Methods comprising iron compositions

Abstract

The present technology relates to the field of iron compositions, feed additives an animal nutrition and growth. In particular, the present technology relates to non-therapeutic method for enhancing the growth performance of an animal, said method comprising the step of administering an iron composition comprising a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of starch; and c) from 0.05 to 5 wt.% of lecithin. Animal feed and methods for preparing animal feed comprising the iron composition disclosed in the present invention are contemplated in the context of the present technology.

Description

4853-01-1-METHODS COMPRISING IRON COMPOSITIONSFIELD OF THE INVENTION

[0001] The present technology relates to the field of iron compositions, feed additives and animal nutrition and growth.BACKGROUND OF THE INVENTION

[0002] In the field of animal nutrition, the growth performance of animals is a crucial factor that determines the efficiency of animal farming. Feed efficiency refers to the amount of feed intake required to produce a specific amount of weight gain. Improved feed efficiency means that animals convert feed into body mass more effectively, thereby reducing feed costs, which represents one of the largest expenses in animal production. Additionally, better feed efficiency is often associated with enhanced overall animal health and growth performance, potentially reducing the need for medical interventions and improving animal welfare. Consequently, it is common practice to supplement animal feed with additives to enhance feed efficiency.

[0003] Iron compounds are commonly used as feed additives across various animal species. Iron is critical to produce hemoglobin, which is essential for oxygen transport in the body. By improving oxygen delivery to tissues, iron supports healthy growth and development in animals, particularly during their early stages. It contributes to the formation of strong muscles, bones, and overall body structure.

[0004] Examples of commonly used iron compounds as feed additives include ferrous carbonate, ferric chloride hexahydrate, ferrous fumarate, ferrous sulfate heptahydrate, ferrous sulfate monohydrate, ferrous chelate of amino acids hydrate, and ferrous chelate of glycine hydrate. Among these, ferrous sulfate is a widely used feed additive that plays a crucial role in animal nutrition. As a rich source of iron, it effectively supplements iron in animal diets, helping to prevent and treat iron deficiency as iron deficiency can lead to anemia, weakness, and reduced productivity.

[0005] However, there are some negative considerations when using ferrous sulfate. One major challenge is the poor absorption and utilization of iron by animals. The potential toxicity of iron can lead to gastrointestinal issues, including stomach irritation and alterations in gastrointestinal transit. Furthermore, ferrous sulfate can negatively affect the palatability of feed, potentially leading to reduced feed intake.

[0006] Despite the benefits of iron supplementation in animal feeds, there remains a need for feed additives that not only can be used for iron supplementation but can also4853-01-2- improve the growth performance of animals while minimizing potential negative side effects.SUMMARY OF THE INVENTION

[0007] The present technology relies on the surprising realization that a specific iron composition can enhance the growth performance of an animal. In particular, it has been found that by carefully selecting the components of an iron composition as disclosed in the present invention it is possible to improve the absorption and utilization of nutrients by animals, and thus feed efficiency can be enhanced compared to a normal diet or diets supplemented with other iron supplements. The present technology is cost effective and allows a widespread use as it represents an effective supplementation without the adverse effect of common iron compositions.

[0008] Thus, in one aspect the present invention relates to a non-therapeutic method of enhancing the growth performance of an animal, said method comprising the step of administering an iron composition comprising: a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of starch; and c) from 0.05 to 5 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%.

[0009] In another aspect, the invention relates to the use of an iron composition for enhancing the growth performance of an animal, said iron composition comprising: a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of starch; and c) from 0.05 to 5 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%.

[0010] In another aspect, the invention relates to a method of preparing an animal feed comprising the step of mixing an animal feed with an iron composition, said iron composition comprising: a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of starch; and c) from 0.05 to 5 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%.

[0011] In another aspect, the invention relates to an animal feed comprising an iron composition, said iron composition comprising: a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of starch; and4853-01-3- c) from 0.05 to 5 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%.

[0012] In the foregoing aspects, the iron composition can particularly be in the form of microcapsules.DETAILED DESCRIPTION OF THE INVENTION

[0013] Various preferred features and embodiments will be described below by way of non-limiting illustration.

[0014] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article unless the context clearly indicates otherwise. By way of example, "an element" means one element or more than one element.

[0015] Except in the Examples, or where otherwise explicitly indicated, all numerical quantities in this description specifying amounts of materials, reaction conditions, molecular weights, number of carbon atoms, and the like, are to be understood as modified by the word "about". The term “about” as used herein, e.g. when referring to a measurable value (such as an amount or weight of a particular component or temperature), refers to variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or, particularly, ±0.1% of the specified amount. Except where otherwise indicated, all numerical quantities in the description specifying amounts or ratios of materials are on a weight basis.

[0016] While overlapping weight ranges for the various components and ingredients that can be contained in the disclosed compositions have been expressed for selected embodiments of the disclosed technology, the amount of each component in the disclosed compositions is selected from its disclosed range such that the sum of all components or ingredients in the composition will total 100 weight percent. The amounts employed will vary with the purpose and character of the desired product and can be readily determined by one skilled in the art.

