Means and methods for modulating behavior

Abstract

The present invention relates to a method for modulating animal behavior, improving animal welfare and increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin to improve animal performance using an oligosaccharide preparation.

Description

[0001] The present invention relates to a method of using an oligosaccharide preparation to modulate animal behavior, improve animal welfare, and increase the production of gamma-aminobutyric acid, kynurenine, and / or serotonin to improve animal performance.

[0002] Secondary metabolites are molecules produced by organisms that play a role in the interaction between the organism and its environment. Some well-known secondary metabolites are caffeine, paclitaxel, nicotine, artemisinin, morphine, botulinum toxin, etc. It is worth noting that many secondary metabolites can be used for multiple purposes. For example, caffeine is thought to be produced by plants as a pesticide, while humans consume caffeine to increase alertness. Other secondary metabolites produced by one organism may act as neurotransmitters in another organism. Neurotransmitters are signaling molecules that allow communication between cells in an organism. Receptors for neurotransmitters can be found, for example, on muscle cells or neurons. Gamma-aminobutyric acid (GABA; CAS-No. 56-12-2; also known as 5-aminobutyric acid) is known to be the main inhibitory neurotransmitter in the brain and spinal cord. GABA slows or blocks specific neural signals in the brain, thereby regulating the anxiety, fear, and / or stress levels perceived by humans and animals. Therefore, an imbalance in GABA activity in the body can lead to related conditions, such as anxiety disorders.

[0003] Kynurenine (KYN) is considered a neuromodulator of stress. In particular, the KYN to tryptophan (TRP) ratio has been found to be particularly correlated with aggressive behaviors (e.g., feather pecking) and social disturbance in laying hens. A lower KYN / TRP ratio was found to be associated with higher social disturbance traits. It is hypothesized that increasing the KYN / TRP ratio may help reduce social disturbance behaviors, thereby improving animal welfare.

[0004] Serotonin in the central nervous system cannot cross the blood-brain barrier, while tryptophan can. Therefore, higher tryptophan levels in the gut mean more tryptophan will cross the blood-brain barrier, where it can be converted into central serotonin. Serotonin is a precursor to melatonin. Therefore, increased serotonin levels will lead to increased melatonin levels. Melatonin and its precursor serotonin are known to influence the production of insulin and glucagon. Increased melatonin concentrations can elevate insulin and glucagon levels in animals. Increased insulin and glucagon levels are also known to enhance fat synthesis. Both insulin and melatonin are further involved in regulating circadian rhythms. Changes in the light cycle affect the levels of insulin and melatonin produced by animals. Altered insulin and melatonin levels, in turn, regulate the animal's physiological response to the change in light cycle. Generally speaking, poultry production and broiler chicken rearing now utilize long daylight hours, up to 23 hours per day. This lighting regimen strongly affects production performance, such as faster fat gain, but it can also be detrimental to animal welfare.

[0005] Tail biting (TB) is an abnormal pathological behavior frequently observed in weaners and growers in commercial pig production and is a major health and welfare issue due to the risk of infection, the pain and stress experienced by the bitten animal, and the additional stress experienced by the entire animal group. Previous studies have shown that pigs that bite their tails have different neurotransmitter concentrations than pigs that do not engage in this aggressive behavior. It is increasingly recognized that neurochemicals or neurotransmitters produced by either or both the animal and / or its microbiome represent a common language for host-microbe communication. Microbial endocrinology has been described to play a role in animal behavior through the microbiota-gut-brain axis (MGBA) (Lyte et al. 2019. Animal. 13(11): 2689-2698). The gut microbiota is known to be able to synthesize neuroactive molecules that are structurally similar to host-derived neurotransmitters, including gamma-aminobutyric acid (GABA), indole derivatives, and catecholamines, such as dopamine and norepinephrine. Many of these co-synthesized neurotransmitters are involved in regulating host mood, behavior, and cognition.

[0006] The GABAergic system is widely distributed in the piglet hippocampus and has an inhibitory effect on hippocampal neurons. In general, GABA is synthesized by decarboxylation of glutamate catalyzed by glutamate decarboxylase (GAD) and is widely distributed in the nervous system to reduce the activity of neurons by binding to GABA receptors. In mice and rats that exhibit aggressive behavior, brain levels of GABA are low, which has been interpreted as consistent with the proposed inhibitory role of GABA in aggression (Clement et al. 1987. Pharmacol Biochem Behav. 26 (1): 83-8). In addition, direct supplementation of GABA can regulate stress, which is reflected in reduced aggressive behavior and improved growth performance. In addition, GABA supplementation can improve transportation stress in growing and finishing pigs (Bi et al. 2020. J Anim Physiol Anim Nutr. 104: 590-596). However, it is worth noting that to date, direct external supply of GABA has produced conflicting results and limited efficacy. Furthermore, GABA has a short half-life in vivo (eg, less than 17 minutes in mice), thus further reducing the efficacy and efficiency of direct GABA administration.

[0007] Despite the undisputed relevance of such secondary metabolites for the health, welfare, and performance of humans and animals, convenient and reliable means and methods for regulating the levels of these secondary metabolites in the body, modulating behavior, and improving welfare, performance, and well-being remain to be identified.

[0008] In view of the prior art as outlined above, it was an object of the present invention to provide means and methods for regulating animal behavior, for improving animal welfare and for improving animal performance in non-stressed as well as stressed animals.

[0009] Surprisingly, this object is achieved by providing a method for: i) regulating animal behavior, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion rate; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion rate; and / or v) reducing the concentration of cortisol in the plasma, saliva and / or hair of animals exposed to stress; the method comprising the steps of: 1) providing an oligosaccharide preparation, An oligosaccharide preparation is produced or capable of being produced by the following steps: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2; 2) feeding the oligosaccharide preparation to the animal; and 3) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal.

[0010] In some embodiments, the method according to the invention for iv) improving animal performance in stressed animals, in particular for improving average weight gain and / or reducing feed conversion ratio, relates to improved animal performance in stressed animals that have received an oligosaccharide preparation compared to stressed animals that have not received an oligosaccharide preparation. In some embodiments, the method according to the invention for v) reducing the concentration of cortisol in the plasma, saliva and / or hair of stressed animals relates to a reduction in cortisol in the plasma, saliva and / or hair of stressed animals that have received an oligosaccharide preparation compared to stressed animals that have not received an oligosaccharide preparation.

[0011] In another aspect, the present invention relates to a method for: i) regulating animal behavior, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobias; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; the method comprising the steps of: 1) providing an oligosaccharide preparation comprising a) reacting at least a first saccharide with a catalyst; at least a first saccharide and a second saccharide, wherein the at least a first saccharide is N-acetylglucosamine; or b) condensing the at least a first saccharide with the at least a second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid; thereby producing the oligosaccharide preparation, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2; 2) feeding the oligosaccharide preparation to the animal; and 3) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal.

[0012] As used herein, the term "condensation" should be interpreted as the steps necessary to allow feed sugars or starting sugars (e.g., simple sugars (monosaccharides)) to combine with each other to form larger sugars (disaccharides, oligosaccharides, or polysaccharides). For example, two monosaccharides can combine with each other to form a disaccharide. Similarly, a monosaccharide can be condensed with a disaccharide to form a trisaccharide, etc. During this condensation reaction, water molecules are released. In this article, condensation is initiated by heating the mixture of feed sugars in the presence of a catalyst.

[0013] In some embodiments, the present invention relates to a method for: i) regulating animal behavior, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or v) reducing cortisol concentrations in the plasma, saliva and / or hair of animals exposed to stress; the method comprising the following The invention relates to a method for producing an oligosaccharide preparation comprising: 1) providing an oligosaccharide preparation, wherein the oligosaccharide preparation is produced or capable of being produced by the following steps: condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2; 2) feeding the oligosaccharide preparation to the animal; and 3) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal.

[0014] In some embodiments, the present invention relates to a method for: i) regulating the behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobias; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in an animal exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or v) reducing the concentration of cortisol in the plasma, saliva and / or hair of an animal exposed to stress; the method comprising the following steps: 1) providing an oligosaccharide preparation produced or capable of being produced by the following steps: condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2; 2) feeding the oligosaccharide preparation to the animal; and 3) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, in particular in the gastrointestinal tract and / or blood of the animal.

[0015] In some embodiments, the present invention relates to a method for: i) regulating animal behavior, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or v) reducing the concentration of cortisol in the plasma, saliva and / or hair of animals exposed to stress; the method comprising the steps of: 1) providing a An oligosaccharide preparation produced or producible by the following steps: 1) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine, and wherein the catalyst is methylphosphonic acid; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2; 2) feeding the oligosaccharide preparation to the animal; and 3) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal.

[0016] In some embodiments, the at least second sugar in a) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the at least first sugar is N-acetylglucosamine) is also N-acetylglucosamine. In other embodiments, the at least second sugar in a) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the at least first sugar is N-acetylglucosamine) is selected from the group consisting of glucose, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably glucose.

