Compositions comprising N-amidino amino acids for drinking water applications

Abstract

The subject matter of the present application is a composition comprising an N-amidino amino acid, a carbon dioxide-generating compound and a gas-releasing compound wherein the equivalence ratio of the gas-releasing compound to the carbon dioxide-generating compound is 1 or more, and the molar ratio of the carbon dioxide-producing compound to the N-amidino amino acid is 1 or more.

Description

Technical Field

[0001] This invention relates to compositions comprising N-amidinyl amino acids ( N The N-guanylamino acid), a carbon dioxide-producing compound and a gas-releasing compound, wherein the equivalent ratio of the gas-releasing compound to the carbon dioxide-producing compound is 1 or greater, and the molar ratio of the carbon dioxide-producing compound to the N-guanylamino acid is 1 or greater, and relates to a diet containing the composition for the treatment and / or prevention of heat stress, transport stress or any other stress-related conditions in poultry. Background Technology

[0002] N-Amino acids are derivatives of amino acids containing a guanidine group, obtained by adding an amino nitrile to the amino acid in question. The most important N-amino amino acid is guanidinoacetic acid (GAA), also known as N-aminoglycine. It is an endogenous substance in animals and humans, playing a central role in the biosynthesis of creatine. Creatine can be obtained through diet and / or may be formed endogenously. Its biosynthesis proceeds from glycine with L-arginine. In mammalian organisms, guanidinoacetic acid is formed primarily in the kidneys by transferring the guanidino group of L-arginine to the amino acid glycine via the enzyme L-Arg:Gly-aminoyltransferase (AGAT). Thus, L-ornithine is produced from L-arginine, which is then metabolized to L-citrulline via methionylation in the urea cycle. In a further step, guanidinoacetic acid is methylated to creatine with S-adenosylmethionine via the enzyme guanidinoacetic acid N-methyltransferase (GAMT). Guanidinoacetic acid was first synthesized in 1861 by Adolph Strecker by the addition of an aminonitrile to glycine in aqueous solution, particularly in a weak ammonia solution (M. Strecker, compt. Rend. 1861, 52, 1212; cited in Ber. Chem. Ges. (now Eur. J. Inorg. Chem.) 1908, 41, 4385). In subsequent publications, guanidinoacetic acid was prepared from an aminonitrile and glycine in isopropanol as a solvent, with sodium hydroxide as the base (CN 102329250 A) or with sodium carbonate as the base (CN101462983 A).

[0003] Alternatively, N-amidinyl amino acids, such as GAA, can also be prepared during fermentation via a transamidination reaction, in which the amidine group of arginine is transferred to various amidine acceptors, such as guanidinoacetate, 3-guanidinopropionic acid, 4-guanidinobutyric acid, 2-guanidinoethanol, hydroxyguanidine, and homoarginine. The fermentation production of guanidinoacetic acid has been extensively studied. For example, WO 2021 / 122400 A1 and WO 2022 / 00828 A1 disclose the fermentation production of GAA using specific developed strains, while Yiwen Zhang et al. disclosed the fermentation production of GAA using a whole-cell catalyst system (Yiwen Zhang, Hang Zhou, Yong Tao, and Baixue Lin, ACS Synth. Biol. 2020, 9, 2066-2075).

[0004] GAA supplementation allows for optimal creatine supply in the organism, which positively influences energy transport in muscle cells. Typically, GAAs can be simply added to the animal's diet. In particular, under certain conditions such as heat stress, transportation stress, and during or after illness, animals often have increased energy requirements but consume less food, while simultaneously having increased demands for water supply (e.g., drinking water). In principle, this increased energy requirement can be met by GAA supplementation. However, compared to creatine, amidoamino acids have the disadvantage of poor water solubility. For example, guanidinoacetic acid has a very poor water solubility of only 1 gram in 278 ml of water at 15°C.

[0005] Several approaches have been taken to address this problem. For example, CN 115137016 A discloses a nutritional preparation for livestock and poultry and a method for preparing said nutritional preparation. This nutritional preparation comprises 40 to 80 parts by weight of guanidinoacetic acid, 8 to 20 parts by weight of anhydrous sodium sulfate, 1 to 15 parts by weight of a sweetener, 3 to 10 parts by weight of citric acid, 10 to 40 parts by weight of glucose, 8 to 20 parts by weight of a beneficial powder, and 1 to 13 parts by weight of ammonium bicarbonate. However, although the preparation is described as water-soluble in CN 115137016, experiments have shown that the preparation in CN 115137016 is insoluble in water without stirring. Therefore, a mechanical stirrer is required to dissolve N-amidinyl amino acids (e.g., guanidinoacetic acid). However, mechanically stirred tanks or troughs are expensive and therefore quite rare on farms. Another disadvantage of stirred tanks or troughs is the higher risk of bacterial contamination.

