Compositions containing N-guanylic acid for drinking water applications

Abstract

The subject matter of this application is a composition comprising an N-guanylic 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 more, and the molar ratio of the carbon dioxide generating compound to the N-guanylic acid is 1 or more.

Description

[Technical Field]

[0001] The present invention relates to a composition comprising an N-guanylic 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 more, and the molar ratio of the carbon dioxide-producing compound to the N-guanylic acid is 1 or more, and to a diet comprising the said composition for use in treating and / or preventing heat stress, transport stress, or any other stress-related condition in poultry.

[0002] N-guanylic acids are derivatives of amino acids containing a guanidine group and can be obtained by the addition of cyanamide to the amino acid in question. The most important N-guanylic acid is guanidinoacetate (GAA), also known as N-guanylglycine. It is an endogenous substance in animals and humans that plays a central role in the biosynthesis of creatine. Creatine can be obtained through diet and / or formed endogenously. Its biosynthesis proceeds from glycine and L-arginine. In mammals, guanidinoacetate is formed mainly in the kidney by the enzyme L-Arg:Gly-amidinotransferase (AGAT), which transfers the guanidine group of L-arginine to the amino acid glycine. Starting from L-arginine, L-ornithine is thus produced, which is then metabolized in the urea cycle and converted to L-citrulline by carbamoylation. In a further step, guanidinoacetic acid is methylated with S-adenosylmethionine by the enzyme guanidinoacetic acid N-methyltransferase (GAMT) to creatine. Guanidinoacetic acid was first synthesized in 1861 by Adolph Strecker by adding cyanamide to glycine in aqueous solution, particularly in dilute aqueous solution of ammonia (cited in M. Strecker, compt. Rend. 1861, 52, 1212; Ber. Chem. Ges. (now Eur. J. Inorg. Chem.) 1908, 41, 4385). In later publications, guanidinoacetic acid was prepared from cyanamide and glycine in isopropanol as a solvent, using sodium hydroxide (CN102329250A) or sodium carbonate (CN101462983A) as a base.

[0003] Alternatively, N-guanylate amino acids, such as GAA, can also be produced in the fermentation process by amidino transfer reactions, which transfer the amidino group of arginine to various amidino group acceptors, such as guanidinoacetic acid, 3-guanidinopropionic acid, 4-guanidinobutyric acid, 2-guanidinoethanol, hydroxyguanidine, and homoarginine. Fermentation production of guanidinoacetic acid has been widely studied. For example, International Publication No. 2021 / 122400(A1) and International Publication No. 2022 / 00828(A1) disclose fermentation production of GAA using specific developmental strains, and Yiwen Zhang et al. disclose fermentation production of GAA using a whole-cell catalytic system (Yiwen Zhang, Hang Zhou, Yong Tao, and Baixue Lin, ACS Synth. Biol. 2020, 9, 2066~2075).

[0004] GAA supplementation allows for an optimal supply of creatine in the organism, which in turn positively impacts energy transport in muscle cells. Typically, GAA can be simply supplemented in the diet given to an animal. In particular, under certain conditions such as heat stress, transport stress, and during and after illness, animals often have increased energy requirements but low food consumption, and at the same time, the animal's need for hydration, such as drinking water, increases. In principle, this increased energy requirement can be compensated for by GAA supplementation. However, compared to creatine, guanylic acid has the disadvantage of very low solubility in water. For example, guanidinoacetic acid has a very low solubility in water, with only 1g soluble in 278ml of water at 15°C.

