Fish feed injected with pest control agent

Abstract

The present disclosure provides a fish feed and a method for preparing a fish feed. The fish feed comprises a basal feed having a porous matrix distributed throughout and a pest control agent composition, wherein the pest control agent composition comprises a pest control agent and one or more oils. The pest control agent composition of the fish feed may be distributed throughout the porous matrix. The pest control agent may comprise an Azadirachtin A enriched Azadirachtin A extract distributed throughout the porous matrix of the basal feed by a vacuum infusion process. Other aspects are also included herein.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 381,362, filed on October 28, 2023, and entitled “PEST CONTROL AGENT—INFUSED FISH FEED,” which is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure relates to a fish feed having one or more pest control agents distributed throughout its porous matrix, and compositions and methods for preparing such fish feed. Background Art

[0004] Aquaculture of various fish species provides an important food source for the growing world population. In 2021, it is estimated that approximately 175 million metric tons of fish were produced for global consumption. The increase in consumer demand for fish has led to an increase in the number of aquaculture farms around the world. With the increase in aquaculture farms, the demand for fish feed to nourish and maintain the large numbers of fish produced, as well as for sustainable technologies for safely and effectively controlling pests in the farmed fish populations, has increased.

[0005] While there are various physical delousing systems and chemical treatments for managing various pests, many undesirable side effects from these systems and agents can result in significant losses to fish farms. Some estimates suggest that the global aquaculture industry loses over $750 million per year due to the need to purchase agents such as parasiticides and expensive equipment, productivity losses from starvation and underfeeding associated with physical delousing systems, excess mortality from physical delousing systems, and the further need to devote staff time to management, control, and research into new parasite management and control methods. In many cases, farmers must monitor their fish to keep sea lice within predetermined thresholds set by each region, and when necessary, they must cull fish to prevent fish suffering and the spread of parasites, resulting in further economic losses.

[0006] One approach to control and prevent pest infestation and infestation is to provide one or more pest control agents in the fish feed for fish, wherein when the pests are fed with fish, they will eventually take in the pest control agents. The manufacture of fish feed by various techniques may require applying high temperature and high shear force to the raw materials before the processing period. Although the raw materials can withstand high processing temperature and shear force, various functional ingredients and activating agents (such as various pest control agents) may be easy to degrade under such temperature and force. Further, many functional ingredients and activating agents are insoluble in various food grade solutions, and therefore need more complicated mechanisms to mix them in the fish feed. Therefore, it is necessary to provide fish feed and a method for mixing these functional ingredients or activating agents in the fish feed using a processing that does not cause the degradation of the functional ingredients or activating agents. Summary of the invention

[0007] The present disclosure provides a fish feed, comprising a base feed having a porous matrix distributed throughout, and a pest control agent composition. The pest control agent composition comprises a pest control agent and one or more oils, wherein the pest control agent composition is distributed throughout the porous matrix of the base feed.

[0008] In one aspect, the pest control agent comprises a neem extract enriched in azadirachtin A.

[0009] In one aspect, the neem extract enriched with azadirachtin A comprises at least 15 wt% to 33 wt% azadirachtin A.

[0010] In one aspect, the one or more oils include one or more of soybean oil, grapeseed oil, canola oil, sunflower oil, safflower oil, rapeseed oil, hydrogenated rapeseed oil, hydropressed linseed oil, camelina oil, flaxseed oil, coconut oil, fish oil, poultry oil or fish oil stearin or fractions thereof.

[0011] In one aspect, the porous matrix is ​​formed by extrusion of a raw material comprising protein and starch.

[0012] In one aspect, the pest control composition is applied to the basal feed via a vacuum infusion process.

[0013] In one aspect, the pest control agent is at least partially distributed throughout the porous matrix of the base feed.

[0014] In one aspect, the pest control agent composition is further coated on the outer surface of the base feed.

[0015] In one aspect, the fish feed comprises between about 15 wt % and about 65 wt % protein and between about 10 wt % and about 45 wt % fat, and wherein the fish feed particles include terrestrial animal protein, fish meal, plant-based protein, or a combination thereof.

[0016] In one aspect, the basal feed comprises one or more of pellets, extruded blocks, steam pellets, flakes, tablets or powder.

[0017] In one aspect, the pest control agent comprises a solid form suspended in one or more oils.

[0018] The present disclosure provides a method for preparing fish feed. The method may include providing a protein and starch mixture and subjecting it to an extrusion process to produce a base feed having a porous matrix distributed throughout. The method may include producing a pest control agent composition by mixing a pest control agent and one or more oils together to produce a suspension. The method may include applying the pest control agent composition to the base feed using a vacuum infusion process to distribute the pest control agent composition throughout the porous matrix of the base feed.

[0019] In one aspect, the pest control agent of the method herein comprises a neem extract enriched in azadirachtin A, the neem extract comprising at least 15 wt% to 33 wt% azadirachtin A.

[0020] In one aspect, the one or more oils used in the methods herein include one or more of soybean oil, grapeseed oil, canola oil, sunflower oil, safflower oil, rapeseed oil, hydrogenated rapeseed oil, hydro-pressed linseed oil, camelina oil, cold-pressed linseed oil, coconut oil, fish oil, poultry oil or fish oil stearin or fractions thereof.

[0021] In one aspect, the extrusion process comprises extrusion cooking a raw material comprising protein and starch.

[0022] In one aspect, a method of producing a base feed by an extrusion process followed by application of the pest control composition by a vacuum infusion process produces a fish feed having a coefficient of variation of less than 10%.

[0023] The present disclosure provides fish feed products according to the methods herein.

[0024] The present disclosure provides a fish feed for reducing, preventing or controlling parasitic infections or infestations in fish, wherein the fish feed comprises: a base feed comprising a porous matrix distributed throughout; and a pest control agent composition comprising a pest control agent and one or more oils; and wherein the pest control agent composition is distributed throughout the porous matrix of the base feed. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The accompanying drawings generally illustrate various aspects described herein by way of example and not limitation. A more complete understanding of the various aspects may be obtained with reference to the following drawings, in which:

[0026] Figure 1 is a schematic diagram of fish feed according to various aspects of this article.

[0027] Figure 2 is a schematic diagram of fish feed according to various aspects of this article.

[0028] Figure 3 is a schematic representation of the life cycle of salmon lice (Lepeophtheirus salmonis) in accordance with various aspects herein.

[0029] Figure 4 is a flow chart of a method according to various aspects of the present invention. DETAILED DESCRIPTION

[0030] Reference will now be made in detail to certain aspects of the presently disclosed subject matter, examples of which are partially illustrated in the accompanying drawings.While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the illustrated subject matter is not intended to limit the claims to the disclosed subject matter.

[0031] As mentioned above, it is necessary to incorporate various functional ingredients or active agents throughout the fish feed. Although such agents can be applied to the outside of the fish feed in some cases, such external application may cause the agent to be lost from the surface during storage, transportation, and direct and long-term exposure to moisture or humidity. Other examples include adding the functional ingredients or active agents directly to the raw materials before forming the fish feed. However, many functional ingredients or active agents are heat-sensitive and easy to degrade under high processing temperatures and shear forces, and further have limited solubility for coating application. Therefore, due to the limitations imposed by the physiochemical properties of such agents, it is not the best choice to incorporate various functional ingredients or active agents into the fish feed during such processing steps. The present disclosure relates to fish feeds with one or more functional ingredients or active agents (such as one or more pest control agents) distributed throughout. The present disclosure also discloses various methods for producing fish feeds, which have one or more functional ingredients or active agents distributed throughout, such as one or more pest control agents, wherein the method for producing such fish feeds does not cause these agents to be significantly degraded during processing.

[0032] As used herein, the term "infection" may refer to a condition in which pests (including various pests as defined elsewhere herein) can invade any internal or external part of a host organism, causing damage to the host organism, and the pests use components of the host organism to maintain themselves, reproduce or colonize the host organism.

[0033] As used herein, the term "infestation" may refer to the presence of an abnormally large number of pests as defined herein, wherein the pests are concentrated in an area in numbers that can cause damage or disease by infecting host organisms.

[0034] As used herein, the term "pest" may refer to any organism that is detrimental to the health, value, or appearance of another organism. The term pest may include, but is not limited to, one or more of various parasites, including worms, helminths, flukes, lice, mites; one or more bacteria; one or more viruses; one or more types of fungi; and various protozoa (e.g., amoebas).

[0035] As used herein, the term "parasite" may refer to one or more external parasites or internal parasites. As used herein, the term "internal parasite" may refer to an organism that inhabits one or more internal microhabitats of another organism. For example, an internal parasite may inhabit one or more of the tissues, organs or systems of a host organism. For example, an internal parasite may inhabit the intestines, blood or both of a host organism. As used herein, the term "external parasite" may refer to an organism that inhabits the external microhabitat of another species or occupies the external microhabitat of another species. For example, an external parasite may inhabit or occupy the surface of a host species. In the case of fish, an external parasite may inhabit the skin of the fish, where they sometimes stay between scales, and they may also feed on mucus, blood, skin, gills, muscle or any combination thereof. An external parasite may include a species belonging to the phylum Arthropoda. Therefore, the term external parasite may also include crustaceans and one or more sea lice that inhabit a fish host. In one aspect, the ectoparasite can include one or more copepods.

[0036] As used herein, the term "pest control agent" may refer to an agent used to reduce, prevent or control an infection or infestation caused or contributed to by one or more pests. In various aspects, the pest control agent described herein may refer to an agent used to reduce, prevent or control an infection or infestation caused by one or more endoparasites or ectoparasites.

[0037] Fish feed

[0038] The present disclosure provides fish feeds for use as fish diets in aquaculture applications. The fish feeds herein may include a base feed. The base feed may include a porous matrix distributed throughout the base feed. In various aspects, the fish feed may also include a pest control agent composition, wherein the pest control agent composition includes one or more pest control agents and one or more oils. The base feed may be formed from various raw materials as described elsewhere herein.

[0039] In various aspects, the pest control agent composition can be distributed throughout the porous matrix of the basic feed. The pest control agent composition is further described elsewhere herein. In some aspects, the basic feed of this article can include a pest control agent composition, which is at least partially distributed throughout the porous matrix of the basic feed, or is completely distributed throughout the porous matrix of the basic feed. In various aspects, the pest control agent composition is further coated on the outer surface of the basic feed. The basic feed can be at least partially coated with the pest control agent composition on the outer surface, or the pest control agent composition is completely coated on the outer surface. In various aspects, the fish feed of this article can be at least partially coated with the pest control agent composition on the outer surface, and can also have the pest control agent composition dispersed at least partially throughout the porous matrix of the basic feed. In various aspects, the pest control agent composition is coated on the outer surface of the basic feed. The pest control agent is partially or completely distributed throughout the porous matrix of the basic feed by a vacuum infusion process. Similarly, the pest control agent of this article is partially or completely coated on the outer surface of the basic feed by a vacuum infusion process. Alternatively, the coating on the outer surface of the base feed may be distributed by a spraying, dipping, roller coating or electrospraying process.

[0040] Reference now Figure 1, a schematic diagram of a fish feed is shown according to various aspects of the present invention. The fish feed 100 can include a porous matrix distributed throughout the fish feed 100, wherein the porous matrix includes a plurality of holes 101. It should be understood that the processing of the fish feed will result in a distribution of pore sizes, so that not all pores are uniform in diameter or distribution throughout the fish feed 100. The fish feed 100 can include a base feed 102 and a pest control agent composition 104 distributed throughout the base feed. The pest control agent composition can also be coated on the outer surface 106 of the base feed 102. It should be understood that the pest control agent composition can include one or more pest control agents and one or more oils, as described elsewhere herein. It should also be understood that the fish feed 100 can include a pest control agent composition 104 distributed throughout the base feed 102 and coated on the outer surface 106 of the base feed 102, and can also include a second pest control agent composition (not shown) coated on the outer surface 108 of the pest control agent composition 104. It is understood that multiple coatings of pest control compositions may be added to the base feed after the initial pest control composition 104 has been added. It is also understood that when multiple pest control compositions are added to the base feed, each pest control composition may contain a different pest control agent and / or a different oil composition, as described elsewhere herein.