[0017] In some embodiments, the at least first sugar and the at least second sugar of b) (condensing at least the first sugar with at least the second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) are not the same sugar as in a) (condensing at least the first sugar with at least the second sugar using a catalyst, wherein the at least first sugar is N-acetylglucosamine). In some embodiments, the at least first sugar and the at least second sugar of b) (condensing at least the first sugar with at least the second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) are both glucose. In some embodiments, the at least first sugar of b) (condensing at least the first sugar with at least the second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) is glucose; and the at least second sugar of b) (condensing at least the first sugar with at least the second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) is selected from N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from arabinose, sucrose, xylose, mannose, lactose, and fructose.

[0018] In some embodiments, the at least first saccharide and the at least second saccharide of a) (condensing at least the first saccharide with at least the second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine) are condensed with at least a third saccharide selected from the group consisting of glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from the group consisting of lactose. In some embodiments, the at least first saccharide and the at least second saccharide of b) (condensing at least the first saccharide with at least the second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid) are condensed with at least a third saccharide selected from the group consisting of glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from the group consisting of lactose. In some embodiments, the method according to the present invention comprises the step of condensing at least a first sugar with at least a second sugar using a catalyst, preferably wherein the catalyst is methylphosphonic acid, wherein the at least first sugar is N-acetylglucosamine, wherein the at least second sugar is glucose, and wherein the at least third sugar is selected from the group consisting of lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose.

[0019] In some embodiments, at least one of the sugars in a) is glucose; wherein the amount of glucose is 50-99% (e.g., wt %) of all sugars used in the condensation step. In some embodiments, the amount of at least the second sugar is 1-50% (e.g., wt %) of all sugars used in the condensation step. In some embodiments, the amount of at least the third sugar is 1-50% (e.g., wt %) of all sugars used in the condensation step. In some embodiments, a) is performed by condensing at least the first sugar with at least the second sugar and at least the third sugar using a catalyst, wherein the at least first sugar, at least the second sugar, and at least the third sugar are used in a ratio of 50-99:1-50:1-50, preferably wherein at least one of the at least first sugar, at least the second sugar, and at least the third sugar is glucose.

[0020] In some embodiments of the method according to the present invention, at least one of the sugars in b) is glucose; wherein the amount of glucose is 50-99% (e.g., weight %) of all sugars used in the condensation step. In some embodiments of the method according to the present invention, the amount of at least the second sugar is 1-50% (e.g., weight %) of all sugars used in the condensation step. In some embodiments of the method according to the present invention, the amount of at least the third sugar is 1-50% (e.g., weight %) of all sugars used in the condensation step. In some embodiments of the method according to the present invention, b) is carried out by condensing at least the first sugar with at least the second sugar and at least the third sugar using a catalyst, wherein the catalyst is methylphosphonic acid, wherein the at least first sugar, at least the second sugar and at least the third sugar are used in a ratio of 50-99:1-50:1-50, preferably wherein at least one of the at least first sugar, at least the second sugar and at least the third sugar is glucose.

[0021] In another aspect, the present invention relates to a method for increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by a microbial community; the method comprising the steps of: 1) providing an oligosaccharide preparation produced or capable of being produced by the steps of: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2; and contacting the microbial community with the oligosaccharide preparation.

[0022] In another aspect, the present invention relates to a method for increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by a microbial community; the method comprising the steps of: 1) providing an oligosaccharide preparation comprising a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2; and contacting the microbial community with the oligosaccharide preparation.

[0023] Thus, the production of gamma-aminobutyric acid, kynurenine and / or serotonin can be achieved independently of the animal hosting the microbial community. In some cases, such methods can be used, for example, in screening studies for substances that can enhance or inhibit the production of gamma-aminobutyric acid, kynurenine and / or serotonin by microorganisms.

[0024] Those skilled in the art will understand that the production of gamma-aminobutyric acid, kynurenine and / or serotonin (e.g., by a microbial consortium) will begin upon contacting one or more cells (e.g., one or more cells of a microbial consortium) with the oligosaccharide preparation, and that the amount of gamma-aminobutyric acid, kynurenine and / or serotonin produced will increase with ongoing incubation or fermentation time.

[0025] In some embodiments, the microbial community is an intestinal microbial community (i.e., contained in or obtainable from an intestinal sample, e.g., from the cecum, ileum), preferably an animal intestinal microbial community, e.g., a pet, dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, wild boar, zoo animals, horse, donkey, poultry, swine, ruminants, chicken, cow, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, clams, trout, tilapia, salmon, carp, catfish, tuna, preferably selected from the group consisting of poultry (such as chicken), pig and ruminants (such as cow).

[0026] In another aspect, the present invention relates to a method for producing an oligosaccharide preparation for: i) regulating the behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobias; iii) increasing the levels of gamma-aminobutyric acid, kynurenine and / or serotonin in an animal, in particular in the gastrointestinal tract and / or blood of the animal; and / or iv) reducing the concentration of cortisol in the plasma, saliva and / or hair of an animal facing stress; the method comprising the steps of: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid; thereby producing the oligosaccharide preparation, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

[0027] In some embodiments, the present invention relates to a method for producing an oligosaccharide preparation for: i) regulating animal behavior, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobias; iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in an animal, in particular in the gastrointestinal tract and / or blood of an animal; and / or iv) reducing the concentration of cortisol in the plasma, saliva and / or hair of an animal facing stress; the method comprising the steps of: condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine, and wherein the catalyst is methylphosphonic acid; thereby producing the oligosaccharide preparation, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2.

[0028] In another aspect, the present invention relates to an oligosaccharide preparation for: i) regulating animal behavior, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobias; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or v) reducing the concentration of cortisol in the plasma, saliva and / or hair of animals exposed to stress; wherein the oligosaccharide preparation is produced or capable of being produced by the following steps: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid; and wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

[0029] In some embodiments, the present invention relates to an oligosaccharide preparation for: i) regulating animal behavior, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobias; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or v) reducing the concentration of cortisol in the plasma, saliva and / or hair of animals exposed to stress; wherein the oligosaccharide preparation is produced or capable of being produced by the following steps: condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine, wherein the catalyst is methylphosphonic acid; and wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2.

[0030] In some embodiments, the at least second sugar in a) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the at least first sugar is N-acetylglucosamine) is also N-acetylglucosamine. In other embodiments, the at least second sugar in a) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the at least first sugar is N-acetylglucosamine) is selected from the group consisting of glucose, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably glucose.

[0031] In some embodiments of the oligosaccharide preparation, the at least first saccharide and the at least second saccharide of b) (condensing at least the first saccharide with at least the second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid) are not the same sugar as in a) (condensing at least the first saccharide with at least the second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine). In some embodiments of the oligosaccharide preparation, the at least first saccharide and the at least second saccharide of b) (condensing at least the first saccharide with at least the second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid) are both glucose. In some embodiments of the oligosaccharide preparation, the at least first saccharide of b) (condensing at least the first saccharide with at least the second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid) is glucose; and the at least second saccharide of b) (condensing at least the first saccharide with at least the second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid) is selected from N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably selected from arabinose, sucrose, xylose, mannose, lactose and fructose.

[0032] In some embodiments of the oligosaccharide preparation, the at least first saccharide and the at least second saccharide of a) (condensing at least the first saccharide with at least the second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine) are condensed with at least a third saccharide selected from the group consisting of glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from the group consisting of lactose. In some embodiments of the oligosaccharide preparation, the at least first saccharide and the at least second saccharide of b) (condensing at least the first saccharide with at least the second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid) are condensed with at least a third saccharide selected from the group consisting of glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from the group consisting of lactose. In some embodiments of the oligosaccharide preparation, the oligosaccharide preparation is produced or producible by condensing at least a first sugar with at least a second sugar using a catalyst, preferably wherein the catalyst is methylphosphonic acid, wherein the at least first sugar is N-acetylglucosamine, wherein the at least second sugar is glucose, and wherein the at least third sugar is selected from the group consisting of lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose.

[0033] In some aspects, the present invention relates to an oligosaccharide preparation for: i) regulating the behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobias; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or v) reducing the concentration of Cortisol in the plasma, saliva and / or hair of animals exposed to stress; wherein the oligosaccharide preparation comprises at least two different monosaccharide units selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably selected from glucose and N-acetylglucosamine; and wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2. In some embodiments, the present invention relates to an oligosaccharide preparation for: i) regulating animal behavior, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobias; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or v) reducing the concentration of cortisol in the plasma, saliva and / or hair of animals exposed to stress; wherein the oligosaccharide preparation comprises at least two different monosaccharide units selected from glucose, N-acetylglucosamine; and at least one third different monosaccharide unit selected from lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably lactose; and wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

[0034] In some embodiments, the oligosaccharide preparation according to the present invention comprises at least 10% by weight (11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, etc.) of said at least first fraction; preferably at least 20% by weight.

[0035] In some embodiments, the oligosaccharide preparation according to the present invention is included in the nutritional composition at an inclusion rate of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, etc.).

[0036] In some embodiments, the oligosaccharide preparation according to the invention comprises at least 4 fractions (e.g., at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 fractions), each having a different DP.

[0037] In some embodiments, the oligosaccharide preparation according to the invention comprises no more than three different monosaccharide units.

[0038] In some embodiments, at least one fraction of the oligosaccharide preparation according to the present invention comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% anhydro subunit-containing oligosaccharides in relative abundance; and and / or wherein each fraction of the oligosaccharide preparation comprises greater than 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, 50%, 60%, 70% or 80% anhydro subunit-containing oligosaccharides in relative abundance.