[0005] Several methods have been employed to solve this problem. For example, CN115137016A discloses a nutritional preparation for livestock and poultry, as well as a method for preparing said nutritional preparation. The nutritional preparation comprises 40-80 parts by weight of guanidinoacetic acid, 8-20 parts by weight of anhydrous sodium sulfate, 1-15 parts of a sweetener, 3-10 parts by weight of citric acid, 10-40 parts by weight of glucose, 8-20 parts of beneficial powder, and 1-13 parts by weight of ammonium bicarbonate. However, although this preparation is referred to as water-soluble in CN115137016, experiments have shown that the preparation according to CN115137016 does not dissolve in water unless stirred. Therefore, a mechanical stirrer is necessary to dissolve N-guanylic acids such as guanidinoacetic acid. However, mechanically stirred tanks or feed troughs are expensive and therefore extremely rare on farms. A further disadvantage of stirred tanks or feed troughs is the high risk of bacterial contamination.

[0006] The liquid application of N-guanylic acids such as guanidinoacetic acid would be a different approach to solving the solubility problems associated with this type of compound. For example, U.S. Patent Application Publication 2009 / 0297656(A1) discloses a liquid formulation for human and animal nutrition comprising an aqueous solution, a guanidinoacetic acid component, and at least one methyl group donor from the group of choline, methionine, and betaine.

[0007] International Publication No. 2021 / 008848(A1) discloses a concentrate for preparing a wet solution containing an aqueous solution of guanidinoacetic acid. More specifically, the concentrate comprises an aqueous solution containing guanidinoacetic acid and at least one salt from the group of calcium chloride and magnesium chloride, the solution containing, based on the total weight of the solution, a) 0.5–4% by weight of guanidinoacetic acid, 10–70% by weight of calcium chloride and / or magnesium chloride, and c) the remainder water, with components a) and b) present in soluble form in water. The large proportion of magnesium chloride or calcium chloride in this concentrate presumably plays a role in improving the solubility of guanidinoacetic acid. This results in a considerably high salt concentration in the tank or feed trough, which is also undesirable on a farm and can poison animals.

[0008] Therefore, there was still a need for a composition containing an N-guanylic acid, such as GAA, that could improve the solubility of N-guanylic acids, such as GAA, in water.

[0009] This problem has been found to be solved by increasing the rate at which N-guanylic acid, e.g., GAA, dissolves in water. More specifically, this problem is solved by including a compound that enables the release of carbon dioxide in the composition containing N-guanylic acid, e.g., GAA. The foaming effect resulting from the release of carbon dioxide is thought to improve the homogeneity of N-guanylic acid, e.g., GAA, in water. This reduces the tendency for aggregates to form and breaks down aggregates that may form and precipitate during N-guanylic acid production.

[0010] Therefore, one object of the present invention is a composition comprising an N-guanylic acid, such as 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 more, and the molar ratio of the carbon dioxide generating compound to the N-guanylic acid is 1 or more.

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

[0012] The carbon dioxide-producing compound reacts with the gas-releasing compound in the presence of water, releasing carbon dioxide. As a result, the dissolution process of N-guanylic acid is significantly accelerated.

[0013] In order to provide or generate carbon dioxide, it is preferable that the carbon dioxide generating compound is a salt of carbonate.

[0014] Therefore, in embodiments of the composition according to the present invention, the carbon dioxide generating compound is an alkali bicarbonate, an alkaline earth bicarbonate, an ammonium bicarbonate, an alkali carbonate, an alkaline earth carbonate, or a mixture thereof.

[0015] In preferred embodiments of the compositions according to the present invention, the carbon dioxide generating compound is sodium bicarbonate, potassium bicarbonate, ammonium bicarbonate, and / or a mixture thereof.

[0016] In this invention, the gas-releasing compound reacts with a carbon dioxide-producing compound, preferably a carbonate salt, in the presence of water to release carbon dioxide. Therefore, in principle, the underlying chemical reaction is an acid-base reaction, and the hydroxonium ion (H3O) provided by the gas-releasing compound is released. + ) under the formation of carbonate (H2CO3) carbonate ions (CO3 2- ) or bicarbonate ions (HCO3) - It reacts with ). The molecule is rapidly converted to water and carbon dioxide in the presence of water at ambient pressure and room temperature, resulting in a foaming or bubbling effect and significantly accelerating the dissolution of N-guanylic acid.