[0017] As used herein, the term “comprising”, which is inclusive or open-ended and does not exclude additional unrecited elements or method steps, is intended to encompass as alternative embodiments, the phrases “consisting essentially of” and “consisting of’ where “consisting of” excludes any element or step not specified and “consisting essentially of” permits the inclusion of additional unrecited elements or steps that do not materially affect the essential or basic and novel characteristics of the composition or method under consideration. The compositions of the disclosed4853-01-4- technology may suitably comprise, consist essentially of, or consist of, the components, elements, and / or ingredients described herein.

[0018] The present method of enhancing the growth performance can be applied to various animal species and growth phases, making it a versatile solution for enhancing the growth performance of animals.

[0019] The term "animal" refers to non-human animals and includes all farm animals, domestic animals and laboratory animals. Examples of farm animals include non-ruminants and ruminants. Ruminants include but are not limited to sheep, goat, cattle and the like; and non-ruminants include but are not limited to horse; rabbit; pig including but not limited to infant pig, piglet, growing-fattening pig, sow and boar; and poultry such as turkey, duck and chicken (including but not limited to broiler chicken, egg-laying chicken) etc. Farm animals also include animals farmed in aquaculture (e.g. salmon, trout, tilapia, catfish, carp, etc). Examples of domestic animals include, but are not limited to, dogs, cats, birds, rabbits, rodents, birds and other domestic pets. Examples of laboratory animals include, but are not limited to, monkey, rats, mice, rabbits, guinea pigs and the like.

[0020] In particular, the animal can be selected from the group consisting of sheep, goat, cattle, horse, rabbit, pig, poultry, laboratory animals, domestica animals or fish. The pig can include an infant pig, piglet, rowing-fattening pig a sow or boar. The poultry can include turkey, duck or chicken. Laboratory animals can include rats, mice, hamster, monkeys, guinea pigs, or fish (e.g. zebrafish). Domestic animals can include dogs, cats, rabbits, rodents or birds.

[0021] The term “animal feed” refers to any compound, preparation, or mixture suitable for, or intended for intake by an animal. Animal feed for a mono-gastric animal typically comprises concentrates as well as vitamins, minerals, enzymes, direct fed microbial, amino acids and / or other feed ingredients (such as in a premix) whereas animal feed for ruminants generally comprises forage (including roughage and silage) and may further comprise concentrates as well as vitamins, minerals, enzymes direct fed microbial, amino acid and / or other feed ingredients (such as in a premix).

[0022] “Animal in need thereof” as used herein includes animals that would benefit from administration of the compositions of the present disclosure.

[0023] The term “body weight gain” as used herein means an increase in live weight of an animal during a given period of time.

[0024] By means of the present invention it is possible to enhance the growth performance of an animal, so that, for example, a given growth rate can be maintained4853-01-5- in an animal on a reduced food intake or an increased growth rate obtained for a given food intake.

[0025] Thus, in one aspect the invention relates to a non-therapeutic method of enhancing the growth performance of an animal, said method comprising the step of administering the iron composition as disclosed herein.

[0026] The invention also includes the use of an iron composition as disclosed herein for enhancing the growth performance of an animal.

[0027] Enhancing the growth performance of an animal can comprise increasing weight gain and / or increasing feed efficiency.

[0028] The term “feed efficiency” as used herein refers the amount of weight gain per unit of feed when the animal is fed ad-libitum or a specified amount of food during a period of time. By “increased feed efficiency” it is meant that the administration of the iron composition or its use of a feed additive results in an increased weight gain per unit of feed intake compared with an animal not receiving the iron composition. In particular, the feed efficiency (%) can be calculated as the quotient between final body weight gain in grams and the total kilocalories consumed during the study and is expressed as a percentage of total caloric intake.

[0029] In particular, the feed efficiency can be increased by the administration of the iron composition disclosed herein by at least 5 %, more particularly at least 10 % c, still more particularly by at least 12 %. More particularly, the feed efficiency can be increased by at least 5 %, more particularly at least 10 %, still more particularly by at least 12 %, over a period of at least 3 weeks. More particularly, the feed efficiency can be increased by at least 5 %, more particularly at least 10 %, still more particularly by at least 12 %, over a period of at least 8 weeks.

[0030] In some embodiments, the increase of body weight gain in animals supplemented and / or feed with the iron composition disclosed herein can increase can by at least 5 %, particularly by at least 10%, more particularly by at least 20% compared to animals not supplemented (i.e. feed with normal diet). More particularly, the increase of body weight can be of 5%, or at least 10%, or at least 20% after a period of at least 3 weeks. Even more particularly, the increase of body weight can be of at least 5%, or at least 10%, or at least 20% after a period of at least 8 weeks.

[0031] The iron composition in the present invention comprises: a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of starch; and c) from 0.05 to 5 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%.4853-01-6-

[0032] The combination of the components and concentration of the foregoing iron composition can synergistically improve growth performance on an animal.

[0033] In particular, the iron pyrophosphate can be in a concentration of from 25 to 70 wt.%, more particularly from 30 to 70 wt.%. The iron pyrophosphate can be in a concentration of from 25 to 59 wt.%, particularly from 30 to 49 wt.%.

[0034] The starch can be native starch or modified starch or mixtures thereof.

[0035] "Native starch" refers to starch that is derived directly from plant sources without undergoing any chemical or physical modification.