[0039] In some embodiments, the weight average molecular weight of the oligosaccharide preparation according to the present invention is 300 g / mol to 5000 g / mol (e.g., about 2000 g / mol to 2800 g / mol, 2100 g / mol to 2700 g / mol, 2200 g / mol to 2600 g / mol, 2300 g / mol to 2500 g / mol, or 2320 g / mol to 2420 g / mol), 500 g / mol to 5000 g / mol, 700 g / mol to 800 g / mol, or 1000 g / mol to 1500 g / mol. 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 300 to 700 g / mol, 400 g / mol to 1300 g / mol, 400 g / mol to 1200 g / mol, 400 g / mol to 1100 g / mol, 500 g / mol g / mol, 500 g / mol to 1200 g / mol, 500 g / mol to 1100 g / mol, 600 g / mol to 1300 g / mol, 600 g / mol to 1200 g / mol, or 600 g / mol to 1100 g / mol); and / or wherein the number average molecular weight of the oligosaccharide preparation is 1000 g / mol to 2000 g / mol (e.g., about 1100 g / mol to 1900 g / mol, 1200 g / mol to 1800 g / mol, mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol, 100 to 1500 g / mol,

[0040] In another aspect, the present invention relates to the use of an oligosaccharide preparation according to the invention and as mentioned herein for: i) regulating the behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobias; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; v) increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by the microflora; and / or vi) reducing cortisol concentrations in the plasma, saliva and / or hair of animals exposed to stress.

[0041] In a particular embodiment, the present invention relates to the use of an oligosaccharide preparation according to the invention and as mentioned herein for: i) regulating the behavior of healthy animals, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia symptoms in healthy animals; iii) improving animal performance, in particular for improving the average weight gain and / or reducing the feed conversion ratio in healthy animals; iv) improving animal performance in animals exposed to stress, in particular for improving the average weight gain and / or reducing the feed conversion ratio in healthy animals exposed to stress; v) increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by the microflora; and / or vi) reducing the concentration of cortisol in the plasma, saliva and / or hair of animals exposed to stress.

[0042] In another aspect, the present invention relates to an oligosaccharide preparation for treating, ameliorating, preventing and / or prophylaxis of one or more conditions associated with an imbalance in gamma-aminobutyric acid, kynurenine and / or cortisol levels; wherein the one or more conditions are selected from anxiety, stress, phobias, systemic inflammation and local inflammation; wherein the oligosaccharide preparation is produced or capable of being produced by the following steps: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid; and wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

[0043] In some embodiments, the present invention relates to an oligosaccharide preparation for treating, ameliorating, preventing, and / or prophylaxis of one or more conditions associated with imbalanced levels of gamma-aminobutyric acid, kynurenine, and / or cortisol in an animal suffering from or at risk of suffering from one or more conditions associated with imbalanced levels of gamma-aminobutyric acid and / or kynurenine, wherein the one or more conditions are selected from anxiety, stress, phobias, systemic inflammation, and local inflammation, wherein the oligosaccharide preparation is produced or capable of being produced by the steps of: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the catalyst is methylphosphonic acid; and wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2. The animal suffering from one or more conditions associated with imbalanced levels of gamma-aminobutyric acid and / or kynurenine can be, for example, an animal (such as a pig) subjected to a situation prone to causing social distress to the animal, such as mixing of individuals in a herd of animals. Similarly, for example, an animal about to experience such a condition may be considered at risk for developing one or more disorders associated with an imbalance in gamma-aminobutyric acid and / or kynurenine levels.

[0044] In some embodiments, the oligosaccharide preparation used for treatment, amelioration, prevention and / or prophylaxis is an oligosaccharide preparation according to the present invention and as described herein.

[0045] In some embodiments, the present invention relates to an oligosaccharide preparation for treating, ameliorating, preventing and / or prophylaxis of one or more conditions associated with an imbalance in gamma-aminobutyric acid, kynurenine and / or cortisol levels; wherein the one or more conditions are selected from anxiety, stress, phobias, systemic inflammation and local inflammation; wherein the oligosaccharide preparation is included in the nutritional composition at an inclusion rate of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm).

[0046] In some embodiments, the animal involved in this article is selected from the group consisting of pets, dogs, cats, canaries, guinea pigs, hamsters, rabbits, mice, rats, deer, wild boars, zoo animals, horses, donkeys, poultry, swine, ruminants, chickens, cattle, sheep, goats, pigs, piglets, turkeys, aquaculture, fish, shrimp, prawns, clams, trout, tilapia, salmon, carp, catfish, tuna; preferably, the animal is selected from the group consisting of poultry (such as chickens), pigs and ruminants (such as cattle); preferably, pigs.

[0047] It has been observed that the health, welfare and / or performance of the host animals are improved in four ways by the means, methods and uses of the present invention. First, the welfare of the production animal population is improved. For monogastric animals such as pigs, chickens or ducks kept in confined spaces, the occurrence of social disturbance behaviors such as feather pecking or tail biting is a common problem. Disruptive behaviors such as these lead to poor welfare of production animals and have therefore been a long-standing problem for animal keepers. The means, methods and uses according to the present invention contribute to improving the welfare of animals.

[0048] Secondly, it can improve the health of the host animal by reducing systemic inflammation. Systemic inflammation results from the release of proinflammatory cytokines by immune-related cells and chronic activation of the innate immune system. It contributes to the development of chronic diseases in animals. The method according to the present invention helps reduce systemic inflammation in animals.

[0049] Third, the method can improve the health of the host animal by reducing local inflammation. Local inflammation occurs in the area affected by a noxious stimulus. Acute local inflammation develops within minutes or hours after the noxious stimulus, is short-lived, and primarily involves the innate immune system. The method according to the present invention helps reduce local inflammation in animals.

[0050] Fourthly, the health of the host animal can be improved by reducing the lighting schedule / duration to the animal's daily circadian rhythm. The circadian rhythm associated with light has a significant impact on the growth of production animals. In the production animal husbandry industry, one way to increase growth rate and meat yield is by extending the lighting. In some extreme cases, lighting for poultry is extended to 23 hours a day, leaving the poultry in darkness for only one hour a day. Although this approach can increase productivity, it has a negative impact on the health and welfare of the animals. It has been observed that the melatonin levels of chickens under 23 hours of light and 1 hour of darkness are reduced to less than half the amount of melatonin in chickens under 16 hours of light and 8 hours of darkness. The means, methods and uses according to the present invention help increase the amount of melatonin and its precursor serotonin, thereby restoring melatonin levels in animals subjected to long periods of light. Since artificially extending the light cycle leads to abnormal behaviors in poultry, such as aggressive interactions (tail biting, feather pecking, activity / motor problems, etc.), restoring melatonin levels in such animals helps improve animal welfare. The inventors of the present application have found that by applying the means, methods and uses according to the present invention to compensate for the production of melatonin, a stronger serotonergic flux into more melatonin can thus be achieved with a reduction in lighting regimes and better animal welfare.

[0051] The means, methods, and uses of the present invention are particularly applicable to production animals in general and can be provided to any suitable animal. In some embodiments, the animal is a monogastric animal. Monogastric animals are generally understood to have a single-chambered stomach. In other embodiments, the animal is a ruminant. Ruminants are generally understood to have a multi-chambered stomach. In some embodiments, the animal is a ruminant in the pre-ruminant stage. Examples of such ruminants in the pre-ruminant stage include brooding calves.

[0052] In some embodiments, the animal is poultry (e.g., chicken, turkey), seafood (e.g., shrimp), sheep, cows, cattle, buffalo, bison, pigs (e.g., starter pigs, growing / finishing pigs), cats, dogs, rabbits, goats, guinea pigs, donkeys, camels, horses, pigeons, ferrets, gerbils, hamsters, mice, rats, birds, or humans.

[0053] In some embodiments, the animal is livestock. In some embodiments, the animal is a companion animal. In some embodiments, the animal is poultry. Examples of poultry include chickens, ducks, turkeys, geese, quail, or Cornish game hens. In one variation, the animal is a chicken. In some embodiments, the poultry is a laying hen, a broiler chicken, or a turkey.

[0054] In other embodiments, the animal is a mammal, including, for example, cattle, pigs, goats, sheep, deer, bison, rabbits, llamas, mules, horses, reindeer, buffalo, yaks, guinea pigs, rats, mice, alpacas, dogs, or cats. In one variation, the animal is a cow. In another variation, the animal is a pig. In another variation, the animal is a sow.

[0055] The present invention is further characterized by the following items:

[0056] Item 1: Method for:

[0057] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0058] ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia;

[0059] iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or

[0060] iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio;

[0061] The method comprises the following steps:

[0062] - providing an oligosaccharide preparation, said oligosaccharide preparation being produced or capable of being produced by the following steps:

[0063] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or

[0064] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0065] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0066] - feeding said oligosaccharide preparation to said animal; and

[0067] - Increase the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal's body, particularly in the animal's gastrointestinal tract and / or blood.