[0017] Therefore, the gas-releasing compound in the composition according to the present invention is preferably an acid, such as an inorganic acid, an organic acid, or a mixture thereof.

[0018] The use of solid organic acids makes it possible to provide the composition according to the invention in solid form. The composition according to the invention being in solid form enables easier handling, easier administration, and more efficient transportation of the composition. Therefore, the composition according to the invention is preferably a solid composition.

[0019] Therefore, in one embodiment of the composition according to the invention, the gas-releasing compound is a solid organic acid.

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

[0021] Therefore, in the context of the present invention, the term equivalent ratio represents the ratio of the gas-releasing compound to the carbon dioxide-generating compound for providing one molecule of carbon dioxide. For example, when the gas-releasing compound is citric acid having three carboxylic acid groups and the carbon dioxide-generating compound is sodium bicarbonate (NaHCO3), 3 equivalents of citric acid react with 3 equivalents of sodium bicarbonate to give 3 equivalents of carbon dioxide. Here, the equivalent ratio of the gas-releasing compound to the carbon dioxide-generating compound is 1:1. In another example, when the gas-releasing compound is citric acid having three carboxylic acid groups and the carbon dioxide-generating compound is sodium carbonate (Na2CO3), 6 equivalents of citric acid react with 3 equivalents of sodium carbonate to give 3 equivalents of carbon dioxide. Here, the equivalent ratio of the gas-releasing compound to the carbon dioxide-generating compound is 2:1.

[0022] Preferably, in the composition according to the invention, the equivalent ratio of the gas-releasing compound to the carbon dioxide-generating compound is in the range of 1 to 3.

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

[0024] The present invention makes it possible to provide a composition containing an N-guanylic acid, e.g., GAA, which contains a higher amount of N-guanylic acid, e.g., GAA, than the prior art composition, e.g., the mixture in International Publication No. 2021 / 008848(A1).

[0025] In another embodiment, the composition according to the present invention comprises more than 4% by weight of an N-guanylic acid, such as GAA, based on the total weight of the composition.

[0026] Preferably, the composition according to the present invention contains 5 to 25% by weight, 5 to 12% by weight, or 5 to 15% by weight of an N-guanylic acid, such as GAA, based on the total weight of the composition.

[0027] In particular, in the field of animal nutrition, the most relevant N-guanylic acid is guanidinoacetic acid.

[0028] Therefore, in further embodiments of the compositions according to the present invention, the N-guanylic acid is guanidinoacetic acid.

[0029] The composition according to the present invention may further contain additional components. A suitable additional component is pyrroloquinoline quinone in a bioavailable form.

[0030] Poultry administered with the composition according to the present invention, further comprising a biologically available form of pyrroloquinoline quinone, showed improved broiler growth performance throughout all periods. Surprisingly, this effect was observed under any conditions, i.e., even when no heat stress was applied to the poultry. Pyrroloquinoline quinone (PQQ) is an important 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 main effect of PQQ is to bind to intracellular proteins and act as an antioxidant, which is recycled intracellularly by glutathione. PQQ 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). Compared to other antioxidants such as ascorbic acid (vitamin C), PQQ has a positive effect on mitochondrial biosynthesis, which directly affects animal energy metabolism, while other antioxidants have no effect 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 broiler performance by increasing intracellular ATP levels through enhanced mitochondrial biosynthesis.

[0031] On the other hand, guanidinoacetate (GAA) improves energy transport from mitochondria to the cytoplasm by increasing the availability of creatine. In mitochondria, creatine binds to the phospho group of adenosine triphosphate (ATP), and in the cytoplasm, phosphocreatine releases the phospho group back to ADP. GAA is also expected to support broilers under oxidative stress, particularly heat stress, through its antioxidant properties, for example, by reducing electron leakage and disrupting the O2-radical mechanism. Since PQQ improves mitochondrial health and ATP production, and GAA improves energy transport from ATP, the combination of PQQ and GAA is thought to interact beneficially at the metabolic level to improve intracellular ATP production and distribution, allowing cells to cope more efficiently with the consequences of heat stress with more available energy and lower oxidative stress, thus this effect is thought to be further enhanced under heat stress conditions.