[0036] The term “modified starch”, as used herein, refers to a starch that has been chemically or physically altered to change its properties. Common methods of modifying starch include physical treatments such as heat, pressure, and mechanical shear, as well as chemical treatments such as acid hydrolysis, cross-linking, oxidation, esterification and enzymatic modification. The modified starch can be selected from the group consisting of thermally treated starch, mechanically treated starch, oxidatively degraded, starch, hydrolytically degraded starch, enzymatically degraded starch, and mixtures thereof.

[0037] In some embodiments, the modified starch can be selected from the group consisting of thermally treated, starch, mechanically treated starch, oxidatively degraded, starch, hydrolytically degraded starch, enzymatically degraded starch and mixtures thereof.

[0038] The starch can be obtained from different natural sources such as, for example, potato starch, corn starch, wheat starch, rice starch, tapioca starch, maize, pea starch, sorghum starch, and mixtures thereof. In particular, the starch can be a maize starch.

[0039] In particular, the starch can be in a concentration of from 50 to 70 wt.%.

[0040] The lecithin can be obtained from different sources. Non-limiting examples of sources of lecithin include soybean, eggs, sunflower seeds, rapeseed, milk, wheat germ, peanuts and corn. In particular, the lecithin can be sunflower lecithin.

[0041] In particular, the lecithin can comprise lecithin comprises sunflower lecithin, soy lecithin and / or egg lecithin.

[0042] In particular, the lecithin can be in a concentration of from 0.1 to 2 wt.%. More particularly, the concentration of lecithin can be from 0.1 to 1 wt.%.

[0043] Some exemplary iron compositions of the invention include:I. An iron composition comprising: a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of modified starch; and4853-01-7- c) from 0.1 to 2 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%II. An iron composition comprising: a) from 30 to 70 wt.% of ferric pyrophosphate; b) from 50 to 70 wt.% of modified starch; and c) from 0.1 to 1 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%

[0044] Advantageously, the composition can be in the form of microcapsules. Microencapsulation with the carefully selected components at the appropriate concentration ensures optimal results helping to control the release of iron in the animal’s digestive system, enhance absorption and utilization of nutrients, and reduce side effects in animal’s health. Another benefit is that the interaction of iron with other nutrients present in the animal feed is avoided and thus stability of other nutrients in the animal feed is not compromised. Thus, for example, degradation of other nutrients that are susceptible to oxidation can be avoided.

[0045] Thus, in one embodiment the intention relates to a method of enhancing growth performance of an animal, wherein iron absorption is enhanced; and / or wherein the adverse effects of iron supplementation are prevented. In particular, the adverse effects of iron supplementation can comprise gastrointestinal issues, more particularly nausea, vomiting, diarrhoea, constipation, and / or abdominal pain are common side effects

[0046] The iron composition can be administered to animals specifically requiring iron supplementation, such as for example animals having iron deficiency, but it is also particularly useful in healthy animals. The term “healthy animal” generally refers to an animal that is free from a disease or condition. Healthy animals show normal physiological function and appropriate growth and development under normal conditions. In the context of the present invention, healthy animal also refers particularly to an animal that does not show nutritional deficiencies, more particularly iron deficiency. In one embodiment, the method of enhancing growth performance of an animal comprises the administration of the iron composition as disclosed herein to a healthy animal.

[0047] The term “microcapsules” as used herein is understood in its broader meaning and refers to minute container that is commonly spherical and includes, without limitation, microspheres, core / shell microcapsules and polynuclear microcapsules.

[0048] In particular, the microcapsule can comprise an internal phase (i.e. core) comprising ferric pyrophosphate and an outer layer (i.e. a shell) comprising modified starch and lecithin.4853-01-8-

[0049] The microcapsules can be obtained by different methods well known in the art of microencapsulation. Useful examples in the context of the present invention include, but are not limited to, spray drying, fluid bed coating, extrusion (e.g. coextrusion), freeze drying, prilling, powder catch, agglomeration, coacervation, liposomes and lipidic particles, electrospraying, solvent evaporation, in situ polymerization, complexation, phase separation, layer-by-layer assembly, nucleate, spray congealing (or spray chilling) and the like. Exemplary microencapsulation methods are also disclosed in “Microencapsulation: Methods and Industrial Applications” (Drugs and The Pharmaceutical Sciences, Volume 158, Edited by Simon Benita, 2006) and Jyothi et al. “Microencapsulation techniques, factors influencing encapsulation efficiency” (J Microencapsul. 2010 May;27(3):187-97), incorporated herein by reference.

[0050] Advantageously, the iron composition in the form of microcapsules can be obtained by spray-drying. Exemplary spray-drying methods are disclosed for example in Masters, K. (1991). "Spray Drying Handbook"; Filkova, I., Huang, L. X., & Mujumdar, A. S. (1992). "Industrial Spray Drying Systems" in "Handbook of Industrial Drying" edited by Mujumdar, A. S.; and Chen, X. D., & Mujumdar, A. S. (2008). "Drying Technologies in Food Processing" incorporated herein by reference.

[0051] The iron composition can be incorporated into an animal feed or can be administrated in liquid formulation.