[0068] Item 2: Method for:

[0069] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0070] ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia;

[0071] iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or

[0072] iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio;

[0073] The method comprises the following steps:

[0074] - providing an oligosaccharide preparation, said oligosaccharide preparation being produced or capable of being produced by the following steps:

[0075] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine;

[0076] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0077] - feeding said oligosaccharide preparation to said animal; and

[0078] - Increase the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal's body, particularly in the animal's gastrointestinal tract and / or blood.

[0079] Item 3: Method for:

[0080] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0081] ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia;

[0082] iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or

[0083] iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio;

[0084] The method comprises the following steps:

[0085] - providing an oligosaccharide preparation, said oligosaccharide preparation being produced or capable of being produced by the following steps:

[0086] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0087] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0088] - feeding said oligosaccharide preparation to said animal; and

[0089] - Increase the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal's body, particularly in the animal's gastrointestinal tract and / or blood.

[0090] Item 4: Method for:

[0091] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0092] ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia;

[0093] iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or

[0094] iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio;

[0095] The method comprises the following steps:

[0096] - providing an oligosaccharide preparation comprising

[0097] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or comprising

[0098] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0099] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0100] - feeding said oligosaccharide preparation to said animal; and

[0101] - Increase the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal's body, particularly in the animal's gastrointestinal tract and / or blood.

[0102] Item 5: Method for:

[0103] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0104] ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia;

[0105] iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or

[0106] iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio;

[0107] The method comprises the following steps:

[0108] - providing an oligosaccharide preparation comprising

[0109] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine;

[0110] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0111] - feeding said oligosaccharide preparation to said animal; and

[0112] - Increase the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal's body, particularly in the animal's gastrointestinal tract and / or blood.

[0113] Item 6: Method for:

[0114] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0115] ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia;

[0116] iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or

[0117] iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio;

[0118] The method comprises the following steps:

[0119] - providing an oligosaccharide preparation comprising

[0120] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0121] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0122] - feeding said oligosaccharide preparation to said animal; and

[0123] - Increase the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal's body, particularly in the animal's gastrointestinal tract and / or blood.

[0124] Item 7: Method for increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by a microbial community, e.g., an intestinal microbial community (i.e., contained in or obtainable from an intestinal sample, e.g., from the cecum, ileum), preferably from an animal intestinal microbial community, e.g., from a pet, dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, wild boar, zoo animal, horse, donkey, poultry, swine, ruminants, chicken, cattle, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, clams, trout, tilapia, salmon, carp, catfish, tuna, preferably selected from the group consisting of poultry (such as chicken), pig and ruminant (such as cattle)); the method comprising the following steps:

[0125] - providing an oligosaccharide preparation, said oligosaccharide preparation being produced or capable of being produced by the following steps:

[0126] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or

[0127] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0128] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0129] - and contacting said microbial community with said oligosaccharide preparation.

[0130] Item 8: Method for increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by a microbial community, e.g., an intestinal microbial community (i.e., contained in or obtainable from an intestinal sample, e.g., from the cecum, ileum), preferably from an animal intestinal microbial community, e.g., from a pet, dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, wild boar, zoo animal, horse, donkey, poultry, swine, ruminants, chicken, cattle, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, clams, trout, tilapia, salmon, carp, catfish, tuna, preferably selected from the group consisting of poultry (such as chicken), pig and ruminant (such as cattle)); the method comprising the following steps:

[0131] - providing an oligosaccharide preparation, said oligosaccharide preparation being produced or capable of being produced by the following steps:

[0132] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine;

[0133] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0134] - and contacting said microbial community with said oligosaccharide preparation.

[0135] Item 9: A method for increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by a microbial community, for example, an intestinal microbial community (i.e., contained in or obtainable from an intestinal sample, for example from the cecum, ileum), preferably from an animal intestinal microbial community, for example from a pet, dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, wild boar, zoo animal, horse, donkey, poultry, swine, ruminants, chicken, cattle, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, clams, trout, tilapia, salmon, carp, catfish, tuna, preferably selected from the group consisting of poultry (such as chicken), pig and ruminant (such as cattle)); the method comprising the following steps:

[0136] - providing an oligosaccharide preparation, said oligosaccharide preparation being produced or capable of being produced by the following steps:

[0137] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0138] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0139] - and contacting said microbial community with said oligosaccharide preparation.

[0140] Item 10: Method for increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by a microbial community, for example, an intestinal microbial community (i.e., contained in or obtainable from an intestinal sample, for example from the cecum, ileum), preferably from an animal intestinal microbial community, for example from a pet, dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, wild boar, zoo animal, horse, donkey, poultry, swine, ruminants, chicken, cattle, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, clams, trout, tilapia, salmon, carp, catfish, tuna, preferably selected from the group consisting of poultry (such as chicken), pig and ruminant (such as cattle)); the method comprising the following steps:

[0141] - providing an oligosaccharide preparation comprising

[0142] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or comprising

[0143] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0144] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0145] - and contacting said microbial community with said oligosaccharide preparation.

[0146] Item 11: Method for increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by a microbial community, for example, an intestinal microbial community (i.e., contained in or obtainable from an intestinal sample, e.g., from the cecum, ileum), preferably from an animal intestinal microbial community, for example from a pet, dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, wild boar, zoo animal, horse, donkey, poultry, swine, ruminants, chicken, cattle, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, clams, trout, tilapia, salmon, carp, catfish, tuna, preferably selected from the group consisting of poultry (such as chicken), pig and ruminant (such as cattle)); the method comprising the following steps:

[0147] - providing an oligosaccharide preparation comprising

[0148] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine;

[0149] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0150] - and contacting said microbial community with said oligosaccharide preparation.

[0151] Item 12: Method for increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by a microbial community, e.g., an intestinal microbial community (i.e., contained in or obtainable from an intestinal sample, e.g., from the cecum, ileum), preferably from an animal intestinal microbial community, e.g., from a pet, dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, wild boar, zoo animal, horse, donkey, poultry, swine, ruminants, chicken, cattle, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, clams, trout, tilapia, salmon, carp, catfish, tuna, preferably selected from the group consisting of poultry (such as chicken), pig and ruminant (such as cattle)); the method comprising the following steps:

[0152] - providing an oligosaccharide preparation comprising

[0153] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0154] wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2;

[0155] - and contacting said microbial community with said oligosaccharide preparation.

[0156] Item 13: Method for producing an oligosaccharide preparation for:

[0157] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0158] ii) improving animal welfare, in particular by reducing anxiety, stress and / or phobias; and / or

[0159] iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal;

[0160] The method comprises the following steps:

[0161] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or

[0162] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0163] Thereby the oligosaccharide preparation is produced, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

[0164] Item 14: Method for producing an oligosaccharide preparation for:

[0165] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0166] ii) improving animal welfare, in particular by reducing anxiety, stress and / or phobias; and / or

[0167] iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal;

[0168] The method comprises the following steps:

[0169] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine;

[0170] Thereby the oligosaccharide preparation is produced, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

[0171] Item 15: Method for producing an oligosaccharide preparation for:

[0172] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0173] ii) improving animal welfare, in particular by reducing anxiety, stress and / or phobias; and / or

[0174] iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal;

[0175] The method comprises the following steps:

[0176] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0177] Thereby the oligosaccharide preparation is produced, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

[0178] Item 16: The method according to any of the preceding items, wherein the at least second sugar of step a) is selected from N-acetylglucosamine, glucose, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably glucose.

[0179] Item 17: The method according to any one of the preceding items, wherein the at least first sugar of step b) is glucose, and wherein the at least second sugar of step b) is selected from N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose.

[0180] Item 18: The method according to any one of the preceding items, wherein the at least first sugar of step a) and the at least second sugar of step a) are condensed with at least a third sugar, the third sugar being selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably lactose.

[0181] Item 19: The method according to any one of the preceding items, wherein the at least first sugar of step b) and the at least second sugar of step b) are condensed with at least a third sugar, the third sugar being selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably lactose.

[0182] Item 20: The process according to any one of the preceding items, wherein the at least second sugar of step a) is glucose; and wherein the catalyst is selected from (+)-camphor-10-sulfonic acid and methylphosphonic acid.

[0183] Item 21: The method according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar of step a) is selected from lactose, mannose, arabinose, fructose, sucrose and xylose; and optionally, wherein the catalyst is selected from (+)-camphor-10-sulfonic acid, methylphosphonic acid and phosphoric acid.

[0184] Item 22: The process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is lactose, and wherein the catalyst is (+)-camphor-10-sulfonic acid.

[0185] Item 23: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is lactose, and wherein the catalyst is methylphosphonic acid.

[0186] Item 24: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is mannose, and wherein the catalyst is methylphosphonic acid.

[0187] Item 25: The process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is arabinose, and wherein the catalyst is (+)-camphor-10-sulfonic acid.

[0188] Item 26: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is arabinose, and wherein the catalyst is methylphosphonic acid.

[0189] Item 27: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is arabinose, and wherein the catalyst is phosphoric acid.

[0190] Item 28: The process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is fructose, and wherein the catalyst is (+)-camphor-10-sulfonic acid.

[0191] Item 29: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is fructose, and wherein the catalyst is methylphosphonic acid.

