Feed compositions containing astaxanthin and methods for making and using the same

Abstract

A feed additive pack includes an antioxidant component with astaxanthin dispersed within the antioxidant component. A fish feeds includes a base feed and a feed additive pack. The antioxidant component comprising particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers. Methods of making feed additive packs and fish feeds, and methods of feeding aquatic animals are also provided.

Description

FEED COMPOSITIONS CONTAINING ASTAXANTHIN AND METHODS FOR MAKING AND USING THE SAMECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 640,991, filed May 1, 2024, the disclosure of which is incorporated by reference herein in its entirety.FIELD

[0002] The present disclosure generally relates to aquaculture feeds, and specifically fish feeds. In particular, the present disclosure relates fish feeds containing astaxanthin and to compositions and methods for making such fish feeds.INTRODUCTION

[0003] Aquaculture of various fish species provides a food source for a growing world population. In 2021, it was estimated that about 175 million metric tons of fish were produced for global consumption. An increase in demand for fish by consumers has resulted in an increase in the number of aquaculture farms around the world. With an increase in aquaculture farms comes an increased need for fish feeds to nourish and maintain the volume of fish being produced and for sustainable technologies for safe and effective pest control in farmed fish populations.

[0004] Astaxanthin is a carotenoid compound included in fish feed, particularly for salmon, to improve the appearance (e.g., color) of the muscle tissue. Since the restrictions on the use of ethoxyquin, the main synthetic antioxidant that was used in fish feed, an increase in the degradation of astaxanthin has been observed during both the manufacturing and the storage of the feed. Although other antioxidants are available on the market, they do not exhibit the same functionality.

[0005] Astaxanthin is susceptible to degradation during processing and / or storage of feed ingredients and the feed. Decrease of astaxanthin in feed ingredients and feed leads to less astaxanthin being available for the fish, resulting in sub-optimal flesh color. The cost of astaxanthin is high and, therefore, preservation of the added astaxanthin in feed ingredients and in feed is desired.SUMMARY

[0006] The present disclosure relates to aquaculture feeds, and specifically fish feeds. In particular, the present disclosure relates to fish feeds including feed additive packs containing astaxanthin, and to compositions and methods for making and using such fish feeds. Theastaxanthin is provided as part of a feed additive pack that may be mixed with a base feed to prepare the fish feed.

[0007] In some embodiments, the feed additive packs include an antioxidant component with astaxanthin dispersed within the antioxidant component. In some embodiments, the fish feeds include a base feed and a feed additive pack. The feed additive pack includes an antioxidant component with astaxanthin dispersed within. The antioxidant component comprising particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers.

[0008] In some embodiments, the antioxidant component in the feed additive packs and fish feeds includes a first particle size population with a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and a second particle size population with a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers. Additionally, D90F is smaller than D90c, and D50F is smaller than D50c.

[0009] In some embodiments, methods for making feed additive packs for adding to a fish feed include grinding an antioxidant mixture to particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers. The methods include mixing the antioxidant mixture particles with particles containing astaxanthin.

[0010] In some embodiments, methods for making feed additive packs for adding to a fish feed include grinding a first amount of an antioxidant mixture to fine particles having a first particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers and grinding a second amount of the antioxidant mixture to coarse particles having a second particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers such that D90F is smaller than D90c and D50F is smaller than D50c. The methods include mixing the coarse particles, the fine particles, and particles containing astaxanthin.

[0011] In some embodiments, the methods for making fish feeds include mixing a feed additive pack with a base feed, where the feed additive pack includes an antioxidant component with astaxanthin dispersed within. The antioxidant component in the feed additive packs and fish feeds includes particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers. The antioxidant component in the feed additive packs and fish feeds may include a first particle size population with a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and a second particle size population with a particle size volume distribution with D50c in a range of 30 to 90micrometers and D90c in a range of 150 to 750 micrometers. Additionally, D90F is smaller than D90c, and D50F is smaller than D50c.

[0012] The antioxidant component particles in the feed additive packs, the fish feeds, the methods for making feed additive packs, and the methods for making fish feeds may have a D80 of 140 micrometers or greater, 145 micrometers or greater, 150 micrometers or greater, 155 micrometers or greater, or 165 micrometers or greater. The D80 of the antioxidant component particles may be 200 micrometers or less, 190 micrometers or less, 185 micrometers or less, or 180 micrometers or less. The antioxidant component particles may have a D10 in a range of 5 to 20 micrometers, 6 to 15 micrometers, or 8 to 12 micrometers. The antioxidant component particles may have a D50 in a range of 40 to 80 micrometers, 45 to 70 micrometers, or 48 to 65 micrometers. The antioxidant component particles may have a D90 in a range of 200 to 400 micrometers, 210 to 350 micrometers, or 220 to 325 micrometers.

[0013] The first particle size population in the feed additive packs, the fish feeds, the methods for making feed additive packs, and the methods for making fish feeds may have a DI OF in a range of 1 to 20 micrometers, a D50F in a range of 35 to 65 micrometers, and a D90F in a range of 120 to 180 micrometers. The second particle size population in the feed additive packs, the fish feeds, the methods for making feed additive packs, and the methods for making fish feeds may have a DIOc in a range of 2 to 20 micrometers, a D50c in a range of 45 to 75 micrometers, and a D90c in a range of 180 to 400 micrometers. The weight ratio of the first particle size population and the second particle size population in the feed additive packs, the fish feeds, the methods for making feed additive packs, and the methods for making fish feeds may be from 0.3 to 3 parts of the first particle size population to every 1 part of the second particle size population.

[0014] In some embodiments, the feed additive packs, the fish feeds, the methods for making feed additive packs, and the methods for making fish feeds may include astaxanthin in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-% and a particle size D90 in a range of about 1 to 10 micrometers. In some embodiments, the feed additive packs, the fish feeds, the methods for making feed additive packs, and the methods for making fish feeds may maintain at least 85 %, at least 90 %, or at least 95 % of the astaxanthin added to the feed additive pack. In some embodiments, the feed additive packs, the fish feeds, the methods for making feed additive packs, and the methods for making fish feeds may maintain 58 % or more of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0015] In some embodiments, the methods of using fish feeds include feeding an aquatic animal by administering an amount of feed to the aquatic animal. The fish feeds include a basefeed and a feed additive pack. The feed additive pack includes an antioxidant component with astaxanthin dispersed within. The antioxidant component in the feed additive packs and fish feeds includes particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers. The antioxidant component may include a first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and a second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers. Additionally, D90F is smaller than D90c, and D50F is smaller than D50c. In some embodiments, the first particle size population may have a DI OF in a range of 1 to 20 micrometers, a D50F in a range of 35 to 65 micrometers, and a D90F in a range of 120 to 180 micrometers. In some embodiments, the second particle size population may have a D 10c in a range of 2 to 20 micrometers, a D50c in a range of 45 to 75 micrometers, and a D90c in a range of 180 to 400 micrometers. In some embodiments, the methods of using fish feeds may include astaxanthin in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-% and a particle size D90 in a range of about 1 to 10 micrometers. In some embodiments, the methods of using fish feeds may maintain at least 85 % of the astaxanthin added to the feed additive pack. In some embodiments, the methods of using fish feeds may maintain 58 % or more of the astaxanthin during processing and storage for at least 3 weeks (21 days).BRIEF DESCRIPTION OF FIGURES

[0016] The drawings illustrate generally, by way of example, but not by way of limitation, various embodiments discussed herein.

[0017] FIG. 1 is a graphical representation of the results of Example 1.

[0018] FIG. 2 is a graphical representation of the results of Example 2.

[0019] It should be understood that numerous other modifications and examples can be devised by those skilled in the art, which fall within the scope and spirit of the principles of this disclosure.DEFINITIONS

[0020] All scientific and technical terms used herein have meanings commonly used in the art unless otherwise specified. The definitions provided herein are to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

[0021] In reference to particle size, the term “particle size volume distribution” is used to indicate particle size distribution measured by volume. Particle size may be measured using anysuitable apparatus, such as the Mastersizer 3000 laser diffraction analyzer, available from Malvern Panalytical Ltd in Malvern, United Kingdom. The term D[X] is used to indicate the size at which X % of the sample (by volume) is smaller than the size. For example, the term DIO is used to indicate the size at which 10 % of the sample (by volume) is smaller than the size; the term D50 is used to indicate the size at which 50 % of the sample (by volume) is smaller than the size; and the term D90 is used to indicate the size at which 90 % of the sample (by volume) is smaller than the size.

[0022] The term “about” is used here in conjunction with numeric values to include normal variations in measurements as expected by persons skilled in the art, and is understood to have the same meaning as “approximately” and to cover a typical margin of error, such as ±5 % of the stated value.

[0023] Terms such as “a,” “an,” and “the” are not intended to refer to only a singular entity, but include the general class of which a specific example may be used for illustration.

[0024] The terms “a,” “an,” and “the” are used interchangeably with the term “at least one.” The phrases “at least one of’ and “comprises at least one of’ followed by a list refers to any one of the items in the list and any combination of two or more items in the list.

[0025] As used here, the term “or” is generally employed in its usual sense including “and / or” unless the content clearly dictates otherwise. The term “and / or” means one or all of the listed elements or a combination of any two or more of the listed elements.

[0026] 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 were 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 subranges (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.

[0027] Unless expressly stated, ppm (parts per million), percentage, and ratios are on a weight basis. Percentage on a weight basis is also referred to as wt-%.

[0028] As used here, “have,” “having,” “include,” “including,” “comprise,” “comprising,” or the like are used in their open-ended sense, and generally mean “including, but not limited to.” It will be understood that “consisting essentially of,” “consisting of,” and the like are subsumed in“comprising” and the like. As used herein, “consisting essentially of,” as it relates to a composition, product, method, or the like, means that the components of the composition, product, method, or the like are limited to the enumerated components and any other components that do not materially affect the basic and novel characteristic(s) of the composition, product, method, or the like.

[0029] The words “preferred” and “preferably” refer to embodiments that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, and is not intended to exclude other embodiments from the scope of the disclosure, including the claims.DETAILED DESCRIPTION

[0030] This disclosure relates to aquaculture feeds, and specifically fish feeds. In particular, the present disclosure relates to fish feeds containing astaxanthin and to compositions and methods for making such fish feeds. The astaxanthin is provided as part of a feed additive pack that may be mixed with a base feed to prepare the fish feed.

[0031] Fish feed used as fish diets in aquaculture applications may be provided in liquid or solid form, or as a combination of liquid and solid forms. Fish feeds in solid form can include pellets, extruded nuggets, flakes, tablets, powders, and the like. In various aspects, the base feed can include a base feed pellet. In some aspects, the base feed pellet can include a porous matrix distributed throughout. Fish feeds in liquid form can include aqueous solutions, oils, oil and water emulsions, slurries, suspensions, and the like. Most commonly, fish feed for use in aquaculture is provided in pelleted form. The fish feeds of the present disclosure are provided in solid form. The fish feed of the present disclosure may be referred to as pellets. However, it is understood that the fish feed may also include kibbles, tablets, granules, and the like.

[0032] The fish feed pellets may be produced using a base feed formulation using raw materials that may be chosen based on the application in which it is to be used and on the fish species. The base feed formulation may include solid raw materials or both solid and liquid raw materials.

[0033] The fish feed may include various ingredients that improve the stability of the feed and / or provide nutritional or health benefits for the fish. In some cases, antioxidants are used to help preserve the quality and nutritional properties of the feed.

[0034] Fish feed, and in particular salmon feed, may include ingredients that improve the color of the muscle tissue. One such ingredient is astaxanthin. However, astaxanthin is sensitive to environmental conditions (e.g., heat) and can break down during processing and storage.Because astaxanthin is an expensive ingredient, it would be desirable to provide a feed additiveformulation that reduces the breakdown of astaxanthin. It would be desirable to provide a feed additive formulation that maintains 65 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days). It would be desirable to provide a feed additive formulation that maintains 58 % or more of the astaxanthin added to the feed additive, during processing and storage for at least 21 days (3 weeks). It would be desirable to provide a feed additive formulation that maintains 50 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days). It would further be desirable to provide a feed formulation that has a desired concentration of astaxanthin when the feed is fed to the animals. It would be desirable to provide a feed formulation that maintains 65 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days). It would further be desirable to provide a feed formulation that maintains 58 % or more of the astaxanthin added to the feed, during processing and storage for 21 days (3 weeks). It would be desirable to provide a feed formulation that maintains 50 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days).

[0035] The present disclosure provides a feed additive formulation that reduces the breakdown of astaxanthin. The present disclosure provides a feed additive formulation that maintains at least 85 %, at least 90 %, or at least 95 % of the astaxanthin added to the feed additive during processing. The present disclosure provides a feed additive formulation that maintains 65 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days). The present disclosure provides a feed additive formulation that maintains 58 % or more of the astaxanthin added to the feed additive, during processing and storage for at least 21 days (3 weeks). The present disclosure provides a feed additive formulation that maintains 50 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days). The present disclosure provides a feed formulation that has a desired concentration of astaxanthin when the feed is fed to the animals. The present disclosure provides a feed formulation that maintains 65 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days). The present disclosure provides a feed formulation that maintains 58 % or more of the astaxanthin during processing and storage for at least 3 weeks (21 days). The present disclosure provides a feed formulation that maintains50 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days).Feed Additive Pack

[0036] The feed additive formulation can be provided as a feed additive pack. The feed additive pack may include a mixture of desired feed additives for mixing into fish feed. Theingredients of the feed additive pack may be included a suitable concentration so that a desired concentration in the fish feed is achieved upon mixing. According to embodiments, the feed additive pack includes astaxanthin. The feed additive pack may further include one or more antioxidants. Sometimes the combination of antioxidants is referred to as the antioxidant pack. The antioxidants may provide beneficial properties to the feed additive pack or the feed. The antioxidants may be selected, at least in part, based on their capability to protect the astaxanthin from degradation during manufacturing and / or storage.

[0037] The feed additive pack of the present disclosure includes astaxanthin and an antioxidant component with one or more antioxidants. The antioxidant component may be provided as antioxidant particles that contain one or more antioxidants. When the feed additive pack is added to (e.g., mixed into) a feed formulation, the astaxanthin and the antioxidant particles may be distributed throughout the feed. The feed may be provided as pellets, extruded nuggets, flakes, tablets, or powder. The astaxanthin and the antioxidant particles may be included in and distributed throughout the pellets, extruded nuggets, flakes, tablets, or powder.

[0038] The one or more antioxidants may protect the astaxanthin from degradation in the feed additive pack and in the feed. It has surprisingly been found that controlling the particle size distribution of the antioxidant component can be used to provide additional protection against degradation of the astaxanthin. The particle size may be controlled by griding the antioxidant component to a desired particle size, or by making the antioxidant component in the desired particle size, or by selecting particles in the desired particle size (e.g., by sieving), or any other method known in the art, or combinations thereof. In some embodiments, the antioxidant component is provided with a bimodal particle size distribution. The bimodal particle size distribution may be achieved, for example, by providing (e.g., by any of the methods mentioned above) one part of the antioxidant component in a first particle size and another part in a second particle size and combining the two parts. Alternatively, the antioxidant component may be made in a manner that results in the bimodal particle size distribution. Thus, according to an embodiment, the feed additive pack includes astaxanthin and an antioxidant component with a first particle size population and a second particle size population. The first particle size population may be referred to as the fine particles, and the second particle size population may be referred to as the coarse particles. Although there may be some overlap in the particle size distribution, the first particle size population generally includes particles that are smaller (finer) than the second particle size population. According to an embodiment, the particle size of the antioxidant particles is controlled to improve the stability of the astaxanthin during manufacturing and storage. The particle size of the antioxidant particles may be controlled toimprove the stability of the astaxanthin in fish feed during manufacturing and storage. The particle size of the antioxidant particles may be controlled to improve the stability of the astaxanthin in feed pellets during manufacturing and storage.