[0052] A "liquid formulation" in the context of the present invention refers to a mixture or solution that contains the iron composition as disclosed herein. Liquid formulations are typically designed to be easily mixed with other feed components or administered directly to animals. The liquid formulation can also include other ingredients such as nutrients, vitamins, minerals, and other additives in a liquid form.

[0053] The invention also relates to a method for preparing an animal feed comprising the step of mixing an animal feed with the iron composition as disclosed herein.

[0054] Included in the present invention is also an animal feed composition comprising the iron composition as disclosed herein.

[0055] Selection of the most appropriate dosages and administration regimes of the iron composition as disclosed herein are within the scope of ordinary person skilled in the art of animal feed and can depend, for example, on the administration route, the desired growth increase, the animal species and its nutritional requirements.

[0056] The iron composition may be administered in daily dosages from 1 to 200 mg, or from 5 to 100 mg, or from 10 to 80 mg of iron composition per kg of animal and day.4853-01-9-

[0057] The animal feed can be diets prepared as mash feed (non-pelleted) or pelleted feed. Typically, the milled animal feed is mixed and sufficient amounts of essential vitamins and minerals are added according to the specifications for the species in question. The iron composition can be added as solid or as liquid formulation (e.g. suspensions). For example, for mash feed a solid or liquid formulation comprising the iron composition may be added before or during the ingredient mixing step. For pelleted feed, the iron composition may also be added before or during the feed ingredient step. Typically, iron composition can be incorporated in a feed additive or premix.

[0058] The pelleted animal feed may be prepared by pelleting the animal feed as described in the paragraph above. Thus, in one embodiment, the invention relates to pelleted animal feed comprising the iron composition described herein.

[0059] The animal feed can also contain plant-based material. Non-limiting examples include legumes, cereals, oats, rye, barley, wheat, maize, corn, sorghum, switchgrass, millet, pearl millet, foxtail millet, soybean, wild soybean, beans, lupin, tepary bean, scarlet runner bean, slimjim bean, lima bean, French bean, Broad bean (fava bean), chickpea, lentil, peanut, Spanish peanut, canola, rapeseed (oilseed rape), rice, beet, cabbage, sugar beet, spinach, quinoa, or pea, in a processed form thereof (such as soybean meal, rapeseed meal) or any combination thereof.

[0060] The animal feed may comprise vegetable proteins. Vegetable proteins may be derived from vegetable protein sources, such as legumes and cereals, for example, materials from plants of the families Fabaceae (Leguminosae), Cruciferaceae, Chenopodiaceae, and Poaceae, such as soy bean meal, lupin meal, rapeseed meal, and combinations thereof. In an embodiment, the vegetable protein source is material from one or more plants of the family Fabaceae, e.g., soybean, lupine, pea, or bean. In another embodiment, the vegetable protein source is material from one or more plants of the family Chenopodiaceae, e.g., beet, sugar beet, spinach or quinoa. Other examples of vegetable protein sources are rapeseed, and cabbage. In another embodiment, soybean is a preferred vegetable protein source. Other examples of vegetable protein sources are cereals such as barley, wheat, rye, oat, maize (corn), rice, and sorghum.

[0061] The animal feed can also contain one or more enzymes. Typically, the aim of the enzymes is to improve feed digestion. Examples of enzymes include, but are not limited to acetylxylan esterase, acylglycerol lipase, alpha-amylase, beta-amylase, arabinofuranosidase, cellobiohydrolases, cellulase, feruloyl esterase, galactanase, alpha-galactosidase, beta-galactosidase, beta-glucanase, beta-glucosidase, triacylglycerol lipase, lysophospholipase, lysozyme, alpha-mannosidase, beta- mannosidase (mannanase), phytase, phospholipase A1 , phospholipase A2,4853-01-10- phospholipase D, protease, pullulanase, pectinesterase, xylanase, beta-xylosidase, or any combination thereof.

[0062] The feed composition may comprise one or more eubiotics. An "eubiotic" refers to a substance or condition that promotes a healthy balance of the microbiota in the gastrointestinal tract. Eubiotics cover a number of different feed additives, such as probiotics, prebiotics, phytogenies (essential oils) and organic acids which are described in more detail below.

[0063] Non-limiting examples of probiotics include bacterium from one or more of the following genera: Lactobacillus, Lactococcus, Streptococcus, Bacillus, Pediococcus, Enterococcus, Leuconostoc, Carnobacterium, Propionibacterium, Bifidobacterium, Clostridium and Megasphaera or any combination thereof. Other examples include, but are not limited to Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus cereus, Bacillus pumilus, Bacillus polymyxa, Bacillus megaterium, Bacillus coagulans, Bacillus circulans, Enterococcus faecium, Enterococcus spp, and Pediococcus spp, Lactobacillus spp, Bifidobacterium spp, Lactobacillus cidophilus, Pediococsus acidilactici, Lactococcus lactis, Bifidobacterium bifidum, Propionibacterium thoenii, Lactobacillus farciminus, Lactobacillus rhamnosus, Clostridium butyricum, Bifidobacterium animalis ssp. animalis, Lactobacillus reuteri, Lactobacillus salivarius ssp. salivarius, Megasphaera elsdenii, Propionibacteria sp.