[0192] Item 30: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is fructose, and wherein the catalyst is methylphosphonic acid.

[0193] Item 31: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is fructose, and wherein the catalyst is phosphoric acid.

[0194] Item 32: The process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is sucrose, and wherein the catalyst is (+)-camphor-10-sulfonic acid.

[0195] Item 33: The process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is sucrose, and wherein the catalyst is (+)-camphor-10-sulfonic acid.

[0196] Item 34: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is sucrose, and wherein the catalyst is methylphosphonic acid.

[0197] Item 35: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is sucrose, and wherein the catalyst is methylphosphonic acid.

[0198] Item 36: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is sucrose, and wherein the catalyst is methylphosphonic acid.

[0199] Item 37: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is sucrose, and wherein the catalyst is phosphoric acid.

[0200] Item 38: Process according to any one of items 1 to 15, wherein the at least second sugar of step a) is glucose; wherein the at least third sugar is xylose, and wherein the catalyst is methylphosphonic acid.

[0201] Item 39: Process according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose and wherein the at least second sugar of step b) is arabinose.

[0202] Item 40: Process according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose and wherein the at least second sugar of step b) is lactose.

[0203] Item 41: Process according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose and wherein the at least second sugar of step b) is N-acetylglucosamine.

[0204] Item 42: Process according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose and wherein the at least second sugar of step b) is xylose.

[0205] Item 43: The process according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is fructose; and wherein the at least third sugar of step b) is arabinose.

[0206] Item 44: The process according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is sucrose; and wherein the at least third sugar of step b) is arabinose.

[0207] Item 45: The process according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is xylose; and wherein the at least third sugar of step b) is arabinose.

[0208] Item 46: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is arabinose; and wherein the at least third sugar of step b) is lactose.

[0209] Item 47: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is fructose; and wherein the at least third sugar of step b) is lactose.

[0210] Item 48: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is N-acetylglucosamine; and wherein the at least third sugar of step b) is lactose.

[0211] Item 49: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is sucrose; and wherein the at least third sugar of step b) is lactose.

[0212] Item 50: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is xylose; and wherein the at least third sugar of step b) is lactose.

[0213] Item 51: The method according to any one of items 1-15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is fructose; and wherein the at least third sugar of step b) is mannose.

[0214] Item 52: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is N-acetylglucosamine; and wherein the at least third sugar of step b) is mannose.

[0215] Item 53: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is sucrose; and wherein the at least third sugar of step b) is mannose.

[0216] Item 54: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is xylose; and wherein the at least third sugar of step b) is mannose.

[0217] Item 55: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is arabinose; and wherein the at least third sugar of step b) is N-acetylglucosamine.

[0218] Item 56: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is fructose; and wherein the at least third sugar of step b) is N-acetylglucosamine.

[0219] Item 57: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is fructose; and wherein the at least third sugar of step b) is N-acetylglucosamine.

[0220] Item 58: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is sucrose; and wherein the at least third sugar of step b) is N-acetylglucosamine.

[0221] Item 59: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is sucrose; and wherein the at least third sugar of step b) is N-acetylglucosamine.

[0222] Item 60: The method according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is sucrose; and wherein the at least third sugar of step b) is N-acetylglucosamine.

[0223] Item 61: Process according to any one of items 1-15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is xylose; and wherein the at least third sugar of step b) is N-acetylglucosamine.

[0224] Item 62: Process according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is fructose; and wherein the at least third sugar of step b) is xylose.

[0225] Item 63: Process according to any one of items 1 to 15, wherein the at least first sugar of step b) is glucose; wherein the at least second sugar of step b) is sucrose; and wherein the at least third sugar of step b) is xylose.

[0226] Item 64: An oligosaccharide preparation for:

[0227] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0228] ii) improving animal welfare, in particular by reducing anxiety, stress and / or phobias; and / or

[0229] iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal;

[0230] The oligosaccharide preparation is produced or can be produced by the following steps:

[0231] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or

[0232] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0233] And wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2.

[0234] Item 65: Oligosaccharide preparation according to Item 64, wherein the at least second sugar of step a) is selected from glucose, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably glucose.

[0235] Item 66: Oligosaccharide preparation according to item 64 or 65, wherein the at least first sugar of step b) is glucose, and wherein the at least second sugar of step b) is selected from N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose.

[0236] Item 67: The oligosaccharide preparation according to any one of items 64 to 66, wherein the at least first saccharide of any one of steps a) or b); and the at least second saccharide of any one of steps a) or b) are condensed with at least a third saccharide selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably selected from lactose.

[0237] Item 68: An oligosaccharide preparation for:

[0238] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0239] ii) improving animal welfare, in particular by reducing anxiety, stress and / or phobias; and / or

[0240] iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal;

[0241] wherein the oligosaccharide preparation comprises at least two different monosaccharide units selected from the group consisting of glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably selected from the group consisting of glucose and N-acetylglucosamine; and wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

[0242] Item 69: An oligosaccharide preparation for:

[0243] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0244] ii) improving animal welfare, in particular by reducing anxiety, stress and / or phobias; and / or

[0245] iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal;

[0246] wherein the oligosaccharide preparation comprises at least two different monosaccharide units selected from glucose, N-acetylglucosamine; and at least one third different monosaccharide unit selected from lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably lactose; and wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

[0247] Item 70: Oligosaccharide preparation according to any of items 64 to 69, wherein the oligosaccharide composition comprises at least 10% by weight (11% by weight, 12% by weight, 13% by weight, 14% by weight, 15% by weight, 16% by weight, 17% by weight, 18% by weight, 19% by weight, 20% by weight, 25% by weight, 30% by weight, 35% by weight, 40% by weight, 45% by weight, 50% by weight, etc.) of said at least first fraction; preferably at least 20% by weight.

[0248] Item 72: Oligosaccharide preparation according to any of items 64-71, wherein the oligosaccharide preparation is included in the nutritional composition at an inclusion rate of at least 50 ppm (e.g. at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm).

[0249] Item 73: An oligosaccharide preparation for:

[0250] i) regulating animal behavior, particularly to reduce aggressive behavior;

[0251] ii) improving animal welfare, in particular by reducing anxiety, stress and / or phobias; and / or

[0252] iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal, particularly in the gastrointestinal tract and / or blood of the animal;

[0253] wherein the oligosaccharide preparation is produced or producible by the method of any one of items 13-63.

[0254] Item 74: The oligosaccharide preparation according to any one of items 64 to 73, wherein the animal is selected from the group consisting of pets, dogs, cats, canaries, guinea pigs, hamsters, rabbits, mice, rats, deer, wild boars, zoo animals, horses, donkeys, poultry, swine, ruminants, chickens, cattle, sheep, goats, pigs, piglets, turkeys, aquaculture, fish, shrimps, prawns, clams, trout, tilapia, salmon, carp, catfish, tuna, preferably, the animal is selected from the group consisting of poultry (such as chickens), pigs and ruminants (such as cattle).

[0255] Item 75: The oligosaccharide preparation according to any one of items 64 to 74, wherein the oligosaccharide preparation comprises at least 4 fractions (e.g., at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 fractions), each having a different DP.

[0256] Item 76: Oligosaccharide preparation according to any of items 64 to 75, wherein the oligosaccharide preparation comprises not more than three different monosaccharide units.

[0257] Item 77: The oligosaccharide preparation according to any one of items 64 to 76, wherein at least one fraction of the oligosaccharide preparation comprises less than 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3% or 2% of oligosaccharides containing oligosaccharides in relative abundance. and / or wherein each fraction of the oligosaccharide preparation comprises greater than 0.2%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, 50%, 60%, 70% or 80% of oligosaccharides containing anhydro subunits in relative abundance.

[0258] Item 78: The oligosaccharide preparation according to any one of items 64 to 77, wherein the weight average molecular weight of the oligosaccharide preparation is 300 g / mol to 5000 g / mol (e.g., about 2000 g / mol to 2800 g / mol, 2100 g / mol to 2700 g / mol, 2200 g / mol to 2600 g / mol, 2300 g / mol to 2500 g / mol, or 2320 g / mol to 2420 g / mol), 500 g / mol to 5000 g / mol, or 2500 g / mol to 2600 g / mol. g / mol, 700 g / mol to 5000 g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 300 to 700 g / mol, 400 g / mol to 1300 g / mol, 400 g / mol to 1200 g / mol, 400 g / mol to 1100 g / mol 1, 500 g / mol to 1300 g / mol, 500 g / mol to 1200 g / mol, 500 g / mol to 1100 g / mol, 600 g / mol to 1300 g / mol, 600 g / mol to 1200 g / mol, or 600 g / mol to 1100 g / mol); and / or wherein the oligosaccharide preparation has a number average molecular weight of 1000 g / mol to 2000 g / mol (e.g., about 1100 g / mol to 1900 g / mol, 120 g / mol, 500 to 1050 g / mol, 500 to 1020 g / mol, 500 to 1010 g / mol, or 1450 to 1550 g / mol).