[0032] In another embodiment, the composition according to the present invention further comprises a bioavailable form of pyrroloquinoline quinone.

[0033] In the context of this invention, the term "biologically active form" is used as is known to those skilled in the art and refers to a form of a compound, in this case PQQ, that is capable of influencing biological processes beyond nutritional value in a manner that affects bodily functions.

[0034] PQQ is a tricarboxylic acid with low solubility in water. Therefore, the present invention uses a biologically active form of PQQ. In principle, the biologically active form of PQQ is not limited in any way. Nevertheless, the biologically active form of pyrroloquinoline quinone is preferably a pyrroloquinoline quinone salt (PQQ salt). This is because the solubility of PQQ is improved in pyrroloquinoline quinone salts, and since it is a biologically active form of PQQ, it dissolves more quickly in aqueous applications, such as drinking water for poultry, which is beneficial.

[0035] In an embodiment of the composition according to the present invention, the biologically active form of pyrroloquinoline quinone is a pyrroloquinoline quinone salt (PQQ salt). In the context of the present invention, the PQQ salt is not subject to any restrictions and can include an inorganic cation or an organic cation. In the context of the present invention, the term inorganic cation is used as known to those skilled in the art and refers to any type of simple cation containing only positively charged ions, for example, monovalent cations such as ammonium cation NH4 + , an alkali metal cation such as Li + , Na + , and K + , or monovalent copper ion Cu + , divalent cations such as alkaline earth metal cations such as Mg 2+ , Ca 2+ , and Ba 2+ , or divalent iron cation Fe 2+ , zinc cation Zn 2+ , or divalent copper ion Cu 2+ , or trivalent cations such as Al 3+ , or trivalent iron cation Fe 3+ is shown. In the context of the present invention, the term organic cation is used as known to those skilled in the art and refers to any type of compound cation, for example, tetramethylammonium cation N[CH3]4 +This shows that a PQQ salt can contain PQQ as a monovalent, divalent, or trivalent anion. Depending on the valence of the PQQ anion, the PQQ salt contains the required number of cations. For example, a PQQ salt containing divalent PQQ may contain two monovalent cations or one divalent cation. For example, if the PQQ salt contains a trivalent PQQ anion, the salt contains two trivalent PQQ anions and three divalent cations. In one embodiment of the composition according to the present invention, the PQQ salt contains an inorganic cation or an organic cation. Preferably, since alkali salts or alkaline earth salts of PQQ have relatively high solubility in water, the PQQ salt contains an inorganic cation, such as an alkali cation or an alkaline earth cation. In another embodiment of the composition according to the present invention, the PQQ salt contains an alkali cation and / or an alkaline earth cation. The disodium salt of PQQ is the most commonly used form because it has high water solubility and is a stable solid with a color ranging from red to brown depending on the level of hydration. PQQ disodium (PQQ*Na2) forms a stable trihydrate (about 12.5% ​​water) and a stable pentahydrate (about 20-21% water). When the anhydrous material is exposed to ambient conditions, it absorbs water to become at least the trihydrate form. In preferred embodiments of the compositions according to the present invention, the PQQ salt comprises a sodium cation and / or a potassium cation.

[0036] It has also been found that only a small amount of biologically active form of PQQ is required to achieve the beneficial effects of the compositions according to the present invention on performance and metabolism in broilers. In further embodiments, the compositions according to the present invention further comprise at least 0.05 ppm of biologically active form of PQQ. Preferably, the compositions according to the present invention further comprise 0.05 ppm to 20 ppm of biologically active form of PQQ. In this case, the recommended PQQ concentration can be easily adjusted by dissolving the composition in a corresponding amount of water.