[0064] Prebiotics are substances that induce the growth or activity of microorganisms (e.g., bacteria and fungi) that contribute to the well-being of their host. Prebiotics are typically non-digestible fiber compounds that pass undigested through the upper part of the gastrointestinal tract and stimulate the growth or activity of beneficial bacteria that colonize the large bowel by acting as substrate for them. Normally, prebiotics increase the number or activity of bifidobacteria and lactic acid bacteria in the Gl tract. Yeast derivatives (inactivated whole yeasts or yeast cell walls) can also be considered as prebiotics. They often comprise mannan-oligosaccharides, yeast betaglucans or protein contents and are normally derived from the cell wall of the yeast, Saccharomyces cerevisiae.

[0065] The animal feed may also include phytogenies, which are a group of natural growth promoters or non-antibiotic growth promoters used as feed additives, derived from herbs, spices or other plants. Phytogenies can be single substances prepared from essential oils / extracts, essential oils / extracts, single plants and mixture of plants (herbal products) or mixture of essential oils / extracts / plants (specialized products). Examples of phytogenies are rosemary, sage, oregano, thyme, clove, and lemongrass. Examples of essential oils are thymol, eugenol, meta-cresol, vaniline, salicylate, resorcine, guajacol,4853-01-11- gingerol, lavender oil, ionones, irone, eucalyptol, menthol, peppermint oil, alpha-pinene; limonene, anethol, linalool, methyl dihydrojasmonate, carvacrol, propionic acid / propionate, acetic acid / acetate, butyric acid / butyrate, rosemary oil, clove oil, geraniol, terpineol, citronellol, amyl and / or benzyl salicylate, cinnamaldehyde, plant polyphenol (tannin), turmeric and curcuma extract.

[0066] The animal feed may also include organic acids (C1-C7). Organic acids are widely distributed in nature as normal constituents of plants or animal tissues. They are also formed through microbial fermentation of carbohydrates mainly in the large intestine. They are often used in swine and poultry production as a replacement of antibiotic growth promoters since they have a preventive effect on the intestinal problems like necrotic enteritis in chickens and Escherichia coli infection in young pigs. Organic acids can be sold as mono component or mixtures of typically 2 or 3 different organic acids. Examples of organic acids are short chain fatty acids (e.g., formic acid, acetic acid, propionic acid, butyric acid), medium chain fatty acids (e.g., caproic acid, caprylic acid, capric acid, lauric acid), di / tri-carboxylic acids (e.g., fumaric acid), hydroxy acids (e.g., lactic acid), aromatic acids (e.g., benzoic acid), citric acid, sorbic acid, malic acid, and tartaric acid or their salt (typically sodium or potassium salt such as potassium diformate or sodium butyrate).

[0067] The composition of the invention may further comprise one or more amino acids. Examples of amino acids which are used in animal feed are lysine, alanine, betaalanine, threonine, methionine and tryptophan. In one embodiment, the amount of amino acid is 0.001% to 10% by weight of the composition.

[0068] The animal feed may include one or more vitamins, such as one or more fatsoluble vitamins and / or one or more water-soluble vitamins. Usually fat- and water- soluble vitamins form part of a so-called premix intended for addition to the feed. Nonlimiting examples of fat-soluble vitamins include vitamin A, vitamin D3, vitamin E, and vitamin K, e.g., vitamin K3. Non-limiting examples of water-soluble vitamins include vitamin C, vitamin B12, biotin and choline, vitamin B1 , vitamin B2, vitamin B6, niacin, folic acid and panthothenate, e.g., Ca-D-panthothenate. In particular, the amount of vitamins can be from 0.001% to 10% by weight of the animal feed.

[0069] The animal feed may optionally include one or more minerals, such as one or more trace minerals and / or one or more macro minerals.

[0070] The animal feed may comprise minerals such as trace minerals or macro minerals. Non-limiting examples of trace minerals include boron, cobalt, chloride, chromium, copper, fluoride, iodine, iron, manganese, molybdenum, iodine, selenium and zinc. Non-limiting examples of macro minerals include calcium, magnesium,4853-01-12- phosphorus, potassium and sodium. In one embodiment, the amount of minerals is 0.001% to 10% by weight of the animal feed.