[0006] Liquid applications of N-amidinyl amino acids (such as guanidinoacetic acid) would be a different way to solve the dissolution problems associated with such compounds. For example, US 2009 / 0297656 A1 discloses a liquid formulation for human and animal nutrition consisting of an aqueous solution, a guanidinoacetic acid component, and at least one methyl donor derived from choline, methionine, and betaine.

[0007] WO 2021 / 008848 A1 discloses a concentrate for preparing a wetting solution comprising an aqueous solution of guanidinoacetic acid. Specifically, the concentrate comprises an aqueous solution containing guanidinoacetic acid and at least one salt derived from calcium chloride and magnesium chloride, wherein the solution, based on the total weight of the solution, comprises: a) 0.5 to 4 wt% guanidinoacetic acid, and b) 10 to 70 wt% calcium chloride and / or magnesium chloride, and c) residual water, wherein components a) and b) are present in dissolved form in the water. The large amounts of magnesium chloride or calcium chloride in this concentrate may be used to improve the solubility of guanidinoacetic acid. This results in a fairly high salt concentration in the tank or vessel, which is undesirable on farms and may cause intoxication in animals. Summary of the Invention

[0008] Therefore, there is still a need for compositions containing N-amidinyl amino acids (e.g., GAA) that allow for improved solubility of N-amidinyl amino acids (e.g., GAA) in water.

[0009] It was discovered that this problem can be solved by improving the dissolution kinetics of N-amidinyl amino acids (e.g., GAA) in water. Specifically, the solution lies in the fact that compositions containing N-amidinyl amino acids (e.g., GAA) also contain compounds that allow carbon dioxide release. It is believed that the sparkling effect caused by this carbon dioxide release results in better homogeneity of the N-amidinyl amino acids (e.g., GAA) in water. This reduces the tendency to form agglomerates, and it breaks down agglomerates that might form during the production of N-amidinyl amino acids, which would otherwise precipitate.

[0010] Therefore, one object of the present invention is to provide a composition comprising an N-amidinyl amino acid (e.g., GAA), a carbon dioxide-generating compound, and a gas-releasing compound, wherein the equivalent ratio of the gas-releasing compound to the carbon dioxide-generating compound is 1 or greater, and the molar ratio of the carbon dioxide-generating compound to the N-amidinyl amino acid is 1 or greater.

[0011] This composition is particularly suitable for drinking water applications.

[0012] In the presence of water, the carbon dioxide-producing compound reacts with the gas-releasing compound to release carbon dioxide. Therefore, the dissolution process of N-amidinyl amino acids is significantly accelerated.

[0013] To provide or generate carbon dioxide, it is preferred that the compound that generates carbon dioxide is a salt of carbonate. Detailed Implementation

[0014] In embodiments of the compositions of the present invention, the compounds that produce carbon dioxide are therefore alkali metal bicarbonates, alkaline earth metal bicarbonates, ammonium bicarbonate, alkali metal carbonates, alkaline earth metal carbonates, or any mixtures of these substances.

[0015] In a preferred embodiment of the composition of the present invention, the compound that produces carbon dioxide is sodium bicarbonate, potassium bicarbonate, ammonium bicarbonate and / or any mixture of these substances.

[0016] In this invention, in the presence of water, the gas-releasing compound reacts with a carbon dioxide-producing compound (preferably a salt of carbonate) to release carbon dioxide. Therefore, in principle, the potential chemical reaction is an acid-base reaction, with hydrated hydrogen ions (H₃O₂) provided by the gas-releasing compound. + ) and carbonate ions (CO3) 2- ) or bicarbonate ions (HCO3) - The reaction forms carbonic acid (H2CO3). This molecule is rapidly converted into water and carbon dioxide in the presence of water under ambient pressure and at room temperature, which causes effervescence or foaming effect and significantly accelerates the dissolution of N-amidinyl amino acids.

[0017] Therefore, it is preferred that the gas-releasing compound in the composition of the present invention is an acid, such as an inorganic acid, an organic acid, or a mixture thereof.

[0018] The use of solid organic acids allows the compositions of the present invention to be provided in solid form. Solid form of the compositions of the present invention allows for easier handling, easier metering, and more efficient transportation. Therefore, it is preferred that the compositions of the present invention be solid compositions.

[0019] In one embodiment of the composition of the present invention, the gas-releasing compound is therefore a solid organic acid.

[0020] In a preferred embodiment of the composition of the present invention, the gas-releasing compound is citric acid, tartaric acid, malic acid and / or any mixture of these substances.