[0039] The antioxidant component may be made up of particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers. The D80 of the antioxidant component particles may be 140 micrometers or greater, 145 micrometers or greater, 150 micrometers or greater, 155 micrometers or greater, or 165 micrometers or greater. The D80 of the antioxidant component particles may be 200 micrometers or less, 190 micrometers or less, 185 micrometers or less, or 180 micrometers or less. The antioxidant component particles may have a D10 of 5 micrometers or greater, 6 micrometers or greater, 7 micrometers or greater, 8 micrometers or greater, 9 micrometers or greater, or 10 micrometers or greater. The D10 may be 20 micrometers or less, 17 micrometers or less, 15 micrometers or less, 14 micrometers or less, 13 micrometers or less, 12 micrometers or less, or 11 micrometers or less. The D10 may be in a range of 5 to 20 micrometers, 6 to 15 micrometers, or 8 to 12 micrometers. The antioxidant component particles may have a D50 of 40 micrometers or greater, 45 micrometers or greater, 48 micrometers or greater, or 50 micrometers or greater. The D50 may be 80 micrometers or less, 75 micrometers or less, 70 micrometers or less, 65 micrometers or less, or 60 micrometers or less. The D50 may be in a range of 40 to 80 micrometers, 45 to 70 micrometers, or 48 to 65 micrometers. The antioxidant component particles may have a D90 of 200 micrometers or greater, 210 micrometers or greater, 220 micrometers or greater, or 230 micrometers or greater. The D90 may be 400 micrometers or less, 375 micrometers or less, 350 micrometers or less, 335 micrometers or less, 330 micrometers or less, or 325 micrometers or less. The D90 may be in a range of 200 to 400 micrometers, 210 to 350 micrometers, or 220 to 325 micrometers.

[0040] In embodiments where the antioxidant pack has a bimodal particle size distribution, the first particle size population may have a particle size volume distribution with DI OF of about 1 micrometer or greater, about 2 micrometers or greater, about 3 micrometers or greater, about 4 micrometers or greater, or about 5 micrometers or greater. The DI OF of the first particle size population may be about 20 micrometers smaller, about 18 micrometers smaller, about 15 micrometers smaller, or about 12 micrometers smaller. The D50F of the first particle size population may be about 20 micrometers or greater, about 25 micrometers or greater, about 30 micrometers or greater, about 35 micrometers or greater, about 40 micrometers or greater, or about 45 micrometers or greater. The D50F of the first particle size population may be about 80 micrometers smaller, about 70 micrometers smaller, about 60 micrometers smaller, or about 50 micrometers smaller. The D90F of the first particle size population may be about 100micrometers or greater, about 110 micrometers or greater, about 120 micrometers or greater, about 130 micrometers or greater, or about 140 micrometers or greater. The D90F of the first particle size population may be about 200 micrometers smaller, about 180 micrometers smaller, about 180 micrometers smaller, or about 160 micrometers smaller. For example, the first particle size population may have a particle size volume distribution with DI OF in a range of 1 to 20 micrometers, D50F in a range of 20 to 80 micrometers, and D90F in a range of 100 to 200 micrometers. The first particle size population may have a particle size volume distribution with D10F in a range of 2 to 15 micrometers, D50F in a range of 30 to 70 micrometers, and D90F in a range of 120 to 180 micrometers. The first particle size population may have a particle size volume distribution with D 10F in a range of 4 to 12 micrometers, D50F in a range of 40 to 60 micrometers, and D90F in a range of 140 to 160 micrometers.

[0041] The second particle size population may have a particle size volume distribution with D 10c of about 1 micrometer or greater, about 2 micrometers or greater, about 3 micrometers or greater, about 4 micrometers or greater, or about 5 micrometers or greater. The D 10c of the second particle size population may be about 20 micrometers smaller, about 18 micrometers smaller, about 15 micrometers smaller, or about 12 micrometers smaller. The D50c of the second particle size population may be about 20 micrometers or greater, about 25 micrometers or greater, about 30 micrometers or greater, about 35 micrometers or greater, about 40 micrometers or greater, or about 45 micrometers or greater. The D50c of the second particle size population may be about 80 micrometers smaller, about 70 micrometers smaller, about 60 micrometers smaller, or about 50 micrometers smaller. The D90c of the second particle size population may be about 100 micrometers or greater, about 110 micrometers or greater, about 120 micrometers or greater, about 130 micrometers or greater, or about 140 micrometers or greater. The D90c of the second particle size population may be about 750 micrometers or smaller, 500 micrometers or smaller, 400 micrometers or smaller, 200 micrometers smaller, about 180 micrometers smaller, about 180 micrometers smaller, or about 160 micrometers smaller. For example, the second particle size population may have a particle size volume distribution with D 10c in a range of 1 to 20 micrometers, D50c in a range of 20 to 80 micrometers, and D90c in a range of 100 to 750 micrometers. The second particle size population may have a particle size volume distribution with D 10c in a range of 2 to 18 micrometers, D50c in a range of 25 to 70 micrometers, and D90c in a range of 120 to 400 micrometers. The second particle size population may have a particle size volume distribution with DIOc in a range of 3 to 15 micrometers, D50c in a range of 30 to 60 micrometers, and D90c in a range of 150 to 200 micrometers.

[0042] In general, within the above particle size distribution ranges, the particle size of the second particle size population is greater than the corresponding particle size distribution of the first particle size population. The lower end, e.g., the D10F and DIOc, of two particle size populations may be close in size, while the middle (e.g., the D50F and D50c) and the upper end (e.g., the D90F and D90c) differ more. For example, DI OF and DIOc may be the same or differ by only about 2 to 5 micrometers, while the D50F and D50c may differ by about 5 to 20 micrometers, and the D90F and D90c may differ by about 30 micrometers or more, such as about 50 to 100 micrometers.

[0043] The two particle size populations may be included in the feed additive pack at a weight ratio of about 0.3 parts or more, about 0.5 parts or more, about 0.8 parts or more, about 1 part or more, about 1.3 parts or more, about 1.5 parts or more, about 1.8 parts or more, about 2 parts or more, about 2.5 parts or more, about 2.8 parts or more, or about 3 parts of the first particle size population for every 1 part of the second particle size population. The first particle size population may be present at about 0.3 to 3 parts for every 1 part of the second particle size population. The first particle size population may be present at about 0.5 to 2 parts for every 1 part of the second particle size population. The first particle size population may be present at about 0.8 to 1.3 parts for every 1 part of the second particle size population. In some embodiments, the first particle size population and the second particle size population are provided at equal amounts (e.g., a weight ratio of 1 : 1).

[0044] The feed additive pack includes astaxanthin. Astaxanthin may be included in the feed additive pack at any suitable concentration that will result in a desired concentration in the feed formulation, based on the inclusion rate of the feed additive pack. For example, in some representative embodiments, the feed additive pack includes about 0.5 wt-% or more, about 1 wt-% or more, about 2 wt-% or more, about 5 wt-% or more, about 8 wt-% or more, about 10 wt-% or more, or about 15 wt-% or more of astaxanthin. The feed additive pack may include about 50 wt-% or less, about 40 wt-% or less, about 30 wt-% or less, about 25 wt-% or less, about 20 wt-% or less, about 15 wt-% or less, or about 12 wt-% or less of astaxanthin. The concentration of astaxanthin in the feed additive pack may be about 0.5 wt-% to about 50 wt-%, from about 2 wt-% to about 25 wt-%, or from about 5 wt-% to about 20 wt-%. Typically, the desired concentration of astaxanthin in the fish feed is about 100 ppm (about 0.01 wt-%).

[0045] The astaxanthin may be a commercially available astaxanthin product. Examples of commercially available astaxanthin products include PANAFERD AX (available from Daesang Corporation, Seoul, Republic of Korea) and CAROPHYLL PINK 10% (available from Royal DSM, Heerlen, Netherlands). Typical astaxanthin products for use in feed applications areprovided as a fine powder that includes astaxanthin and excipients. The excipients may coat astaxanthin crystals. The particle size (D90) of the astaxanthin may be about 1 to 10 micrometers, or about 4 micrometers.

[0046] The antioxidant component of the feed additive pack may include any suitable mixture of antioxidants. The antioxidant component may further other ingredients, such as carriers, binders, emulsifiers, chelators, and combinations of two or more thereof. Some of the components may serve more than one function. For example, some components may act as antioxidants and emulsifiers, or antioxidants and chelators.

[0047] The antioxidant component of the feed additive pack may include one or more primary antioxidants and one or more secondary antioxidants. In some cases, the secondary antioxidant exhibits a synergistic effect with the primary antioxidant. The primary antioxidant may be a free radical scavenger. The primary antioxidant may include butylated hydroxyanisole (BHA), a carotenoid (for example, beta carotene, canthaxanthin, or the like), a flavonoid (for example, theaflavin, quercetin, or the like), phenolic acid (for example, carnosic acid, gallic acid, or the like), a vitamin (for example, tocopherol, tocotrienol, or the like), tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof. The amount of the primary antioxidant(s) may be adjusted based on the intended inclusion rate of the feed additive pack in the feed such that the desired concentration is achieved in the resulting feed. For example, in some representative embodiments, the primary antioxidant(s) may be present in the feed additive pack at a total concentration of about 1 wt-% or greater, about 2 wt-% or greater, about 3 wt-% or greater, about 4 wt-% or greater, or about 5 wt-% or greater. The primary anti oxi dant(s) may be present in the feed additive pack at a total concentration of about 10 wt-% or less, about 8 wt-% or less, or about 6 wt-% or less. The primary antioxidant(s) may be present in the feed additive pack at a total concentration of about 1 wt-% to 10 wt-%, or from about 4 wt-% to about 8 wt-%.

[0048] The secondary antioxidant may be or may include an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof. Examples of suitable oxygen scavengers include ascorbic acid and salts thereof, erythorbic acid, sulfites, alphatocopherol, and combinations of two or more thereof. Examples of suitable emulsifiers include powder lecithin, polysorbate (e.g., polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, or the like), and combinations of two or more thereof. Examples of suitable chelators include citric acid, EDTA, glycine, and combinations thereof. The secondary antioxidant may include ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol, polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof. The amount of the secondary antioxidant(s) may beadjusted based on the intended inclusion rate of the feed additive pack in the feed such that the desired concentration is achieved in the resulting feed. For example, in some representative embodiments, the secondary antioxidant(s) may be present in the feed additive pack at a total concentration of about 15 wt-% or greater, about 18 wt-% or greater, about 20 wt-% or greater, or about 22 wt-% or greater. The secondary antioxidant(s) may be present in the feed additive pack at a total concentration of about 50 wt-% or less, about 40 wt-% or less, about 30 wt-% or less, or about 25 wt-% or less. The secondary anti oxi dant(s) may be present in the feed additive pack at a total concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%. In a preferred embodiment, the secondary antioxidant includes citric acid. The citric acid may make up the majority or all of the secondary antioxidant. The citric acid may present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0049] The feed additive pack may include a carrier. Any suitable carriers may be used. The carrier may include flour, meal, or starch prepared from a grain or a legume. Examples of suitable carriers include wheat, barley, soybean meal, dehulled fava bean, pea starch, dehulled pea, and the like, and combinations of two or more thereof.

[0050] According to an embodiment, the desired concentration of astaxanthin is maintained or substantially maintained throughout the manufacturing and storage of the feed additive pack and feed. Manufacturing may include various processes, including mixing, grinding, and pelleting steps. At least some of the processing steps may include application of heat and / or steam to the feed composition. In some cases, processing of the feed composition that includes the feed additive includes extruding the feed composition into pellets. The storage conditions may be typical storage conditions for a feed additive or a fish feed. For example, the storage conditions may include temperature from about -10 degrees Celsius to about 35 degrees Celsius, more typically temperature from about 10 degrees Celsius to about 20 degrees Celsius, and average humidity from about 80 % to about 95%. Storage conditions may include protection from direct sunlight. Fish feed or the feed additive pack may be stored under such typical storage conditions for up to one year or more. The feed additive pack maintains at least 85 %, at least 90 %, or at least 95 % of the astaxanthin added to the feed additive during processing. The amount of astaxanthin lost during feed manufacturing may be 11 % or less, 10 % or less, 9 % or less, 8 % or less, 7 % or less, 6 % or less, 5 % or less, 4.9 % or less, 4.75 % or less, 4.5 % or less, 4 % or less, 3.5 % or less, 3 % or less, 2.5 % or less, or 2 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). In some embodiments, no astaxanthin is lost during processing.The amount of astaxanthin after processing may be about 85 % or more, about 89 % or more, about 90 % or more, about 91 % or more, about 92 % or more, about 93 % or more, about 94 % or more, about 95 % or more, about 95.1 % or more, about 95.25 % or more, about 95.5 % or more, about 96 % or more, about 96.5 % or more, about 97 % or more, about 97.5 % or more, about 98 % or more, or about 99 % of the initial amount of astaxanthin (astaxanthin added to the feed additive and / or feed composition). The feed formulation has a desired concentration of astaxanthin when the feed is fed to the animals. The present disclosure provides a feed additive pack that maintains about 60 % or more, about 62 % or more, about 65 % or more, about 67 % or more, or about 70 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days). The amount of astaxanthin lost during combined processing and storage for 2 weeks (14 days) may be 40 % or less, 38 % or less, 35 % or less, or 30 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). The present disclosure provides a feed additive pack that maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days). The amount of astaxanthin lost during combined processing and storage for 3 weeks (21 days) may be 42 % or less, 40 % or less, 35 % or less, or 32 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). The present disclosure provides a feed additive pack that maintains about 50 % or more, about 52 % or more, about 60 % or more, or about 62 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days). The amount of astaxanthin lost during combined processing and storage for 4 weeks (28 days) may be 48 % or less, 45 % or less, 40 % or less, or 38 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). Although there is no desired minimum loss, in practice, some astaxanthin is lost during processing and storage. For example, at least 20 % or at least 25 % of the astaxanthin that was added to the feed composition prior to processing may be lost during processing and storage. The feed additive pack may maintain about 78 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage of the feed for at least 1 week (7 days). The feed additive pack may maintain about 65 % or more, 76 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage of the feed for at least 2 weeks (14 days).Fish Feeds

[0051] The feed additive pack of the present disclosure may be included in a fish feed, to be used as fish diets in aquaculture applications. According to an embodiment, the feed additive pack is capable of decreasing the degradation of astaxanthin during processing and / or storage. The fish feeds disclosed herein can include a base feed. The feed additive pack may be mixed with the base feed. The feed additive pack may be mixed with the base feed prior to manufacturing of feed pellets from the base feed.

[0052] It will be appreciated that the fish feeds herein can include suitable types of fish feed specific for a given fish species. The fish feeds may be used as a component of a diet fed to any species belonging to the families Cyprinidae, Cichlidae, Pangasiidae, Sciaenidae, Serranidae, Carangidae, Sparidae, Lateolabracidae, Moronidae, Mugilidae, Cypriniformes, Latidae, Eleotridae, Tilapiini and Salmonidae. In various aspects, this disclosure provides a fish feed or fish feed diet for species within the family Salmonidae. The fish feeds provided herein may be used to feed wild fish or farmed fish. In various aspects, both wild fish and farmed fish may be fed simultaneously. Further, the fish feed may be used to feed freshwater fish or salt water (e.g., marine) fish, or both.

[0053] The fish feeds herein can include a number of different ingredients or raw materials that can sustain life, growth, and reproduction of the fish. The fish feeds can include any substrate that is edible to fish. For example, an edible substrate can provide a source of nutrition to the fish or may be an inert substrate with no nutritive value to the fish. In various aspects, the fish feeds herein can include feeds that are either nutritional fish feeds or non-nutritional feeds. Nutritional fish feeds can include a nutritional food stuff formulated for fish as part of its diet as the main source of nutrition, growth, and reproduction. Suitable nutritional fish feeds can include one or more of proteinaceous material as a source of proteins, peptides, and amino acids; carbohydrates; and fats, as described below. Non-nutritional fish feeds can include any substrate that is edible to fish but does not provide nutrition to sustain life, growth, or reproduction. In various aspects, the nutritional or non-nutritional fish feeds herein can include one or more compounds designed to alter the quality, quantity, or appearance of a fish and fish tissue. For example, a nutritional or non-nutritional fish feed can include a carotenoid compound to improve the appearance (e.g., color) of the muscle tissue. By way of example, the carotenoid compound can include compounds such as astaxanthin. The astaxanthin may be added to the feed as part of the feed additive pack of the present disclosure.