[0071] The animal feed may further comprise coloring agents, stabilizers, growth improving additives and aroma compounds / flavorings, polyunsaturated fatty acids (PLIFAs); reactive oxygen generating species, antioxidants, anti-microbial peptides, antifungal polypeptides and mycotoxin management compounds. Examples of coloring agents are carotenoids such as beta-carotene, astaxanthin, and lutein. Examples of aroma compounds / flavourings are creosol, anethol, deca-, undeca- and / or dodecalactones, ionones, irone, gingerol, piperidine, propylidene phatalide, butylidene phatalide, capsaicin and tannin. Examples of antimicrobial peptides (AMP's) are CAP18, Leucocin A, Tritrpticin, Protegrin-1 , Thanatin, Defensin, Lactoferrin, Lactoferricin, and Ovispirin such as Novispirin (Robert Lehrer, 2000), Plectasins, and Statins. Examples of antifungal polypeptides (AFP's) are the Aspergillus giganteus, and Aspergillus niger peptides, as well as variants and fragments thereof which retain antifungal activity. Examples of polyunsaturated fatty acids are C18, C20 and C22 polyunsaturated fatty acids, such as arachidonic acid, docosohexaenoic acid, eicosapentaenoic acid and gamma-linoleic acid. Examples of reactive oxygen generating species are chemicals such as perborate, persulphate, or percarbonate; and enzymes such as an oxidase, an oxygenase or a synthetase. Antioxidants can be used to limit the number of reactive oxygen species which can be generated such that the level of reactive oxygen species is in balance with antioxidants. Mycotoxins, such as deoxynivalenol, aflatoxin, zearalenone and fumonisin can be found in animal feed and can result in negative animal performance or illness. Compounds which can manage the levels of mycotoxin, such as via deactivation of the mycotoxin or via binding of the mycotoxin, can be added to the feed to ameliorate these negative effects. Examples of mycotoxin management compounds are Vitafix®, Vitafix® Ultra (Nuscience), Mycofix®, Mycofix® Secure, FUMzyme®, Biomin® BBSH, Biomin® MTV (Biomin), Mold-Nil®, Toxy-Nil® and Unike® Plus (Nutriad).

[0072] The present invention will be better understood by reference to the following examples, which serve to illustrate the invention, but not to limit the same.EXAMPLESEXAMPLE 1

[0073] A homogenous dispersion of the ingredients as set forth in Table 1 was prepared in water and the dispersion was then dried using a spray dryer to generate the iron microcapsules.4853-01-13-Table 1EXAMPLE 2

[0074] The concentration of ferric pyrophosphate was increased to 53% and hydroxypropyl methyl cellulose (HPMC) was added as a processing aid (Table 2). The ingredients were added to water to form a homogenous dispersion and then spray dried to generate the iron microcapsules.Table 2EXAMPLE 3

[0075] The efficacy of the composition of EXAMPLE 1 was evaluated in laboratory rats and compared with a common source of iron, namely ferrous sulphate. The study was carried out following the Guide for the Care and Use of Laboratory Animals and in accordance with the European provisions of Directive 86 / 609 / EEC on the protection of animals used for experimental and other scientific purposes.

[0076] Four-week-old, male Wistar rats were used for the study. Rats were individually caged in the animal facility at 22°C with a 12-h light / 12-h dark cycle and were fed ad libitum a standard chow diet. Firstly, after 7 days of acclimation, rats were exposed to an iron depletion period (Period 1- depletion period). For that, all thirty animals were randomly distributed in 2 different groups: a Fe-deficient group (n=20), which received a Fe-deficient diet for 24 days in order to induce iron deficiency; and a control group (n=10), which received a Fe-sufficient diet. At the end of the depletion period, rats with iron4853-01-14- deficiency (determined by hemoglobin (Hb) levels quantification) were assigned to 2 groups of 10 animals. Then, rats were exposed to an iron repletion period (Period 2- Repletion period), in which animals were supplemented with the ferrous sulfate or the composition of EXAMPLE 1 and fed the Fe-deficient diet for 21 days. All iron supplements were administered after a 5-hour fasting period to enhance its absorption.

[0077] Finally, at the end of the repletion period, rats were exposed to iron supplementation and Fe-normal diet for 9 additional weeks (Period 3- follow up period) in order to evaluate the gastrointestinal tolerability profile of each formulation as well as their effect on body weight gain and food efficiency.

[0078] Animals in the control group were fed a Fe-normal diet during all the study and did not receive iron supplements.

[0079] The study design is summarized in Table 3:Table 3

[0080] Hemoglobin levels were determined at the end of the depletion period (Period 1) and at the end of the repletion period (Period 2). Results in Table 4 demonstrate hemoglobin levels were lower in animals feed with iron deficient diets after the depletion period, but all animals recovered from anemia after supplementation with either ferrous sulfate or the composition of EXAMPLE 1 (results were not statistically different between groups).4853-01-15-Table 4

[0081] Body weight and food intake were recorded once each week during the three periods of the study.

[0082] Daily average food intake during the study was of 19.7, 18.9 and 19.5 g per day for Control groups, Supplemented with Ferrous Sulfate and EXAMPLE 1. Thus, while food consumption was slightly lower for ferrous sulfate group did not significantly differ between control and Example 1 groups. These results denote that EXAMPLE 1 was unlikely to affect food acceptance.

[0083] Weight increase for the different study periods is summarized in Table 5.Table 5

[0084] During the repletion period the weight increase in animals in control group and those supplemented with the composition of EXAMPLE 1 was similar.

[0085] Remarkably, those animals supplemented with the composition of EXAMPLE 1 , showed a higher weight increase than the other groups during the whole supplementation period (Period 2 + 3), specially compared to control group (188.5 vs 150.9).

[0086] Surprisingly, the highest increase in body weight in those animals supplemented with the composition of EXAMPLE 1 was observed during the follow up period (Period 3). This is the highest increase was observed once all the animals had already recovered from anemia and considered healthy.4853-01-16-

[0087] Feed efficiency (%) was calculated as the quotient between final body weight gain in grams and the total kilocalories consumed during the study and is expressed as a percentage of total caloric intake.