[0021] Therefore, in the context of this invention, the term "equivalent ratio" refers to the ratio of the gas-releasing compound to the carbon dioxide-producing compound to obtain one carbon dioxide molecule. For example, in the case where the gas-releasing compound is citric acid having three carboxylic acid groups and the carbon dioxide-producing compound is sodium bicarbonate (NaHCO3), 3 equivalents of citric acid react with 3 equivalents of sodium bicarbonate to obtain 3 equivalents of carbon dioxide. Here, the equivalent ratio of the gas-releasing compound to the carbon dioxide-producing compound is 1:1. In another example, in the case where the gas-releasing compound is citric acid having three carboxylic acid groups and the carbon dioxide-producing compound is sodium carbonate (Na2CO3), 6 equivalents of citric acid react with 3 equivalents of sodium carbonate to obtain 3 equivalents of carbon dioxide. Here, the equivalent ratio of the gas-releasing compound to the carbon dioxide-producing compound is 2:1.

[0022] Preferably, in the composition of the present invention, the equivalence ratio of the gas-releasing compound to the carbon dioxide-producing compound is in the range of 1 to 3.

[0023] To ensure that carbon dioxide is produced or provided as completely as possible, it is preferred that the equivalence ratio of the gas-releasing compound to the gas-producing compound is at least 1.05:1.

[0024] The present invention allows for the provision of compositions containing N-amidinyl amino acids (e.g., GAA) having a higher amount of N-amidinyl amino acids (e.g., GAA) compared to prior art compositions (e.g., mixtures of WO 2021 / 008848 A1).

[0025] In another embodiment, the composition of the present invention contains more than 4% by weight of N-amidinyl amino acids, such as GAA, based on the total weight of the composition.

[0026] Preferably, based on the total weight of the composition, the composition of the present invention comprises 5 to 25 wt%, 5 to 12 wt%, or 5 to 15 wt% of an N-amidinyl amino acid, such as GAA.

[0027] The N-amidinyl amino acid with the highest relevance, especially in the field of animal nutrition, is guanidinoacetic acid.

[0028] Therefore, in another embodiment of the composition of the present invention, the N-amidinyl amino acid is guanidinoacetic acid.

[0029] The compositions of the present invention may further comprise additional components. Suitable additional components are pyrroloquinoline quinones in a biologically available form.

[0030] Poultry receiving the compositions of the present invention, which additionally contain a bioavailable form of pyrroloquinoline quinone, exhibited improved broiler growth performance at all stages. Surprisingly, this effect was observed under all conditions, i.e., even when no heat stress was applied to the poultry. Pyrroloquinoline quinone (PQQ) is a key redox cofactor in animal and human nutrition (Mitchell et al., Analytical Biochemistry 1999, 269, 317; Noji et al., Journal of Agricultural and Food Chemistry 2007, 55, 7258). The primary function of PQQ is to bind to intracellular proteins and act as an antioxidant, circulating within the cells via glutathione. It is considered a vitamin-like substance (Kasahara et al., Nature 2003, 422, 832; Felton et al., Nature 2005, 433, E10; Rucker et al., Nature 2005, 433, E10-11; Ames et al., PNAS 2018, 115, 10836), and compared to other antioxidants such as ascorbic acid (vitamin C), PQQ exerts a positive effect on mitochondrial biogenesis, which directly affects energy metabolism in animals, while other antioxidants have no effect on mitochondrial function or show a decrease in mitochondrial function (Harris et al., Journal of Nutritional Biochemistry 2013, 24, 2076). Therefore, the use of PQQ aims to improve the aforementioned performance of meat-type poultry by increasing intracellular ATP levels through enhanced mitochondrial biogenesis.

[0031] On the other hand, guanidinoacetic acid (GAA) improves energy transport from mitochondria to the cytoplasm by increasing creatine availability. In the mitochondria, creatine binds to the phosphate group of adenosine triphosphate (ATP), and in the cytoplasm, phosphocreatine releases it back to ADP. It is expected that GAA should also support meat-type poultry suffering from oxidative stress (particularly heat stress) through its antioxidant properties, such as reducing electron leakage and interference in the O2-free radical mechanism. Since PQQ improves mitochondrial health and ATP production, and GAA improves energy transport from ATP, the combination of PQQ and GAA is believed to interact beneficially at the metabolic level, improving intracellular ATP production and distribution, and this effect is even enhanced under heat stress conditions, as higher energy availability and lower oxidative stress allow cells to respond more effectively to the consequences of heat stress.

[0032] In another embodiment, the composition of the present invention further comprises a biologically available form of pyrroloquinoline quinone.

[0033] In the context of this invention, the term "bioactive form" is used as is known to those skilled in the art and refers to a form of compound (here, PQQ) that allows for the influence of biological processes beyond nutritional value in a manner that affects bodily functions.