[0259] Item 79: Use of the oligosaccharide preparation according to any one of items 64 to 78:

[0260] i) regulating the behavior of animals (e.g. healthy animals), in particular for reducing aggressive behavior;

[0261] ii) improving animal welfare (e.g. of healthy animals), in particular for reducing anxiety, stress and / or phobia symptoms;

[0262] iii) improving animal performance (e.g. of healthy animals), in particular for improving average weight gain and / or reducing feed conversion ratio;

[0263] iv) improving animal performance in animals exposed to stress (e.g. healthy animals), in particular for improving average weight gain and / or reducing feed conversion ratio; and / or

[0264] v) increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by the microbial community.

[0265] Item 80: An oligosaccharide preparation for use in treating, ameliorating, preventing and / or prophylaxis of one or more conditions associated with an imbalance in gamma-aminobutyric acid and / or kynurenine levels; wherein the one or more conditions are selected from anxiety, stress, phobias, systemic inflammation and local inflammation, wherein the oligosaccharide preparation is produced or capable of being produced by the following steps:

[0266] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or

[0267] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0268] And wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2.

[0269] Item 81: An oligosaccharide preparation for use in treating, ameliorating, preventing and / or prophylaxis of one or more conditions associated with an imbalance in GABA and / or kynurenine levels in an animal suffering from or at risk of suffering from one or more conditions associated with an imbalance in GABA and / or kynurenine levels, wherein the one or more conditions are selected from the group consisting of anxiety, stress, phobias, systemic inflammation and local inflammation, wherein the oligosaccharide preparation is produced or capable of being produced by the following steps:

[0270] a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or

[0271] b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid;

[0272] And wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2.

[0273] Item 82: The oligosaccharide preparation for treatment, improvement, prevention and / or prophylaxis according to item 80 or 81, wherein the oligosaccharide preparation is the oligosaccharide preparation according to any one of items 64 to 78.

[0274] Item 83: An oligosaccharide preparation for use in treatment, improvement, prevention and / or prophylaxis according to any one of items 80-82, wherein the oligosaccharide preparation is contained in the nutritional composition at an inclusion rate of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm).

[0275] It is also contemplated that the oligosaccharide preparation may be provided in the form of a powder formulation comprising at least 20% (w / w) of the oligosaccharide preparation mentioned herein; at least 25% (wt / wt) of a silica-based adsorbate (e.g., diatomaceous earth, amorphous precipitated silica) having an average particle size D of less than or equal to 3000 μm (e.g., 100-500 μm, 200-500 μm, 200-300 μm); and optionally 0-25% (wt / wt) of water and / or auxiliary substances; wherein the % is based on the total weight of the powder formulation. For example, such a powder formulation may comprise 30-70% (wt / wt) of the oligosaccharide preparation mentioned herein; 30-70% (wt / wt) of a silica-based adsorbate (e.g., having an average particle size of at least 50 μm); and 0-21% (wt / wt) of water; wherein the % is based on the total weight of the powder formulation. In some embodiments, the oligosaccharide preparation is formulated similarly to the protocol described in any one of Examples 22 to 26 and 33 of WO 2020 / 097458.

[0276] As used herein, " oligosaccharide preparation " can comprise one or more monosaccharides or sugar, and / or dehydrated monosaccharide, and / or one or more compound molecules, for example disaccharide, trisaccharide, tetrasaccharide or the sugar of higher polymerization, it contains two or more monosaccharide subunits, and monosaccharide subunits are connected to form oligosaccharide by glycosidic bond." oligosaccharide " can also refer to the compound molecule containing two or more monosaccharide subunits, wherein at least one monosaccharide unit is replaced by dehydrated subunit. " oligosaccharide " can optionally be functionalized. When used in this article, the term " oligosaccharide " covers all kinds of oligosaccharides, wherein each monosaccharide subunit in oligosaccharide is independently and optionally functionalized and / or replaced by its corresponding dehydrated monosaccharide subunit.

[0277] The "anhydro subunit" can be a reversible thermal dehydration product of a monosaccharide or a monosaccharide subunit or a sugar caramelization product. For example, the "anhydro subunit" can be an anhydromonosaccharide, such as anhydroglucose. As another example, the "anhydro subunit" can be linked to one or more conventional monosaccharide subunits and / or to one or more anhydromonosaccharide subunits via a glycosidic bond.

[0278] Oligosaccharides may be characterized by containing two or more monosaccharide subunits linked by glycosidic bonds. In this regard, the term "glucose oligosaccharide" may refer to a compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds. "Glucose oligosaccharide" may also refer to a compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds, wherein at least one monosaccharide subunit is replaced by an anhydroglucose subunit. Similarly, "galactose oligosaccharide" may refer to a compound containing two or more galactose monosaccharide subunits linked by glycosidic bonds. "Galactose oligosaccharide" may also refer to a compound containing two or more galactose monosaccharide subunits linked by glycosidic bonds, wherein at least one monosaccharide subunit is replaced by an anhydrogalactose subunit. Similarly, glucose-galactose-oligosaccharide may be a compound comprising one or more glucose monosaccharide subunits and one or more galactose monosaccharide subunits linked by glycosidic bonds. Glucose-galactose oligosaccharide may also refer to a compound in which at least one monosaccharide subunit is replaced by its corresponding anhydroglucose subunit. Glucose-galactose-xylose-oligosaccharide may refer to a compound produced by the condensation reaction of glucose, galactose and xylose. The oligosaccharide preparation comprising glucose-galactose-xylose-oligosaccharide may comprise glucose-galactose-oligosaccharide, glucose-xylose-oligosaccharide, galactose-xylose-oligosaccharide, glucose-galactose-xylose-oligosaccharide, and a compound comprising one or more glucose monosaccharide subunits, one or more xylose monosaccharide subunits and one or more galactose monosaccharide subunits and their anhydromonosaccharide subunits linked by glycosidic bonds.

[0279] As used herein, unless otherwise indicated, the terms "monosaccharide unit" and "monosaccharide subunit" are used interchangeably. A "monosaccharide subunit" may refer to a monosaccharide monomer in an oligosaccharide. For an oligosaccharide with a degree of polymerization (DP) of 1, the oligosaccharide may be referred to as a monosaccharide subunit or a monosaccharide. For an oligosaccharide with a degree of polymerization greater than 1, the monosaccharide subunits are linked via glycosidic bonds.

[0280] As used herein, the term "conventional monosaccharide" may refer to a monosaccharide that does not contain an anhydro subunit. The term "conventional disaccharide" may refer to a disaccharide that does not contain an anhydro subunit. Thus, the term "conventional subunit" may refer to a subunit that is not an anhydro subunit.

[0281] As used herein, the term "relative abundance" or "abundance" may refer to the abundance of a substance in terms of how common or rare the substance is. For example, a DP1 fraction (i.e., a fraction having a DP of 1) containing 10% anhydro subunit-containing oligosaccharides in relative abundance may refer to a plurality of DP1 oligosaccharides, wherein 10% of the DP1 oligosaccharides by number are anhydromonosaccharides.

[0282] Degree of polymerization (DP) distribution:The degree of polymerization distribution of the oligosaccharide preparation can be determined by any suitable analytical method and instrument, including but not limited to end group method, osmotic pressure method (osmotic pressure determination method), ultracentrifugation, viscosity measurement, light scattering method, size exclusion chromatography (SEC), SEC-MALLS, field flow fractionation (field flow fractionation, FFF), asymmetric flow field flow fractionation (asymmetric flow field flow fractionation, A4F), high-performance liquid chromatography (high-performance liquid chromatography, HPLC) and mass spectrometry (mass spectrometry, MS). For example, the distribution of the degree of polymerization can be determined and / or detected by mass spectrometry (such as MALDI-MS, LC-MS or GC-MS). For another example, the degree of polymerization distribution can be determined and / or detected by SEC, such as gel permeation chromatography (gel permeation chromatography, GPC). As another example, the degree of polymerization distribution can be determined and / or detected by HPLC, FFF or A4F. In another example, the degree of polymerization of the oligosaccharide preparation can be determined based on its molecular weight and molecular weight distribution (for a more detailed description, see WO 2020 / 097458).

[0283] Anhydrosubunit level : The oligosaccharide preparation may comprise n different fractions, wherein each fraction has a different DP, and wherein n is any integer, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc. In some embodiments, each of the n oligosaccharide fractions of the oligosaccharide preparation described herein independently comprises anhydro subunit levels. For example, in some embodiments, the DP1 fraction comprises 10% anhydro subunit-containing oligosaccharides in relative abundance, and the DP2 fraction comprises 15% anhydro subunit-containing oligosaccharides in relative abundance. For another example, in some embodiments, the DP1 fraction, the DP2 fraction, and the DP3 fraction each comprise 5%, 10%, and 2% anhydro subunit-containing oligosaccharides in relative abundance, respectively. In other embodiments, two or more oligosaccharide fractions may comprise similar levels of anhydro subunit-containing oligosaccharides. For example, in some embodiments, the DP1 fraction and the DP3 fraction each comprise approximately 5% anhydro subunit-containing oligosaccharides in relative abundance.

[0284] The dehydrated subunit level can be determined by any suitable analytical method, such as nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry, HPLC, FFF, A4F, or any combination thereof. In some embodiments, the dehydrated subunit level is at least partially determined by mass spectrometry such as MALDI-MS. In some embodiments, the dehydrated subunit level can be determined at least in part by NMR. In some embodiments, the dehydrated subunit level can be determined at least in part by HPLC. For example, in some embodiments, the level of the dehydrated subunit can be determined by MALDI-MS, as described in more detail in WO 2020 / 097458.