[0041] In various aspects, the fish food 100 may also include one or more coating compositions that lack a pest control agent and are distributed on the outer surface 108 of the pest control agent composition 104. Figure 2 , a schematic diagram of a fish feed 200 is shown according to various aspects of the present invention. The fish feed 200 contains similar numbered features as the fish feed 100, such as Figure 1 As shown, and additionally comprising a coating composition 202 lacking a pest control agent. The coating composition 202 may include an oil-based composition. The coating composition may include one or more oils and one or more solid particles. The oil in the coating composition may include, but is not limited to, vegetable oil, canola oil, poultry oil, rapeseed oil, fish oil, soybean oil, water-pressed linseed oil, camelina oil, fish oil stearin, coconut oil, hydrogenated rapeseed oil, and any combination or fraction thereof. The solid particles may include, but are not limited to, wheat gluten, soy protein concentrate, pea protein concentrate, fish meal, concentrated fish soluble protein, feather meal, bentonite, diatomaceous earth, zeolite, ammonium phosphate, dicalcium phosphate, monocalcium phosphate, algae oil, and any combination thereof.

[0042] In various aspects, the coating composition is applied to the outer surface of the pest control agent composition. The fish feed can be at least partially coated with the coating composition on the outer surface of the pest control agent composition, or the coating composition can be completely coated with the coating composition on the outer surface of the pest control agent composition. In various aspects, the coating composition can form at least two layers around the fish feed. The coating composition can be in the form of a slurry. In various aspects, the coating composition slurry can be prepared by adding one or more solid particles to a certain volume of oil to form a coating composition slurry. The coating composition slurry can be used to coat the fish feed or fish feed product with at least two layers, at least three layers, at least four layers or more layers. In various aspects, the coating composition can be used for the purpose of preventing leakage from the fish feed, including preventing the pest control agent composition from leaking from the fish feed.

[0043] It should be understood that the fish feed herein can include fish feed of the appropriate type specific to a given fish species. The fish feed can be used as a component of a diet of any species belonging to the following families: Cyprinidae, Cichlidae, Panfish, Sciaenidae, Serranidae, Carpidae, Sparidae, Perchidae, Perchidae, Mugilidae, Cypriniformes, Perchidae, Channaidae, Tilapia and Salmonidae. In various fields, the present disclosure provides a fish feed or fish feed diet for species in the Salmonidae. The fish feed provided herein can be used for feeding wild fish or cultured fish. In various fields, both wild fish and cultured fish can be fed simultaneously. Further, the fish feed can be used for feeding freshwater fish or saltwater (e.g., seawater) fish or both.

[0044] Fish feed of the present disclosure can be produced by using raw materials to produce basic feed, and these raw materials can be selected based on their application and fish species. In various aspects, fish feed is solid fish feed. In other aspects, fish feed can include both solid fish feed components and liquid fish feed components. Basic feed can include particles, extruded blocks, steam particles, flakes, tablets, powders, etc. In various aspects, basic feed can include basic feed particles. In various aspects, fish feed can include extruded, compressed or granular basic feed. In some aspects, basic feed can include porous matrix distributed everywhere. Fish feed containing liquid fish feed components can include aqueous solution, oil, oil and water emulsion, slurry, suspension, etc. In various aspects, solid fish feed can also include one or more oils, which are arranged on the porous matrix present in the fish feed or distributed throughout the porous matrix. In various aspects, one or more pest control agents can be suspended in one or more oils and arranged on the porous matrix present in the fish feed or distributed throughout the porous matrix.

[0045] The fish feed herein may include a variety of different ingredients or raw materials that can maintain the life, growth and reproduction of fish. The fish feed may include any substrate that is edible to fish. For example, an edible substrate may provide a source of nutrition for fish, or may be an inert substrate that has no nutritional value to fish. In various aspects, the fish feed herein may include feed as a nutritional fish feed or a non-nutritional feed. The nutritional fish feed may include a nutritional food formulated for fish as part of its diet as the main source of nutrition, growth and reproduction. Suitable nutritional fish feeds may include one or more of the following: proteinaceous substances as a source of protein, peptides and amino acids; carbohydrates; and fats, as described below. Non-nutritional fish feeds may include any substrate that is edible to fish but does not provide nutrition to maintain life, growth or reproduction. In various aspects, the nutritional or non-nutritional fish feed herein may include one or more compounds intended to change the quality, quantity or appearance of fish and fish tissues. For example, nutritional or non-nutritional fish feeds may include carotenoid compounds to improve the appearance (e.g., color) of muscle tissue. For example, carotenoid compounds may include compounds such as astaxanthin.

[0046] Fish feeds herein may include complete fish feeds. Complete fish feeds may include feeds for fish that are compounded to feed as the sole ration and can maintain life, promote growth and maintain reproduction without taking in any additional substances except water. Complete fish feeds may include a complex mixture containing various energy sources such as carbohydrates, proteins and fats. In various aspects, fish feeds herein may include at least protein and starch. Additional ingredients necessary to support the life, growth and reproduction of fish, such as vitamins and minerals, may be included. 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, starch, tapioca starch, wheat, wheat gluten, guar meal, guar protein concentrate, peas, pea protein concentrate, pea starch, beans, broad beans, sunflower meal, vegetable oil, canola oil, poultry oil, rapeseed oil, fish oil, soybean oil, hydro-pressed linseed oil, camelina oil, lecithin, macromolecular minerals, minerals, vitamins, amino acids, pigments, and any combination thereof. It should be understood that the fish feed herein can include fish meal, which can include materials of plant or animal origin. Any animal-derived materials present in the fish meal can be derived from fish of the same species or fish of a different species (e.g., heterologous species or non-fish species).

[0047] The total protein content in the fish feed can be 10% to 70% by weight, 15% to 65% by weight, 20% to 60% by weight, or 25% to about 55% by weight. The total protein in the fish feed can be at least 10%, 15%, 20%, 25%, 30%, 35%, 38%, 40%, 42%, 44%, 46%, 48%, 50%, 52%, 55%, 60%, 65%, or at least 70% by weight, or any amount within any of the foregoing ranges. The total protein in the fish feed can vary depending on the formulation, species, age, and intended use of the feed. It should be understood that the various protein requirements of the species of fish receiving the fish feed can be adjusted to meet the protein requirements of the species.

[0048] The protein in the fish feed herein can come from any suitable source, including but not limited to one or more of the following: fish meal, terrestrial animal protein (e.g., poultry meal), plant-based protein (e.g., vegetable meal), or any combination thereof. The fish feed can contain 0% to 80%, 10% to 80%, 20% to 75%, 30% to 70%, 60% to 80%, or 10% to 30%, or at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 75% fish meal by weight, or any amount within any of the foregoing ranges. The fish feed may comprise 0% to 80%, 10% to 80%, 20% to 75%, 30% to 70%, 60% to 80%, or 10% to 30%, or at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 75% by weight of terrestrial animal protein, or any amount within any of the foregoing ranges. The fish feed may comprise between 0% to 80%, 10% to 80%, 20% to 75%, 30% to 70%, 60% to 80%, or 10% to 30%, or at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 75% by weight of plant-based protein, or any amount within any of the foregoing ranges.

[0049] The total fat (e.g., oil, fat and / or lipid) in the fish feed can be 5% to 50%, 10% to 45%, 15% to 40% or 20% to 35%. The total fat in the fish feed can be at least 5% by weight, 10% by weight, 15% by weight, 20% by weight, 25% by weight, 30% by weight, 35% by weight, 40% by weight, 45% by weight or at least 50% by weight, or any amount within any of the foregoing ranges. The total fat in the fish feed can vary depending on the formulation, the target fish species and the intended use of the feed. It should be understood that the various requirements of the species of fish receiving the fish feed for fat can be adjusted to meet the requirements of the species for fat. Fats suitable for use herein can include, but are not limited to, fats provided by canola oil, poultry oil, rapeseed oil, fish oil, soybean oil, hydro-pressed linseed oil, camelina oil, palm oil, lecithin or any combination or fraction thereof.

[0050] The moisture content of the fish feed herein can vary depending on the content and preparation method of the feed. In various aspects, the moisture content can be 1 wt % to 20 wt %, 2 wt % to 18 wt %, 5 wt % to 15 wt % or 6 wt % to 12 wt %.

[0051] In addition to the porous matrix arrangement throughout the fish feed or the pest control agent coated on the outer surface of the fish feed, the fish feed herein may also include one or more pest control agents added to the basal feed, provided that these pest control agents can withstand various processing temperatures and shear forces. In various fields, the one or more pest control agents may be present in the basal feed in an amount that effectively produces an inhibitory effect on one or more pests, as described elsewhere herein. Therefore, the basal feed herein may also include a pest control agent for preventing and treating an infection or infestation caused by one or more pests. In various fields, the basal feed herein may include a pest control agent for preventing and treating an infection or infestation of an endoparasite or an ectoparasite. In many aspects, parasitic infection or infestation is a copepod infection or infestation. In some aspects, parasitic infection or infestation is a sea louse infection or infestation. Various pests suitable as targets of the pest control agent herein are described elsewhere.

[0052] Each pest control agent contained in the base feed or fish feed herein can be individually capable of preventing and treating one or more of parasites, bacteria, viruses, fungi or protozoa infections or infestations. Therefore, it should be understood that any given pest control agent used in many aspects described herein can be referred to as exhibiting one or more inhibitory effects, including antiparasitic effects (e.g., anti-ectoparasites, anti-endoparasites), antibacterial effects, antiviral effects, antifungal effects or antiprotozoal effects. In various aspects, the inhibitory effect may include an antiparasitic effect, wherein the antiparasitic effect may also include an anti-ectoparasitic effect, an anti-endoparasitic effect or both. It should be understood that the inhibitory effect may lead to reducing, preventing or controlling the concentration and spread of various parasites, bacteria, viruses, fungi or protozoa described herein. In various aspects, the pest control agent of the present disclosure may produce an inhibitory effect on one or more pests, including one or more effects for reducing, preventing or controlling the concentration and spread of various endoparasites or ectoparasites. Reducing, preventing or controlling parasites may include completely preventing infection or infestation in a fish population or on each individual fish, reducing the total number of parasites present in a fish population or on each individual fish, or controlling how many parasites are present in a fish population or on each individual fish in accordance with local regulatory requirements. It should be understood that the inhibitory effects described herein may be measured on a population of fish infected or infested with one or more pests that is fed a diet lacking azadirachtin A-enriched neem extract.