[0034] PQQ is a tricarboxylic acid with poor solubility in water. Therefore, the present invention uses a bioactive form of PQQ. In principle, this bioactive form of PQQ is not limited in any way. Nevertheless, it is preferred that the bioactive form of pyrroloquinoline quinone is a pyrroloquinoline quinone salt (PQQ salt) because the pyrroloquinoline quinone salt provides improved water solubility for the PQQ, which is beneficial to the bioactive form of PQQ and to faster dissolution in aqueous applications (e.g., in poultry drinking water).

[0035] In embodiments of the compositions of the present invention, the bioactive form of pyrroloquinolinequinone is a pyrroloquinolinequinone salt (PQQ salt). In the context of the present invention, the PQQ salt is not limited in any way and may contain inorganic or organic cations. Throughout the present invention, the term "inorganic cation" is used as is known to those skilled in the art and refers to any type of simple cation containing only positively charged ions, such as monovalent cations, such as ammonium cations NH4+. + Alkali metal cations (e.g., Li) + Na + and K + ), or monovalent copper ions Cu + Divalent cations, such as alkaline earth metal cations (e.g., Mg); 2+ Ca 2+ and Ba2 + ), or ferrous cations Fe 2+ Zinc cation Zn 2+ or divalent copper ions Cu 2+ Or trivalent cations, such as Al 3+ Or trivalent iron cation Fe 3+ In the context of this invention, the term "organic cation" is used as is known to those skilled in the art and refers to any type of compound cation, such as tetramethylammonium cation N[CH3]4. + The PQQ salt may contain PQQ as a monovalent, divalent, or trivalent anion. Depending on the valence of the PQQ anion, the PQQ salt contains a necessary number of cations. For example, a PQQ salt containing divalent PQQ may contain two monovalent cations or one divalent cation. For example, when the PQQ salt contains trivalent PQQ anions, the salt contains two trivalent PQQ anions and three divalent cations. In one embodiment of the composition of the present invention, the PQQ salt contains inorganic or organic cations. Preferably, the PQQ salt contains inorganic cations, such as alkali metal cations or alkaline earth metal cations, because alkali metal or alkaline earth metal salts of PQQ have relatively high solubility in water. In another embodiment of the composition of the present invention, the PQQ salt contains alkali metal cations and / or alkaline earth metal cations. Disodium salt of PQQ is the most commonly used form because it has higher water solubility and is a stable solid at 40°C, with a color ranging from red to brown depending on the hydration level. Disodium PQQ (PQQ) Na2) forms a stable trihydrate (about 12.5% ​​water) and a stable pentahydrate (about 20 to 21% water). If exposed to environmental conditions, the anhydrous material will absorb water to at least the form of the trihydrate. In a preferred embodiment of the composition of the invention, the PQQ salt comprises sodium cations and / or potassium cations.

[0036] It has also been found that only a small amount of the bioactive form of PQQ is required to achieve the beneficial effects of the composition of the present invention on the performance and metabolism of meat-type poultry. In another embodiment, the composition of the present invention further comprises at least 0.05 ppm of the bioactive form of PQQ. Preferably, the composition of the present invention further comprises 0.05 ppm to 20 ppm of the bioactive form of PQQ. The recommended PQQ concentration can then be easily adjusted by dissolving the composition in a corresponding amount of water.

[0037] It is believed that N-amidinyl amino acids, such as guanidinoacetic acid, have a positive effect on the efficacy of prebiotics and / or probiotics. Therefore, it is beneficial to combine N-amidinyl amino acids with prebiotics and / or probiotics in the compositions of the present invention.

[0038] In one embodiment, the composition of the present invention further comprises prebiotics and / or probiotics.

[0039] Preferably, the probiotics include certain species of the genus Bacillus (…). Bacillus spp. strains, especially Bacillus subtilis ( B. subtilis (e.g., DSM 32315 or DSM 32540), Bacillus amyloliquefaciens ( B. starch liquefier (e.g., CECT 5940) and / or any mixture of these strains.

[0040] In another embodiment, the composition of the present invention further comprises probiotics, said probiotics including strains selected from Bacillus subtilis (e.g., DSM 32315 or DSM 32540), Bacillus amyloliquefaciens (e.g., CECT 5940) and / or any mixture thereof.

[0041] Bacillus subtilis DSM 32315 has been identified through screening of naturally occurring isolates. It was deposited in DSMZ on May 12, 2016, under the aforementioned Accession Number, in the name of Evonik Degussa Ltd., in accordance with the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purpose of Patent Procedure. Bacillus subtilis DSM 32315 is from Evonik's GutCare... ® Ingredients. GutCare ® It is a direct-feeding microbial solution based on the spore-forming Bacillus subtilis DSM 32315 strain, which has the inherent ability to produce a variety of secondary metabolites.