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

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

[0039] Preferably, the probiotics include strains of the genus Bacillus, particularly strains of B. subtilis (e.g., DSM 32315 or DSM 32540), B. amyloricephaciens (e.g., CECT 5940), and / or mixtures of any of these.

[0040] In another embodiment, the composition according to the present invention further comprises a probiotic containing a strain selected from B. subtilis (e.g., DSM 32315 or DSM 32540), B. amyloricephaciens (e.g., CECT 5940), and / or a mixture of any of these.

[0041] Bacillus subtilis DSM 32315 was identified through screening of naturally occurring isolates. DSM 32315 was deposited with the DSMZ on May 12, 2016, under the aforementioned accession number, in the name of Evonik Degussa GmbH, in accordance with the Budapest Convention for the International Recognition of the Deposit of Microorganisms in Patent Proceedings. Bacillus subtilis DSM 32315 is the raw material for Evonik's GutCare registered trademark. GutCare is a direct-feed live bacterial solution based on the spore-forming Bacillus subtilis DSM 32315 strain, which possesses the unique ability to produce a variety of secondary metabolites.

[0042] Bacillus subtilis DSM 32540 was identified by targeted screening of naturally occurring isolates. DSM 32540 was deposited with the DSMZ on June 14, 2017, under the aforementioned accession number, in the name of Evonik Degussa GmbH, in accordance with the Budapest Convention for the International Recognition of the Deposit of Microorganisms in Patent Proceedings. Bacillus subtilis DSM 32540 is a raw material for Evonik's GutPlus registered trademark, a probiotic for gut microbiota management.

[0043] The Bacillus amyloricephasiens strain is deposited in the Spanish Type Culture Collection under accession number CECT 5940. CECT 5940 is unmodified and does not contain plasmids. The Ecobiol registered trademark is a feed additive consisting of naturally growing, fast-growing Bacillus amyloricephasiens CECT 5940. The Ecobiol registered trademark can improve animal health and production conditions and can help growers address quality, profitability, and sustainability challenges.

[0044] Furthermore, the compositions according to the present invention may further contain 5-aminolevulinic acid, a precursor of heme synthesis. A higher heme content in the organism in question means that more oxygen is present in the animal, leading to increased agility. The compositions according to the present invention may also contain derivatives of 5-aminolevulinic acid. Suitable salts of 5-aminolevulinic acid are, for example, alkali metal salts or alkaline earth metal salts, ammonium salts or acid addition salts of 5-aminolevulinic acid, such as 5-aminolevulinic acid hydrochloride salts. Suitable derivatives of 5-aminolevulinic acid are, for example, 5-aminolevulinic acid having a masking or protecting group on the hydroxyl group, such as esters, or 5-aminolevulinic acid having a masking or protecting group on the amino group, such as tert-butyloxycarbonyl, also known as a Boc group. Preferably, one or more masking groups can be cleaved from 5-aminolevulinic acid under physiological conditions. If a 5-aminolevulinic acid derivative has two masking or protecting groups, it is preferable that they can be cleaved simultaneously under the same conditions.

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

[0046] The composition according to the present invention is not limited in any way by the number of the one or more aforementioned additional components and their individual amounts, provided that the one or more additional components do not adversely affect the beneficial effects of the composition according to the present invention.

[0047] The presence of carbon dioxide generating or providing compounds, and gas-releasing compounds, has the effect of releasing carbon dioxide when the compounds according to the present invention are dissolved in water. Therefore, the compositions according to the present invention can also be considered as effervescent compositions. The compositions are not limited in any way with respect to their physical appearance. In the simplest case, the compositions are powders and can therefore already be used as supplements to animal diets. Nevertheless, it is also possible to further process the compositions according to the present invention into any conceivable form or physical appearance, for example, tablets, specifically effervescent tablets or effervescent tablets.

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

[0049] The compositions according to the present invention are suitable for use in the treatment and / or prevention of heat stress, transport stress, or any other stress-related conditions in poultry and / or livestock. For example, administration of GAA to poultry reduces the mortality rate of poultry during transport.