[0053] In other aspects, the inhibitory effect on pests may include one or more of anti-feeding, anti-molting, anti-fertility and anti-reproductive effects or anti-parasitic effects. As used herein, "anti-feeding" may refer to the effect of one or more pest control agents that stops or inhibits pest feeding, thereby causing malnutrition, stunted development, blocked or delayed molting, and death. Malnourished sea lice are also unlikely to effectively immunomodulate their hosts, so their ability to escape host immunity is weak. As used herein, the term "anti-molting" may refer to the effect of one or more pest control agents that prevent or delay the molting process of pests. The molting process occurs when pests grow and shed their exoskeleton from one life stage to the next life stage, and is controlled by hormones and neurons, and the pest control agents herein may play one or more anti-molting effects on pests. As used herein, the terms "anti-fertility effect" and "anti-reproductive effect" are collectively referred to as "anti-fertility and anti-reproductive effects", and may include one or more effects on male or female reproduction. It is understood that the term "fertility" can refer to the actual number of offspring produced by a female or eggs released from a female, and the term "fecundity" can refer to the biological potential for reproduction, and because of their close relationship, the two terms as used herein can be used interchangeably unless otherwise indicated. Antifertility and anti-fertility effects can include reducing total gamete production in males and females, completely or partially inhibiting viable egg production, changing the anatomy and morphology of male or female gametes, changing the potential for fertilization of eggs, and reducing the total number of pregnant female pests.

[0054] In the application form, the amount of the pest control agent included in the fish feed herein can be about 0.01 grams to 100 grams of pest control agent / kilogram of fish feed (g / kg), about 90g / kg fish feed, about 80g / kg fish feed, about 70g / kg fish feed, about 60g / kg fish feed, about 50g / kg fish feed, about 40g / kg fish feed, about 30g / kg fish feed, about 20g / kg fish feed, about 1g to 10g / kg fish feed, about 2g to 9g / kg fish feed, about 3g to 7g / kg fish feed, about 4g to 6g / kg fish feed, or about 5g / kg fish feed. The total amount of the pest control agent can include the pest control agent found in the base feed, the pest control agent found in one or more pest control agent compositions injected into the porous matrix of the base feed, the pest control agent composition coated on the outer surface of the base feed, or any combination thereof.

[0055] In various aspects, the fish feed herein can contain an amount of pest control agent and / or active ingredient effective to produce an inhibitory effect on one or more pests, including about 0.01 g / kg fish feed, 0.05 g / kg fish feed, 0.1 g / kg fish feed, 0.2 g / kg fish feed, 0.3 g / kg fish feed, 0.4 g / kg fish feed, 0.5 g / kg fish feed, 0.6 g / kg fish feed, 0.7 g / kg fish feed, 0.8 g / kg fish feed, 0.9 g / kg fish feed, 1.0 g / kg fish feed. / kg fish feed, 1.25g / kg fish feed, 1.5g / kg fish feed, 1.75g / kg fish feed, 2.0g / kg fish feed, 2.25g / kg fish feed, 2.5g / kg fish feed, 2.75g / kg fish feed, 3.0g / kg fish feed, 5.0g / kg fish feed, 5.25g / kg fish feed, 5.5g / kg fish feed, 5.75g / kg fish feed, 6.0g / kg fish feed, 6.25g / kg fish feed, 6.5g / kg fish feed, 6.75g / kg / kg fish feed, 7.0g / kg fish feed, 7.0g / kg fish feed, 7.25g / kg fish feed, 7.5g / kg fish feed, 7.75g / kg fish feed, 8.0g / kg fish feed, 8.25g / kg fish feed, 8.5g / kg fish feed, 8.75g / kg fish feed, 9.0g / kg fish feed, 9.25g / kg fish feed, 9.5g / kg fish feed, 9.75g / kg fish feed, 10.0g / kg fish feed, 15g / kg fish feed, 20g / kg The total amount of the pest control agent may include a concentration of 100 g / kg fish feed, 25 g / kg fish feed, 30 g / kg fish feed, 35 g / kg fish feed, 40 g / kg fish feed, 45 g / kg fish feed, 50 g / kg fish feed, 55 g / kg fish feed, 60 g / kg fish feed, 65 g / kg fish feed, 70 g / kg fish feed, 75 g / kg fish feed, 80 g / kg fish feed, 85 g / kg fish feed, 90 g / kg fish feed, 95 g / kg fish feed, or 100 g / kg fish feed, or any amount within any of the foregoing concentration ranges. The total amount of the pest control agent may include the pest control agent found in the base feed, the pest control agent found in one or more pest control agent compositions injected into the porous matrix of the base feed, the pest control agent composition coated on the outer surface of the base feed, or any combination thereof.

[0056] It should be understood that the above concentrations correspond to an amount of total pest control agent of about 0.001 wt% to 10 wt% (% w / w) relative to the fish feed. In various aspects, the fish feed herein can contain an amount of pest control agent effective to produce an inhibitory effect on one or more pests, including 0.001% w / w, 0.002% w / w, 0.003% w / w, 0.004% w / w, 0.005% w / w, 0.006% w / w, 0.007% w / w, 0.008% w / w, 0.009% w / w, 0.010% w / w, 0.020% w / w, 0.030% w / w, 0.040% w / w, 0.050% w / w, 0.060% w / w, 0.070% w / w, 0.080% w / w, 0.090% w / w, 0.10% w / w, 0.11% w / w, 0.12% w / w, 0.13% w / w, 0.14% w / w, 0.15% w / w, 0.16% w / w, 0.17% w / w, 0.18% w / w, 0.19% w / w, 0.20% w / w, 0.21% w / w, 0.22% w / w, 0.23% w / w, 0.24% w / w, 0.25% w / w, 0.26% w / w, 0.27% w / w, 0.28% w / w, 0.29% w / w, 0.30% w / w, 0.31% w / w, 0.32% w / w, 0.33% w / w, 0.34% w / w, 0.35% w / w, 0.36% w / w, 0.37% w / w, 0.38% w / w, 0.39% w / w, 0.40% w / w, 0.41% w / w, 0.42% w / w, 0.43% w / w, 0.44% w / w, 0.45% w / w, 0.46% w / w, 0.47% w / w, 0.48% w / w, 0.49% w / w, 0.50% w / w, 0.51% w / w, 0.52% w / w, 0.53% w / w, 0.54% w / w, 0.55% w / w, 0.56% w / w, 0.57% w / w, 0.58% w / w, 0.59% w / w, 0.60% w / w, 0.61% w / w, 0.62% w / w, 0.63% w / w, 0.64% w / w, 0.65% w / w, 0.66% w / w, 0.67% w / w, 0.68% w / w, 0.69% w / w, 0.70% w / w, 0.71% w / w, 0.72% w / w, 0.73% w / w, 0.74% w / w, 0.75% w / w, 0.76% w / w, 0.77% w / w, 0.78% w / w, 0.79% w / w, 0.80% w / w, 0.81% w / w, 0.82% w / w, 0.83% w / w, 0.84% w / w, 0.85% w / w, 0.86% w / w, 0.87% w / w, 0.88% w / w, 0.89% w / w, 0.90% w / w, 0.91% w / w, 0.92% w / w, 0.93% w / w, 0.94% w / w, 0.95% w / w, 0.96% w / w, 0.97% w / w, 0.98% w / w, 0.99% w / w, 1.0% w / w, 2.0% w / w, 3.0% w / w, 4.0% w / w, 5.0% w / w, 6.0% w / w, 7.0% w / w, 8.0% w / w, 9.0% w / w, or 10.0% w / w, or any amount within any range of the foregoing values. The total amount of pest control agent may include pest control agent found in the base feed, pest control agent found in one or more pest control agent compositions injected into the porous matrix of the base feed, pest control agent compositions coated on the outer surface of the base feed, or any combination thereof.

[0057] In various aspects herein, fish feeds may be administered to fish having a concentration of the pest control agent to fish feed selected from the group consisting of 0.05% w / w, 0.06% w / w, 0.07% w / w, 0.08% w / w, 0.09% w / w, 0.10% w / w, 0.11% w / w, 0.12% w / w, 0.13% w / w, 0.14% w / w , 0.15% w / w, 0.16% w / w, 0.17% w / w, 0.18% w / w, 0.19% w / w, 0.20% w / w, 0.30% w / w, 0.40% w / w, 0.50% w / w, 0.60% w / w, 0.70% w / w, 0.80% w / w, 0.90% w / w or 1.0% w / w, or a range within any of the foregoing concentrations.

[0058] Pest control agents

[0059] Pest control agents suitable for use in fish feed and the pest control agent compositions herein can have an adverse effect on pests that feed on the host. It should be understood that when the host fish ingests the pest control agent as a component of its daily diet within a given duration, the pest control agent can be transferred to the body of the pest when the pest is fed from the host. Pest control agents can include any efficacy ingredients or active agents that affect, promote or contribute to eradicating or reducing pest infections or pest infestations of fish or fish schools. In addition, as a result of reducing, preventing or controlling infection or infestation, suitable pest control agents can alleviate or improve one or more symptoms associated with pest infection or pest infestation. Pest control agents used herein can have biological activity to one or more fish pests and one or more fish species.

[0060] It should be understood that when a fish ingests a pest control agent as described herein, the pest control agent is distributed systemically throughout the tissues and body fluids of the fish. Pests may be exposed to the pest control agent after ingesting the pest control agent through the skin, meat, blood, mucus, mucous membranes, or other tissues of the host organism. Regulation of pests (such as regulation of pest behavior and life cycle) occurs to reduce, prevent, or control infection or infestation of pests in fish. In various aspects, pests can be repelled or killed by the pest control agent herein. Therefore, the pest control agent herein can be provided to pests in fish feed or pest control agent compositions in an amount sufficient to regulate pest behavior.

[0061] Regulation of pests can have many effects on pest colonies, including ultimately reducing the number of live pests that can infect or infest host fish. Regulation of pests can include regulation of pest mortality. It should be understood that regulation of pest mortality can include reducing the number of live pests present on fish or in fish habitats. Regulation of pests can also include regulation of pest behavior, including changes in feeding habits, changes in feeding patterns, changes in appetite, changes in movement patterns, changes in swimming and migration patterns, changes in mating patterns, changes in development, changes in fertility, or any combination thereof compared to the pests found on control fish not fed with pest control agents. Changes in feeding patterns can include a reduction in feeding patterns. Changes in appetite can include a reduction in appetite. Changes in movement can include a reduction in movement. Changes in swimming and migration patterns can include a reduction in swimming and migration due to lethargy and lack of energy. Changes in mating patterns can include a reduction in mating patterns, which in turn can result in a reduction in the development or production of offspring. Alterations in development may include inhibition of development due to inhibition of the molting process resulting in a reduction in pest development or a delay in pest development. Alterations in fertility may include inhibition or delay in egg laying, failure to produce viable eggs, or a reduction in the total number of pregnant female pests.

[0062] In various aspects, the regulation of pests can include changes in the development of pests in their life cycle, including the regulation of growth or progress in a specific life stage, the regulation of growth or progress from one life stage to the next life stage (e.g., regulating molting), the regulation of laying eggs, the regulation of fertility, or any combination thereof. The regulation of growth or progress in a specific life stage can include stopping the growth of the organism and preventing further physical development, including the reduction of body size or sexual development. In some aspects, the regulation of growth or progress from one life stage to the next life stage can include preventing pests from transitioning from one life stage to the next life stage by suppressing the molting process. The regulation of laying eggs can include reducing female laying eggs, which can further cause the fertility of adult females to decrease. The regulation of fertility can include reducing the fertility of female and male pests.

[0063] In various aspects, applying a pest control agent to fish as described herein can also have a beneficial effect on the fish. The application of a pest control agent can impart a beneficial effect by improving the welfare of the fish by reducing the parasite load or total number of parasites in a given environment surrounding the fish. The application of a pest control agent can impart a beneficial effect by reducing the overall mortality rate within a fish population by alleviating or reducing the impact of parasitic infections or infestations on the fish population. The application of a pest control agent can impart a beneficial effect by minimizing or completely eliminating the impact on the quality and quantity of fish flesh within a fish population.