[0042] Bacillus subtilis DSM 32540 was identified through targeted screening of naturally occurring isolates. It was deposited in DSMZ on June 14, 2017, under the aforementioned accession number and in the name of Evonik Degussa Ltd, in accordance with the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purpose of Patent Procedure. Bacillus subtilis DSM 32540 is from Evonik's GutPlus... ® Ingredients. GutPlus ® It is a probiotic used for gut microbiota management.

[0043] A strain of *Bacillus amyloliquefaciens*, with accession number CECT 5940, has been deposited in the Spanish Type Culture Collection. It is not genetically modified and does not carry a plasmid. Ecobiol ® It is a feed additive composed of naturally occurring, fast-growing Bacillus amyloliquefaciens CECT 5940. It can improve animal health and production conditions, and help producers address challenges related to quality, profitability, and sustainability.

[0044] Furthermore, the compositions of the present invention may additionally contain 5-aminolevulinic acid, a precursor for heme synthesis. Increased heme in the organism under discussion implies the presence of more oxygen in the animal, resulting in increased agility. It is also conceivable to include derivatives of 5-aminolevulinic acid in the compositions of the present invention. Suitable salts of 5-aminolevulinic acid are, for example, alkali metal or alkaline earth metal salts, ammonium salts of 5-aminolevulinic acid, or acid addition salts, such as 5-aminolevulinic acid salts. Suitable derivatives of 5-aminolevulinic acid are, for example, 5-aminolevulinic acid with a masking or protecting group at a hydroxyl group (e.g., an ester) or an amino group (e.g., a tert-butoxycarbonyl group, also known as a Boc group). Preferably, the one or more masking groups will be cleavable from the 5-aminolevulinic acid under physiological conditions. In the case where the 5-aminolevulinic acid derivative has two masking or protecting groups, it is preferred that they be cleavable simultaneously under the same conditions.

[0045] In another embodiment, the composition of the present invention further comprises 5-aminolevulinic acid, its derivatives, its salts, and / or any mixture thereof.

[0046] The compositions of the present invention are not limited in any way in terms of the quantity and amount of one or more of the above-mentioned additional components, provided that the one or more additional components do not adversely affect one or more of the beneficial effects of the compositions of the present invention.

[0047] The presence of compounds that produce or provide carbon dioxide and compounds that release gas has the effect of releasing carbon dioxide when the compounds of the present invention dissolve in water. Therefore, the compositions of the present invention can also be considered fizzy compositions. The composition is not limited in any way regarding its physical appearance. In its simplest form, it is a powder, and therefore it can already be used as a supplement to animal diets. However, the compositions of the present invention can also be further processed into any conceivable form or physical appearance, for example, into tablets, particularly effervescent or effervescent tablets.

[0048] In another embodiment, the composition of the present invention is an effervescent tablet.

[0049] The compositions of this invention are suitable for treating and / or preventing heat stress, transport stress, or any other stress-related diseases in poultry and / or livestock. For example, administration of GAA to poultry can reduce poultry mortality during transport.

[0050] The diets of the present invention are suitable for the treatment and / or prevention of heat stress, transportation stress or any other stress-related diseases in poultry and / or livestock, wherein the diets comprise the compositions of the present invention and are applied to poultry and / or livestock.

[0051] In the context of this invention, the term "poultry" is used to refer to any kind of domesticated bird that is kept in captivity for its practical use. Examples of poultry are domestic fowls, including chickens (or meat-type and egg-laying birds), turkeys, geese, quails, and ducks, which are raised to produce meat or eggs. Preferably, in the context of this invention, the term "poultry" refers to chickens or meat-type birds.

[0052] In the context of this invention, the term "livestock" is used to refer to domesticated animals raised in an agricultural environment to provide labor and produce a variety of consumer products, such as meat, eggs, milk, fur, leather, and wool. Specifically, the term "livestock" is used to refer to animals raised for consumption, particularly domesticated ruminants such as cattle, sheep, goats, and pigs.