[0050] The diet according to the present invention is suitable for use in treating and / or preventing heat stress, transport stress, or any other stress-related conditions in poultry and / or livestock, and the diet comprises the composition according to the present invention and is administered to poultry and / or livestock.

[0051] In the context of this invention, the term poultry is used to refer to any type of domesticated bird that is raised in a rearing environment for its usefulness. Examples of poultry include domesticated birds, including chickens (i.e., broilers and laying hens), turkeys, geese, quail, and ducks, which are raised to produce meat or eggs. Preferably, in the context of this invention, the term poultry refers to chickens or broilers.

[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 products for consumption, such as meat, eggs, milk, fur, leather, and wool. In particular, the term "livestock" is used to refer to animals raised for consumption, specifically 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 high ambient temperatures. Heat stress can be chronic or acute. In the context of this invention, the term chronic heat stress is used to determine the duration of high ambient temperatures. In contrast, acute heat stress is understood in the context of this invention to determine sudden and short-term extremely high ambient temperatures. As a rule, the diet according to this invention is not limited to chronic or acute heat stress. In the context of this invention, the term high ambient temperature is used to determine temperatures above the comfortable temperature of poultry, particularly in the context of the term chronic heat stress. As already mentioned above, the temperature comfort zone of birds depends on the bird's age. Older birds are more sensitive to high temperatures. Typically, birds feel comfortable at about 21-24°C (70-75°F) and function normally up to about 27°C (80°F). However, above 27°C (up to about 30°C), feed consumption decreases, but water intake increases. In broiler birds, feed conversion ratio (FCR) and weight gain decrease, and in laying hens and breeding groups, egg production decreases. At temperatures of 30–32°C (86–95°F), a significant decrease in egg production and eggshell quality is observed. In laying hens, as ambient temperature rises, FCR based on egg mass and FCR per 12 eggs increase. When temperatures exceed approximately 35–37°C (96–100°F), birds attempt to lower their body temperature by vigorously shaking their throats, but temperatures in this range result in some mortality. Preferably, temperatures above 27°C, especially at least 30°C, are considered high ambient temperatures in the context of this invention. In particular, temperatures in the range of 30–40°C or 34–40°C are considered high temperatures.

[0054] The diet according to the present invention can be administered to poultry exposed to temperatures exceeding 27°C.

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

[0056] Typically, the high temperatures to which poultry are exposed follow a cyclical pattern. The temperature has a minimum value at night, then gradually rises during the day until it reaches a maximum value, and then decreases again at night from this maximum value back to the minimum value. When the described temperature pattern lasts for several days or weeks, it is also referred to as chronic cyclical heat stress in the context of this invention.

[0057] The diet according to the present invention can be administered to poultry that are chronically subjected to heat stress.

[0058] The duration of temperature for chronic heat stress is preferably at least 5 hours per day. Preferably, the duration of temperature or high ambient temperature is 5 to 24 hours per day, in particular, at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 hours, or even up to 24 hours per day.

[0059] The diet according to the present invention can be administered to poultry that are exposed to temperatures above 27°C for at least 5 hours a day.

[0060] In many cases, specifically in southern countries, particularly subtropical and tropical countries, and generally in countries near the equator, high temperatures are associated with high relative humidity. In the context of this invention, the term high relative humidity is used to indicate a relative humidity of at least 40% on average. Preferably, in the context of this invention, the relative humidity is in the range of 40-70% on average, and more particularly 50-60% on average.

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

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

[0063] In principle, the diet according to the present invention can be administered to poultry at any stage, multiple stages, or all stages of their life cycle.

[0064] The life cycle of a laying hen can be divided into the pre-laying period and three production phases or three production periods: the early production phase, the growth / juvenile production phase, and the nesting phase, sometimes referred to as the pre-laying period and phases I-III.