[0054] The fish feed may be a complete fish feed. A complete fish feed may be a feed for fish that is compounded to be fed as the sole ration and that can maintain life, promote growth, andsustain reproduction without any additional substances being consumed except water. Complete fish feeds can include compounded mixtures containing various energy sources such as carbohydrates, proteins, and fats. In various aspects, the fish feeds herein can include at least a protein and a starch. Additional ingredients may be included, such as vitamins and minerals as necessary to support the life, growth, and reproduction of fish. A complete fish feed can include ingredients such as, but not limited to, fish meal, poultry meal, plant meal, vegetable meal, corn meal, corn gluten meal, soy meal, soy protein concentrate, single cell protein, insect meal, algae meal, algae oil, krill meal, krill oil, meat meal, blood meal, feather meal, starches, tapioca starch, wheat, wheat gluten, guar meal, guar protein concentrate, peas, pea protein concentrate, pea starch, beans, fava beans, sunflower meal, vegetable oil, canola oil, poultry oil, rapeseed oil, fish oil, soy oil, linseed oil, camelina oil, lecithin, macro-minerals, minerals, vitamins, amino acids, pigments, and any combinations thereof. It will be appreciated that the fish feeds herein can include fish meal that can include plant or animal derived matter. Any animal derived matter present in the fish meal may be derived from the same species of fish or a different species of fish (e.g., heterologous species or non-fish species).

[0055] The fish feed may include any suitable amount of protein. In some representative embodiments, the total protein content in the fish feed may be about 10 wt-% or greater, about 20 wt-% or greater, about 25 wt-% or greater, about 30 wt-% or greater, about 35 wt-% or greater, about 40 wt-% or greater, about 45 wt-% or greater, about 50 wt-% or greater, about 55 wt-% or greater, about 60 wt-% or greater, or about 65 wt-% or greater. In some representative embodiments, the total protein content in the fish feed may be about 70 wt-% or lower, about 65 wt-% or lower, about 60 wt-% or lower, about 55 wt-% or lower, about 50 wt-% or lower, about 45 wt-% or lower, about 40 wt-% or lower, about 35 wt-% or lower, about 30 wt-% or lower, or about 25 wt-% or lower. In some representative embodiments, the total protein content in the fish feed may be from about 10 wt-% to about 70 wt-%, from about 20 wt-% to about 60 wt-%, from about 30 wt-% to about 50 wt-%, or from about 35 wt-% to about 45 wt-%. Typically, the desired concentration of protein in the fish feed is about 40 wt-%. The total protein content in the fish feed may be variable depending on the formulation, species, age, and intended use of the feed. It will be appreciated that the various protein requirements of the species of fish receiving the fish feed may be adjusted to meet the protein requirements of that species.

[0056] The protein in the fish feeds herein may be from any suitable source including, but not limited to, one or more of fish meal, land-animal protein (e.g., poultry meal), plant-based protein (e.g., vegetable meal), or any combinations thereof.

[0057] In some representative embodiments, the fish meal may be included in the fish feed at a concentration of about 0 wt-% or greater, about 5 wt-% or greater, about 10 wt-% or greater, about 20 wt-% or greater, about 25 wt-% or greater, about 30 wt-% or greater, about 35 wt-% or greater, about 40 wt-% or greater, about 45 wt-% or greater, about 50 wt-% or greater, about 55 wt-% or greater, about 60 wt-% or greater, about 65 wt-% or greater, about 70 wt-% or greater, or about 75 wt-% or greater. In some representative embodiments, the fish meal may be included in the fish feed at a concentration of about 80 wt-% or lower, about 75 wt-% or lower, about 70 wt-% or lower, about 65 wt-% or lower, about 60 wt-% or lower, about 55 wt-% or lower, about 50 wt-% or lower, about 45 wt-% or lower, about 40 wt-% or lower, about 35 wt-% or lower, about 30 wt-% or lower, about 25 wt-% or lower, about 20 wt-% or lower, about 15 wt-% or lower, about 10 wt-% or lower, or about 5 wt-% or lower. In some representative embodiments, the fish meal may be included in the fish feed at a concentration of about 0 wt-% to about 80 wt-%, from about 10 wt-% to about 70 wt-%, from about 20 wt-% to about 60 wt-%, from about 30 wt-% to about 50 wt-%, or from about 35 wt-% to about 45 wt-%.

[0058] In some representative embodiments, the land-animal protein may be included in the fish feed at a concentration of about 0 wt-% or greater, about 5 wt-% or greater, about 10 wt-% or greater, about 20 wt-% or greater, about 25 wt-% or greater, about 30 wt-% or greater, about 35 wt-% or greater, about 40 wt-% or greater, about 45 wt-% or greater, about 50 wt-% or greater, about 55 wt-% or greater, about 60 wt-% or greater, about 65 wt-% or greater, about 70 wt-% or greater, or about 75 wt-% or greater. In some representative embodiments, the land-animal protein may be included in the fish feed at a concentration of about 80 wt-% or lower, about 75 wt-% or lower, about 70 wt-% or lower, about 65 wt-% or lower, about 60 wt-% or lower, about 55 wt-% or lower, about 50 wt-% or lower, about 45 wt-% or lower, about 40 wt-% or lower, about 35 wt-% or lower, about 30 wt-% or lower, about 25 wt-% or lower, about 20 wt-% or lower, about 15 wt-% or lower, about 10 wt-% or lower, or about 5 wt-% or lower. In some representative embodiments, the land-animal protein may be included in the fish feed at a concentration of from about 0 wt-% to about 80 wt-%, from about 10 wt-% to about 70 wt-%, from about 20 wt-% to about 60 wt-%, from about 30 wt-% to about 50 wt-%, or from about 35 wt-% to about 45 wt-%.

[0059] In some representative embodiments, the plant-based protein may be included in the fish feed at a concentration of about 0 wt-% or greater, about 5 wt-% or greater, about 10 wt-% or greater, about 20 wt-% or greater, about 25 wt-% or greater, about 30 wt-% or greater, about 35 wt-% or greater, about 40 wt-% or greater, about 45 wt-% or greater, about 50 wt-% or greater, about 55 wt-% or greater, about 60 wt-% or greater, about 65 wt-% or greater, about 70 wt-% or greater, or about 75 wt-% or greater. In some representative embodiments, the plant-based proteinmay be included in the fish feed at a concentration of about 80 wt-% or lower, about 75 wt-% or lower, about 70 wt-% or lower, about 65 wt-% or lower, about 60 wt-% or lower, about 55 wt-% or lower, about 50 wt-% or lower, about 45 wt-% or lower, about 40 wt-% or lower, about 35 wt-% or lower, about 30 wt-% or lower, about 25 wt-% or lower, about 20 wt-% or lower, about 15 wt-% or lower, about 10 wt-% or lower, or about 5 wt-% or lower. In some representative embodiments, the plant-based protein may be included in the fish feed at a concentration of from about 0 wt-% to about 80 wt-%, from about 10 wt-% to about 70 wt-%, from about 20 wt-% to about 60 wt-%, from about 30 wt-% to about 50 wt-%, or from about 35 wt-% to about 45 wt-%.

[0060] The fish feed may include any suitable amount of total fat (e.g., oil, fat, and / or lipids). In some representative embodiments, the total fat content in the fish feed may be about 5 wt-% or greater, about 10 wt-% or greater, about 20 wt-% or greater, about 25 wt-% or greater, about 30 wt-% or greater, about 35 wt-% or greater, about 40 wt-% or greater, or about 45 wt-% or greater. In some representative embodiments, the total fat content in the fish feed may be about 50 wt-% or lower, about 45 wt-% or lower, about 40 wt-% or lower, about 35 wt-% or lower, about 30 wt-% or lower, about 25 wt-% or lower, about 20 wt-% or lower, about 15 wt-% or lower, or about 10 wt-% or lower. In some representative embodiments, the total fat content in the fish feed may be from about 5 wt-% to about 50 wt-%, from about 10 wt-% to about 40 wt-%, from about 20 wt-% to about 30 wt-%, from about 30 wt-% to about 50 wt-%, or from about 35 wt-% to about 45 wt-%. Typically, the desired concentration of fat in the fish feed is about 35 wt-%. The total fat content in the fish feed may be variable depending on the formulation, target fish species, and intended use of the fish feed. It will be appreciated that the various fat requirements of the species of fish receiving the fish feed and may be adjusted to meet the fat requirements of that species. Suitable fats for use herein may include, but are not to be limited to, those provided by canola oil, poultry oil, rapeseed oil, fish oil, soy oil, linseed oil, camelina oil, palm oil, lecithin, or any combinations or fractions thereof.

[0061] EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) are omega-3 fatty acids that are components of the total fatty acid content in certain foods or supplements. The term “EPA / DHA of total fatty acids” refers to the combined percentage or amount of EPA and DHA present relative to the total amount of fatty acids in the product. The fish feed may include any suitable amount of EPA / DHA of total fatty acids. In some representative embodiments, the EPA / DHA of total fatty acids content in the fish feed may be about 2 wt-% or greater, about 2.5 wt-% or greater, about 3 wt-% or greater, about 4 wt-% or greater, about 5 wt-% or greater, about 7.5 wt-% or greater, about 10 wt-% or greater, about 12.5 wt-% or greater, about 15 wt-% or greater, about 17.5 wt-% or greater, about 20 wt-% or greater, or about 22.5 wt-% or greater. Insome representative embodiments, the EPA / DHA of total fatty acids may be included in the fish feed at a concentration of about 22.5 wt-% or lower, about 20 wt-% or lower, about 17.5 wt-% or lower, about 15 wt-% or lower, about 12.5 wt-% or lower, about 10 wt-% or lower, about 7.5 wt-% or lower, about 5 wt-% or lower, about 4 wt-% or lower, about 3 wt-% or lower, or about2.5 wt-% or lower. In some representative embodiments, the EPA / DHA of total fatty acids may be included in the fish feed at a concentration of from about 2 wt-% to about 25 wt-%, from about 5 wt-% to about 20 wt-%, from about 7.5 wt-% to about 17.5 wt-%, or from about 10 wt-% to about 15 wt-%. Typically, the desired EPA / DHA of total fatty acids in the fish feed is about 7 wt-%.

[0062] The fish feed may include appropriate amount of starch. In some representative embodiments, the starch content may be included in the fish feed at a concentration of about 2 wt-% or greater, about 2.5 wt-% or greater, about 3 wt-% or greater, about 4 wt-% or greater, about 5 wt-% or greater, about 7.5 wt-% or greater, about 10 wt-% or greater, about 12.5 wt-% or greater, about 15 wt-% or greater, about 17.5 wt-% or greater, about 20 wt-% or greater, or about22.5 wt-% or greater. In some representative embodiments, the starch content may be included in the fish feed at a concentration of about 22.5 wt-% or lower, about 20 wt-% or lower, about 17.5 wt-% or lower, about 15 wt-% or lower, about 12.5 wt-% or lower, about 10 wt-% or lower, about7.5 wt-% or lower, about 5 wt-% or lower, about 4 wt-% or lower, about 3 wt-% or lower, or about2.5 wt-% or lower. In some representative embodiments, the starch content may be included in the fish feed at a concentration of from about 2 wt-% to about 25 wt-%, from about 5 wt-% to about 20 wt-%, from about 7.5 wt-% to about 17.5 wt-%, or from about 10 wt-% to about 15 wt-%. Typically, the desired starch content in the fish feed is about 8 wt-%.

[0063] The fish feed may include appropriate amount of dry matter. In some representative embodiments, the dry matter content may be included in the fish feed at a concentration of about 80 wt-% or greater, about 85 wt-% or greater, about 90 wt-% or greater, or about 95 wt-% or greater. In some representative embodiments, the dry matter content may be included in the fish feed at a concentration of about 99 wt-% or lower, about 95 wt-% or lower, about 90 wt-% or lower, or about 85 wt-%. In some representative embodiments, the dry matter content may be included in the fish feed at a concentration of from about 80 wt-% to about 99 wt-%, from about 85 wt-% to about 95 wt-%, or from about 87.5 wt-% to about 90 wt-%. Typically, the dry matter content in the fish feed is about 95 wt-%.

[0064] The moisture content of the fish feeds herein may vary depending on the contents and preparation method of the feed. The fish feed may include appropriate amount of moisture content. In some representative embodiments, the moisture content may be included in the fish feed at aconcentration of 1 wt-% or greater, about 1.25 wt-% or greater, about 1.5 wt-% or greater, about1.75 wt-% or greater, about 2 wt-% or greater, about 2.5 wt-% or greater, about 3 wt-% or greater, about 4 wt-% or greater, about 5 wt-% or greater, about 7.5 wt-% or greater, about 10 wt-% or greater, about 12.5 wt-% or greater, about 15 wt-% or greater, or about 17.5 wt-% or greater. In some representative embodiments, the moisture content may be included in the fish feed at a concentration of about 20 wt-% or lower, about 17.5 wt-% or lower, about 15 wt-% or lower, about 12.5 wt-% or lower, about 10 wt-% or lower, about 7.5 wt-% or lower, about 5 wt-% or lower, about 4 wt-% or lower, about 3 wt-% or lower, about 2.5 wt-% or lower, about 2 wt-% or lower, about 1.75 wt-% or lower, about 1.5 wt-% or lower, about 1.25 wt-% or lower. In some representative embodiments, the moisture content may be included in the fish feed at a concentration of from about 1 wt-% to about 20 wt-%, from about 5 wt-% to about 15 wt-%, or from about 7.5 wt-% to about 12.5 wt-%.

[0065] According to an embodiment, the desired concentration of astaxanthin is maintained or substantially maintained throughout the manufacturing and storage of the feed. The storage conditions may be typical storage conditions for feed additive or fish feed. For example, the storage conditions temperature from about -10 degrees Celsius to about 35 degrees Celsius, more typically temperature from about 10 degrees Celsius to about 20 degrees Celsius, and with average humidity from about 80 % to about 95%. Storage conditions may include protection from direct sunlight. Fish feed or the feed additive pack may be stored under such typical storage conditions for up to one year or more. According to an embodiment, the methods of the disclosure provide for the particle size of the antioxidant particles to be controlled to improve the stability of the astaxanthin during processing and storage. The feed additive pack maintains at least 85 %, at least 90 %, or at least 95 % of the astaxanthin added to the feed additive during manufacturing. The amount of astaxanthin lost during feed manufacturing may be 11 % or less, 10 % or less, 9 % or less, 8 % or less, 7 % or less, 6 % or less, 5 % or less, 4.9 % or less,4.75 % or less, 4.5 % or less, 4 % or less, 3.5 % or less, 3 % or less, 2.5 % or less, or 2 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). In some embodiments, no astaxanthin is lost during processing. The amount of astaxanthin after processing may be about 85 % or more, about 89 % or more, about 90 % or more, about 91 % or more, about 92 % or more, about 93 % or more, about 94 % or more, about 95 % or more, about 95.1 % or more, about 95.25 % or more, about 95.5 % or more, about 96 % or more, about 96.5 % or more, about 97 % or more, about 97.5 % or more, about 98 % or more, or about 99 % of the initial amount of astaxanthin (astaxanthin added to the feed additive and / or feed composition. The feed formulation has adesired concentration of astaxanthin when the feed is fed to the animals. The feed formulation maintains about 60 % or more, about 62 % or more, about 65 % or more, about 67 % or more, or about 70 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days). The amount of astaxanthin lost during combined processing and storage for 2 weeks (14 days) may be 40 % or less, 38 % or less, 35 % or less, or 30 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). The feed formulation maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days). The amount of astaxanthin lost during combined processing and storage for 3 weeks (21 days) may be 42 % or less, 40 % or less, 35 % or less, or 32 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). The feed formulation maintains about 50 % or more, about 52 % or more, about 60 % or more, or about 62 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days). The amount of astaxanthin lost during combined processing and storage for 4 weeks (28 days) may be 48 % or less, 45 % or less, 40 % or less, or 38 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). Although there is no desired minimum loss, in practice, some astaxanthin is lost during storage. For example, at least 20 % or at least 25 % of the astaxanthin that was added to the feed composition prior to processing may be lost during processing and storage. The feed formulation may maintain about 78 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 1 week (7 days). The feed formulation may maintain about 76 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 2 weeks (14 days).Methods of Making

[0066] The present disclosure provides methods for making fish feed. In some embodiments, the present disclosure provides methods for making feed additive packs for adding to fish feed. The present disclosure provides methods for making fish feed, and feed additive packs for adding to the fish feed, which include antioxidant component and astaxanthin. The present disclosure provides methods for making fish feed, and feed additive packs for adding to the fish feed, with formulations that reduce the breakdown of astaxanthin. By controlling the particle size distribution of the antioxidant component in the feed, the astaxanthin contained in the feedadditive pack can be further protected against degradation. The desired particle size of the antioxidant component provides additional protection against degradation of the astaxanthin. The desired particle size of the antioxidant component provides additional protection against degradation of the astaxanthin during processing. The particle size may be controlled to a desired particle size as discussed above with regard to the feed additive pack. Control of the particle size may be achieved by any suitable method, including making the antioxidant component in the desired particle size or by reducing the particle size by known methods, such as grinding, sieving, and the like. In some embodiments, the desired particle size of the antioxidant component is achieved by griding the antioxidant component. In some embodiments, the desired particle size of the antioxidant component is achieved by making the antioxidant component in the desired particle size. In some embodiments, the desired particle size of the antioxidant component is achieved by selecting particles in the desired particle size (e.g., by sieving).