[0053] In the context of this invention, the term "heat stress" is used to determine exposure to elevated ambient temperatures. Heat stress can be chronic or acute. In the context of this invention, the term "chronic heat stress" is used to determine an extended period of elevated ambient temperature. In contrast, in the context of this invention, "acute heat stress" is understood to determine a sudden and short period of extremely high ambient temperature. In principle, the diet of this invention is not limited to chronic or acute heat stress. In the context of this invention, particularly in the context of the term "chronic heat stress," the term "elevated ambient temperature" is used to determine temperatures above the comfort temperature for poultry. As mentioned above, the temperature comfort zone for birds depends on their age. Older birds are more sensitive to high temperatures. Generally, birds are comfortable at temperatures of about 21 to 24°C (70 to 75°F) and function normally at temperatures of up to about 27°C (80°F). However, above 27°C (up to about 30°C), feed consumption decreases while water intake increases. Reduced feed conversion ratio (FCR) and weight gain in broiler birds, and decreased egg production in both layer and breeder flocks. Significant decreases in egg production and eggshell quality were observed at temperatures between 30 and 32°C (86 to 95°F). In broiler birds, both the FCR based on egg mass and the FCR per dozen eggs increased with increasing ambient temperature. When temperatures exceeded approximately 35 to 37°C (96 to 100°F), birds attempted to reduce body heat through vigorous gular fluttering; however, temperatures within this range resulted in a degree of mortality. Preferably, in the context of this invention, temperatures exceeding 27°C, particularly at least 30°C, are considered elevated ambient temperatures. Specifically, temperatures in the range of 30 to 40°C or 34 to 40°C are considered elevated temperatures.

[0054] The diet of the present invention can be applied to poultry exposed to temperatures above 27°C.

[0055] The diet of the present invention can be applied to poultry exposed to temperatures of at least 30°C, for example, at least 34°C.

[0056] Typically, the elevated temperatures exposed to poultry follow a cyclical pattern: the temperature reaches its lowest point at night, then rises steadily during the day until it reaches its highest point, and from that highest point, the temperature again decreases at night until it reaches its lowest point. If this temperature pattern continues for several days or weeks, it is also referred to in the context of this invention as chronic cyclic heat stress.

[0057] The diet of the present invention can be applied to poultry suffering from chronic heat stress.

[0058] The extended temperature period of chronic heat stress is preferably at least 5 hours per day. Preferably, this extended temperature period or the extended period of elevated ambient temperature represents a period of 5 to 24 hours per day, particularly at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 hours per day, or even up to 24 hours.

[0059] The diet of the present invention can be applied to poultry exposed to temperatures exceeding 27°C for at least 5 hours per day.

[0060] In many cases, particularly in southern countries, especially in subtropical, tropical, and generally equatorial countries, rising temperatures are accompanied by higher relative humidity. In the context of this invention, the term "higher relative humidity" is used to mean an average relative humidity of at least 40%. Preferably, in the context of this invention, the range of relative humidity is from 40% to 70% on average, and particularly from 50% to 60% on average.

[0061] The diet of the present invention can be applied to poultry exposed to an average relative humidity of at least 40%.

[0062] Poultry can be exposed to an average relative humidity of at least 45%.

[0063] In principle, the diet of the present invention can be applied to poultry at any stage, multiple stages, or all stages of feeding poultry, that is, at any stage or multiple stages of their lifespan.

[0064] The lifespan of a laying hen can be divided into a pre-layer phase and three production phases or three production periods, namely the initial production phase, the grower juvenile production phase, and the nesting phase, sometimes also referred to as the pre-layer phase and phases I to III.

[0065] The lifespan of poultry raised for meat production can be divided into three stages: starter, grower, and finisher. For example, the entire lifespan of a chicken can be considered as 39 days, with days (d-0) to d-10 called the starter stage, days (d-10) to d-21 called the grower stage, and days (d-21) to d-39 called the finisher stage. Poultry raised for egg production also have different growth and feeding periods. In principle, the diet of the present invention applied to poultry with chronic heat stress is not limited to any specific stage or period of the poultry's lifespan. Therefore, the diet can be applied to poultry with chronic heat stress at any conceivable point in time during or within any of the following stages: the starter, grower, and / or finisher stages. Nevertheless, it is preferred to apply the diet to poultry with chronic heat stress during or within the finisher stage. In the context of this invention, the term "in" is used to refer to a selected point in time, such as an hour, day, or week, which need not be a continuous period of time. In contrast, in the context of this invention, the term "during" is used to refer to a continuous period of time, such as an hour, day, or week.

[0066] The diet of the present invention can be applied to poultry at any, multiple, or all stages of their lifespan.

[0067] The diet of the present invention can be applied to poultry from the beginning of the brooding stage to slaughter, or from the beginning of the growing stage to slaughter.

[0068] Example

[0069] 1. Embodiment of the present invention: Determination of the dissolution time of a foaming mixture containing GAA

[0070] In an intermittent mill (TubeMill, IKA), 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9 g (94 mmol) of sodium bicarbonate (analytical grade, purchased from Merck) and 19.1 g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer) were mixed together for 3 minutes to obtain a foaming composition containing GAA.