[0285] glycosidic bond :In some embodiments, the oligosaccharide preparations described herein comprise a variety of glycosidic bonds. The type and distribution of glycosidic bonds can depend on the source and manufacturing method of the oligosaccharide preparation. In some embodiments, the type and distribution of various glycosidic bonds can be measured and / or detected by any suitable method known in the art (e.g., NMR). For example, in some embodiments, glycosidic bonds are measured and / or detected by proton NMR, carbon NMR, 2D NMR (e.g., 2D NMR, HSQC, HMBC, DOSY, COSY, ECOSY, TOCSY, NOESY, or ROESY) or any combination thereof. In some embodiments, glycosidic bonds are measured and / or detected at least in part by proton NMR. In some embodiments, glycosidic bonds are measured and / or detected at least in part by carbon NMR. In some embodiments, glycosidic bonds are measured and / or detected at least in part by 2D HSQC NMR.

[0286] In some embodiments, the oligosaccharide preparation may comprise one or more α-(1,2) glycosidic bonds, α-(1,3) glycosidic bonds, α-(1,4) glycosidic bonds, α-(1,6) glycosidic bonds, β-(1,2) glycosidic bonds, β-(1,3) glycosidic bonds, β-(1,4) glycosidic bonds, β-(1,6) glycosidic bonds, α-(1,1)-α glycosidic bonds, α-(1,1)-β glycosidic bonds, β-(1,1)-β glycosidic bonds, or any combination thereof.

[0287] In some embodiments, the glycosidic bond type distribution of the oligosaccharide preparation can be about 0 mol% to 60 mol%, 5 mol% to 55 mol%, 5 mol% to 50 mol%, 5 mol% to 45 mol%, 5 mol% to 40 mol%, 5 mol% to 35 mol%, 5 mol% to 30 mol%, 5 mol% to 25 mol%, 10 mol% to 60 mol%, 10 mol% to 55 mol%, 10 mol% to 50 mol%, 10 mol% to 45 mol%, 10 mol% to 40 mol%, 10 mol% to 35 mol%, 15 mol% to 60 mol%, 15 mol% to 55 mol%, % to 45 mol%, 15 mol% to 40 mol%, 15 mol% to 35 mol%, 20 mol% to 60 mol%, 20 mol% to 55 mol%, 20 mol% to 50 mol%, 20 mol% to 45 mol%, 20 mol% to 40 mol%, 20 mol% to 35 mol%, 25 mol% to 60 mol%, 25 mol% to 55 mol%, 25 mol% to 50 mol%, 25 mol% to 45 mol%, 25 mol% to 40 mol%, or 25 mol% to 35 mol% of α-(1,6) glycosidic bonds.

[0288] Molecular weight The molecular weight and molecular weight distribution of the oligosaccharide preparation can be determined by any suitable analytical means and instrument, such as end group method, osmotic pressure (osmotic pressure determination method), ultracentrifugation, viscosity measurement, light scattering method, SEC, SEC-MALLS, FFF, A4F, HPLC and mass spectrometry. In some embodiments, molecular weight and molecular weight distribution are measured by mass spectrometry, such as MALDI-MS, LC-MS or GC-MS. In some embodiments, molecular weight and molecular weight distribution are measured by size exclusion chromatography (SEC), such as gel permeation chromatography (GPC). In other embodiments, molecular weight and molecular weight distribution are measured by HPLC. In some embodiments, molecular weight and molecular weight distribution are measured by MALDI-MS.

[0289] In some embodiments, the weight average molecular weight of the oligosaccharide preparation is 300 g / mol to 5000 g / mol (e.g., about 2000 g / mol to 2800 g / mol, 2100 g / mol to 2700 g / mol, 2200 g / mol to 2600 g / mol, 2300 g / mol to 2500 g / mol, or 2320 g / mol to 2420 g / mol), 500 g / mol to 5000 g / mol, 700 g / mol to 5000 g / mol, g / mol, 500 g / mol to 2000 g / mol, 700 g / mol to 2000 g / mol, 700 g / mol to 1500 g / mol, 300 g / mol to 1500 g / mol, 300 g / mol to 2000 g / mol, 300 to 700 g / mol, 400 g / mol to 1300 g / mol, 400 g / mol to 1200 g / mol, 400 g / mol to 1100 g / mol, 500 g / mol to g / mol, 500 g / mol to 1200 g / mol, 500 g / mol to 1100 g / mol, 600 g / mol to 1300 g / mol, 600 g / mol to 1200 g / mol, or 600 g / mol to 1100 g / mol); and / or wherein the number average molecular weight of the oligosaccharide preparation is 1000 g / mol to 2000 g / mol (e.g., about 1100 g / mol to 1900 g / mol, 1200 to 1800 g / mol, g / mol, 1300 to 1700 g / mol, 1400 to 1600 g / mol, 300 to 1050 g / mol, 300 to 1020 g / mol, 300 to 1010 g / mol, 400 to 1050 g / mol, 400 to 1020 g / mol, 400 to 1010 g / mol, 500 to 1050 g / mol, 500 to 1020 g / mol, 500 to 1010 g / mol, or 1450 to 1550 g / mol).

[0290] Types of oligosaccharides :In some embodiments, the type of oligosaccharides present in the oligosaccharide preparations mentioned herein may depend on the type of feed sugar. For example, in some embodiments, when the feed sugar includes glucose, the oligosaccharide preparation comprises glucose-oligosaccharides. For example, in some embodiments, when the feed sugar includes fructose, the oligosaccharide preparation comprises fructose-oligosaccharides. For another example, in some embodiments, when the feed sugar includes fructose and glucose, the oligosaccharide preparation comprises glucose-fructose-oligosaccharides.

[0291] In some embodiments, the oligosaccharide preparation comprises one or more types of monosaccharide subunits. In some embodiments, the oligosaccharide preparation can comprise oligosaccharides having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more different types of monosaccharide subunits.

[0292] Method for producing oligosaccharide preparation Oligosaccharide preparations can be made by heating an aqueous composition comprising one or more feed sugars and a catalyst to a temperature and for a time sufficient to induce polymerization, wherein the catalyst is selected from the group consisting of: (+)-camphor-10-sulfonic acid; 2-pyridinesulfonic acid; 3-pyridinesulfonic acid; 8-hydroxy-5-quinolinesulfonic acid hydrate; α-hydroxy-2-pyridinemethanesulfonic acid; (β)-camphor-10-sulfonic acid; butylphosphonic acid; diphenylphosphinic acid; hexylphosphonic acid; methylphosphonic acid ; Phenylphosphinic acid; Phenylphosphonic acid; Tert-butylphosphonic acid; SS)-VAPOL hydrogen phosphate; 6-quinolinesulfonic acid, 3-(1-pyridyl)-1-propanesulfonate; 2-(2-pyridyl)ethanesulfonic acid; 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine-p,p'-disulfonic acid monosodium salt hydrate; 1,1'-binaphthyl-2,2'-diyl-hydrogen phosphate; Bis(4-methoxyphenyl)phosphinic acid; Phenyl(3,5- dimethylphenyl)phosphinic acid; L-cysteic acid monohydrate; poly(styrenesulfonic acid-co-divinylbenzene); lysine; ethanedisulfonic acid; ethanesulfonic acid; isethionic acid; homocysteine; HEPBS (N-(2-hydroxyethyl)piperazine-N'-(4-butanesulfonic acid)); HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid); 2-hydroxy-3-morpholinopropanesulfonic acid; 2-(N-morpholino)ethanesulfonic acid; methanesulfonic acid; formic hydrazide ; naphthalene-1-sulfonic acid; naphthalene-2-sulfonic acid; perfluorobutanesulfonic acid; 6-sulfoquinovose; trifluoromethanesulfonic acid; 2-aminoethanesulfonic acid; benzoic acid; chloroacetic acid; trifluoroacetic acid; hexanoic acid; heptanoic acid; octanoic acid; nonanoic acid; lauric acid; palmitic acid; stearic acid; arachidic acid; aspartic acid; glutamic acid; serine; threonine; glutamine; cysteine; glycine; proline; alanine; valine; isoleucine; leucine; methionine; phenylalanine; tyrosine; tryptophan.

[0293] In some embodiments, polymerization of the feed sugar is achieved by step-growth polymerization. In some embodiments, polymerization of the feed sugar is achieved by condensation polymerization.

[0294] Feed sugarThe one or more feed sugars used in the method for making the oligosaccharide preparation described herein may comprise one or more types of sugar. In some embodiments, the one or more feed sugars include monosaccharides, disaccharides, trisaccharides, tetrasaccharides, or any mixture thereof. The feed sugar can be, for example, one or more selected from glucose, lactose, galactose, mannose, glucosamine, N-acetylglucosamine, arabinose, xylose, sucrose, and fructose.

[0295] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "an agent" includes a plurality of such agents, reference to "the oligosaccharide" includes reference to one or more oligosaccharides (or oligosaccharides) and equivalents thereof known to those skilled in the art, and so forth.