[0067] In some embodiments, the antioxidant component is provided with a bimodal particle size distribution. The bimodal particle size distribution may be achieved, for example, by providing (e.g., by any of the methods mentioned above) one part of the antioxidant component in a first particle size and another part in a second particle size and combining the two parts. Alternatively, the antioxidant component may be made in a manner that results in the bimodal particle size distribution. A bimodal particle size distribution of the antioxidant component provides additional protection against degradation of the astaxanthin. The bimodal particle size distribution of the antioxidant component provides additional protection against degradation of the astaxanthin during processing. The bimodal particle size distribution of the antioxidant component provides additional protection against degradation of the astaxanthin during storage. Thus, in some embodiments, the disclosure provides methods for making a feed additive pack that includes astaxanthin and an antioxidant component with a first particle size population and a second particle size population. The first particle size population may be referred to as the fine particles, and the second particle size population may be referred to as the coarse particles. Although there may be some overlap in the particle size distribution, the first particle size population generally includes particles that are smaller (finer) than the second particle size population.

[0068] The particle size of the antioxidant particles may be controlled to improve the stability of the astaxanthin in fish feed during processing and storage. The particle size of the antioxidant particles may be controlled to improve the stability of the astaxanthin in feed pellets during processing and storage. According to an embodiment, the methods of the disclosure provide forthe particle size of the antioxidant particles to be controlled to improve the stability of the astaxanthin during processing and storage. The present disclosure provides a feed additive pack that maintains at least 85 %, at least 90 %, or at least 95 % of the astaxanthin added to the feed additive during processing. During manufacturing, the feed composition is processed into a feed. Manufacturing may include various processes, including mixing, grinding, and pelleting steps. At least some of the processing steps may include application of heat and / or steam to the feed composition. In some cases, the feed composition that includes the feed additive may be extruded into pellets during manufacturing. The amount of astaxanthin lost during feed manufacturing (e.g., processing of the feed composition into the feed) may be 11 % or less, 10 % or less, 9 % or less, 8 % or less, 7 % or less, 6 % or less, 5 % or less, 4.9 % or less, 4.75 % or less, 4.5 % or less, 4 % or less, 3.5 % or less, 3 % or less, 2.5 % or less, or 2 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). In some embodiments, no astaxanthin is lost during processing. The amount of astaxanthin after processing may be about 85 % or more, about 89 % or more, about 90 % or more, about 91 % or more, about 92 % or more, about 93 % or more, about 94 % or more, about 95 % or more, about 95.1 % or more, about 95.25 % or more, about 95.5 % or more, about 96 % or more, about 96.5 % or more, about 97 % or more, about 97.5 % or more, about 98 % or more, or about 99 % of the initial amount of astaxanthin (astaxanthin added to the feed additive and / or feed composition. The present disclosure provides a feed formulation that has a desired concentration of astaxanthin when the feed is fed to the animals. The feed formulation maintains about 60 % or more, about 62 % or more, about 65 % or more, about 67 % or more, or about 70 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days). The amount of astaxanthin lost during combined processing and storage for 2 weeks (14 days) may be 40 % or less, 38 % or less, 35 % or less, or 30 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). The feed formulation maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days). The amount of astaxanthin lost during combined processing and storage may be 42 % or less, 40 % or less, 35 % or less, or 32 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). The feed formulation maintains about 50 % or more, about 52 % or more, about 60 % or more, or about 62 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days). The amount of astaxanthin lost during combined processing andstorage for 4 weeks (28 days) may be 48 % or less, 45 % or less, 40 % or less, or 38 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). Although there is no desired minimum loss, in practice, some astaxanthin is lost during processing and storage. For example, at least 20 % or at least 25 % of the astaxanthin that was added to the feed composition prior to processing may be lost during processing and storage. The feed formulation may maintain about 78 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 1 week (7 days). The feed formulation may maintain about 76 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 2 weeks (14 days).

[0069] The method for making fish feed may include the preparation of raw materials and further processing to make the fish feed. The method includes mixing of the various components to prepare a feed composition. The method may further include processing the feed composition into the feed. The feed composition includes the feed additive pack and a base feed. The method includes mixing the feed additive pack and the base feed. The method may involve mixing the components to improve nutrient distribution and consistency in the production process.

[0070] The preparation of the raw materials may include various mixing and grinding steps. For example, certain raw materials may be mixed together and ground to a desired particle size. In some embodiments, raw materials may be ground to a desired particle size separately, prior to being mixed together. Any suitable means to grind material may be used, for example, a hammer mill.

[0071] A base feed may be formed from various raw materials described elsewhere herein. The base feed may include macro-ingredients and numerous micro-ingredients. In some embodiments, base feed may include all macro-ingredients and micro-ingredients. The base feed ingredients may be loaded into a mixer, where it undergoes pre-grind mixing. Pre-grind mixing may be done to improve homogeneity of the base feed. After pre-grind mixing, the base feed may be subjected to grinding to refine the particle size. Mixing and grinding may be used to improve the feed’s texture and digestibility and its suitability for consumption by the fish. In some embodiments, additional micro-ingredients may be added to the base feed prior to the pregrind mixing. In some embodiments, additional micro-ingredients may be added to the base feed after the pre-grind mixing but prior to the grinding. In some embodiments, the additional microingredients may be ground prior to their addition to the base feed. In some embodiments, the additional micro-ingredients may not be ground prior to the addition to the base feed.

[0072] The feed additive pack may be added to the base feed. The feed additive pack may be added to the base feed after the pre-grind mixing and grinding. In some embodiments, all macroingredients, along with micro-ingredients, and feed additive pack, including astaxanthin, may be loaded into a mixer, where they undergo pre-grind mixing. The mixture may then be ground to obtain the desirable particle size. The mixture may again be mixed after being ground.

[0073] The feed additive pack may be made by mixing the components of the feed additive pack. Some of the components of the feed additive pack may be pre-mixed and ground prior to mixing with other components. In some embodiments, the feed additive pack includes an antioxidant component that is mixed and ground prior to addition of the astaxanthin. The antioxidant component may be ground to two separate particle size populations, a first particle size population and a second particle size population. The first particle size population may be referred to as the fine particles, and the second particle size population may be referred to as the coarse particles.

[0074] The antioxidant component may be ground to particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers. The D80 of the antioxidant component particles may be 140 micrometers or greater, 145 micrometers or greater, 150 micrometers or greater, 155 micrometers or greater, or 165 micrometers or greater. The D80 of the antioxidant component particles may be 200 micrometers or less, 190 micrometers or less, 185 micrometers or less, or 180 micrometers or less. The antioxidant component particles may have a D10 of 5 micrometers or greater, 6 micrometers or greater, 7 micrometers or greater, 8 micrometers or greater, 9 micrometers or greater, or 10 micrometers or greater. The D10 may be 20 micrometers or less, 17 micrometers or less, 15 micrometers or less, 14 micrometers or less, 13 micrometers or less, 12 micrometers or less, or 11 micrometers or less. The D10 may be in a range of 5 to 20 micrometers, 6 to 15 micrometers, or 8 to 12 micrometers. The antioxidant component particles may have a D50 of 40 micrometers or greater, 45 micrometers or greater, 48 micrometers or greater, or 50 micrometers or greater. The D50 may be 80 micrometers or less, 75 micrometers or less, 70 micrometers or less, 65 micrometers or less, or 60 micrometers or less. The D50 may be in a range of 40 to 80 micrometers, 45 to 70 micrometers, or 48 to 65 micrometers. The antioxidant component particles may have a D90 of 200 micrometers or greater, 210 micrometers or greater, 220 micrometers or greater, or 230 micrometers or greater. The D90 may be 400 micrometers or less, 375 micrometers or less, 350 micrometers or less, 335 micrometers or less, 330 micrometers or less, or 325 micrometers or less. The D90 may be in a range of 200 to 400 micrometers, 210 to 350 micrometers, or 220 to 325 micrometers.

[0075] In embodiments where the antioxidant pack has a bimodal particle size distribution, the antioxidant component may be ground to a first particle size population having a particle size volume distribution with DI OF of about 1 micrometer or greater, about 2 micrometers or greater, about 3 micrometers or greater, about 4 micrometers or greater, or about 5 micrometers or greater. The DI OF of the first particle size population may be about 20 micrometers smaller, about 18 micrometers smaller, about 15 micrometers smaller, or about 12 micrometers smaller. The D50F of the first particle size population may be about 20 micrometers or greater, about 25 micrometers or greater, about 30 micrometers or greater, about 35 micrometers or greater, about 40 micrometers or greater, or about 45 micrometers or greater. The D50F of the first particle size population may be about 80 micrometers smaller, about 70 micrometers smaller, about 60 micrometers smaller, or about 50 micrometers smaller. The D90F of the first particle size population may be about 100 micrometers or greater, about 110 micrometers or greater, about 120 micrometers or greater, about 130 micrometers or greater, or about 140 micrometers or greater. The D90F of the first particle size population may be about 200 micrometers smaller, about 180 micrometers smaller, about 180 micrometers smaller, or about 160 micrometers smaller. For example, the first particle size population may have a particle size volume distribution with DI OF in a range of 1 to 20 micrometers, D50F in a range of 20 to 80 micrometers, and D90F in a range of 100 to 200 micrometers. The first particle size population may have a particle size volume distribution with DI OF in a range of 2 to 15 micrometers, D50F in a range of 30 to 70 micrometers, and D90F in a range of 120 to 180 micrometers. The first particle size population may have a particle size volume distribution with DI OF in a range of 4 to 12 micrometers, D50F in a range of 40 to 60 micrometers, and D90F in a range of 140 to 160 micrometers.

[0076] The antioxidant component may be ground to a second particle size population having a particle size volume distribution with D 10c of about 1 micrometer or greater, about 2 micrometers or greater, about 3 micrometers or greater, about 4 micrometers or greater, or about 5 micrometers or greater. The D 10c of the second particle size population may be about 20 micrometers smaller, about 18 micrometers smaller, about 15 micrometers smaller, or about 12 micrometers smaller. The D50c of the second particle size population may be about 20 micrometers or greater, about 25 micrometers or greater, about 30 micrometers or greater, about 35 micrometers or greater, about 40 micrometers or greater, or about 45 micrometers or greater. The D50c of the second particle size population may be about 80 micrometers smaller, about 70 micrometers smaller, about 60 micrometers smaller, or about 50 micrometers smaller. The D90c of the second particle size population may be about 100 micrometers or greater, about 110micrometers or greater, about 120 micrometers or greater, about 130 micrometers or greater, or about 140 micrometers or greater. The D90c of the second particle size population may be about 750 micrometers or smaller, 500 micrometers or smaller, 400 micrometers or smaller, 200 micrometers or smaller, about 180 micrometers or smaller, about 175 micrometers or smaller, or about 160 micrometers or smaller. For example, the second particle size population may have a particle size volume distribution with D 10c in a range of 1 to 20 micrometers, D50c in a range of 20 to 80 micrometers, and D90c in a range of 100 to 750 micrometers. The second particle size population may have a particle size volume distribution with D 10c in a range of 2 to 18 micrometers, D50c in a range of 25 to 70 micrometers, and D90c in a range of 120 to 400 micrometers. The second particle size population may have a particle size volume distribution with DIOc in a range of 3 to 15 micrometers, D50c in a range of 30 to 60 micrometers, and D90c in a range of 150 to 200 micrometers.

[0077] In general, within the above particle size distribution ranges, the particle size of the second particle size population is greater than the corresponding particle size distribution of the first particle size population.

[0078] According to an embodiment, the first and second particle size populations are mixed together. The first and second particle size populations may be mixed together at a weight ratio of about 0.3 parts or more, about 0.5 parts or more, about 0.8 parts or more, about 1 part or more, about 1.3 parts or more, about 1.5 parts or more, about 1.8 parts or more, about 2 parts or more, about 2.5 parts or more, about 2.8 parts or more, or about 3 parts of the first particle size population for every 1 part of the second particle size population. The first and second particle size populations may be mixed together at a weight ratio of 0.3 to 3 parts for every 1 part of the second particle size population. The first and second particle size populations may be mixed together at a weight ratio of about 0.5 to 2 parts for every 1 part of the second particle size population. The first and second particle size populations may be mixed together at a weight ratio of about 0.8 to 1.3 parts for every 1 part of the second particle size population. In some embodiments, the first and second particle size populations may be mixed together in equal amounts (e.g., a weight ratio of 1 : 1).

[0079] The astaxanthin may be provided as particles. The astaxanthin particles may be mixed into the antioxidant component having a mixture of first and second particle size populations. The astaxanthin particles may be dispersed within the antioxidant component. The astaxanthin particles may be substantially evenly dispersed throughout the antioxidant component. The astaxanthin particles may have a particle size (D90) in a range of about 1 to 10 micrometers, orabout 4 micrometers. If the astaxanthin particles are larger, the particles size may be reduced to a desired size range.

[0080] The prepared feed additive pack may be mixed into the base feed for preparing the feed composition. The feed composition may also include additional ingredients in addition to the base feed and the feed additive pack. The base feed or any additional ingredients may undergo further grinding to facilitate their incorporation into the feed. In some embodiments, the base feed or any additional ingredients do not undergo additional grinding. After the feed additive pack is added to the base feed, the mixture may undergo mixing to distribute the feed additive pack throughout the base feed. After mixing of the based feed and feed additive pack, the mixture may be preconditioned to adjust its moisture content. The moisture content of the mixture may be brought to about 10 wt-% or more, about 15 wt-% or more, about 18 wt-% or more, about 20 wt-% or more, about 25 wt-% or more, about 30 wt-% or more, about 35 wt-% or more, about 40 wt-% or more, or about 45 wt-% or more. The moisture content of the mixture may be brought to about 50 wt-% or lower, about 45 wt-% or lower, about 40 wt-% or lower, about 35 wt-% or lower, about 30 wt-% or lower, about 25 wt-% or lower, about 20 wt-% or lower, about 15 wt-% or lower, or about 10 wt-% or lower. The moisture content of the mixture may be brought to from about 18 wt-% to about 40 wt-%.

[0081] The mixture may be then processed through extrusion to form pellets or other desired shapes, such as nuggets, steam pellets, or tablets. Extrusion may be followed by drying to remove excess moisture. The extruded feed may be cooled to ambient temperature. The extruded feed may be dried to a desired final moisture content to improve the stability and shelf-life of the final product. One or more additives may be included in or on the pellets. To further enhance the quality of the feed, the feed pellets may be coated. The pellets may be coated by one or more coating agents or oils. By way of example, a layer or coating of agent may be sealed by the application of a layer or coating of oil, such as fish oil. In various aspects, one or more further layers or coatings of fish feed may be applied to the (optionally sealed) coating or layer of fish feed. In this way, any given fish feed flake or pellet may include multiple layers of fish feed and / or sealing substance layers. In some embodiments, a thin layer of coating may be applied to improve appearance and nutritional content. Examples of suitable coating methods include vacuum coating, spray coating, dip coating, and the like. Alternatively, the feed mixture may be provided as a powder or as flakes. Finally, the feed may be packed into appropriate packaging materials to help preserve their freshness and quality until use.