[0064] The fish feed described herein may contain or be supplemented with one or more pest control agents. When the fish feed contains at least two or more different pest control agents, each pest control agent may be individually active (or biologically active) and capable of regulating one or more of the behavior, development or fertility of the pest. Alternatively, the pest control agent may be a component of a pest control agent composition that can be fed to fish alone. Each pest control agent may be individually effective against one or more different pests as described herein.

[0065] Pest control agents suitable for use in the fish feed and pest control agent compositions herein may include one or more active agents, including synthetic or natural agents. The one or more synthetic or natural agents may include agents classified as active pharmaceutical ingredients, veterinary products, etc. In some aspects, active agents for the pest control agents herein may be obtained from plants belonging to the genus Azadirachta. Pest control agents may be obtained or extracted from Azadirachta indica (a tree commonly referred to as "Azadirachta indica"). Extracts prepared from plants belonging to the genus Azadirachta (e.g., Azadirachta indica) may include potent terpenoids, including one or more azadirachtinoid compounds. Azadirachtin compounds include azadirachtin compounds such as azadirachtin A, azadirachtin B, azadirachtin D, azadirachtin E, azadirachtin F, azadirachtin G, azadirachtin H, azadirachtin I, azadirachtin K and / or other azadirachtin variants. Extracts from plants belonging to the genus Azadirachtin may also include many other components in varying amounts. In some aspects, the extract may include additional compounds such as the limonoid compounds salannin, nimbin, deacetyl salannin and 6-deacetyl azadirachtin. In various aspects, the extract may also include one or more azadirachtin compounds.

[0066] As used herein, the term "azadirachtin" may refer to a collective term applicable to a large group of active compounds, and is intended to cover not only all naturally occurring variants or derivatives of azadirachtin, including but not limited to azadirachtin A, B, D, E, F, G, H, I, K, but also all synthetic variants, fragments, analogs and derivatives thereof. In this regard, it should be understood that any azadirachtin variant, fragment, derivative or analog used herein should be functional in that they exhibit at least one inhibitory effect as described.

[0067] Azadirachtin can be obtained or extracted from any part of the neem plant (including, for example, leaves, stems, bark, fruits, seeds, or any combination thereof) by one or more extraction processes. Suitable extraction methods may include techniques utilizing mechanical pressing of neem seeds (i.e., kernels) and the use of non-polar solvents. Various solvent extraction techniques utilizing alcohol or water extraction processes, mechanical pressing, and non-polar extraction methods can be used to produce azadirachtin A-rich pest control agents for use herein, and are described in U.S. Pat. Nos. 4,556,562, 5,695,763, and 11,096,404, the contents of which are incorporated herein by reference in their entirety.

[0068] For example, azadirachtin can be effectively recovered from the seeds of the neem tree. An exemplary method for recovering azadirachtin from neem seeds can include providing neem seeds, crushing the neem seeds, extracting azadirachtin from the crushed seeds with water, and then extracting azadirachtin from the water by adding a second extraction solvent, the second extraction solvent comprising a non-aqueous solvent that is immiscible with water and has a higher solubility of azadirachtin than water or a surfactant having a turbidity temperature between 20°C and 80°C. Concentrated azadirachtin can be recovered from the second extraction solution and exhibits high activity as an insecticide and parasiticide. Extraction methods using polar solvents (e.g., water) produce extracts rich in polar components, such as azadirachtin compounds.

[0069] In various aspects, azadirachtin suitable for use herein includes azadirachtin A, whose scientific name is dimethyl[2aR-[2aα,3β,4β(1aR*,2S*,3aS*,6aS*,7S*,7aS*),4aβ,5α,7aS*,8β(E),10β,10aα,10bβ]]-10-(acetyloxy)octahydro-3,5-dihydroxy-4-methyl-8-[(2-methyl-1-oxo-2-butenyl)oxy]-4-(3a,6a,7,7a)-tetrahydro-6a-hydroxy-7a-methyl-2,7-methanofurano[2,3-b]epoxy[e]oxepin -1a(2H)-yl)-1H,7H-naphtho-[1,8-bc:4,4a-c']difuran-5,10a(8H)-dicarboxylate.

[0070] Azadirachtin A is the most abundant of a group of azadirachtin compounds. Azadirachtin A accounts for about 80% of the azadirachtin compounds in the neem seed kernel. The structural formula of azadirachtin A is:

[0071]

[0072] The pest control agent herein can include a neem extract, which is an aqueous extract. In various aspects, the neem extract can include an aqueous extract of neem seeds. The aqueous extract of neem seeds can include an aqueous extract of neem seed kernels. The aqueous extract of neem seed kernels can include an aqueous extract of whole neem seeds (including neem seed kernels and neem seed coatings). The aqueous extract of neem seeds can be in liquid form, or it can be dried to remove water to produce a powder form. For example, the neem extract herein can include an aqueous extract of neem seeds or an aqueous extract of neem seed kernels that have been dried into a powder.

[0073] It should be understood that the pest control agents described herein are not the same as neem extracts described as neem oil or solvent-first neem extracts. In various aspects, the pest control agents containing azadirachtin herein contain more abundant azadirachtin active ingredients, and in particular azadirachtin A, compared to neem oil and other oil-based formulations. This is because azadirachtin compounds (such as azadirachtin A) are relatively polar complex terpenoids with a large number of oxygen functional groups, which makes the molecule moderately water-soluble (e.g., a solubility of about 2 g / L). Therefore, compared to neem oil or solvent-first neem extracts, azadirachtin compounds (such as azadirachtin A) are present in extracts obtained using polar solvents at much higher concentrations. Without wishing to be bound by any particular theory, it is believed that the bioavailability of the active ingredients in the water-based extract of the extract enriched with azadirachtin A of the present disclosure to the target parasite can be greater than the bioavailability of the active ingredients in neem oil to the target parasite due to the increased solubility and miscibility of the water-based extract in water. Therefore, the pest control agent herein does not contain, consist of, or consist essentially of neem oil. The pest control agent of the fish feed provided herein may also contain, consist of, or consist essentially of azadirachtin A.

[0074] The pest control agent comprising a neem extract rich in azadirachtin A may include a pest control agent having at least 15% to 33% by weight of azadirachtin A. In various aspects, the pest control agent comprising a neem extract rich in azadirachtin A may include a pest control agent having at least 20% to 26% by weight of azadirachtin A. In various aspects, the pest control agent comprising a neem extract rich in azadirachtin A may include a pest control agent having at least 28% to 31% by weight of azadirachtin A. In some aspects, the pest control agent comprising a neem extract rich in azadirachtin A may include a pest control agent having at least 29% to 30% by weight of azadirachtin A. In other aspects, the pest control agent comprising a neem extract rich in azadirachtin A may include a pest control agent having at least 34% to 40% by weight of azadirachtin A. In various aspects, the pest control agent rich in azadirachtin A can include a pest control agent having 30±1% by weight of azadirachtin A. In various aspects, the pest control agent rich in azadirachtin A can include a pest control agent having 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, or 33%, or any amount falling within any of the foregoing ranges. In yet other aspects, the pest control agent comprising a neem extract rich in azadirachtin A can include a pest control agent having at least 34% to 45% by weight or at least 38% to 43% by weight of azadirachtin A. As used herein, unless otherwise specified, the terms "neem extract enriched with azadirachtin A" and "azadirachtin A enriched composition" are used interchangeably. Exemplary neem extract compositions suitable for use as azadirachtin A enriched pest control agent herein may include the formulations as summarized in Table 1.

[0075] Table 1. Exemplary Azadirachtin A-enriched pest control formulations

[0076] Azadirachtin composition %w / w Azadirachtin A 15-33 Azadirachtin B ≤19.0 Azadirachtin D ≤13.0 Azadirachtin E ≤5.0 Azadirachtin F ≤5.0 Azadirachtin G ≤5.0 Azadirachtin H ≤5.0 Azadirachtin I ≤5.0 Azadirachtin K and other azadirachtins ≤5.0 Total Azadirachtin 52.0

[0077] The pest control agent comprising the neem extract enriched with azadirachtin A may also contain other azadirachtin compounds in various concentrations. The azadirachtin compound may include an azadirachtin compound such as ≤19.0% w / w, or ≤6.0% w / w, or 4.0 to 6.0% w / w, or 5.6% w / w to 6.0% w / w of azadirachtin B; ≤6.0% w / w, or ≤13.0% w / w, or 2.5 to 5.0, or 4.0% w / w to 5.0% w / w of azadirachtin D; ≤5.0% w / w, or 1.0% w / w to 5.0% w / w, or 1.5% w / w to 2.0% w / w of azadirachtin E; ≤5.0% w / w, or 1.0% w / w to 5.0% w / w, or 1.5% w / w to 2.0% w / w of azadirachtin E; w / w of azadirachtin F; ≤5.0% w / w, or 1.0% w / w to 5.0% w / w, or 1.5% w / w to 2.0% w / w of azadirachtin G; ≤5.0% w / w, or 1.0% w / w to 5.0% w / w, or 2.5% w / w to 4.0% w / w of azadirachtin H; ≤5.0% w / w, or 1.0% w / w to 4.0% w / w, or 1.5% w / w to 2.5% w / w of azadirachtin I; and ≤5.0% w / w, or 1.0% w / w to 5.0% w / w, or 2.5% w / w to 4.0% w / w of azadirachtin K and / or other azadirachtin variants. The extract may also comprise ≤ 5.0% w / w, or from 1.0% w / w to 5.0% w / w, or from 2.5% w / w to 4.0% w / w of azadirachtin.

[0078] Exemplary pest control agents suitable for use herein may include an aqueous extract of neem seeds that have been dried into a powder. The powder may include the appearance of a fine white powder. Exemplary pest control agents may include azadirachtin A at a concentration of 17% to 37% by weight, azadirachtin B at a concentration of 0% to 19% by weight, and azadirachtin D at a concentration of 0% to 13% by weight. Exemplary pest control agents may also include trace amounts of other limonoid compounds, including azadirachtin and saranlin.

[0079] Exemplary pest control agents comprising azadirachtin A-enriched compositions include, but are not limited to (Coromandel, Intl. Ltd., Telangana, India) or Technical (Coromandel International Ltd., Telangana, India) or any derivatives thereof.

[0080] Method for preparing fish feed

[0081] The disclosure herein provides a method for preparing a fish feed comprising one or more pest control agents. Figure 3 , a method 300 for preparing fish feed as described is shown according to various aspects of the present invention. The method 300 may include preparing a fish feed injected with a certain amount of a pest control agent composition. The method 300 may include preparing the fish feed by providing at least a protein and starch mixture at 302. The method 300 may include subjecting the protein and starch mixture to an extrusion process at 304 to produce a base feed having a porous matrix distributed throughout. The method may include producing a pest control agent composition by mixing the pest control agent and one or more oils together to produce a suspension at 306. The method may include applying the pest control agent composition to the base feed using a vacuum infusion process at 308 to distribute the pest control agent composition throughout the porous matrix of the base feed. The disclosure herein may also include fish feed manufactured according to the method described herein. The method may also include a step of sealing the fish feed to prevent leakage of nutrients and pest control agents by applying a fish oil coating to the fish feed after the vacuum infusion process. Any sealing material for sealing the pest control agent composition may be applied so that it covers all or part of the surface of the fish feed. In various aspects, the extrusion process comprises an extrusion cooking process. In various aspects, the methods herein are suitable for preparing fish feed products according to the steps described herein.