[0065] The lifespan of birds raised in a rearing environment for meat production can be divided into three phases: the initiation phase, the rearing phase, and the finishing phase. For example, the entire lifespan of a chicken may be 39 days, of which the days from d-0 to d-10 (d) are called the initiation phase, the days from d-10 to d-21 are called the rearing phase, and the days from d-21 to d-39 are called the finishing phase. Birds raised for egg production also have different growth and feeding periods. In principle, the administration of the diet according to the present invention to poultry that is chronically subjected to heat stress is not limited to any particular phase or period in the life of the poultry. Therefore, the diet can be administered to poultry that is chronically subjected to heat stress at any conceivable point in the middle or between any of the phases, i.e., during or between the initiation phase, rearing phase and / or finishing phase. Nevertheless, it is preferable to administer the diet to poultry that is chronically subjected to heat stress during or between the finishing phase. In the context of this invention, the term "during" in relation to a period is used to indicate a selected point in time, such as an hour, day, or week, which are not necessarily consecutive periods. In contrast, the term "between" in relation to a period is used in the context of this invention to indicate a consecutive period of hours, days, or weeks.

[0066] The diet according to the present invention can be administered to poultry at any stage, multiple stages, or all stages of their life.

[0067] The diet according to the present invention can be administered to poultry from the beginning of the rearing period until slaughter, or from the beginning of the rearing period until slaughter.

[0068] Examples: 1. Example according to the present invention: Determination of dissolution time of a foaming mixture containing GAA A foaming composition containing GAA was obtained by mixing 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9 g (94 mmol) of sodium bicarbonate (PA grade, purchased from Merck), and 19.1 g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer) together in a batch mill (TubeMill, IKA) for 3 minutes.

[0069] The resulting solid mixture was added to 10 liters of tap water in a beaker. The solid mixture dissolved almost completely as it sank. The mixture was not stirred or moved. After 3 minutes, there was no solid mixture left at the bottom of the beaker.

[0070] 2. Example not based on the present invention: Determination of dissolution time of GAA (without foaming mixture) A beaker containing 10 liters of tap water was prepared, and 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China) was added. The powder settled at the bottom of the beaker. The mixture was not stirred or moved. After 4 days, no solid material was visible at the bottom of the beaker.

[0071] 3. Example not based on the present invention: Solubility test of four preparations of CN115137016A containing GAA Preparations containing GAA according to CN115137016A (see paragraph

[0029] of CN115137016A) were prepared. Specifically, four preparations containing GAA according to CN115137016A were prepared using GAA as a powder (purchased from Gendone, China) and GAA as granular material (purchased from Gendone, China). Other chemicals used were sodium sulfate (anhydrous, purchased from Merck), citric acid (monohydrate, purchased from Merck), glucose (purchased from ChemPur), 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. The compositions of the four preparations for GAA powder and GAA granular material are summarized in Table 3.

[0072] [Table 1]

[0073] A total of eight preparations were subjected to dissolution tests to determine the dissolution time. The results are summarized in Table 2.

[0074] [Table 2]

[0075] In eight solubility tests with stirring, the four preparations containing GAA powder gave faster dissolution times than the other four preparations containing GAA granules. However, all eight comparative preparations gave longer dissolution times than the mixture according to the present invention. In eight solubility tests without stirring, there was no significant difference between the preparations containing GAA powder and those containing GAA granules. Rather, all eight preparations according to CN115137016A, whether containing GAA powder or GAA granules, are unsuitable for use in unstirred water tanks.

[0076] 4. Example according to the present invention: Solubility test of a foaming composition containing GAA and the registered trademark GutCare (without maltodextrin) 3g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9g (94 mmol) of sodium bicarbonate (PA grade, purchased from Merck), 19.1g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer), and 97.8 mg of dried biomass GutCare registered trademark (maltodextrin-free, 0.92 × 10⁻¹⁶) 11 The compounds (calculated in CFU / g, manufactured by Evonik Espana y Portugal, SAU, Leon) were mixed together in a batch mill (TubeMill, IKA) for 3 minutes to obtain a foaming composition containing GAA and the registered trademark GutCare.