[0082] Ordinarily, astaxanthin is susceptible to degradation during processing and / or storage of feed ingredients and the feed. It has surprisingly been found that including a bimodal particlesize distribution of the antioxidant component of the feed additive pack provides additional protection against degradation of the astaxanthin. According to an embodiment, at least 85 %, at least 90 %, or at least 95 % of the astaxanthin added to the feed additive pack during processing is preserved during the method of making the feed additive pack and the feed. The amount of astaxanthin lost during feed manufacturing (e.g., processing of the feed composition into the feed) may be 11 % or less, 10 % or less, 9 % or less, 8 % or less, 7 % or less, 6 % or less, 5 % or less, 4.9 % or less, 4.75 % or less, 4.5 % or less, 4 % or less, 3.5 % or less, 3 % or less, 2.5 % or less, or 2 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). In some embodiments, no astaxanthin is lost during processing. The amount of astaxanthin after processing may be about 85 % or more, about 89 % or more, about 90 % or more, about 91 % or more, about 92 % or more, about 93 % or more, about 94 % or more, about 95 % or more, about 95.1 % or more, about 95.25 % or more, about 95.5 % or more, about 96 % or more, about 96.5 % or more, about 97 % or more, about 97.5 % or more, about 98 % or more, or about 99 % of the initial amount of astaxanthin (astaxanthin added to the feed additive and / or feed composition. About 60 % or more, about 62 % or more, about 65 % or more, about 67 % or more, or about 70 % or more of the astaxanthin may be preserved in the feed during processing and storage for at least 2 weeks (14 days). The amount of astaxanthin lost during combined processing and storage for 2 weeks (14 days) may be 40 % or less, 38 % or less, 35 % or less, or 30 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). About 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin may be preserved in the feed during processing and storage for at least 3 weeks (21 days). The amount of astaxanthin lost during combined processing and storage may be 42 % or less, 40 % or less, 35 % or less, or 32 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). About 50 % or more, about 52 % or more, about 60 % or more, or about 62 % or more of the astaxanthin may be preserved in the feed during processing and storage for at least 4 weeks (28 days). The amount of astaxanthin lost during combined processing and storage for 4 weeks (28 days) may be 48 % or less, 45 % or less, 40 % or less, or 38 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additional grinding, and / or pelleting or other processing). Although there is no desired minimum loss, in practice, some astaxanthin is lost during processing and storage. For example, at least 20 % or at least 25 % of the astaxanthin that was added to the feed composition prior to processing may belost during processing and storage. The feed formulation may maintain about 78 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 1 week (7 days). The feed formulation may maintain about 76 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 2 weeks (14 days).Methods of Using

[0083] The present disclosure provides methods of using the fish feed. The present disclosure provides methods of feeding the fish feed to fish. The fish feed includes a feed additive pack that includes an antioxidant component and astaxanthin. The fish feed is formulated to reduce the breakdown or degradation of astaxanthin. A bimodal particle size distribution of the antioxidant component in the feed additive pack included in the fish feed provides additional protection against degradation of the astaxanthin. The bimodal particle size distribution of the antioxidant component provides additional protection against degradation of the astaxanthin during manufacturing (e.g., processing of the feed composition into the feed). The bimodal particle size distribution of the antioxidant component provides additional protection against degradation of the astaxanthin during storage. Thus, in some embodiments, the disclosure provides methods of using a fish feed that includes astaxanthin and an antioxidant component with a first particle size population and a second particle size population. The first particle size population may be referred to as the fine particles, and the second particle size population may be referred to as the coarse particles. Although there may be some overlap in the particle size distribution, the first particle size population generally includes particles that are smaller (finer) than the second particle size population.

[0084] The method of using the fish feed includes administering the feed to fish, where prior to administering (e.g., during processing) the feed additive pack maintains at least 85 %, at least 90 %, or at least about 95 % of the astaxanthin added to the feed additive. Preferably, the fish feed has a desired concentration of astaxanthin when the feed is fed to the fish. The feed may maintain at least about 50 % of the astaxanthin during processing and storage for at least 2 weeks (14 days). The feed may maintain at least about 58 % of the astaxanthin during processing and storage for at least 3 weeks (21 days). The feed may maintain at least about 50 % of the astaxanthin during processing and storage for at least 4 weeks (28 days). The amount of astaxanthin lost during combined processing and storage may be about 50 % or less, about 45 % or less, about 42 % or less, about 40 % or less, about 35 % or less, or about 32 % or less of the astaxanthin added to the feed composition prior to processing (e.g., prior to mixing, additionalgrinding, and / or pelleting or other processing). Thus, the feed administered to the fish may include about 50% or more, about 55% or more, about 60 % or more, about 65 % or more, or about 70 % or more of the astaxanthin that was added to the feed during manufacturing.

[0085] In some embodiments, the feed is manufactured by adding 100 ppm of astaxanthin to the feed. At the time of feeding the feed to an animal, the feed may have about 50 ppm or more, about 65 ppm or more, about 68 ppm or more, or about 70 ppm or more of astaxanthin.

[0086] In some embodiments, the present disclosure provides methods for feeding an aquatic animal. The method of feeding an aquatic animal may include administering a specific amount of feed. Further, the method may include administering a specific amount of astaxanthin to the fish. The fish feeds can be formulated such that the concentration of the astaxanthin / antioxidant component administered to the fish through the fish feed can be approximately from 1 mg to 120 mg per kg body weight / day (mg / kg / day), 1 to 90 mg / kg / day, 1 to 80 mg / kg / day, 1 to 70 mg / kg / day, 1 to 60 mg / kg / day, 5 to 50 mg / kg / day, 10 to 40 mg / kg / day, 15 to 35 mg / kg / day, 20 to 30 mg / kg / day, 0.01 to 10 mg / kg / day, or 0.01 to 5.0 mg / kg / day.EMBODIMENTS

[0087] The following is a list of exemplary embodiments according to the present disclosure:Embodiment Set 1

[0088] Embodiment l is a feed additive pack comprising: an antioxidant component; and astaxanthin dispersed within the antioxidant component, the antioxidant component comprising a first particle size population and a second particle size population, the first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and the second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90F is smaller than D90c and D50F is smaller than D50c.

[0089] Embodiment 2 is the feed additive pack of embodiment 1, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers.

[0090] Embodiment 3 is the feed additive pack of embodiment 1 or 2, wherein the first particle size population has a D50F in a range of 35 to 65 micrometers.

[0091] Embodiment 4 is the feed additive pack of any one of embodiments 1 to 3, wherein the first particle size population has a D90F in a range of 120 to 180 micrometers.

[0092] Embodiment 5 is the feed additive pack of any one of embodiments 1 to 4, wherein the second particle size population has a DIOc in a range of 2 to 20 micrometers.

[0093] Embodiment 6 is the feed additive pack of any one of embodiments 1 to 5, wherein the second particle size population has a D50c in a range of 45 to 75 micrometers.

[0094] Embodiment 7 is the feed additive pack of any one of embodiments 1 to 6, wherein the second particle size population has a D90c in a range of 180 to 400 micrometers.

[0095] Embodiment 8 is the feed additive pack of any one of embodiments 1 to 7, wherein a weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part of the second particle size population.

[0096] Embodiment 9 is the feed additive pack of any one of embodiments 1 to 8, wherein a weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0097] Embodiment 10 is the feed additive pack of any one of embodiments 1 to 9, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-%.

[0098] Embodiment 11 is the feed additive pack of any one of embodiments 1 to 10, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers.

[0099] Embodiment 12 is the feed additive pack of any one of embodiments 1 to 11, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0100] Embodiment 13 is the feed additive pack of any one of embodiments 1 to 12, wherein the primary antioxidant comprises a free radical scavenger.

[0101] Embodiment 14 is the feed additive pack of any one of embodiments 1 to 13, wherein the primary antioxidant comprises butylated hydroxyanisol (BHA), a carotenoid, a flavonoid, phenolic acid, a tocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0102] Embodiment 15 is the feed additive pack of any one of embodiments 1 to 14, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%.

[0103] Embodiment 16 is the feed additive pack of any one of embodiments 1 to 15, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0104] Embodiment 17 is the feed additive pack of any one of embodiments 1 to 16, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol, polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0105] Embodiment 18 is the feed additive pack of any one of embodiments 1 to 17, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0106] Embodiment 19 is the feed additive pack of any one of embodiments 1 to 18, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0107] Embodiment 20 is the feed additive pack of any one of embodiments 1 to 19, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0108] Embodiment 21 is the feed additive pack of any one of embodiments 1 to 20, wherein during processing the feed additive pack maintains at least 85 % of the astaxanthin added to the feed additive.

[0109] Embodiment 22 is the feed additive pack of any one of embodiments 1 to 21, wherein the feed additive pack maintains 58 % or more of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0110] Embodiment 22A is the feed additive pack of any one of embodiments 1 to 22, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0111] Embodiment 22B is the feed additive pack of any one of embodiments 1 to 22A, wherein the feed additive pack maintains about 76 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 2 weeks (14 days).

[0112] Embodiment 22C is the feed additive pack of any one of embodiments 1 to 22B, wherein the feed additive pack maintains about 78 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 1 week (7 days).

[0113] Embodiment 23 is a fish feed comprising:a base feed; and a feed additive pack comprising: an antioxidant component; and astaxanthin dispersed within the antioxidant component, the antioxidant component comprising a first particle size population and a second particle size population, the first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and the second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90F is smaller than D90c and D50F is smaller than D50c.

[0114] Embodiment 24 is the fish feed of embodiment 23, wherein the astaxanthin is added to the fish feed at a first concentration, and wherein the astaxanthin is present in the fish feed at a second concentration after processing and storage for 21 days or longer, and wherein the second concentration is at least 66 % of the first concentration.

[0115] Embodiment 25 is the fish feed of embodiments 23 or 24, wherein the fish feed is a complete fish feed.

[0116] Embodiment 26 is the fish feed of any one of embodiments 23 to 25, wherein the base feed comprises protein, starch, and fat.

[0117] Embodiment 27 is the fish feed of any one of embodiments 23 to 26, wherein the fish feed comprises from 28 wt-% to 60 wt-% protein, from 6 wt-% to 15 wt-% of starch, and from 18 wt-% to 40 wt-% fat.

[0118] Embodiment 28 is the fish feed of any one of embodiments 23 to 27, wherein the fish feed comprises land-animal protein, fish meal, plant-based protein, or a combination of two or more thereof.

[0119] Embodiment 29 is the fish feed of any one of embodiments 23 to 28, wherein the fish feed comprises one or more of pellets, extruded nuggets, steam pellets, flakes, tablets, granules, or powders.

[0120] Embodiment 30 is the fish feed of any one of embodiments 23 to 29, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers.

[0121] Embodiment 31 is the fish feed of any one of embodiments 23 to 30, wherein the first particle size population has a D50F in a range of 35 to 65 micrometers.

[0122] Embodiment 32 is the fish feed of any one of embodiments 23 to 31, wherein the first particle size population has a D90F in a range of 120 to 180 micrometers.

[0123] Embodiment 33 is the fish feed of any one of embodiments 23 to 32, wherein the second particle size population has a DIOc in a range of 2 to 20 micrometers.

[0124] Embodiment34 is the fish feed of any one of embodiments 23 to 33, wherein the second particle size population has a D50c in a range of 45 to 75 micrometers.

[0125] Embodiment 35 is the fish feed of any one of embodiments 23 to 34, wherein the second particle size population has a D90c in a range of 180 to 400 micrometers.

[0126] Embodiment 36 is the fish feed of any one of embodiments 23 to 34, wherein a weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part of the second particle size population.

[0127] Embodiment 37 is the fish feed of any one of embodiments 23 to 36, wherein a weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0128] Embodiment 38 is the fish feed of any one of embodiments 23 to 37, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-%.

[0129] Embodiment 39 is the fish feed of any one of embodiments 23 to 38, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers.

[0130] Embodiment 40 is the fish feed of any one of embodiments 23 to 39, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0131] Embodiment 41 is the fish feed of any one of embodiments 23 to 40, wherein the primary antioxidant comprises a free radical scavenger.

[0132] Embodiment 42 is the fish feed of any one of embodiments 23 to 41, wherein the primary antioxidant comprises butylated hydroxyanisol (BHA), a carotenoid, a flavonoid, phenolic acid, a tocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0133] Embodiment 43 is the fish feed of any one of embodiments 23 to 42, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%.

[0134] Embodiment 44 is the fish feed of any one of embodiments 23 to 43, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0135] Embodiment 45 is the fish feed of any one of embodiments 23 to 44, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol, polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0136] Embodiment 46 is the fish feed of any one of embodiments 23 to 45, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0137] Embodiment 47 is the fish feed of any one of embodiments 23 to 46, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0138] Embodiment 48 is the fish feed of any one of embodiments 23 to 47, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0139] Embodiment 49 is the fish feed of any one of embodiments 23 to 48, wherein during processing the feed additive pack maintains at least 85 % of the astaxanthin added to the feed additive pack.

[0140] Embodiment 50 is the fish feed of any one of embodiments 23 to 49, wherein the feed additive pack maintains 58 % or more of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0141] Embodiment 50A is the fish feed of any one of embodiments 23 to 50, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0142] Embodiment 50B is the fish feed of any one of embodiments 23 to 50A, wherein the feed additive pack maintains about 76 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 2 weeks (14 days).

[0143] Embodiment 50C is the fish feed of any one of embodiments 23 to 50B, wherein the feed additive pack maintains about 78 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 1 week (7 days).

[0144] Embodiment 51 is a method for making a feed additive pack for adding to a fish feed, the method comprising: grinding a first amount of an antioxidant mixture to fine particles having a first particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers;grinding a second amount of the antioxidant mixture to coarse particles having a second particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90F is smaller than D90c and D50F is smaller than D50c; and mixing the coarse particles, the fine particles, and particles containing astaxanthin.

[0145] Embodiment 52 is the method of embodiment 51, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers.

[0146] Embodiment 53 is the method of embodiment 51 or 52, wherein the first particle size population has a D50F in a range of 35 to 65 micrometers.

[0147] Embodiment 54 is the method of any one of embodiments 51 to 53, wherein the first particle size population has a D90F in a range of 120 to 180 micrometers.

[0148] Embodiment 55 is the method of any one of embodiments 51 to 54, wherein the second particle size population has a DIOc in a range of 2 to 20 micrometers.

[0149] Embodiment 56 is the method of any one of embodiments 51 to 55, wherein the second particle size population has a D50c in a range of 45 to 75 micrometers.

[0150] Embodiment 57 is the method of any one of embodiments 51 to 56, wherein the second particle size population has a D90c in a range of 180 to 400 micrometers.

[0151] Embodiment 58 is the method of any one of embodiments 51 to 57, wherein a weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part of the second particle size population.

[0152] Embodiment 59 is the method of any one of embodiments 51 to 58, wherein a weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0153] Embodiment 60 is the method of any one of embodiments 51 to 59, wherein the astaxanthin is present in the method at a concentration of about 0.5 wt-% to about 50 wt-%.

[0154] Embodiment 61 is the method of any one of embodiments 51 to 60, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers.

[0155] Embodiment 62 is the method of any one of embodiments 51 to 61, wherein the method comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0156] Embodiment 63 is the method of any one of embodiments 51 to 62, wherein the primary antioxidant comprises a free radical scavenger.

[0157] Embodiment 64 is the method of any one of embodiments 51 to 63, wherein the primary antioxidant comprises butylated hydroxyanisol (BHA), a carotenoid, a flavonoid, phenolic acid, atocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0158] Embodiment 65 is the method of any one of embodiments 51 to 64, wherein the primary antioxidant is present in the feed additive pack ant at a concentration of about 1 wt-% to about 10 wt-%.

[0159] Embodiment 66 is the method of any one of embodiments 51 to 65, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0160] Embodiment 67 is the method of any one of embodiments 51 to 66, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol, polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0161] Embodiment 68 is the method of any one of embodiments 51 to 67, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0162] Embodiment 69 is the method of any one of embodiments 51 to 68, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0163] Embodiment 70 is the method of any one of embodiments 51 to 69, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0164] Embodiment 71 is the method of any one of embodiments 51 to 70, wherein during processing the feed additive pack maintains at least 85 % of the astaxanthin added to the feed additive.