[0071] The resulting solid mixture was added to 10 L of tap water provided in the beaker. During settling, the solid mixture almost completely dissolved. The mixture was neither stirred nor moved. After 3 minutes, no solid mixture remained at the bottom of the beaker.

[0072] 2. Examples not of this invention: Determination of the dissolution time of GAA (excluding foaming mixtures)

[0073] A beaker containing 10 L of tap water was provided, and 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China) was added. The powder settled to the bottom of the beaker. The mixture was not stirred or moved. After 4 days, no solid matter was detectable at the bottom of the beaker.

[0074] 3. Examples not of this invention: Solubility tests of four GAA-containing preparations according to CN 115137016 A

[0075] Preparations containing GAA according to CN 115137016 A (see paragraph

[0029] of CN 115137016 A) were prepared. Specifically, four preparations containing GAA according to CN 115137016 A were prepared using GAA as powder (purchased from Gendone, China) and GAA as microparticles (purchased from Gendone, China). Other chemicals used were sodium sulfate (anhydrous, purchased from Merck), citric acid (monohydrate, purchased from Merck), glucose (purchased from ChemPur), and ammonium bicarbonate (purchased from Roth), saccharin sodium salt, sodium cyclamate, and xylitol as sweeteners, and ascorbic acid as a beneficial powder. A total of eight mixtures were prepared. Table 3 summarizes the composition of the four preparations, respectively, for GAA powder and GAA microparticles:

[0076] Table 1: Composition of the eight comparative preparations

[0077] A total of eight preparations were subjected to dissolution tests to determine the appropriate dissolution time. The results are summarized in Table 2.

[0078] Table 2: Overview of Solubility Test

[0079] In the eight stirred solubility tests, the four preparations containing GAA powder provided faster dissolution times compared to the other four preparations containing GAA microparticles. However, all eight comparative preparations provided longer dissolution times compared to the mixtures of the present invention. In the eight unstirred solubility tests, there was no significant difference between the preparations containing GAA powder and the preparations containing GAA microparticles. Specifically, all eight preparations according to CN115137016 A, regardless of whether they contain GAA powder or GAA microparticles, are unsuitable for use in unstirred water tanks.

[0080] 4. Embodiments of the present invention: including GAA and GutCare ® Solubility test of foaming compositions (excluding maltodextrin)

[0081] In a batch mill (TubeMill, IKA), 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9 g (94 mmol) of sodium bicarbonate (analytical grade, purchased from Merck), 19.1 g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer) and 97.8 mg of dried biomass GutCare were added. ® (Excluding maltodextrin, at 0.92×10) 11 CFU / g calculation, from Evonik Espanyol Portugal, SAU, Leon) mixed together for 3 minutes to obtain a mixture containing GAA and GutCare. ® A foaming composition.

[0082] The resulting foaming mixture was added to 10 L of tap water provided in a beaker. After 10 minutes, samples were taken from the surface, middle, and bottom, and the spore distribution in these samples was determined. The result was 5.2 × 10⁻⁶. 5 CFU / mL (surface sample), 3.6 × 10 5 CFU / mL (middle sample) and 3.1×10 5 CFU / mL (bottom sample).

[0083] 5. Embodiments of the present invention: including GAA and GutCare ® Solubility test of foaming compositions (containing maltodextrin)

[0084] In a batch mill (TubeMill, IKA), 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9 g (94 mmol) of sodium bicarbonate (analytical grade, purchased from Merck), 19.1 g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer) and 81.1 mg of dried biomass GutCare were added. ® (Contains maltodextrin, at 1.1×10) 11 CFU / g calculation, from Evonik Espanyol Portugal, SAU, Leon) mixed together for 3 minutes to obtain a mixture containing GAA and GutCare. ® A foaming composition.

[0085] The resulting foaming mixture was added to 10 L of tap water provided in a beaker. After 10 minutes, samples were taken from the surface, middle, and bottom, and the spore distribution in these samples was determined. The result was 6.6 × 10⁻⁶. 5 CFU / mL (surface sample), 1.1 × 10 6 CFU / mL (middle sample) and 7.2×10 5 CFU / mL (bottom sample).

[0086] 6. Embodiments of the present invention: comprising GAA and Ecobiol ® Solubility test of foaming composition

[0087] In an intermittent mill (TubeMill, IKA), 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9 g (94 mmol) of sodium bicarbonate (analytical grade, purchased from Merck), 19.1 g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer) and 22.5 mg of dried biomass Ecobiol were added. ® (according to 4×10) 11 CFU / g calculation, from Evonik Espanyol Portugal, SAU, Leon) mixed together for 3 minutes to obtain a mixture containing GAA and Ecobiol. ® A foaming composition.