[0082] In various aspects, the basal feed herein can be prepared using an extrusion process or a pressing process. It should be understood that processing the basal feed can include cooking the raw materials at a temperature and shear force that exceeds the stability of the pest control agent described herein. In many aspects, since the compound degrades at high processing temperatures and high shear forces, it is not desirable to include a thermally unstable pest control agent during the extrusion process or the pressing process. Therefore, the present disclosure provides the use of a separate vacuum infusion step to incorporate the pest control agent composition into the basal feed after processing. The vacuum infusion step can be performed after the basal feed is initially extruded or pressed. In some aspects, the vacuum infusion step can be performed immediately after the basal feed is extruded or pressed. In other aspects, the vacuum infusion step can be performed at a predetermined time after the extrusion or pressing process. It should be understood that the vacuum infusion step allows the oil and pest control agent suspension to be incorporated into the pores of the basal feed. The oil and pest control agent suspension can be maintained in the pores via various forces (including spatial or geometric restrictions, Kepler forces, capillary action, etc.). As a result of using an oil and pest control agent suspension in the infusion process, evaporation from the pores of the basal feed is minimized, thus less pest control agent is lost during storage, transportation and handling.

[0083] One advantage of performing the vacuum infusion step after the extrusion process or the compression process is that there is limited or no degradation of the functional or active agent when it is incorporated into the base feed as a separate and distinct processing step. Another important advantage of performing the vacuum infusion step after the extrusion process or the compression process is that the efficacy ingredient or active agent can be added to the base feed using equipment separate from the main manufacturing equipment used to prepare the base feed. This advantage allows for a more streamlined manufacturing process, customized temperature control, dedicated or isolated processing lines for adding the pest control agent composition, reduced or eliminated cross-contact of the manufacturing equipment used to prepare the base feed with any functional or active agent, and more simplified cleaning routines for both the base feed manufacturing equipment and the equipment used for the vacuum infusion process.

[0084] The method herein can generate a fish feed containing one or more pest control agents, which are distributed throughout the porous matrix of the basic feed or distributed around the outer surface of the basic feed. It should be understood that the fish feed described herein can include a pest control agent, which is at least partially dispersed throughout the porous matrix of the basic feed. In various aspects, the fish feed herein can include a pest control agent, which is at least completely dispersed throughout the porous matrix of the basic feed. In some aspects, the fish feed herein can be at least partially coated with a pest control agent on the outer surface of the basic feed, or a pest control agent composition can be completely coated on the outer surface of the basic feed. In various aspects, the fish feed herein can include a pest control agent, which is at least partially coated with a pest control agent on the outer surface of the basic feed, and is at least partially dispersed throughout the porous matrix of the basic feed.

[0085] Methods for incorporating a pest control agent into a fish feed may include incorporating the pest control agent so that the final concentration of the pest control agent in the fish feed includes about 0.01 grams of pest control agent per kilogram of fish feed (g / kg) to about 1000 g / kg, or about 0.01 g / kg, 0.1 g / kg, 1 g / kg, 2 g / kg, 3 g / kg, 4 g / kg, 5 g / kg, 6 g / kg, 7 g / kg, 8 g / kg, 9 g / kg, or 10 g / kg, 20 g / kg, 30 g / kg, 40 g / kg, 50 g / kg, 60 g / kg, 70 g / kg, 80 g / kg, 90 g / kg, 100 g / kg, 250 g / kg, 500 g / kg, 750 g / kg, or 1000 g / kg, or any amount within any of the foregoing ranges. Concentrations are described in more detail herein with reference to fish feed.

[0086] Methods for incorporating a pest control agent into fish feed may include determining the final concentration of the pest control agent incorporated as part of the fish feed. Determination of the final concentration may include sampling the fish feed using various quantitative analytical methods. For example, the fish feed sample may be extracted by overnight protein precipitation in methanol. After extraction, the sample may be analyzed using Supel TM The samples were cleaned with QuE Z-Sep+ (Sigma Aldrich, St. Louis, Missouri, USA) adsorbent, which is a silica-based material with an activated zirconium oxide phase, a particle size of approximately 50 μm, and a pore size of 70 Å. The resulting extraction solution may be filtered through a polytetrafluoroethylene filter having a pore size of 0.2 μm or greater. Analysis of the final concentration of the pest control agent in the fish feed sample may be performed using high performance liquid chromatography with ultraviolet detection (HPLC-UV).

[0087] The methods herein may include extracting a neem extract enriched in azadirachtin A, including extracts having at least 15% w / w to 33% w / w azadirachtin A, at least 28% w / w to 31% w / w azadirachtin A, and at least 29% w / w to 30% w / w azadirachtin A.

[0088] The uniformity of the amount of the pest control agent in the fish feed of this article can be measured by the analytical method of this article and by evaluating the coefficient of variation. Without wishing to be bound by any particular theory, it should be understood that the coefficient of variation (CV) is the ratio of the standard deviation to the mean value, and shows the degree of variation relative to the population mean value. The coefficient of variation is expressed as a percentage (%) and can be used as a standardized measure of the variability within a population or data set. A higher CV represents a greater variability between the data, and a lower CV represents a lower variability between the data. It should be understood that in the case of mixing ingredients for food products, a CV of less than 10% represents excellent mixing, and the method does not need to be further optimized. See, for example, Feed Manufacturing, Testing Mixer Performance, Bulletin M1172, Kansas State University Agriculture Experiment Station and Cooperative Extension Service. September 2017).

[0089] In a specific example herein, various batches of fish feed are prepared by providing a protein and starch mixture and subjecting it to an extrusion process to produce a base feed with a porous matrix distributed throughout. The pest control agent composition is produced by mixing a composition rich in azadirachtin A and one or more oils together to produce a suspension of a composition rich in azadirachtin A and oil. The composition rich in azadirachtin A and the oil suspension are applied to the base feed using a vacuum infusion process to distribute the composition rich in azadirachtin A and the oil suspension throughout the porous matrix of the base feed. The coefficient of variation of the fish feed is measured from the initial processing time to eight weeks after processing. Using the method herein and pest control rich in azadirachtin A, CV values ​​of less than 10% (i.e., <10%) are consistently reported. The coefficient of variation of the feed batches measured is shown in Table 2. The average CV of all batches during eight weeks was 2.35%.

[0090] Table 2. Relationship between coefficient of variation of fish feed and storage time

[0091] Storage time (weeks) n Average azadirachtin A (mg / g) Standard Deviation Coefficient of variation (%) 0 6 2.01 0.09 4.38 1 3 1.90 0.02 1.26 2 3 1.88 0.05 2.61 3 3 1.87 0.03 1.34 4 3 1.92 0.07 3.42 8 3 1.97 0.02 1.12

[0092] Fish feed containing azadirachtin A

[0093] The fish feed herein may include a fish feed supplemented with the pest control agent azadirachtin A. The fish feed may be applied to various fish as part of a fish feed diet to control arthropod pests in an aquaculture environment. The fish feed may be at least partially coated with a composition rich in azadirachtin A on the outer surface or completely coated with a composition rich in azadirachtin A on the outer surface. In some aspects, the fish feed herein may include a composition rich in azadirachtin A at least partially dispersed throughout the fish feed. In various aspects, a solid feed (such as a base feed particle) may also include a composition rich in azadirachtin A disposed on the surface or distributed throughout the fish feed, such as oil disposed in a porous matrix on the outer surface of the base feed particle. In various aspects, the fish feed herein may be at least partially coated with a composition rich in azadirachtin A on the outer surface, and may also have a composition rich in azadirachtin A at least partially dispersed throughout the fish feed. In various aspects, the fish feed may include one or more layers of a composition rich in azadirachtin A on the outer surface.

[0094] The fish feed herein may comprise a composition enriched with azadirachtin A at a concentration of about 0.01 to 100 grams per kilogram (g / kg) of fish feed, about 90 g / kg of fish feed, about 80 g / kg of fish feed, about 70 g / kg of fish feed, about 60 g / kg of fish feed, about 50 g / kg of fish feed, about 40 g / kg of fish feed, about 30 g / kg of fish feed, about 20 g / kg of fish feed, about 0.01 to 10 g / kg of fish feed, about 1 g to 10 g / kg of fish feed, about 2 g to 9 g / kg of fish feed, about 3 g to 7 g / kg of fish feed, about 4 g to 6 g / kg of fish feed, or about 5 g / kg of fish feed.

[0095] In various aspects, the fish feed herein may comprise a composition rich in azadirachtin A at a concentration of about 0.01 g of the composition rich in azadirachtin A per kilogram of fish feed (g / kg), 0.05 g / kg of fish feed, 0.1 g / kg of fish feed, 0.2 g / kg of fish feed, 0.3 g / kg of fish feed, 0.4 g / kg of fish feed, 0.5 g / kg of fish feed, 0.6 g / kg of fish feed, 0.7 g / kg of fish feed, 0.8 g / kg of fish feed, 0.9 g / kg of fish feed, 1.0 g / kg of fish feed. , 1.25g / kg fish feed, 1.5g / kg fish feed, 1.75g / kg fish feed, 2.0g / kg fish feed, 2.25g / kg fish feed, 2.5g / kg fish feed, 2.75g / kg fish feed, 3.0g / kg fish feed, 5.0g / kg fish feed, 5.25g / kg fish feed, 5.5g / kg fish feed, 5.75g / kg fish feed, 6.0g / kg fish feed, 6.25g / kg fish feed, 6.5g / kg fish feed, 6.75g / kg fish feed feed, 7.0g / kg fish feed, 7.0g / kg fish feed, 7.25g / kg fish feed, 7.5g / kg fish feed, 7.75g / kg fish feed, 8.0g / kg fish feed, 8.25g / kg fish feed, 8.5g / kg fish feed, 8.75g / kg fish feed, 9.0g / kg fish feed, 9.25g / kg fish feed, 9.5g / kg fish feed, 9.75g / kg fish feed, 10.0g / kg fish feed, 15g / kg fish feed, 20g / kg fish feed The present invention relates to a concentration of 1g / kg fish feed, 25g / kg fish feed, 30g / kg fish feed, 35g / kg fish feed, 40g / kg fish feed, 45g / kg fish feed, 50g / kg fish feed, 55g / kg fish feed, 60g / kg fish feed, 65g / kg fish feed, 70g / kg fish feed, 75g / kg fish feed, 80g / kg fish feed, 85g / kg fish feed, 90g / kg fish feed, 95g / kg fish feed, 100g / kg fish feed, or any amount within any of the foregoing concentration ranges.

