine.

[0126] Aspect 59 provides a fish feed for feeding to salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment, the fish feed comprising: the fish feed additive of Aspect 58; and a base feed component.

[0127] Aspect 60 provides a method of feeding salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment, the method comprising: feeding the salmonids residing in the low oxygen environment the fish feed of Aspect 59, the fish feed additive of Aspect 58, or a combination thereof.

[0128] Aspect 61 provides a fish feed additive for feeding to salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment, the fish feed additive comprising creatine and / or phosphocreatine.

[0129] Aspect 62 provides a fish feed for feeding to salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment, the fish feed comprising: the fish feed additive of Aspect 61; and a base feed component

[0130] Aspect 63 provides a method of feeding salmonids residing in a low oxygen environment or in preparation for residing in a low oxygen environment, the method comprising: feeding the salmonids residing in the low oxygen environment the fish feed of Aspect 61, the fish feed additive of Aspect 62, or a combination thereof.

[0131] Aspect 61 provides the fish feed additive, fish feed, method of forming the fish feed, or method of feeding fish of any one or any combination of Aspects 1-60 optionally configured such that all elements or options recited are available to use or select from.

Claims

CLAIMSWhat is claimed is:

1. A fish feed additive comprising: guanadinoacetic acid.

2. The fish feed additive of claim 1, wherein when fed to fish residing in a low oxygen environment or in preparation for residing in the low oxygen environment having an oxygen saturation of 10% to 95%, the fish feed additive is effective to reduce or eliminate one or more symptoms of the low oxygen environment on the fish.

3. The fish feed additive of claim 1, wherein the guanadinoacetic acid is 10 wt% to 100 wt% of the fish feed additive.

4. The fish feed additive of claim 1, further comprising one or more secondary components, wherein the one or more secondary components are 1 wt% to 90 wt% of the fish feed additive.

5. The fish feed additive of claim 4, wherein the secondary component comprises arginine, creatine, phosphocreatine, or a combination thereof.6 A fish feed comprising: the fish feed additive of claim 1 ; and a base feed component.

7. The fish feed of claim 6, wherein the fish feed additive is 0.01 wt% to 5 wt% of the fish feed.

8. The fish feed of claim 6, wherein the base feed component is 95 wt% to 99.99 wt% of the fish feed.

9. The fish feed of claim 6, wherein arginine, creatine, phosphocreatine, or a combination thereof, from the fish feed additive is 0.001 wt% to 4 wt% of the fish feed.

10. The fish feed of claim 6, wherein the fish feed additive is vacuum coated on the base feed component, or wherein the fish feed is an extruded mixture of the fish feed additive and the base feed component.

11. The fish feed of claim 6, wherein the fish feed is for feeding to salmonids residing in a low' oxygen environment or in preparation for residing in the low oxygen environment12. A method of forming the fish feed of claim 6, the method comprising: combining the fish feed additive of claim 1 and the base feed component to form the fish feed.

13. A method of feeding fish, the method comprising: feeding the fish the fish feed additive of claim 1 and / or the fish feed of claim 6.

14. The method of claim 13, wherein the fish feed additive is 0.01 wt% to 5 wt% of a diet of the fish.

15. The method of claim 13, wherein the fish are salmonids.

16. The method of claim 13, wherein the fish reside in, or are preparing to reside in, a low oxygen environment having an oxygen saturation of 30% to 85%17. The method of claim 16, wherein the method is effective to reduce or eliminate one or more symptoms of the low oxygen environment on the fish.

18. The method of claim 13, wherein: the fish is a salmonid; feeding the fish comprises feeding the salmonid a feed composition comprising a base feed and comprising a feed additive that is 0.01 wt% to 5 wt% of the fish feed, wherein the feed additive is 50 wt% to 100 wt% guanadinoacetic acid; the salmonid resides in, or is preparing to reside in, an environment having an oxygen saturation of 30% to 85%; andthe method is effective to improve to maintain overall fish health, growth rate, survival rate, rate of feed consumption, gill efficiency, rate of damaged gill cell repair, gill cell energy production, gill cell energy' storage, gill processes including oxygen absorption, gill processes including ionic homeostasis, gill processes including osmoregulation, gill processes including pH control, blood anion gap, blood pH, blood sodium ion concentration, or a combination thereof, as compared to the same salmonid in the same environment and fed a diet free of the fish feed additive.

19. A method of feeding salmonids residing in a low oxygen environment or in preparation for residing in the low oxygen environment, the method comprising: feeding the salmonids a fish feed comprising arginine, a fish feed additive comprising arginine, or a combination thereof.

20. A method of feeding salmonids residing in a low oxygen environment or in preparation for residing in the low oxygen environment, the method comprising: feeding the salmonids a fish feed comprising creatine and / or phosphocreatme, a fish feed additive comprising creatine and / or phosphocreatine, or a combination thereof.

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