[0088] Feed efficiency at the end of the study for the different groups is summarized in Table 6.Table 6

[0089] Surprisingly a significant higher feed efficiency was observed for animals supplemented with the composition of EXAMPLE 1 , indicating an increased conversion of energy intake into body weight in this group of animals.EXAMPLE 4

[0090] In vitro dissolution profiles provide important information on the bioequivalence of different formulations and are valuable tools when multiple products are being compared. Dissolution tests can be used to discriminate between different formulations and identify characteristics that may have an effect on the bioavailability of the compound of interest.

[0091] The in vitro dissolution profile of composition of EXAMPLE 1 was compared with other commercially available compositions of iron pyrophosphate as set out in Table 7. Non-encapsulated ferric pyrophosphate (246 mg / g) from Dr. Paul Lohmann, was used as control as is the same source of iron as the microencapsulated samples. Ferrous sulphate (300 mg / ml) from Dr. Paul Lohmann was used as a positive control as it is largely recognised as being the most bioavailable source of elemental iron.4853-01-17-Table 7

[0092] The amount of iron released from each sample was evaluated over a 4 hour period using the Electrolab dissolution system (TDL08L) with a USP Type II Paddle replicating gastric juices with a pH value of 1.0. The test was performed by transferring 900 mL of 0.1 N HCI to six dissolution vessels and allowing the temperature to stabilise at 37°C ± 0.5 °C. A quantity of sample containing 14 mg elemental iron was added to the dissolution vessel and stirred at 50 rpm. Aliquots were taken after 0.5, 2, 4 hours.

[0093] Iron was quantified in each aliquot following a previously stablished method [P. Niedzielski, et al. Determination of Iron Species in Samples of Iron-Fortified Food, Food Anal Methods, 2014, 7, 2023-32], Briefly, aliquots at each time point were first filtered using a micron filter to remove undissolved iron. Test and standard solutions were then prepared by sequential addition of Sodium Acetate, Ascorbic Acid, and either filtered Ferrous Sulphate or dissolution aliquots, followed by 1 ,10-Phenanthroline. The reaction mixture was allowed to stand for 10 minutes to complete complexation, forming an orange-colored complex. The absorbance of the resulting solution was then measured using a Shimadzu LIV spectrophotometer (LIV1800) at a wavelength of 510 nm. The iron content was then calculated using the following formula:Where,Aspi = Absorbance of sampleAstd = Absorbance of standardVSpi = Dilution of sampleVstd = Dilution of standardWavg = Average weight of sampleWstd = Weight of standardWspi = Actual Weight of sampleDissolution profiles are summarized in Table 8.4853-01-18-Table 8Iron Release (%)Time (h) 0.5 2 4CTRL 25.3 ±1.0 55.1 ±2.6 65.8±3.1EX.1 66.1 ±2.0 87.5±0.7 92.6±1.4MSU 24.7±2.3 64.2±5.1 78.6±5.9MUL 30.3±2.0 52.6±3.2 61.1±3.3Results expressed as means ± standard deviation for n=6.

[0094] Example 1 performed the best among the samples in the study with 66.2+2.2% of iron released after 30 minutes. The amount of iron released gradually in Example 1 and increased over 4 hours, resulting in 92.5±0.7% iron released. Both the control (iron pyrophosphate) and MUL performed worst with 25.0±5.9% and 30.8±1.9% released after 0.5 hours, respectively. After 4 hours maximum release was again lowest at 75.0±6.8% for control and 64.8±3.1% for MUL.EXAMPLE 5

[0095] Iron absorption was determined in CACO-2 / Raji cocultured intestinal M-cell model. Same samples as in Table 7 were first subjected to an in vitro digestion to simulate the biological fate of the samples following the INFOGEST standardized methodology previously described [A. Brodkorb, et al. INFOGEST static in vitro simulation of gastrointestinal food digestion, Nature Protocols 2019 14:4, 2019, 14, 991- 1014], Digestion of the samples covered three stages: oral, gastric, and intestinal. In the simulated in vitro digestion, the process is divided into three key steps, which are the oral phase (mouth), gastric phase (stomach), and small intestine (jejunum). In the oral phase, the macrostructure breakdown and bolus formation take place due to the mechanical action and the enzyme a-amylase activity within the food matrix. The gastric phase occurs in an acidic environment with the presence of the pepsin enzyme. And finally, in the small intestine step, the pancreatin enzyme and bile extract finished the digestion of the samples. Finally, after the 2h intestinal phase, the digested sample (DS) was obtained. Part of the DS was centrifuged at 3200 g for 60 minutes at 4°C to obtain the soluble phase (SP), which is the fraction with the iron potentially available for absorption, and the insoluble phase (IP), which is the phase with the iron that potentially passes to the colon. Once obtained, the samples were collected and immediately put in ice to stop the enzymatic reactions. The samples were stored at -80°C until further analysis.4853-01-19-

[0096] The Caco-2 cell line (ATCC® HTB-37 ™, Manassas, VA, USA) was cultured in growth medium containing Eagle Minimum Essential Medium (EMEM) supplemented with 20% fetal bovine serum (FBS, Gibco-Thermo Fisher Scientific) and antibiotics (penicillin 100 U / mL, streptomycin 30 pg / mL, PAN-Biotech, Aidenbach, Germany), following ATCC recommendations. The Raji cell line of lymphoblast-like cells (ATCC® CCL-86™) was cultured in growth medium containing RPMI-1640 supplemented with 10% FBS and antibiotics (penicillin 100 U / mL, streptomycin 30 pg / mL, PAN-Biotech, Aidenbach, Germany), following ATCC recommendations. Both cell lines were incubated in a CO2 incubator at 37 °C in a humidified atmosphere containing 5% CO2. All reagents were from ATCC otherwise stated.