[0096] It should be understood that the concentration of the above-mentioned azadirachtin A-rich composition is equivalent to about 0.001% w / w (weight percentage) to 10% w / w of the azadirachtin A-rich composition for fish feed. In various aspects, the fish feed herein may comprise an azadirachtin A-rich composition that is 0.001% w / w, 0.002% w / w, 0.003% w / w, 0.004% w / w, 0.005% w / w, 0.006% w / w, 0.007% w / w, 0.008% w / w, 0.009% w / w, 0.010% w / w, 0.020% w / w, 0.030% w / w, 0.040% w / w, 0.050% w / w, 0.060% w / w, 0.070% w / w, 0.080% w / w, 0.090% w / w, 0.10% w / w, 0.11% w / w, 0.12% w / w, 0.13% w / w, 0.14% w / w, 0.15% w / w, 0.16% w / w, 0.17% w / w, 0.18% w / w, 0.19% w / w, 0.20% w / w, 0.21% w / w, 0.22% w / w, 0.23% w / w, 0.24% w / w, 0.25% w / w, 0.26% w / w, 0.27% w / w, 0.28% w / w, 0.29% w / w, 0.30% w / w, 0.31% w / w, 0.32% w / w, 0.33% w / w, 0.34% w / w, 0.35% w / w, 0.36% w / w, 0.37% w / w, 0.38% w / w, 0.39% w / w, 0.40% w / w, 0.41% w / w, 0.42% w / w, 0.43% w / w, 0.44% w / w, 0.45% w / w, 0.46% w / w, 0.47% w / w, 0.48% w / w, 0.49% w / w, 0.50% w / w, 0.51% w / w, 0.52% w / w, 0.53% w / w, 0.54% w / w, 0.55% w / w, 0.56% w / w, 0.57% w / w, 0.58% w / w, 0.59% w / w, 0.60% w / w, 0.61% w / w, 0.62% w / w, 0.63% w / w, 0.64% w / w, 0.65% w / w, 0.66% w / w, 0.67% w / w, 0.68% w / w, 0.69% w / w, 0.70% w / w, 0.71% w / w, 0.72% w / w, 0.73% w / w, 0.74% w / w, 0.75% w / w, 0.76% w / w, 0.77% w / w, 0.78% w / w, 0.79% w / w, 0.80% w / w, 0.81% w / w, 0.82% w / w, 0.83% w / w, 0.84% w / w, 0.85% w / w, 0.86% w / w, 0.87% w / w, 0.88% w / w, 0.89% w / w, 0.90% w / w, 0.91% w / w, 0.92% w / w, 0.93% w / w, 0.94% w / w, 0.95% w / w, 0.96% w / w, 0.97% w / w, 0.98% w / w, 0.99% w / w, 1.0% w / w, 2.0% w / w, 3.0% w / w, 4.0% w / w, 5.0% w / w, 6.0% w / w, 7.0% w / w, 8.0% w / w, 9.0% w / w or 10.0% w / w (i.e., the weight of the azadirachtin A-enriched composition to the weight of the fish feed), or any amount within any range of the foregoing values.

[0097] In various aspects herein, a fish feed having a composition enriched with azadirachtin A at a concentration selected from the group consisting of 0.05% w / w, 0.06% w / w, 0.07% w / w, 0.08% w / w, 0.09% w / w, 0.10% w / w, 0.11% w / w, 0.12% w / w, 0.13% w / w, 0.14% w / w, 0.15% w / w , 0.16% w / w, 0.17% w / w, 0.18% w / w, 0.19% w / w, 0.20% w / w, 0.30% w / w, 0.40% w / w, 0.50% w / w, 0.60% w / w, 0.70% w / w, 0.80% w / w, 0.90% w / w or 1.0% w / w (i.e., the weight of the azadirachtin A-enriched composition to the weight of the fish feed), or a range within any of the foregoing concentrations.

[0098] Azadirachtins may be relatively unstable in water, however, when they are components of fish feed, such as when dispersed throughout or coated on fish feed, azadirachtins including azadirachtin A are at least temporarily stable, allowing them to exhibit their full biological activity during feeding. Any fish feed that falls to the seafloor will degrade after prolonged exposure to water. In addition, it should be noted that azadirachtin A or any neem extract agent disclosed herein presents minimal risk of toxic effects on fish or humans, and is therefore safe for use in wild and farmed fish. Arthropods and other invertebrates are sensitive to the active ingredient (i.e., azadirachtin A), while higher organisms including mammals are not affected. In addition, since azadirachtin A or any neem extract as described herein is readily soluble in water, they do not reside and accumulate in fish. In contrast, once application is stopped, the pest control agent may quickly lose its effectiveness because it is metabolized, degraded and / or excreted from the body. In the case of farmed fish stocks, this ensures that there is little to no harvest withdrawal period following administration of the fish feed or composition as described herein.

[0099] The fish feed provided herein can include a composition rich in azadirachtin A and one or more other agents. The one or more or other agents can include anti-ectoparasitic agents, antimicrobial agents (e.g., antibiotics, antibacterial agents, antifungal agents, antiviral agents), antiparasitic agents (e.g., anti-endoparasitic agents or anti-ectoparasitic agents) or antiprotozoal agents. The one or more or other agents can be mixed with the fish feed or coated on the fish feed, or layered in the fish feed. The one or more or other agents can be provided separately (e.g., in liquid or solid form) and can be applied separately (e.g., before or after the fish feed) or simultaneously with the fish feed (e.g., during the fish feed).

[0100] Pest control composition

[0101] The present disclosure also provides a pest control agent composition for application to fish, wherein the pest control agent composition may include one or more pest control agents. It should be understood that the pest control agent composition is not a fish feed, but is intended to be applied to fish separately or in addition to fish feed. The pest control agent composition can be provided separately for application before, during or after the application of fish feed. Therefore, in various aspects, the pest control agent composition herein may be suitable for use as a veterinary product or dietary supplement for reducing, preventing or controlling pest infections or pest infestations in fish in some aspects. The pest control agent composition may be applied to fish at a concentration described elsewhere herein. For example, the pest control agent composition herein may be applied in an amount of about 0.01 grams of pest control agent / kilogram of fish feed (g / kg) to about 100g / kg, as described elsewhere herein. In various aspects, the pest control agent composition comprises a neem extract rich in azadirachtin A.

[0102] The pest control agent composition herein may include an azadirachtin extract rich in azadirachtin A. The pest control agent composition may include a liquid, solid or semisolid form, and may also include one or more of an excipient, a diluent, a carrier, a vitamin, a mineral or a combination thereof. The pest control agent composition may be in the form of a dietary supplement provided as any one of granules, flakes, pellets, powders, tablets, pills, capsules, etc. In various aspects, the pest control agent composition herein may be formed into many shapes and sizes. In various aspects, the fish feed herein may be in the shape of a triangle, square, rectangle, sphere, rhombus, cylinder, spherical, clover-shaped, amorphous, etc. The fish feed may be formed by a method including one or more of extrusion, distillation, cold pressing, high pressure processing, etc.

[0103] Alternatively, the pest control agent composition can be provided in a form that can be eaten by fish but does not provide nutrition to the fish. For example, the pest control agent composition can include a veterinary product that can contain a substance or combination of substances for managing or preventing fish diseases. The pest control agent composition can also be formulated for parenteral administration. Therefore, the pest control agent composition can include a pharmaceutically acceptable carrier, diluent or excipient or a combination thereof. In addition, the pest control agent composition can be sterile.

[0104] The pest control agent composition herein can be included in one or more types of fish feeds intended to be mixed with another composition (such as a base feed). The pest control agent composition can be in the form of a premix, a concentrate, a base mix, a supplement, a surface feed, a liquid drenching agent, or a combination thereof.

[0105] The pest control agent in the pest control agent composition herein may include one or more agents for reducing, preventing or controlling infections or infestations caused or contributed to by one or more endoparasites or ectoparasite pests (including any type of worms, worms, trematodes, lice, mites, bacteria, viruses, fungi and protozoa, as described elsewhere). Each pest control agent contained in the pest control agent composition may be individually capable of reducing, preventing or controlling one or more of parasitic, bacterial, viral, fungal or protozoan infections or infestations. For example, the pest control agent composition herein may include a pest control agent composition that exhibits one or more inhibitory effects, including antiparasitic effects, antibacterial effects, antiviral effects, antifungal effects, antiprotozoal effects, or any combination thereof.

[0106] The pest control agent composition can be applied before, during or after applying any fish feed. In some respects, the pest control agent composition can be applied together with the fish feed that does not contain the pest control agent. In some respects, the pest control agent composition can be co-applied with the fish feed containing the pest control agent. When co-applied with the fish feed containing the pest control agent, the independent pest control agent composition can include the pest control agent identical with the pest control agent in the fish feed, or it can be the pest control agent different from the pest control agent in the fish feed. When co-applied with the fish feed containing the pest control agent, the concentration of the independent pest control agent composition can be identical with the concentration of the pest control agent in the fish feed, or it can be different from the concentration of the pest control agent in the fish feed. The pest control agent composition herein can be included in the fish feed in the form of a veterinary product or any complete and balanced dietary supplement for fish feed, or can be provided as a component of a complete fish feed.

[0107] Target fish and pests

[0108] The fish feed provided herein can be fed as a fish feed diet, or used to feed any fish susceptible to infection or infestation by one or more pests. For example, the fish feed can be used as a component of a diet fed to any farmed fish (including, for example, commercially relevant fish species) in aquaculture. For example, the fish feed provided herein can form a part of a diet fed to any freshwater fish, saltwater fish, or marine fish. The fish feed can be used as a component of a diet fed to any species belonging to the following families: Cyprinidae, Cichlidae, Panfish, Sciaenidae, Serranidae, Carpidae, Sparidae, Perchidae, Perchidae, Mugilidae, Cypriniformes, Perchidae, Channaidae, Tilapia, and Salmonidae. Therefore, the fish feed herein can be used to feed species belonging to any genus within these families, particularly those species that are cultured for human or animal consumption. For example, the fish feed described herein can be used to feed species belonging to Salmo and / or Oncorhynchus, but is not limited thereto. In particular, the fish feed herein can be used to control pests in populations of wild or farmed salmon or trout species, including, for example, any of Atlantic salmon (Salmo salar), Pacific salmon, Arctic char (Char), or rainbow trout. In addition, the fish feed can be used as a pest control agent for other fish species within the aquaculture industry, such as sea bass, porgy, grouper, pompano, and tuna, as well as for the pet and ornamental fish industry, for example, for goldfish (Carassius auratus) pest control.

[0109] Pests of target fish herein may include ectoparasites, including species belonging to the phylum Arthropoda. Susceptible arthropods include various copepods, including many sea lice species that inhabit fish hosts. For example, in the case where fish belong to the salmonid family (e.g., Salmo salar and / or Pacific salmon), the pest control agent of fish feed provided herein can prevent and control sea lice infection, sea lice infestation, copepod infection, copepod infestation or any combination thereof. The pest control agent present in the fish feed provided herein can prevent and control ectoparasites, including parasitic crustaceans, also referred to as copepods, which belong to the genus Argulusssp. or the genus Caligus ssp. In particular, the pest control agent used in the fish feed described herein can effectively prevent and control the copepod infection and infestation of one or more types of farmed fish. As used herein, the term "copepod" refers to a group of crustaceans found in fresh water and in seawater, and it has one or more parasitic stages of its life cycle. Unless otherwise specified, the term copepod may refer to any of the various sea lice species described herein.

[0110] The copepod species that are ectoparasites of Atlantic salmon belong to the genus Lepeophtheirus and are referred to as salmon lice. As used herein, it is understood that the term "sea louse" refers to the singular form, and the term "sea lice" refers to the plural form. Unless otherwise specified, each term can be used interchangeably. Salmon lice (Lepeophtheirus salmonis) is a copepod ectoparasite species that lives primarily on salmon (including Atlantic salmon and Pacific salmon) and sometimes on sea trout. Other types of copepod ectoparasites that infect fish belonging to the Salmonidae family include sea lice (Caligus clemensi), long-spined fish lice (Caligus elongatus) and Chilean fish lice (Caligus rogercresseyi). Sea lice, including those from the genus Lepeophtheirus and the genus Lepeophtheirus, are ectoparasites that feed on the blood, mucus, muscle and skin of various salmon species.