[0088] The resulting foaming mixture was added to 10 L of tap water provided in a beaker. After 10 minutes, samples were taken from the surface, middle, and bottom, and the spore distribution in these samples was determined. The result was 2.2 × 10⁻⁶. 6 CFU / mL (surface sample), 1.3 × 10 6CFU / mL (middle sample) and 9.3×10 5 CFU / mL (bottom sample).

[0089] 7. Embodiments of the present invention: Solubility test of foaming compositions containing GAA and 5-aminolevulinic acid.

[0090] In an intermittent mill (TubeMill, IKA), 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9 g (94 mmol) of sodium bicarbonate (analytical grade, purchased from Merck), 19.1 g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer) and 0.16 g (0.96 mmol) of 5-aminolevulinic acid (5-Ala, as hydrochloride, purchased from Haihang Industries, China) were mixed together for 3 minutes to obtain a foaming composition containing GAA and 5-aminolevulinic acid.

[0091] The resulting foaming mixture was added to 10 L of tap water provided in a beaker. After 10 minutes, samples were taken from the surface, middle, and bottom, and the amount of 5-Ala was determined by HPLC. The results are summarized in Table 1:

[0092] Table 3: Results of HPLC determination of 5-Ala.

[0093] 8. Embodiments of the present invention: Solubility test of foaming compositions containing GAA and PQQ

[0094] In an intermittent mill (TubeMill, IKA), 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9 g (94 mmol) of sodium bicarbonate (analytical grade, purchased from Merck), 19.1 g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer) and 1.9 g (0.004 mmol) of disodium phosphate pentahydrate (PentaQQ, Anthem Biosciences Pvt., Ltd.) were mixed together for 3 minutes to obtain a foaming mixture containing GAA and PQQ.

[0095] The resulting foaming mixture was added to 10 L of tap water provided in a beaker. After 10 minutes, samples were taken from the surface, middle, and bottom, and the amount of PQQ was determined by HPLC. The results are summarized in Table 2:

[0096] Table 4: Results of HPLC determination of PQQ.

[0097]

Claims

1. A composition comprising an N-amidinyl amino acid, a carbon dioxide-generating compound, and a gas-releasing compound, wherein the equivalent ratio of the gas-releasing compound to the carbon dioxide-generating compound is 1 or greater, and the molar ratio of the carbon dioxide-generating compound to the N-amidinyl amino acid is 1 or greater.

2. The composition according to claim 1, wherein the carbon dioxide-generating compound is an alkali metal bicarbonate, an alkaline earth metal bicarbonate, ammonium bicarbonate, an alkali metal carbonate, an alkaline earth metal carbonate, or any mixture thereof.

3. The composition according to claim 1 or 2, wherein the carbon dioxide-generating compound is sodium bicarbonate, potassium bicarbonate, ammonium bicarbonate and / or any mixture of these substances.

4. The composition according to any one of claims 1 to 3, wherein the gas-releasing compound is a solid organic acid.

5. The composition according to any one of claims 1 to 4, wherein the gas-releasing compound is citric acid, tartaric acid, malic acid and / or any mixture of these substances.

6. The composition according to any one of claims 1 to 5, wherein the equivalent ratio of the gas-releasing compound to the carbon dioxide-generating compound is in the range of 1 to 3.

7. The composition according to any one of claims 1 to 6, wherein the equivalent ratio of the gas-releasing compound to the carbon dioxide-generating compound is at least 1.05:

1.

8. The composition according to any one of claims 1 to 7, wherein the composition comprises more than 4% by weight of the N-amidinyl amino acid based on the total weight of the composition.

9. The composition according to any one of claims 1 to 8, wherein the N-amidinyl amino acid is guanidinoacetic acid.

10. The composition according to any one of claims 1 to 9, wherein the composition further comprises pyrroloquinoline quinone salt (PQQ salt).

11. The composition according to any one of claims 1 to 10, wherein the composition further comprises a PQQ salt, the PQQ salt comprising an alkali metal cation and / or an alkaline earth metal cation.

12. The composition according to any one of claims 1 to 11, wherein the composition further comprises prebiotics and / or probiotics.

13. The composition according to any one of claims 1 to 12, wherein the composition further comprises probiotics, said probiotics including those selected from Bacillus subtilis (… B. subtilis (e.g., DSM 32315 or DSM 32540), Bacillus amyloliquefaciens ( B. amyloliquefaciens (e.g., CECT 5940) and / or any mixture of these strains.

14. The composition according to any one of claims 1 to 13, wherein the composition further comprises 5-aminolevulinic acid, its derivatives, its salts and / or any mixture of these substances.

15. The composition according to any one of claims 1 to 14, wherein the composition is an effervescent tablet.

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