[0077] The resulting effervescent composition was added to 10 L of tap water in a beaker. After 10 minutes, samples were taken from the surface, center, 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 (central sample), and 3.1 × 10 5 This is CFU / mL (bottom sample).

[0078] 5. Example according to the present invention: Solubility test of a foaming composition containing GAA and the registered trademark GutCare (containing maltodextrin) 3g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9g (94 mmol) of sodium bicarbonate (PA grade, purchased from Merck), 19.1g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer), and 81.1 mg of dried biomass GutCare registered trademark (containing maltodextrin, 1.1 × 10⁻¹⁶). 11The compounds (calculated in CFU / g, manufactured by Evonik Espana y Portugal, SAU, Leon) were mixed together in a batch mill (TubeMill, IKA) for 3 minutes to obtain a foaming composition containing GAA and the registered trademark GutCare.

[0079] The resulting effervescent composition was added to 10 L of tap water in a beaker. After 10 minutes, samples were taken from the surface, center, 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 (central sample), and 7.2 × 10 5 This is CFU / mL (bottom sample).

[0080] 6. Example according to the present invention: Solubility test of a foaming composition containing GAA and Ecobiol registered trademarks 3g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9g (94 mmol) of sodium bicarbonate (PA grade, purchased from Merck), 19.1g (99.4 mmol) of citric acid (anhydrous, purchased from Jungbunzlauer), and 22.5 mg of dried biomass Ecobiol registered trademark (4 × 10⁻¹⁰). 11 The mixture was calculated in CFU / g and mixed together in a batch mill (TubeMill, IKA) for 3 minutes to obtain a foaming composition containing GAA and the registered trademark Ecobiol.

[0081] The resulting effervescent composition was added to 10 L of tap water in a beaker. After 10 minutes, samples were taken from the surface, center, 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 6 CFU / mL (central sample), and 9.3 × 10 5 This is CFU / mL (bottom sample).

[0082] 7. Example according to the present invention: Solubility test of a foaming composition containing GAA and 5-aminolevulinic acid 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9 g (94 mmol) of sodium bicarbonate (PA 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 in a batch mill (TubeMill, IKA) for 3 minutes to obtain a foaming composition containing GAA and 5-aminolevulinic acid.

[0083] The resulting effervescent composition was added to 10 L of tap water in a beaker. After 10 minutes, samples were taken from the surface, center, and bottom, and the amount of 5-ala was determined by HPLC. The results are summarized in Table 1.

[0084] [Table 3]

[0085] 8. Example according to the present invention: Solubility test of a foaming composition containing GAA and PQQ A foaming composition containing GAA and PQQ was obtained by mixing 3 g (25.3 mmol) of guanidinoacetic acid (GAA, powder, purchased from Gendone, China), 7.9 g (94 mmol) of sodium bicarbonate (PA 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 PQQ disodium salt pentahydrate (PentaQQ, Anthem Biosciences Pvt., Ltd.) together in a batch mill (TubeMill, IKA) for 3 minutes.

[0086] The resulting effervescent composition was added to 10 L of tap water in a beaker. After 10 minutes, samples were taken from the surface, center, and bottom, and the amount of PQQ was determined by HPLC. The results are summarized in Table 2.

[0087] [Table 4]

Claims

1. A composition comprising an N-guanylic 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 more, and the molar ratio of the carbon dioxide generating compound to the N-guanylic acid is 1 or more.

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

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 a mixture thereof.

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-producing 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-producing compound is at least 1.05:

1.

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

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

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

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

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

13. The composition according to any one of claims 1 to 12, further comprising a probiotic containing a strain selected from B. subtilis (e.g., DSM 32315 or DSM 32540), B. amyloricephaciens (e.g., CECT 5940), and / or a mixture thereof.

14. The composition according to any one of claims 1 to 13, further comprising 5-aminolevulinic acid, its derivatives, its salts and / or mixtures thereof.

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

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