[0111] Reference now Figure 4 , the life cycle of the salmon lice (ie, L. salmonis) is shown according to various aspects herein. Figure 1Aspects of the have been adapted from Sea Lice Research Centre, 2020, "SLRC-Life cycle of the salmon louse (Lepeophtheirus salmonis)", https: / / doi.org / 10.18710 / GQTYYL, Dataverse NO, V1. The life cycle of salmon lice is complex and consists of eight different life stages. The larval stage begins after the eggs hatch and includes two different stages, including larval 1 stage and larval 2 stage, shown as 400 and 402, respectively. The larval 1 stage 400 and the larval 2 stage 402 of the salmon louse passively drift in the water, wherein, typically, both stages are about 0.5 mm to 0.7 mm in length and their color is relatively translucent. At temperatures up to about 15°C, the larval 1 stage 400 and the larval 2 stage 402 exist for about 52 hours and 175 hours, respectively. After molting from the larval 2 stage 402, the salmon louse transitions to the infective copepod stage 404 at about 10 days of age, and it should be understood that development at any life stage may depend on a variety of environmental factors, including but not limited to temperature, salinity, light hours, etc. The length of the salmon louse in the copepod stage 404 is about 0.7mm to 1.0mm. In the copepod stage 404, the salmon louse is usually attached to the fish along the fins or scales of the fish. After attachment, the salmon louse further molts to the larval 1 stage 406. In the larval 1 stage 406, the salmon louse is more firmly attached to the host fish with the help of the frontal filaments. The larval 1 life stage usually lasts about 10 days. The salmon louse life cycle continues to molt from the larval 1 stage 406 to the larval 2 stage 408, while remaining firmly attached to the fish. During the larval 1 stage 406 and the larval 2 stage 408, the average length of the salmon louse is 1.0mm to 2.5mm. The larval 2 life stage usually lasts about 10 days.

[0112] After the two larval stages, the salmon lice further molts into the pre-adult stage, in which the salmon lice become active and can swim or move around the surface of the fish. In the pre-adult stage, the sexual development of females and males begins to differentiate. The pre-adult stage consists of 2 different stages, wherein males develop from the larval 2 stage 408 to the pre-adult 1 male stage 410, and females develop to the pre-adult 1 female stage 412. Females spend about 10 days in the pre-adult 1 female stage 412 and about 12 days in the pre-adult 2 female stage 416. Males spend about 8 days in the pre-adult 1 male stage 410 and about 9 days in the pre-adult 2 male stage 414. From the pre-adult 1 stage, the sex continues to develop to the pre-adult 2 male stage 414 and the pre-adult 2 female stage 416. During the pre-adult stage, the salmon lice grow to an average length of about 2.5mm to 3.5mm. At the final adult stage, adult males 418 and adult females 420 can be distinguished by physical and phenotypic characteristics. That is, adult females 420 are 8 mm to 20 mm in length, including two egg mass bags 422 visible from the rear of the organism. In contrast, adult males 418 are 5 mm to 7 mm in length.

[0113] On average, salmon lice can live up to about 215 days throughout their life cycle. It should be understood that each stage of the salmon lice life cycle may depend on temperature, water salinity, water currents, pollution levels, and various additional environmental factors. Thus, the complete life cycle of a salmon lice may vary from 32 days to 215 days, depending on fluctuations in such external factors.

[0114] Thus, in various aspects, the disclosure herein provides a fish feed comprising a pest control agent that can achieve any of the reduction, prevention, or control of scab louse and fish lice infestation or infestation. Scab louse infestation or infestation can be caused or contributed to by salmon lice (Lepeophtheirus salmonis). Fish lice infestation or infestation can be caused or contributed to by sea lice (sea lice, long-spined fish lice, and Chilean fish lice). In various aspects, the fish feed can be used for species belonging to the Salmonidae family. For example, the fish feed herein can be a salmon genus and / or a Pacific salmon genus fish feed. However, it should be understood that the fish feed herein can be given to any fish susceptible to scab louse and fish lice infestation or infestation.

[0115] Method of applying pest control agents to fish

[0116] The pest control agent herein can be applied to fish in fish feed and pest control agent compositions. The management method using the pest control agent in fish feed can be referred to as an in-feed agent delivery method. Therefore, the present disclosure provides an in-feed agent delivery method for reducing, preventing or controlling pests. It should be understood that the in-feed agent delivery method is not a method for topically applying the pest control agent to the target pest. The in-feed agent delivery method herein can also utilize a pest control agent in a non-feed form, such as a veterinary product or a dietary supplement. Therefore, the present disclosure also provides a veterinary product or a dietary supplement as a medicament for reducing, preventing or controlling pests.

[0117] Pest control agents not included in fish feed can be applied to fish as supplements to fish feed in separate pest control agent compositions, such as in the form of veterinary products or dietary supplements. Fish feed can be applied simultaneously or separately with the pest control agent composition. It should be noted that various pest control agents herein can be applied to sick fish, fish infested with parasites, otherwise healthy fish (to prevent parasitic infection), or fish with weaker aggressiveness due to different infections or conditions unrelated to pest infection or infestation. It should be understood that sick or less aggressive fish may eat less, and therefore may ingest lower concentrations of pest control agents. Therefore, management methods based on the use of both fish feed and pest control agent compositions may be particularly useful for managing fish whose appetite is affected by disease, infection, infestation (or fish with weaker aggressiveness that usually eat less fish feed). In addition, for fish with weaker aggressiveness or low food intake, the simultaneous use of pest control agent compositions and fish feed supplemented with pest control agents can promote or ensure that the correct concentration of pest control agents is applied to fish.

[0118] The concentration of the pest control agent added to the fish feed or pest control composition herein can be an amount effective to achieve the desired regulation of the behavior, development or mortality of the pest as discussed elsewhere herein. It should be understood that the exact amount of the pest control agent to be added to the fish feed or pest control composition herein can vary depending on, for example, the species of fish, the number of fish to be fed, the degree of infection or infestation, etc. Other factors that affect the amount of the pest control agent added to the fish feed or pest control composition include, for example, the presence of possible competitors for the feed (i.e., other non-target animal species that may eat the fish feed), the type of pest to be controlled, the age and maturity of the pest, the age and maturity of the fish, the season, the type of water (e.g., pH, salinity, purity, temperature), and the aggressiveness of the fish. It should be understood that the concentration of the pest control agent added to the fish feed or pest control composition herein can include an amount effective to achieve the desired effect of regulating the behavior and development of the pest, wherein the effective amount includes one or more concentrations or concentration ranges as described herein. It will be appreciated that an effective amount may be determined by comparison to a control fish or fish population not fed the pest control agent.

[0119] The fish feed and the pest control agent composition can be formulated so that the concentration of the pest control agent applied to the fish via the fish feed or the pest control agent composition can be about 0.01 mg to 100 mg of the pest control agent / kg body weight / day (mg / kg / day), 1 mg to 90 mg / kg / day, 1 mg to 80 mg / kg / day, 1 mg to 70 mg / kg / day, 1 mg to 60 mg / kg / day, 5 mg to 50 mg / kg / day, 10 mg to 40 mg / kg / day, 15 mg to 35 mg / kg / day, 20 mg to 30 mg / kg / day, 0.01 mg to 10 mg / kg / day, or 0.01 mg to 5.0 mg / kg / day. In various aspects, the pest control agent is applied to the fish in the fish feed at a target concentration of 0.01 mg to 5.0 mg of azadirachtin A / kg body weight / day. In various aspects, the pest control agent is applied to fish in fish feed at a target concentration of 1.5 mg to 2.5 mg azadirachtin A / kg body weight / day. In some aspects, the pest control agent is applied to fish in fish feed at a target concentration of 2.6 mg to 5.0 mg azadirachtin A / kg body weight / day.

[0120] It should be understood that the amount of pest control agent applied to fish can include an amount effective to produce an inhibitory effect on one or more pests, which is in the range of about 0.01 mg to 100 mg / kg / day (e.g., mg pest control agent / kg body weight / day), including at least 0.01 mg / kg / day, 0.02 mg / kg / day, 0.03 mg / kg / day, 0.04 mg / kg / day, 0.05 mg / kg / day, 0.06 mg / kg / day, 0.07 mg / kg / day, 0.08 mg / kg / day, 0.09 mg / kg / day, 10 ... .09mg / kg / day, 0.10mg / kg / day, 0.20mg / kg / day, 0.30mg / kg / day, 0.40mg / kg / day, 0.50mg / kg / day, 0.60mg / kg / day, 0.70mg / kg / day, 0.80mg / kg / day, 0.90mg / kg / day, 1.0mg / kg / day, 2.0mg / kg / day, 3.0mg / kg / day, 4.0mg / kg / day, 5.0mg / kg / day, 6.0mg / kg / day, 7.0mg / kg / day .0mg / kg / day, 8.0mg / kg / day, 9.0mg / kg / day, 10.0mg / kg / day, 11.0mg / kg / day, 12.0mg / kg / day, 13.0mg / kg / day, 14.0mg / kg / day, 15.0mg / kg / day, 16.0mg / kg / day, 17.0mg / kg / day, 18.0mg / kg / day, 19.0mg / kg / day, 20.0mg / kg / day, 25.0mg / kg / day, 30.0mg / kg / day In some embodiments, the present invention relates to an inhibitory effect of at least one embodiment of the present invention. The inhibitory effect of at least one embodiment of the present invention may be at least one embodiment of the present invention. The inhibitory effect of at least one embodiment of the present invention may be at least one embodiment of the present invention. The inhibitory effect of at least one embodiment of the present invention may be at least one embodiment of the present invention.

[0121] Fish feed or pest control agent composition can be applied for a period of time, as long as the desired inhibitory effect is reached. For example, the pest control agent composition or fish feed can be applied in about 10 to 20 days or about 14 to 18 days. It should be understood that the pest control agent composition, fish feed or both can be applied continuously for 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days or 20 days, or any number of days falling within any of the aforementioned ranges. In various fields, the pest control agent composition or fish feed herein can be applied for a longer period of time, such as more than 20 days. In various fields, fish feed or pest control agent can be applied for at least 11 days. In various fields, fish feed or pest control agent can be applied for at least 14 days. It should be understood that the time required for applying the pest control agent composition or fish feed herein can have a variable length, so as to target the developmental life stage of the pests present in the school of fish, for water temperature, pest control agent concentration or any combination thereof. In some aspects, the pest control agents herein can be applied prophylactically in the diet of fish in an amount effective to prevent pest infection or infestation from occurring within the fish population. It should also be understood that the pest control agents herein can be applied prophylactically in the diet at any time during the life cycle of the fish, such as from stocking to harvest, seasonally, or during an outbreak of infection or infestation within a population or a nearby farm.

[0122] In various management methods, the pest control agent of this paper can be applied for several days non-continuously, wherein the pest control agent is applied after a period of rest in the predetermined time period, and then again after a period of rest in the predetermined time period, and so on. For example, in some aspects, the pest control agent can be applied for three days every 10 days. In other aspects, the pest control agent can be applied for seven days every 14 days. The method of applying the pest control agent and then resting for a period of time within the predetermined time can be reused as required, or until reducing, preventing or controlling pests to infect or infest. It should be understood that the pest control agent can be applied for a predetermined time period, including applying 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days every 5 days to 30 days of rest between application.

[0123] During application, the pest control agent composition or fish feed can be applied multiple times a day as needed to achieve an inhibitory effect. For example, the pest control agent composition or fish feed described herein can be applied about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more times a day.

[0124] It should be understood that the definitions described herein apply to all aspects described unless otherwise stated.

[0125] In this document, the terms "a", "an" or "the" are used to include one or more than one, unless the context clearly dictates otherwise. The term "or" is used to refer to a non-exclusive "or" unless otherwise indicated. All publications, patents, and patent documents cited in this document are incorporated herein by reference in their entirety, as if individually incorporated by reference. In the event of an inconsistency between the usages of this document and those documents so incorporated by reference, the usage in the incorporated references should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.

[0126] Values ​​expressed in range format should be interpreted in a flexible manner to include not only the values ​​explicitly listed as the limits of the range, but also all individual values ​​or subranges covered within the range, as if each value and subrange were explicitly listed. For example, a range of "about 0.1% to about 5%" or "about 0.1% to 5%" should be interpreted to include not only about 0.1% to about 5%, but also individual values ​​(e.g., 1%, 2%, 3% and 4%) and subranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the specified range. Unless otherwise indicated, the statement "about X to Y" has the same meaning as "about X to about Y". Similarly, unless otherwise indicated, the statement "about X, Y or about Z" has the same meaning as "about X, about Y or about Z".