[0165] Embodiment 72 is the method of any one of embodiments 51-71, wherein the feed additive pack maintains 58 % or more of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0166] Embodiment 72A is the method of any one of embodiments 51 to 72, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0167] Embodiment 72B is the method of any one of embodiments 51 to 72A, wherein the feed additive pack maintains about 76 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 2 weeks (14 days).

[0168] Embodiment 72C is the method of any one of embodiments 51 to 72B, wherein the feed additive pack maintains about 78 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 1 week (7 days).

[0169] Embodiment 73 is a method for making a fish feed, the method comprising: mixing a feed additive pack with a base feed, the feed additive pack comprising: an antioxidant component; and astaxanthin dispersed within the antioxidant component, the antioxidant component comprising a first particle size population and a second particle size population, the first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and the second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90F is smaller than D90c and D50F is smaller than D50c.

[0170] Embodiment 74 is the method of embodiment 73, wherein the astaxanthin is added to the fish feed at a first concentration, and wherein the astaxanthin is present in the fish feed at a second concentration after processing and storage for 21 days or longer, and wherein the second concentration is at least 66 % of the first concentration.

[0171] Embodiment 75 is the method of embodiment 73 or 74, wherein the fish feed is a complete fish feed.

[0172] Embodiment 76 is the method of any one of embodiments 73 to 75, wherein the base feed comprises protein, starch, and fat.

[0173] Embodiment 77 is the method of any one of embodiments 73 to 76, wherein the fish feed comprises from 28 wt-% to 60 wt-% protein, from 6 wt-% to 15 wt-% of starch, and from 18 wt-% to 40 wt-% fat.

[0174] Embodiment 78 is the method of any one of embodiments 73 to 77, wherein the fish feed comprises land-animal protein, fish meal, plant-based protein, or a combination of two or more thereof.

[0175] Embodiment 79 is the method of any one of embodiments 73 to 78, wherein the fish feed comprises one or more of pellets, extruded nuggets, steam pellets, flakes, tablets, granules, or powders.

[0176] Embodiment 80 is the method of any one of embodiments 51 to 79, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers.

[0177] Embodiment 81 is the method of any one of embodiments 51 to 80, wherein the first particle size population has a D50F in a range of 35 to 65 micrometers.

[0178] Embodiment 82 is the method of any one of embodiments 51 to 81, wherein the first particle size population has a D90F in a range of 120 to 180 micrometers.

[0179] Embodiment 83 is the method of any one of embodiments 51 to 82, wherein the second particle size population has a DIOc in a range of 2 to 20 micrometers.

[0180] Embodiment 84 is the method of any one of embodiments 51 to 83, wherein the second particle size population has a D50c in a range of 45 to 75 micrometers.

[0181] Embodiment 85 is the method of any one of embodiments 51 to 84, wherein the second particle size population has a D90c in a range of 180 to 400 micrometers.

[0182] Embodiment 86 is the method of any one of embodiments 51 to 85, wherein a weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part of the second particle size population.

[0183] Embodiment 87 is the method of any one of embodiments 51 to 86, wherein a weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0184] Embodiment 88 is the method of any one of embodiments 51 to 87, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-%.

[0185] Embodiment 89 is the method of any one of embodiments 51 to 88, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers.

[0186] Embodiment 90 is the method of any one of embodiments 51 to 89, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0187] Embodiment 91 is the method of any one of embodiments 51 to 90, wherein the primary antioxidant comprises a free radical scavenger.

[0188] Embodiment 92 is the method of any one of embodiments 51 to 91, wherein the primary antioxidant comprises butylated hydroxyanisol (BHA), a carotenoid, a flavonoid, phenolic acid, a tocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0189] Embodiment 93 is the method of any one of embodiments 51 to 92, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%.

[0190] Embodiment 94 is the method of any one of embodiments 51 to 93, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0191] Embodiment 95 is the method of any one of embodiments 51 to 94, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol, polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0192] Embodiment 96 is the method of any one of embodiments 51 to 95, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0193] Embodiment 97 is the method of any one of embodiments 51 to 96, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0194] Embodiment 98 is the method of any one of embodiments 51 to 97, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0195] Embodiment 99 is the method of any one of embodiments 51 to 98, wherein during processing the feed additive pack maintains at least 85 % of the astaxanthin added to the feed additive.

[0196] Embodiment 100 is the method of any one of embodiments 51 to 99, wherein the fish feed maintains 58 % or more of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0197] Embodiment 100A is the method of any one of embodiments 51 to 100, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0198] Embodiment 100B is the method of any one of embodiments 51 to 100A, wherein the feed additive pack maintains about 76 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 2 weeks (14 days).

[0199] Embodiment 100C is the method of any one of embodiments 51 to 100B, wherein the feed additive pack maintains about 78 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 1 week (7 days).

[0200] Embodiment 101 is the method of any one of embodiments 48 to 100C, further comprising extruding the fish feed composition to form pellets.

[0201] Embodiment 102 is a method of feeding an aquatic animal, the method comprising administering an amount of feed to the aquatic animal, the feed comprising: a base feed; and a feed additive pack, the feed additive pack comprising: an antioxidant component; and astaxanthin dispersed within the antioxidant component, the antioxidant component comprising a first particle size population and a second particle size population, the first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and the second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90F is smaller than D90c and D50F is smaller than D50c.

[0202] Embodiment 103 is the method of embodiment 102, wherein the astaxanthin is added to the fish feed at a first concentration, and wherein the astaxanthin is present in the fish feed at a second concentration after processing and storage for 21 days or longer, and wherein the second concentration is at least 66 % of the first concentration.

[0203] Embodiment 104 is the method of embodiment 102 or 103, wherein the fish feed is a complete fish feed.

[0204] Embodiment 105 is the method of any one of embodiments 102 to 104, wherein the base feed comprises protein, starch, and fat.

[0205] Embodiment 106 is the method of any one of embodiments 102 to 105, wherein the fish feed comprises from 28 wt-% to 60 wt-% protein, from 6 wt-% to 15 wt-% of starch, and from 18 wt-% to 40 wt-% fat.

[0206] Embodiment 107 is the method of any one of embodiments 102 to 106, wherein the fish feed comprises land-animal protein, fish meal, plant-based protein, or a combination of two or more thereof.

[0207] Embodiment 108 is the method of any one of embodiments 102 to 107, wherein the fish feed comprises one or more of pellets, extruded nuggets, steam pellets, flakes, tablets, granules, or powders.

[0208] Embodiment 109 is the method of any one of embodiments 102 to 108, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers.

[0209] Embodiment 110 is the method of any one of embodiments 102 to 109, wherein the first particle size population has a D50F in a range of 35 to 65 micrometers.

[0210] Embodiment 111 is the method of any one of embodiments 102 to 110, wherein the first particle size population has a D90F in a range of 120 to 180 micrometers.

[0211] Embodiment 112 is the method of any one of embodiments 102 to 111, wherein the second particle size population has a DIOc in a range of 2 to 20 micrometers.

[0212] Embodiment 113 is the method of any one of embodiments 102 to 112, wherein the second particle size population has a D50c in a range of 45 to 75 micrometers.

[0213] Embodiment 114 is the method of any one of embodiments 102 to 113, wherein the second particle size population has a D90c in a range of 180 to 400 micrometers.

[0214] Embodiment 115 is the method of any one of embodiments 102 to 114, wherein weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part of the second particle size population.

[0215] Embodiment 116 is the method of any one of embodiments 102 to 115, wherein weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0216] Embodiment 117 is the method of any one of embodiments 102 to 116, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50

[0217] Embodiment 118 is the method of any one of embodiments 102 to 117, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers.

[0218] Embodiment 119 is the method of any one of embodiments 102 to 118, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0219] Embodiment 120 is the method of embodimentl l9, wherein the primary antioxidant comprises a free radical scavenger.

[0220] Embodiment 121 is the method of embodiment 119 or 120, wherein the primary antioxidant comprises butylated hydroxyanisol (BHA), butylated hydroxytoluene (BHT), propylgallate (PG), a carotenoid, a flavonoid, a phenolic acid, a tocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0221] Embodiment 122 is the method of any one of embodiments 119 to 121, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%.

[0222] Embodiment 123 is the method of any one of embodiments 119 to 122, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0223] Embodiment 124 is the method of any one of embodiments 119 to 123, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol, polysorbate, butylated hydroxytoluene (BEIT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0224] Embodiment 125 is the method of any one of embodiments 119 to 124, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0225] Embodiment 126 is the method of any one of embodiments 119 to 125, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

[0226] Embodiment 127 is the method of any one of embodiments 119 to 126, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0227] Embodiment 128 is the method of any one of embodiments 102 to 127, wherein during processing the fish feed maintains at least 85 % of the astaxanthin added to the feed additive pack.

[0228] Embodiment 129 is the method of any one of embodiments 102 to 128, wherein the fish feed maintains 58 % or more of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0229] Embodiment 129A is the method of any one of embodiments 102 to 129, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0230] Embodiment 129B is the method of any one of embodiments 102 to 129A, wherein the feed additive pack maintains about 76 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 2 weeks (14 days).

[0231] Embodiment 129C is the method of any one of embodiments 102 to 129B, wherein the feed additive pack maintains about 78 % or more, about 80 % or more, about 82 % or more, about 84 % or more, or about 86 % of the astaxanthin during storage for at least 1 week (7 days).

[0232] Embodiment 130 is the method of any one of embodiments 102 to 129C, further comprising extruding the fish feed composition to form pellets

[0233] Embodiment 131 is the method of any one of embodiments 102 to 130, wherein the fish feed comprises an amount of astaxanthin that is at least 85 % of the astaxanthin added to the fish feed prior to mixing.

[0234] Embodiment 132 is the method of any one of embodiments 102 to 131, wherein the feed comprises an amount of astaxanthin that is 58 % or more of the astaxanthin added to the feed.

[0235] The technology of the present disclosure 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.Embodiment Set 2

[0236] Embodiment 2-1 is a feed additive pack comprising: an antioxidant component; and astaxanthin dispersed within the antioxidant component, the antioxidant component comprising particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers.The D80 of the antioxidant component particles may be 140 micrometers or greater, 145 micrometers or greater, 150 micrometers or greater, 155 micrometers or greater, or 165 micrometers or greater. The D80 of the antioxidant component particles may be 200 micrometers or less, 190 micrometers or less, 185 micrometers or less, or 180 micrometers or less.

[0237] Embodiment 2-2 is the feed additive pack of embodiment 2-1, wherein the particles have a D10 in a range of 5 to 20 micrometers, 6 to 15 micrometers, or 8 to 12 micrometers.

[0238] Embodiment 2-3 is the feed additive pack of embodiment 2-1 or 2-2, wherein the particles have a D50 in a range of 40 to 80 micrometers, 45 to 70 micrometers, or 48 to 65 micrometers.

[0239] Embodiment 2-4 is the feed additive pack of any one of the preceding embodiments, wherein the particles have a D90 in a range of 200 to 400 micrometers, 210 to 350 micrometers, or 220 to 325 micrometers.

[0240] Embodiment 2-5 is the feed additive pack of any one of the preceding embodiments, wherein the antioxidant component comprises a first particle size population and a second particle size population, the first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and the second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90p is smaller than D90c and D50F is smaller than D50c.

[0241] Embodiment 2-6 is the feed additive pack of embodiment 2-5, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers, 2 to 15 micrometers, or 4 to 12 micrometers.

[0242] Embodiment 2-7 is the feed additive pack of embodiments 2-5 or 2-6, wherein the first particle size population has a D50F in a range of 20 to 80 micrometers, 30 to 70 micrometers, 35 to 65 micrometers, or 40 to 60 micrometers.

[0243] Embodiment 2-8 is the feed additive pack of any one of embodiments 2-5 to 2-7, wherein the first particle size population has a D90F in a range of 100 to 200 micrometers, 120 to 180 micrometers, 140 to 160 micrometers.

[0244] Embodiment 2-9 is the feed additive pack of any one of embodiments 2-5 to 2-8, wherein the second particle size population has a DIOc in a range of 1 to 20 micrometers or 20 to 80 micrometers or 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0245] Embodiment 2-10 is the feed additive pack of any one of embodiments 2-5 to 2-9, wherein the second particle size population has a D50c in a range of 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0246] Embodiment 2-11 is the feed additive pack of any one of embodiments 2-5 to 2-10, wherein the second particle size population has a D90c in a range of 100 to 400 micrometers, 110 to 200 micrometers, or 120 to 180 micrometers.

[0247] Embodiment 2-12 is the feed additive pack of any one of embodiments 2-5 to 2-11, wherein weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part, about 0.5 to 2 parts of the first particle size population for every 1 part, or about 0.8 to 1.3 parts of the first particle size population for every 1 part of the second particle size population.

[0248] Embodiment 2-13 is the feed additive pack of any one of embodiments 2-5 to 2-12, wherein weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0249] Embodiment 2-14 is the feed additive pack of any one of the preceding embodiments, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-%, about 2 wt-% to about 25 wt-%, or about 5 wt-% to about 20 wt-%.

[0250] Embodiment 2-15 is the feed additive pack of any one of the preceding embodiments, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers, or about 4 micrometers.

[0251] Embodiment 2-16 is the feed additive pack of any one of the preceding embodiments, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0252] Embodiment 2-17 is the feed additive pack of embodiment 2-16, wherein the primary antioxidant comprises a free radical scavenger.

[0253] Embodiment 2-18 is the feed additive pack of embodiment 2-16 or 2-17, wherein the primary antioxidant comprises butylated hydroxyanisol (BHa), butylated hydroxytoluene (BHT), propyl gallate (PG), a carotenoid, a flavonoid, a phenolic acid, a tocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0254] Embodiment 2-19 is the feed additive pack of any one of embodiments 16 to 18, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%.

[0255] Embodiment 2-20 is the feed additive pack of any one embodiments 2-16 to 2-19, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0256] Embodiment 2-21 is the feed additive pack of any one of embodiments 2-16 to 2-20, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alphatocopherol, polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0257] Embodiment 2-22 is the feed additive pack of any one of embodiments 2-16 to 2-21, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0258] Embodiment 2-23 is the feed additive pack of any one of embodiments 2-16 to 2-22, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present inthe feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0259] Embodiment 2-24 is the feed additive pack of any one embodiments 2-16 to 2-23, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0260] Embodiment 2-25 is the feed additive pack of any one of the preceding embodiments, wherein during processing the feed additive pack maintains at least 85 %, at least 90 %, or at least 95 % of the astaxanthin added to the feed additive pack.

[0261] Embodiment 2-26 is the feed additive pack of any one of the preceding embodiments, wherein the feed additive pack maintains about 60 % or more, about 62 % or more, about 65 % or more, about 67 % or more, or about 70 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days).

[0262] Embodiment 2-27 is the feed additive pack of any one of the preceding embodiments, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0263] Embodiment 2-28 is the feed additive pack of any one of the preceding embodiments, wherein the feed additive pack maintains about 50 % or more, about 52 % or more, about 60 % or more, or about 62 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days).

[0264] Embodiment 2-29 is a fish feed comprising: a base feed; and a feed additive pack comprising: an antioxidant component; and astaxanthin dispersed within the antioxidant component, the antioxidant component comprising particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers. The D80 of the antioxidant component particles may be 140 micrometers or greater, 145 micrometers or greater, 150 micrometers or greater, 155 micrometers or greater, or 165 micrometers or greater. The D80 of the antioxidant component particles may be 200 micrometers or less, 190 micrometers or less, 185 micrometers or less, or 180 micrometers or less.

[0265] Embodiment 2-30 is the fish feed of embodiment 2-29, wherein the astaxanthin is added to the fish feed at a first concentration, and wherein the astaxanthin is present in the fish feed at a second concentration after processing and storage for 21 days or longer, and wherein the second concentration is at least 66 % of the first concentration.

[0266] Embodiment 2-31 is the fish feed of embodiment 2-29 or 2-30, wherein the fish feed is a complete fish feed.

[0267] Embodiment 2-32 is the fish feed of any one of embodiments 2-29 to 2-31, wherein the base feed comprises protein, starch, and fat.