[0097] The Caco-2 / Raji co-culture (M-cell model) of the gastrointestinal epithelium was cultivated by modifying the protocol previously described [T. Ahmad, et al. A comparison of three Peyer’s patch “M-like” cell culture models: particle uptake, bacterial interaction, and epithelial histology, Eur J Pharm Biopharm, 2017, 119, 426-36; A. des Rieux, et al. An improved in vitro model of human intestinal follicle- associated epithelium to study nanoparticle transport by M cells, Eur J Pharm Sci, 2007, 30, 380-91], Briefly, 5 x 105cells / cm2of Caco-2 cells were seeded into the apical compartment of 3.0 pm pore polycarbonate transwell inserts (Corning, Somerville, MA, USA) in a 6-well plate and grown for 15 days at 37 °C, 5% CO2. The medium on both the apical and basolateral compartments was changed every other day. On the 15th day, 4 x 105cells / cm2of Raji B cells were seeded into the basolateral compartment. The co-culture grew for 5 days under standard incubation conditions, and the medium was changed only in the apical compartment every day.

[0098] Samples (digested samples diluted 1 / 50) were added on the apical side of the insert and incubated for 1 h. The integrity of the cell monolayers was determined by measuring transepithelial electrical resistance (TEER) using Millicell ERS-2 (Millipore Corporation, Burlington, MA, USA) before and after sample incubation. Samples collected from the basolateral side of the insert, postincubation, were used to determine the transported Fe by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Transported Fe (%) was determined as follows: pg of Fe in basolateral samplesIron transport (%) = - pg of l F= —e : i -n ap :ica ;l - samp ;l -esx 1004853-01-20-

[0099] Percentage of iron transported is summarized in Table 9.Table 9Values represent mean ± standard deviation (S.D) (n=5)***p < 0.001 , two-tailed Student's t-test vs control.

[0100] These results show a significant increase in iron transportation through the epithelial intestinal barrier using Example 1 compared to the control sample. However, none of the other samples showed this result.

[0101] The invention has been explained in relation to its preferred embodiments, it is to be understood that various modifications thereof will become apparent to those skilled in the art upon reading the specification. Therefore, it is to be understood that the invention disclosed herein is intended to cover such modifications as fall within the scope of the appended claims.

Claims

4853-01-21-CLAIMS:

1. A non-therapeutic method of enhancing the growth performance of an animal, said method comprising the step of administering an iron composition comprising: a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of starch; and c) from 0.05 to 5 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%.

2. The method of claim 1 , wherein enhancing the growth performance of an animal comprises increasing weight gain and / or increasing feed efficiency.

3. The method of claim 1 or claim 2, wherein the ferric pyrophosphate is in a concentration of from 25 to 70 wt.%.

4. The method of any one of the previous claims, wherein the starch is a native starch or a modified starch, preferably a modified starch selected from the group consisting of native starch, thermally treated starch, mechanically treated starch, oxidatively degraded, starch, hydrolytically degraded starch, enzymatically degraded starch, and mixtures thereof.

5. The method of any one of the previous claims, wherein the starch is in a concentration of from 50 to 70 wt.%.

6. The method of any one of the previous claims, wherein the lecithin comprises sunflower lecithin, soy lecithin and / or egg lecithin.

7. The method of any one of the previous claims, wherein the lecithin is in a concentration of from 0.1 to 2 wt.%.

8. The method of any one of the previous claims, wherein the iron composition is in the form of microcapsules.

9. The method of claim 8, wherein the microcapsules comprise an internal phase comprising the iron pyrophosphate and an outer layer comprising the starch and the lecithin.4853-01-22-10. The method of any one of the previous claims, wherein the animal is selected from the group consisting of sheep, goat, cattle, horse, rabbit, pig, poultry and laboratory animals.

11. The method of any one of the previous claims, wherein the iron composition is incorporated into an animal feed.

12. The method of any one of previous claims, wherein the iron is administrated in the form of a liquid formulation.

13. Use of an iron composition for enhancing the growth performance of an animal, said iron composition comprising: d) from 25 to 80 wt.% of ferric pyrophosphate; e) from 40 to 80 wt.% of starch; and f) from 0.05 to 5 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%.

14. A method of preparing an animal feed comprising the step of mixing an animal feed with an iron composition, said iron composition comprising: a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of starch; and c) from 0.05 to 5 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%.

15. Animal feed comprising an iron composition, said iron composition comprising: a) from 25 to 80 wt.% of ferric pyrophosphate; b) from 40 to 80 wt.% of starch; and c) from 0.05 to 5 wt.% of lecithin; wherein the sum of the components does not exceed 100 wt.%.

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