[0127] Unless expressly stated, ppm (parts per million), percentages and ratios are based on weight. Percentages based on weight (%w / w or w / w%) are also referred to herein as weight percent (wt%) or weight percent (%wt.).

[0128] Exemplary embodiments of the present invention are as follows;

[0129] Embodiment 1: A fish feed, comprising:

[0130] a basal feed comprising a porous matrix distributed throughout; and

[0131] a pest control agent composition comprising a pest control agent and one or more oils;

[0132] wherein the pest control agent composition is distributed throughout the porous matrix of the basal feed.

[0133] Embodiment 2: The fish feed according to embodiment 1, wherein the pest control agent comprises a neem extract enriched in azadirachtin A.

[0134] Embodiment 3: The fish feed according to embodiment 2, wherein the neem extract enriched with azadirachtin A comprises at least 15 wt% to 33 wt% azadirachtin A.

[0135] Embodiment 4: The fish feed according to any one of embodiments 1 to 3, wherein the one or more oils comprise one or more of the following: soybean oil, grape seed oil, canola oil, sunflower oil, safflower oil, rapeseed oil, hydrogenated rapeseed oil, hydro-pressed linseed oil, camelina oil, cold-pressed linseed oil, coconut oil, fish oil, poultry oil or fish oil stearin or fractions thereof.

[0136] Embodiment 5: The fish feed according to any one of embodiments 1 to 4, wherein the porous matrix is ​​formed by extrusion of a raw material comprising protein and starch.

[0137] Embodiment 6: The fish feed according to any one of Embodiments 1 to 5, wherein the pest control agent composition is applied to the base feed via a vacuum infusion process.

[0138] Embodiment 7: The fish feed according to any one of Embodiments 1 to 6, wherein the pest control agent is at least partially distributed throughout the porous matrix of the base feed.

[0139] Embodiment 8: The fish feed according to any one of Embodiments 1 to 7, wherein the pest control agent composition is further coated on the outer surface of the base feed.

[0140] Embodiment 9: The fish feed of any one of Embodiments 1 to 8, wherein the fish feed comprises between about 15% and about 65% by weight protein and between about 10% and about 45% by weight fat; and

[0141] The fish feed pellets include terrestrial animal protein, fish meal, plant-based protein, or a combination thereof.

[0142] Embodiment 10: The fish feed according to any one of embodiments 1 to 9, wherein the base feed comprises one or more of pellets, extruded blocks, steam pellets, flakes, tablets or powder.

[0143] Embodiment 11: The fish feed of any one of Embodiments 1 to 10, wherein the pest control agent comprises a solid form suspended in one or more oils.

[0144] Embodiment 12: A method for preparing fish feed, the method comprising:

[0145] providing a protein and starch mixture and subjecting it to an extrusion process to produce a base feed having a porous matrix distributed throughout;

[0146] producing a pest control agent composition by mixing the pest control agent and one or more oils together to produce a suspension;

[0147] The pest control composition is applied to the basal feed using a vacuum infusion process to distribute the pest control composition throughout the porous matrix of the basal feed.

[0148] Embodiment 13: The method according to Embodiment 12, wherein the pest control agent comprises a neem extract enriched in azadirachtin A, the neem extract enriched in azadirachtin A comprising at least 15% to 33% azadirachtin A by weight.

[0149] Embodiment 14: The method according to any one of Embodiments 12 or 13, wherein the one or more oils include one or more of the following: soybean oil, grapeseed oil, canola oil, sunflower oil, safflower oil, rapeseed oil, hydrogenated rapeseed oil, hydro-pressed linseed oil, camelina oil, cold-pressed linseed oil, coconut oil, fish oil, poultry oil or fish oil stearin or their fractions.

[0150] Embodiment 15: The method according to any one of Embodiments 12 to 14, wherein the extrusion process comprises extrusion cooking of a raw material comprising protein and starch.

[0151] Embodiment 16: The method according to any one of embodiments 12 to 15, wherein the method of producing the base feed by an extrusion process and subsequently applying the pest control composition by a vacuum infusion process produces a fish feed with a coefficient of variation of less than 10%.

[0152] Embodiment 17: A fish feed product according to the method of any one of Embodiments 12 to 16.

[0153] Embodiment 18: A fish feed for reducing, preventing or controlling parasitic infections or infestations in fish, wherein the fish feed comprises:

[0154] a basal feed comprising a porous matrix distributed throughout; and

[0155] a pest control agent composition comprising a pest control agent and one or more oils;

[0156] wherein the pest control agent composition is distributed throughout the porous matrix of the basal feed.

[0157] Embodiment 19: The fish feed according to Embodiment 18, wherein the pest control agent comprises a neem extract enriched in azadirachtin A.

[0158] Embodiment 20: The fish feed according to embodiment 19, wherein the neem extract enriched with azadirachtin A comprises at least 15% to 33% azadirachtin A by weight.

Claims

1. A fish feed, comprising: a basal feed comprising a porous matrix distributed throughout; and a pest control agent composition comprising a pest control agent and one or more oils; wherein the pest control agent composition is distributed throughout the porous matrix of the basal feed.

2. The fish feed according to claim 1, wherein the pest control agent comprises a neem extract enriched in azadirachtin A.

3. The fish feed as claimed in claim 2, wherein the neem extract enriched with azadirachtin A comprises at least 15 wt% to 33 wt% of azadirachtin A.

4. The fish feed according to any one of claims 1 to 3, wherein the one or more oils comprise one or more of the following: soybean oil, grape seed oil, canola oil, sunflower seed oil, safflower oil, rapeseed oil, hydrogenated rapeseed oil, hydro-pressed linseed oil, camelina oil, cold-pressed linseed oil, coconut oil, fish oil, poultry oil or fish oil stearin or fractions thereof.

5. The fish feed according to any one of claims 1 to 4, wherein the porous matrix is ​​formed by extruding a raw material comprising protein and starch.

6. The fish feed according to any one of claims 1 to 5, wherein the pest control agent composition is applied to the base feed via a vacuum infusion process.

7. The fish feed according to any one of claims 1 to 6, wherein the pest control agent is at least partially distributed throughout the porous matrix of the base feed.

8. The fish feed according to any one of claims 1 to 7, wherein the pest control agent composition is further coated on the outer surface of the base feed.

9. The fish feed of any one of claims 1 to 8, wherein the fish feed comprises between about 15% and about 65% by weight protein and between about 10% and about 45% by weight fat; and The fish feed pellets include terrestrial animal protein, fish meal, plant-based protein, or a combination thereof.

10. A method for preparing fish feed, the method include: providing a protein and starch mixture and subjecting it to an extrusion process to produce a base feed having a porous matrix distributed throughout; producing a pest control agent composition by mixing the pest control agent and one or more oils together to produce a suspension; The pest control composition is applied to the basal feed using a vacuum infusion process to distribute the pest control composition throughout the porous matrix of the basal feed.

11. The method of claim 10, wherein the pest control agent comprises azadirachtin A-rich neem extract comprising at least 15 to 33 wt% azadirachtin A.

12. The method of any one of claims 10 or 11, wherein the one or more oils comprise one or more of soybean oil, grape seed oil, canola oil, sunflower seed oil, safflower oil, rapeseed oil, hydrogenated rapeseed oil, hydro-pressed linseed oil, camelina oil, cold-pressed linseed oil, coconut oil, fish oil, poultry oil or fish oil stearin or fractions thereof.

13. The method according to any one of claims 10 to 12, wherein the extrusion process comprises extrusion cooking of a raw material comprising protein and starch.

14. A method according to any one of claims 10 to 13, wherein the process of producing a base feed by an extrusion process followed by application of the pest control agent composition by a vacuum infusion process produces a fish feed having a coefficient of variation of less than 10%.

15. A fish feed product according to the method of any one of claims 10 to 14.

16. A fish feed, comprising: a basal feed comprising a porous matrix distributed throughout; and a pest control agent composition comprising a pest control agent and one or more oils; wherein the pest control agent composition is distributed throughout the porous matrix of the basal feed.

17. The fish feed according to claim 16, wherein the pest control agent comprises a neem extract enriched in azadirachtin A.

18. The fish feed as claimed in claim 17, wherein the neem extract enriched with azadirachtin A comprises at least 15 wt% to 33 wt% of azadirachtin A.

19. The fish feed of claim 16, wherein the one or more oils comprise one or more of soybean oil, grape seed oil, canola oil, sunflower oil, safflower oil, rapeseed oil, hydrogenated rapeseed oil, hydro-pressed linseed oil, camelina oil, cold-pressed linseed oil, coconut oil, fish oil, poultry oil or fish oil stearin or fractions thereof.

20. The fish feed of claim 16, wherein the porous matrix is ​​formed by extruding a raw material comprising protein and starch.

21. The fish feed of claim 16, wherein the pest control agent composition is applied to the base feed via a vacuum infusion process.

22. The fish food of claim 16, wherein the pest control agent is at least partially distributed throughout the porous matrix of the base food.

23. The fish feed according to claim 16, wherein the pest control agent composition is further coated on an outer surface of the base feed.

24. The fish feed of claim 16, wherein the fish feed comprises between about 15% and about 65% protein by weight and between about 10% and about 45% fat by weight; and The fish feed pellets include terrestrial animal protein, fish meal, plant-based protein, or a combination thereof.

25. The fish feed of claim 16, wherein the base feed comprises one or more of pellets, extruded blocks, steam pellets, flakes, tablets or powder.

26. The fish feed of claim 16, wherein the pest control agent comprises a solid form suspended in one or more oils.

27. A method for preparing fish feed, the method include: providing a protein and starch mixture and subjecting it to an extrusion process to produce a base feed having a porous matrix distributed throughout; producing a pest control agent composition by mixing the pest control agent and one or more oils together to produce a suspension; The pest control composition is applied to the basal feed using a vacuum infusion process to distribute the pest control composition throughout the porous matrix of the basal feed.

28. The method of claim 27, wherein the pest control agent comprises azadirachtin A-rich neem extract comprising at least 15 wt% to 33 wt% azadirachtin A.

29. The method of claim 27, wherein the one or more oils comprise one or more of soybean oil, grape seed oil, canola oil, sunflower oil, safflower oil, rapeseed oil, hydrogenated rapeseed oil, hydro-pressed linseed oil, camelina oil, cold-pressed linseed oil, coconut oil, fish oil, poultry oil or fish oil stearin or fractions thereof.

30. The method of claim 27, wherein the extrusion process comprises extrusion cooking of a raw material comprising protein and starch.

31. The method of claim 27, wherein the method of producing a base feed by an extrusion process followed by application of the pest control agent composition by a vacuum infusion process produces a fish feed having a coefficient of variation of less than 10%.

32. A fish feed product according to the method of any one of claims 16 to 31.

33. A fish feed for reducing, preventing or controlling parasitic infection or infestation in fish, wherein the fish feed comprises: a basal feed comprising a porous matrix distributed throughout; and a pest control agent composition comprising a pest control agent and one or more oils; wherein the pest control agent composition is distributed throughout the porous matrix of the basal feed.

34. The fish feed of claim 33, wherein the pest control agent comprises a neem extract enriched in azadirachtin A.

35. The fish feed as claimed in claim 33, wherein the neem extract enriched with azadirachtin A comprises at least 15 wt% to 33 wt% azadirachtin A.

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