[0268] Embodiment 2-33 is the fish feed of any one of embodiments 2-29 to 2-32, wherein the fish feed comprises from 28 wt-% to 60 wt-% protein, from 6 wt-% to 15 wt-% of starch, and from 18 wt-% to 40 wt-% fat.

[0269] Embodiment 2-34 is the fish feed of any one of embodiments 2-29 to 2-33, wherein the fish feed comprises land-animal protein, fish meal, plant-based protein, or a combination of two or more thereof.

[0270] Embodiment 2-35 is the fish feed of any one of embodiments 2-29 to 2-34, wherein the fish feed comprises one or more of pellets, extruded nuggets, steam pellets, flakes, tablets, granules, or powders.

[0271] Embodiment 2-36 is the feed additive pack of embodiment 1, wherein the particles have a D10 in a range of 5 to 20 micrometers, 6 to 15 micrometers, or 8 to 12 micrometers.

[0272] Embodiment 2-37 is the feed additive pack of embodiment 1 or 2, wherein the particles have a D50 in a range of 40 to 80 micrometers, 45 to 70 micrometers, or 48 to 65 micrometers.

[0273] Embodiment 2-38 is the feed additive pack of any one of the preceding embodiments, wherein the particles have a D90 in a range of 200 to 400 micrometers, 210 to 350 micrometers, or 220 to 325 micrometers.

[0274] Embodiment 2-39 is the fish feed of any one of the preceding embodiments, wherein the antioxidant component comprises a first particle size population and a second particle size population, the first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and the second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90p is smaller than D90c and D50F is smaller than D50c.

[0275] Embodiment 2-40 is the fish feed of embodiment 2-39, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers, 2 to 15 micrometers, or 4 to 12 micrometers.

[0276] Embodiment 2-41 is the fish feed of embodiment 2-39 or 2-40, wherein the first particle size population has a D50F in a range of 20 to 80 micrometers, 30 to 70 micrometers, 35 to 65 micrometers, or 40 to 60 micrometers.

[0277] Embodiment 2-42 is the fish feed of any one of embodiments 2-39 to 2-41, wherein the first particle size population has a D90F in a range of 100 to 200 micrometers, 120 to 180 micrometers, 140 to 160 micrometers.

[0278] Embodiment 2-43 is the fish feed of any one of embodiments 2-39 to 2-42, wherein the second particle size population has a DIOc in a range of 1 to 20 micrometers or 20 to 80 micrometers or 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0279] Embodiment 2-44 is the fish feed of any one of embodiments 2-39 to 2-43, wherein the second particle size population has a D50c in a range of 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0280] Embodiment 2-45 is the fish feed of any one of embodiments 2-39 to 2-44, wherein the second particle size population has a D90c in a range of 100 to 400 micrometers, 110 to 200 micrometers, or 120 to 180 micrometers.

[0281] Embodiment 2-46 is the fish feed of any one of embodiments 2-39 to 2-45, wherein weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part, about 0.5 to 2 parts of the first particle size population for every 1 part, or about 0.8 to 1.3 parts of the first particle size population for every 1 part of the second particle size population.

[0282] Embodiment 2-47 is the fish feed of any one of embodiments 2-39 to 2-46, wherein weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0283] Embodiment 2-48 is the fish feed of any one of embodiments 2-29 to 2-47, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-%, about 2 wt-% to about 25 wt-%, or about 5 wt-% to about 20 wt-%.

[0284] Embodiment 2-49 is the fish feed of any one of embodiments 2-29 to 2-48, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers, or about 4 micrometers.

[0285] Embodiment 2-50 is the fish feed of any one of embodiments 2-29 to 2-49, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0286] Embodiment 2-51 is the fish feed of embodiment 2-50, wherein the primary antioxidant comprises a free radical scavenger.

[0287] Embodiment 2-52 is the fish feed of embodiment 2-50 or 2-51, wherein the primary antioxidant comprises butylated hydroxyanisol (BHa), butylated hydroxytoluene (BHT), propyl gallate (PG), a carotenoid, a flavonoid, a phenolic acid, a tocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0288] Embodiment 2-53 is the fish feed of any one of embodiments 2-50 to 2-52, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%.

[0289] Embodiment 2-54 is the fish feed of any one of embodiments 2-50 to 2-53, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0290] Embodiment 2-55 is the fish feed of any one of embodiments 2-50 to 2-54, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol, polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0291] Embodiment 2-56 is the fish feed of any one of embodiments 2-50 to 2-55, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0292] Embodiment 2-57 is the fish feed of any one of embodiments 2-50 to 2-56, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0293] Embodiment 2-58 is the fish feed of any one of embodiments 2-50 to 2-57, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0294] Embodiment 2-59 is the fish feed of any one of embodiments 2-29 to 2-58, wherein during processing the feed additive pack maintains at least 85 %, at least 90 %, or at least 95 % of the astaxanthin added to the feed additive pack.

[0295] Embodiment 2-60 is the fish feed of any one of embodiments 2-29 to 2-59, wherein the feed additive pack maintains about 60 % or more, about 62 % or more, about 65 % or more, about67 % or more, or about 70 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days).

[0296] Embodiment 2-61 is the fish feed of any one of embodiments 2-29 to 2-60, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more, about68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0297] Embodiment 2-62 is the fish feed of any one of embodiments 2-29 to 2-61, wherein the feed additive pack maintains about 50 % or more, about 52 % or more, about 60 % or more, or about 62 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days).

[0298] Embodiment 2-63 is a method for making a feed additive pack for adding to a fish feed, the method comprising: grinding an antioxidant mixture to particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers; and mixing the particles with particles containing astaxanthin.

[0299] Embodiment 2-64 is the method of embodiment 2-63, wherein the particles have a D10 in a range of 5 to 20 micrometers, 6 to 15 micrometers, or 8 to 12 micrometers.

[0300] Embodiment 2-65 is the method of embodiment 2-63 or 2-64, wherein the particles have a D50 in a range of 40 to 80 micrometers, 45 to 70 micrometers, or 48 to 65 micrometers.

[0301] Embodiment 2-66 is the method of any one of embodiments 2-63 to 2-65, wherein the particles have a D90 in a range of 200 to 400 micrometers, 210 to 350 micrometers, or 220 to 325 micrometers.

[0302] Embodiment 2-67 is the method of any one of embodiments 2-63 to 2-66, wherein the grinding comprises grinding a first amount of the antioxidant mixture to fine particles having a first particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers; and grinding a second amount of the antioxidant mixture to coarse particles having a second particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90p is smaller than D90c and D50F is smaller than D50c.

[0303] Embodiment 2-68 is the method of embodiment 2-67, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers, 2 to 15 micrometers, or 4 to 12 micrometers.

[0304] Embodiment 2-69 is the method of embodiment 2-67 or 2-68, wherein the first particle size population has a D50F in a range of 20 to 80 micrometers, 30 to 70 micrometers, 35 to 65 micrometers, or 40 to 60 micrometers.

[0305] Embodiment 2-70 is the method of any one of embodiments 2-67 to 2-69, wherein the first particle size population has a D90F in a range of 100 to 200 micrometers, 120 to 180 micrometers, 140 to 160 micrometers.

[0306] Embodiment 2-71 is the method of any one of embodiments 2-67 to 2-70, wherein the second particle size population has a DIOc in a range of 1 to 20 micrometers or 20 to 80 micrometers or 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0307] Embodiment 2-72 is the method of any one of embodiments 2-67 to 2-71, wherein the second particle size population has a D50c in a range of 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0308] Embodiment 2-73 is the method of any one of embodiments 2-67 to 2-72, wherein the second particle size population has a D90c in a range of 100 to 400 micrometers, 110 to 200 micrometers, or 120 to 180 micrometers.

[0309] Embodiment 2-74 is the method of any one of embodiments 2-67 to 2-73, wherein weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part, about 0.5 to 2 parts of the first particle size population for every 1 part, or about 0.8 to 1.3 parts of the first particle size population for every 1 part of the second particle size population.

[0310] Embodiment 2-75 is the method of any one of embodiments 2-67 to 2-74, wherein weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0311] Embodiment 2-76 is the method of any one of embodiments 2-63 to 2-75, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-%, about 2 wt-% to about 25 wt-%, or about 5 wt-% to about 20 wt-%.

[0312] Embodiment 2-77 is the method of any one of embodiments 2-63 to 2-76, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers, or about 4 micrometers..

[0313] Embodiment 2-78 is the method of any one of embodiments 2-63 to 2-77, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0314] Embodiment 2-79 is the method of embodiment 2-78, wherein the primary antioxidant comprises a free radical scavenger.

[0315] Embodiment 2-80 is the method of embodiment 2-78 or 2-79, wherein the primary antioxidant comprises butylated hydroxyanisol (BHa), butylated hydroxytoluene (BHT), propyl gallate (PG), a carotenoid, a flavonoid, a phenolic acid, a tocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0316] Embodiment 2-81 is the method of any one of embodiments 2-78 to 2-80, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%.

[0317] Embodiment 2-82 is the method of any one of embodiments 2-78 to 2-81, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0318] Embodiment 2-83 is the method of any one of embodiments 2-78 to 2-82, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol,polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0319] Embodiment 2-84 is the method of any one of embodiments 2-78 to 2-83, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0320] Embodiment 2-85 is the method of any one of embodiments 2-78 to 2-84, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0321] Embodiment 2-86 is the method of any one of embodiments 2-78 to 2-85, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0322] Embodiment 2-87 is the method of any one of embodiments 2-78 to 2-86, wherein during processing the feed additive pack maintains at least 85 % of the astaxanthin added to the feed additive pack.

[0323] Embodiment 2-88 is the method of any one of embodiments 2-78 to 2-87, wherein the feed additive pack maintains about 60 % or more, about 62 % or more, about 65 % or more, about67 % or more, or about 70 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days).

[0324] Embodiment 2-89 is the method of any one of embodiments 2-78 to 2-88, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more, about68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0325] Embodiment 2-90 is the method of any one of embodiments 2-78 to 2-89, wherein the feed additive pack maintains about 50 % or more, about 52 % or more, about 60 % or more, or about 62 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days).

[0326] Embodiment 2-91 is a method for making a fish feed, the method comprising: mixing a feed additive pack with a base feed, the feed additive pack comprising: an antioxidant component; and astaxanthin dispersed within the antioxidant component, the antioxidant component comprising particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers. The D80 of the antioxidant component particles may be 140 micrometers or greater, 145 micrometers or greater, 150 micrometers or greater, 155 micrometers or greater, or 165 micrometers or greater. The D80 of the antioxidant component particles may be 200 micrometers or less, 190 micrometers or less, 185 micrometers or less, or 180 micrometers or less.

[0327] Embodiment 2-92 is the method of embodiment 2-91, wherein the particles have a DIO in a range of 5 to 20 micrometers, 6 to 15 micrometers, or 8 to 12 micrometers.

[0328] Embodiment 2-93 is the method of embodiment 2-91 or 2-92, wherein the particles have a D50 in a range of 40 to 80 micrometers, 45 to 70 micrometers, or 48 to 65 micrometers.

[0329] Embodiment 2-94 is the method of any one of embodiments 2-91 to 2-93, wherein the particles have a D90 in a range of 200 to 400 micrometers, 210 to 350 micrometers, or 220 to 325 micrometers.

[0330] Embodiment 2-95 is the method of any one of embodiments 2-91 to 2-94, wherein the astaxanthin is added to the fish feed at a first concentration, and wherein the astaxanthin is present in the fish feed at a second concentration after processing and storage for 21 days or longer, and wherein the second concentration is at least 66 % of the first concentration.

[0331] Embodiment 2-96 is the method of any one of embodiments 2-91 to 2-95, wherein the fish feed is a complete fish feed.

[0332] Embodiment 2-97 is the method of any one of embodiments 2-91 to 2-96, wherein the base feed comprises protein, starch, and fat.

[0333] Embodiment 2-98 is the method of any one of embodiments 2-91 to 2-97, wherein the fish feed comprises from 28 wt-% to 60 wt-% protein, from 6 wt-% to 15 wt-% of starch, and from 18 wt-% to 40 wt-% fat.

[0334] Embodiment 2-99 is the method of any one of embodiments 2-91 to 2-98, wherein the fish feed comprises land-animal protein, fish meal, plant-based protein, or a combination of two or more thereof.

[0335] Embodiment 2-100 is the method of any one of embodiments 2-91 to 2-99, wherein the fish feed comprises one or more of pellets, extruded nuggets, steam pellets, flakes, tablets, granules, or powders.

[0336] Embodiment 2-101 is the method of any one of embodiments 2-91 to 2-100, wherein the antioxidant component comprises a first particle size population and a second particle size population, the first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and the second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90p is smaller than D90c and D50F is smaller than D50c.

[0337] Embodiment 2-102 is the method of embodiment 2-101, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers, 2 to 15 micrometers, or 4 to 12 micrometers.

[0338] Embodiment 2-103 is the method of embodiment 2-101 or 2-102, wherein the first particle size population has a D50F in a range of 20 to 80 micrometers, 30 to 70 micrometers, 35 to 65 micrometers, or 40 to 60 micrometers.

[0339] Embodiment 2-104 is the method of any one of embodiments 2-101 to 2-103, wherein the first particle size population has a D90F in a range of 100 to 200 micrometers, 120 to 180 micrometers, 140 to 160 micrometers.

[0340] Embodiment 2-105 is the method of any one of embodiments 2-101 to 2-104, wherein the second particle size population has a DIOc in a range of 1 to 20 micrometers or 20 to 80 micrometers or 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0341] Embodiment 2-106 is the method of any one of embodiments 2-101 to 2-105, wherein the second particle size population has a D50c in a range of 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0342] Embodiment 2-107 is the method of any one of embodiments 2-101 to 2-106, wherein the second particle size population has a D90c in a range of 100 to 400 micrometers, 110 to 200 micrometers, or 120 to 180 micrometers.

[0343] Embodiment 2-108 is the method of any one of embodiments 2-101 to 2-107, wherein weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part, about 0.5 to 2 parts of the first particle size population for every 1 part, or about 0.8 to 1.3 parts of the first particle size population for every 1 part of the second particle size population.

[0344] Embodiment 2-109 is the method of any one of embodiments 2-101 to 2-108, wherein weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0345] Embodiment 2-110 is the method of any one of embodiments 2-91 to 2-109, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-%, about 2 wt-% to about 25 wt-%, or about 5 wt-% to about 20 wt-%.

[0346] Embodiment 2-111 is the method of any one of embodiments 2-91 to 2-110, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers, or about 4 micrometers..

[0347] Embodiment 2-112 is the method of any one of embodiments 2-91 to 2-111, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0348] Embodiment 2-113 is the method of embodiment 2-112, wherein the primary antioxidant comprises a free radical scavenger.

[0349] Embodiment 2-114 is the method of embodiment 2-112 or 2-113, wherein the primary antioxidant comprises butylated hydroxyanisol (BHa), butylated hydroxytoluene (BHT), propyl gallate (PG), a carotenoid, a flavonoid, a phenolic acid, a tocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0350] Embodiment 2-115 is the method of any one of embodiments 2-112 to 2-114, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%.

[0351] Embodiment 2-116 is the method of any one of embodiments 2-112 to 2-115, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0352] Embodiment 2-117 is the method of any one of embodiments 2-112 to 2-116, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol, polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0353] Embodiment 2-118 is the method of any one of embodiments 2-112 to 2-117, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0354] Embodiment 2-119 is the method of any one of embodiments 2-112 to 2-118, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0355] Embodiment 2-120 is the method of any one of embodiments 2-112 to 2-119, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0356] Embodiment 2-121 is the method of any one of embodiments 2-91 to 2-120, wherein during processing the feed additive pack maintains at least 85 % of the astaxanthin added to the feed additive pack.

[0357] Embodiment 2-122 is the method of any one of embodiments 2-91 to 2-121, wherein the feed additive pack maintains about 60 % or more, about 62 % or more, about 65 % or more, about 67 % or more, or about 70 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days).

[0358] Embodiment 2-123 is the method of any one of embodiments 2-91 to 2-122, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more,about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0359] Embodiment 2-124 is the method of any one of embodiments 2-91 to 2-123, wherein the feed additive pack maintains about 50 % or more, about 52 % or more, about 60 % or more, or about 62 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days).

[0360] Embodiment 2-125 is the method of any one of embodiments 2-91 to 2-124, further comprising extruding the fish feed composition to form pellets.

[0361] Embodiment 2-126 is a method of feeding an aquatic animal, the method comprising administering an amount of feed to the aquatic animal, the feed comprising: a base feed; and a feed additive pack, the feed additive pack comprising: an antioxidant component; and astaxanthin dispersed within the antioxidant component, the antioxidant component comprising particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers. The D80 of the antioxidant component particles may be 140 micrometers or greater, 145 micrometers or greater, 150 micrometers or greater, 155 micrometers or greater, or 165 micrometers or greater. The D80 of the antioxidant component particles may be 200 micrometers or less, 190 micrometers or less, 185 micrometers or less, or 180 micrometers or less.

[0362] Embodiment 2-127 is the method of embodiment 2-126, wherein the particles have a D10 in a range of 5 to 20 micrometers, 6 to 15 micrometers, or 8 to 12 micrometers.

[0363] Embodiment 2-128 is the method of embodiment 2-126 or 2-127, wherein the particles have a D50 in a range of 40 to 80 micrometers, 45 to 70 micrometers, or 48 to 65 micrometers.

[0364] Embodiment 2-129 is the method of any one of embodiments 2-126 to 2-128, wherein the particles have a D90 in a range of 200 to 400 micrometers, 210 to 350 micrometers, or 220 to 325 micrometers.

[0365] Embodiment 2-130 is the method of any one of embodiments 2-126 to 2-129, wherein the astaxanthin is added to the fish feed at a first concentration, and wherein the astaxanthin is present in the fish feed at a second concentration after processing and storage for 21 days or longer, and wherein the second concentration is at least 66 % of the first concentration.

[0366] Embodiment 2-131 is the method of any one of embodiments 2-126 to 2-130, wherein the fish feed is a complete fish feed.

[0367] Embodiment 2-132 is the method of any one of embodiments 2-126 to 2-131, wherein the base feed comprises protein, starch, and fat.

[0368] Embodiment 2-133 is the method of any one of embodiments 2-126 to 2-132, wherein the fish feed comprises from 28 wt-% to 60 wt-% protein, from 6 wt-% to 15 wt-% of starch, and from 18 wt-% to 40 wt-% fat.

[0369] Embodiment 2-134 is the method of any one of embodiments 2-126 to 2-133, wherein the fish feed comprises land-animal protein, fish meal, plant-based protein, or a combination of two or more thereof.

[0370] Embodiment 2-135 is the method of any one of embodiments 2-126 to 2-134, wherein the fish feed comprises one or more of pellets, extruded nuggets, steam pellets, flakes, tablets, granules, or powders.

[0371] Embodiment 2-136 is the method of any one of embodiments 2-126 to 2-135, wherein the antioxidant component comprises a first particle size population and a second particle size population, the first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and the second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90p is smaller than D90c and D50F is smaller than D50c.

[0372] Embodiment 2-137 is the method of embodiment 2-136, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers, 2 to 15 micrometers, or 4 to 12 micrometers.

[0373] Embodiment 2-138 is the method embodiment 2-136 or 2-137, wherein the first particle size population has a D50F in a range of 20 to 80 micrometers, 30 to 70 micrometers, 35 to 65 micrometers, or 40 to 60 micrometers.

[0374] Embodiment 2-139 is the method of any one of embodiments 2-136 to 2-138, wherein the first particle size population has a D90F in a range of 100 to 200 micrometers, 120 to 180 micrometers, 140 to 160 micrometers.

[0375] Embodiment 2-140 is the method of any one of embodiments 2-136 to 2-139, wherein the second particle size population has a DIOc in a range of 1 to 20 micrometers or 20 to 80 micrometers or 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0376] Embodiment 2-141 is the method of any one of embodiments 2-136 to 2-140, wherein the second particle size population has a D50c in a range of 20 to 80 micrometers, 45 to 75 micrometers, or 30 to 60 micrometers.

[0377] Embodiment 2-142 is the method of any one of embodiments 2-136 to 2-141, wherein the second particle size population has a D90c in a range of 100 to 400 micrometers, 110 to 200 micrometers, or 120 to 180 micrometers.

[0378] Embodiment 2-143 is the method of any one of embodiments 2-136 to 2-142, wherein weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part, about 0.5 to 2 parts of the first particle size population for every 1 part, or about 0.8 to 1.3 parts of the first particle size population for every 1 part of the second particle size population.

[0379] Embodiment 2-144 is the method of any one of embodiments 2-136 to 2-143, wherein weight ratio of the first particle size population and the second particle size population is 1 part of first particle size population to every 1 part of second particle size population.

[0380] Embodiment 2-145 is the method of any one of embodiments 2-126 to 2-144, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-%, about 2 wt-% to about 25 wt-%, or about 5 wt-% to about 20 wt-%.

[0381] Embodiment 2-146 is the method of any one of embodiments 2-126 to 2-145, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers, or about 4 micrometers.

[0382] Embodiment 2-147 is the method of any one of embodiments 2-126 to 2-146, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

[0383] Embodiment 2-148 is the method of embodiment 2-147, wherein the primary antioxidant comprises a free radical scavenger.

[0384] Embodiment 2-149 is the method of embodiment 2-147 or 2-148, wherein the primary antioxidant comprises butylated hydroxyanisol (BHa), butylated hydroxytoluene (BHT), propyl gallate (PG), a carotenoid, a flavonoid, a phenolic acid, a tocopherol, a tocotrienol, tertiary butylhydroquinone (TBHQ), or a combination of two or more thereof.

[0385] Embodiment 2-150 is the method of any one of embodiments 2-147 to 2-149, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%.

[0386] Embodiment 2-151 is the method of any one of embodiments 2-147 to 2-150, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof.

[0387] Embodiment 2-152 is the method of any one of embodiments 2-147 to 2-151, wherein the secondary antioxidant comprises ascorbic acid, erythorbic acid, a sulfite, alpha-tocopherol, polysorbate, butylated hydroxytoluene (BHT), propyl gallate (PG), citric acid, ethylenediamine tetraacetic acid (EDTA), monoglyceride, or a combination of two or more thereof.

[0388] Embodiment 2-153 is the method of any one of embodiments 2-147 to 2-152, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0389] Embodiment 2-154 is the method of any one of embodiments 2-147 to 2-153, wherein the secondary antioxidant comprises citric acid, and wherein the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 50 wt-%, about 15 wt-% to about 40 wt-%, or from about 18 wt-% to about 30 wt-%.

[0390] Embodiment 2-155 is the method of any one of embodiments 2-147 to 2-154, wherein the carrier comprises a flour, meal, or starch prepared from a grain or a legume.

[0391] Embodiment 2-156 is the method of any one of embodiments 2-126 to 2-155, wherein during processing the feed additive pack maintains at least 85 % of the astaxanthin added to the feed additive pack.

[0392] Embodiment 2-157 is the method of any one of embodiments 2-126 to 2-156, wherein the feed additive pack maintains about 60 % or more, about 62 % or more, about 65 % or more, about 67 % or more, or about 70 % or more of the astaxanthin during processing and storage for at least 2 weeks (14 days).

[0393] Embodiment 2-158 is the method of any one of embodiments 2-126 to 2-157, wherein the feed additive pack maintains about 58 % or more, about 60 % or more, about 64 % or more, about 68 % or more, about 72 % or more, or about 70 % of the astaxanthin during processing and storage for at least 3 weeks (21 days).

[0394] Embodiment 2-159 is the method of any one of embodiments 2-126 to 2-158, wherein the feed additive pack maintains about 50 % or more, about 52 % or more, about 60 % or more, or about 62 % or more of the astaxanthin during processing and storage for at least 4 weeks (28 days).

[0395] Embodiment 2-160 is the method of any one of embodiments 2-126 to 2-159, further comprising extruding the fish feed composition to form pellets.

[0396] Embodiment 2-161 is the method of any one of embodiments 2-126 to 2-160, wherein the fish feed comprises an amount of astaxanthin that is at least 85 % of the astaxanthin added to the fish feed prior to mixing.

[0397] Embodiment 2-162 is the method of any one of embodiments 2-126 to 2-161, wherein the feed comprises an amount of astaxanthin that is 58 % or more of the astaxanthin added to the feed.EXAMPLESExample 1

[0398] Fish feed samples were prepared using various particle sizes of antioxidant particles. The effect of the particle size of the antioxidant particles on the stability of the astaxanthin in fish feed samples during processing and storage was studied. The studied test samples included samples with coarse particle sizes (Coarse), with finely ground particle sizes (Fine), and a mixture of the two (Hybrid).

[0399] An antioxidant pack was ground to a coarse particle size using a 1.5 mm screen and to a fine particle size using a 0.75 mm screen on a hammer mill. A 50:50 mixture (by weight) of the coarse and fine particles was prepared. The particle size distribution of each sample was measured using a Mastersizer 3000. The particle size distribution (by volume) is shown below in TABLE 1A.TABLE 1 A. Antioxidant pack particle size distribution

[0400] The concentration of astaxanthin (mg / kg) in the raw ingredient was obtained from the manufacturer. The feed composition included about 37 wt-% protein, 35 wt-% fat, 8 wt-% starch, and about 6 wt-% of moisture. The percentage of EPA / DHA of total fatty acids was about 7 wt-%. The level of astaxanthin in the feed composition was calculated based on inclusion level of the ingredient. The astaxanthin level in the prepared feed was measured after processing (day 0) and during storage after 1 week, 2 weeks, and 3 weeks. The astaxanthin level was measured by HPLC. All tests were performed in triplicates.

[0401] The astaxanthin concentrations (average of three samples) and losses during processing and storage are shown in TABLE IB below.TABLE IB. Astaxanthin losses during processing and storage

[0402] It was observed that the astaxanthin loss during the process was lower in both the fine and the mixed (hybrid) particle size than in the coarse ground feed. Although the losses during storage were greater in the hybrid than in the coarse grind, the overall losses in the hybrid were significantly lower than in the coarse, and also lower than in the fine grind.Example 2

[0403] Fish feed samples were prepared similar to Example 1, using different particle sizes of the antioxidant component. The effect of the particle size of the antioxidant component on the stability of the astaxanthin in fish feed samples during processing and storage was studied.

[0404] The feeds were formulated to contain 100 mg / kg of astaxanthin. The average measured astaxanthin level in the mix was 99.3 mg / kg. The feed was made using a single screw extruder (WENGER X85) with a target bulk density of 510 g / L.

[0405] An antioxidant pack was ground on a hammer mill using 0.5 mm, 0.75 mm, 1 mm, 1.5 mm, and 2 mm screens. Samples were prepared with antioxidant pack samples ground with each screen size, as well as by combining 0.75 mm and 1.5 mm screen ground samples at ratios of 25:75, 50:50, and 75:25 (by weight). The sample ground using the 1.5 mm screen was used as the reference sample. The particle size distribution of each sample was measured using a Mastersizer 3000. The particle size distribution (by volume) is shown below in TABLE 2A.TABLE 2A. Antioxidant pack particle size distribution

[0406] The initial level of astaxanthin in the feed composition was calculated based on inclusion level of the ingredient. The astaxanthin level in the prepared feed was measured after processing (day 0) and during storage after 2 weeks and 4 weeks. The samples were stored as whole pellets in a feed bag at room temperature. The astaxanthin level was measured by HPLC. All tests were performed in triplicate.

[0407] The astaxanthin losses (average of three samples) during processing and storage are shown in TABLE 2B below. The results are graphically presented in FIG. 2.TABLE 2B. Antioxidant losses

[0408] It was observed that astaxanthin losses during the process and during storage were lower in the samples that included particles ground with a screen size of 1 mm or 0.75 mm. The overall loss was smallest in the sample that included an equal amount of 1.5 mm and 0.75 mm screen ground antioxidant particles and the sample that was ground using the 1 mm screen.

[0409] All references and publications cited herein are expressly incorporated herein by reference in their entirety into this disclosure, except to the extent they may directly contradict this disclosure. Although specific embodiments have been illustrated and described herein, it will be appreciated by those of ordinary skill in the art that a variety of alternate and / or equivalent implementations maybe substituted for the specific embodiments shown and described without departing from the scope of the present disclosure. It should be understood that this disclosure is not intended to be unduly limited by the illustrative embodiments and examples set forth herein and that such examples and embodiments are presented by way of example only with the scope of the disclosure intended to be limited only by the claims set forth here.

Claims

CLAIMS1. A feed additive pack comprising: an antioxidant component; and astaxanthin dispersed within the antioxidant component, the antioxidant component comprising particles having a particle size volume distribution with D80 in a range of 140 to 200 micrometers.

2. The feed additive pack of claim 1, wherein the particles have a DIO in a range of 5 to 20 micrometers.

3. The feed additive pack of claim 1 or 2, wherein the particles have a D50 in a range of 40 to 80 micrometers.

4. The feed additive pack of any one of the preceding claims, wherein the particles have a D90 in a range of 150 to 400 micrometers.

5. The feed additive pack of any one of the preceding claims, wherein the antioxidant component comprises a first particle size population and a second particle size population, the first particle size population having a particle size volume distribution with D50F in a range of 20 to 80 micrometers and D90F in a range of 100 to 200 micrometers, and the second particle size population having a particle size volume distribution with D50c in a range of 30 to 90 micrometers and D90c in a range of 150 to 750 micrometers, wherein D90F is smaller than D90c and D50F is smaller than D50c.

6. The feed additive pack of claim 5, wherein the first particle size population has a DI OF in a range of 1 to 20 micrometers, a D50F in a range of 35 to 65 micrometers, and a D90F in a range of 120 to 180 micrometers.

7. The feed additive pack of claim 5 or 6, wherein the second particle size population has a DIOc in a range of 2 to 20 micrometers, a D50c in a range of 45 to 75 micrometers, and a D90c in a range of 180 to 400 micrometers.

8. The feed additive pack of any one of claims 5 to 7, wherein weight ratio of the first particle size population and the second particle size population is from 0.3 to 3 parts of the first particle size population to every 1 part of the second particle size population.

9. The feed additive pack of any one of the preceding claims, wherein the astaxanthin is present in the feed additive pack at a concentration of about 0.5 wt-% to about 50 wt-%.

10. The feed additive pack of any one of the preceding claims, wherein the astaxanthin has a particle size D90 in a range of about 1 to 10 micrometers.

11. The feed additive pack of any one of the preceding claims, wherein the feed additive pack comprises a primary antioxidant, a secondary antioxidant, and a carrier, wherein the primary antioxidant is present in the feed additive pack at a concentration of about 1 wt-% to about 10 wt-%, wherein the secondary antioxidant is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%, optionally wherein the secondary antioxidant exhibits a synergistic effect with the primary antioxidant.

12. The feed additive pack of claim 11, wherein the primary antioxidant comprises a free radical scavenger.

13. The feed additive pack of claim 11 or 12, wherein the secondary antioxidant comprises an oxygen scavenger, emulsifier, chelator capable of chelating iron, or a combination of two or more thereof, optionally wherein the secondary antioxidant comprises citric acid, and the citric acid is present in the feed additive pack at a concentration of about 15 wt-% to about 40 wt-%.

14. The feed additive pack of any one of the preceding claims, wherein during processing the feed additive pack maintains 85 % or more of the astaxanthin added to the feed additive pack.

15. The feed additive pack of any one of the preceding claims, wherein the feed additive pack maintains 58 % or more of the astaxanthin during processing and storage for at least 3 weeks (21 days).

16. A fi sh feed compri sing : a base feed; and the feed additive pack of any one of the preceding claims.

17. The fish feed of claim 16, wherein the fish feed is a complete fish feed, comprising from 28 wt-% to 60 wt-% protein, from 6 wt-% to 15 wt-% of starch, and from 18 wt-% to 40 wt-% fat.

18. A method of feeding an aquatic animal, the method comprising administering an amount of feed to the aquatic animal, the feed comprising:a base feed; and the feed additive pack of any one of claims 1 to 15.

19. The method of claim 18, wherein the fish feed is a complete fish feed, comprising from28 wt-% to 60 wt-% protein, from 6 wt-% to 15 wt-% of starch, and from 18 wt-% to 40 wt-% fat.

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