[0296] When ranges are used herein for physical properties (e.g., molecular weight) or chemical properties (e.g., chemical formulae), all combinations and subcombinations of ranges and specific embodiments thereof are intended to be included. When referring to a number or a numerical range, the term "about" means that the number or numerical range referred to is an approximation within the experimental variability (or statistical experimental error), and thus, in some cases, the number or numerical range will vary between 1% and 15% of the number or numerical range.

[0297] In the following, the present invention is further described by way of non-limiting examples. Example

[0298] The present invention as disclosed herein is not limited to the particular implementations, figures, methodologies, examples, schemes, etc. described herein, but only by the claims.

[0299] Example 1

[0300] A library of synthetic oligosaccharide preparations is prepared using different combinations of feed sugars and catalysts. The feed sugars are selected from the group consisting of glucose, lactose, galactose, mannose, glucosamine, N-acetylglucosamine, arabinose, xylose, sucrose, and fructose. The catalyst is selected from sulfuric acid, acetic acid, propionic acid, butyric acid, L-glutamic acid, L-lysine, phosphoric acid, (+)-camphor-10-sulfonic acid, methylphosphonic acid, L-cysteine, hydrochloric acid, citric acid, lactic acid, 3-(1-pyridyl)-1-propanesulfonate, 2-pyridinesulfonic acid, 3-pyridinesulfonic acid, α-hydroxy-2-pyridinemethanesulfonic acid, (β)-camphor-10-sulfonic acid, butylphosphonic acid, diphenylphosphonic acid, hexylphosphonic acid, phenylphosphonic acid, phenylphosphonic acid, tert-butylphosphonic acid, 2-(2-pyridyl)ethanesulfonic acid, 3-(2-pyridyl)-5,6-diphenyl-5,6-diphenyl-1,2,4-triazine-p,p'-disulfonic acid monosodium salt hydrate, 1,1'-binaphthyl-2,2'-diyl hydrogenphosphate, bis(4-methoxyphenyl)phosphinic acid and phenyl(3,5-ditolyl)phosphinic acid.

[0301] In principle, the reaction on a 10 g scale is carried out by charging a corresponding mass of powdered sugar (on a dry solid basis) into a 20 mL scintillation vial, melting the contents by heating to 135° C. under stirring, and allowing water to drain from the reaction vessel for several hours by adding the catalyst and continuing heating until a sufficient conversion is achieved, as indicated by an increase in viscosity, a brown or amber color. Table 1 below shows exemplary combinations of feed sugars and corresponding catalysts for the synthesis of oligosaccharide preparations. OP IDs listed in the order of Table 1 below The feed sugars of 1-61 were used in the following ratios: 98:2, 75:5:20, 85:5:10, 93:5:2, 97:3, 90:5:5, 90:5:5, 68:2:30, 90:5:5, 90:10, 65:5:30, 95:5, 90:5:5, 67:3:30, 95:5, 94:5:1, 95:5, 92:5:3, 90:5:5, 94:5:1, 90:10, 90:5:5, 90:5:5, 95:5, 95:5, 95:5, 85:5:10, 94:5:1, 85:5 :10, 94:1:5, 92:5:3, 98:2, 85:5:10, 90:5:5, 90:5:5, 97:3, 95:5, 90:5:5, 75:5:20, 90:5:5, 98:2, 99:1, 94:5:1, 93:5:2, 85:5:10, 92:5:3, 90:5:5, 92:5:3, 92:5:3, 98:2, 99:1, 90:5:5, 88:2:10, 94:5:1, 92:5:3, 92:5:3, 90:5:5, 93:2:5, 85:5:10 and 94:5:1.

[0302] Table 1: Feed sugars and catalysts used to prepare synthetic oligosaccharide preparations with the indicated oligosaccharide preparation identifiers (OP IDs).

[0303]

[0304]

[0305]

[0306] Example 2

[0307] The amount of catalyst present in the oligosaccharide preparation after production was quantified by anion chromatography. The analysis was performed on a Thermo-Fisher ICS-3000 ion chromatography system equipped with an EG-3000 eluent generator, an AS autosampler, and a CD-3000 conductivity detector. Separation was achieved on an AS19 column (4.0 mm × 250 mm) protected by an AG19 precolumn (4.0 mm × 50 mm). The mobile phase was supplied by an eluent generator (EGCII). A KOH gradient (0-55 mM) was generated using a HPLC-MS / MS / MS cartridge. Chromatographic parameters were: flow rate 1.0 mL / min, injection volume 25 μL, column temperature 30°C. Calculations were based on peak areas. Quantification was achieved by comparing peak areas to an external standard calibration curve. All catalysts used were quantifiable and contained less than 0.1%, or 1000 ppm, after the condensation reaction.

[0308] A sample of the oligosaccharide preparation prepared as in Example 1 was further analyzed by gel permeation chromatography on an Agilent 1260 Infinity system comprising a degasser, a binary pump and a thermostat column compartment equipped with an Agilent 1200 refractive index detector. The columns used were PL aquagel-OH guard columns, 7.5 x 50 mm, 5 μM (Agilent part number: PL1149-1530) and PL aquagel-OH, 7.5 x 300 mm, 5 μM (Agilent part number: PL1120-6520), while the solvent used was 200 mM sodium nitrate in water, the flow rate was 0.9 mL / min, and the column temperature was 40° C. Glucose (for quantification of total sugar content) and pullulan standards (for size distribution) were used as standards. Oligosaccharide preparations with OP IDs 50, 43, 57, and 54 were found to have DP1 contents (DP: degree of polymerization; DP1: degree of polymerization of 1) of 29.5%, 27.3%, 11.6%, and 50.5% by weight, respectively; and DP2+ contents (oligosaccharides with a degree of polymerization of 2 or greater) of 42.2%, 39.2%, 45.0%, and 22.1% by weight, respectively. Additional analysis of OPID 54 by liquid chromatography with an evaporative light scattering detector coupled to high-resolution mass spectrometry (LC-ELSD-HRMS) confirmed a DP1 content of 69.22 area%, a DP2 content of 24.64 area%, and a DP3+ content of 6.14 area%. Furthermore, Mn (number average molecular weight) and Mw (weight average molecular weight) were determined. OP IDs 50, 43, 57, and 54 had Mn values ​​of 477, 574, 710, and 690 g / mol, respectively; and Mw values ​​of 578, 779, 1059, and 1008 g / mol, respectively. OP ID 57 had higher Mn and Mw values ​​than the other OD IDs tested, consistent with the higher DP2+ content of this oligosaccharide preparation. The total dry polysaccharide contents were 71.7 wt%, 66.5 wt%, 56.6 wt%, and 72.6 wt%, respectively.

[0309] In addition, for oligosaccharide preparations with OP IDs 50, 43, 57, and 54, the refractive index was measured at 78.9, 79.6, 77.0, and 73.6 degrees Brix, respectively.

[0310] Example 3

[0311] To test the increase in GABA production by the intestinal porcine microbiome, the following assay was performed under anaerobic conditions. To this end, a minimal medium solution, a polysaccharide solution (containing the oligosaccharide preparation to be tested), a feed extract solution, and a porcine cecal slurry (containing the intestinal microbiota) were prepared.

[0312] The minimal culture medium solution was prepared to contain 900 mg / L sodium chloride, 26 mg / L calcium chloride dihydrate, 20 mg / L magnesium chloride hexahydrate, 40 mg / L ammonium sulfate, 300 mg / L potassium dihydrogen phosphate, 1500 mg / L sodium dihydrogen phosphate, 5000 mg / L sodium bicarbonate, 150 mg / L histidine, 75 mg / L glycine, 150 mg / L tryptophan, 150 mg / L arginine, 150 mg / L methionine, 150 mg / L glutamine, 10 .... L threonine, 225 mg / L valine, 225 mg / L isoleucine, 300 mg / L leucine, 400 mg / L cysteine, 450 mg / L proline, 75 mg / L phenylalanine, 75 mg / L tyrosine, 75 mg / L glutamic acid, 10 mg / L manganese chloride solution, 4 mg / L ferric sulfate solution, 1 mg / L cobalt chloride solution, 1 mg / L pyridoxine solution, 1 mg / L pantothenic acid solution, and 0.125 mg / L biotin solution.

[0313] A polysaccharide solution was prepared at a concentration of 6 Brix for each individual oligosaccharide preparation to be tested, diluted with distilled water. For this purpose, the oligosaccharide preparation OP ID 1-61 prepared in Example 1 was used.

[0314] The feed extract solution was prepared by mixing ground pig feed (as detailed in Table 2) with 20 mL of distilled water, followed by incubation at 125° C. for 15 minutes. Thereafter, the mixture was pelleted by centrifugation and the supernatant was incubated at 121° C. for 15 minutes. Thereafter, sterile distilled water was added to a final volume of 20 mL.

[0315] Table 2: Pig feed composition. * Premixed vitamin and mineral content per kg of diet: Vitamin A 6510 IU; Vitamin E 75 mg; Vitamin K 2 mg; Vitamin D3 2007 IU; Vitamin B1 0.99 mg; Vitamin B2 5.02 mg; Vitamin B6 2.01 mg; Vitamin B12 0.03 mg; Pantothenic acid 17.5 mg; Folic acid 0.51 mg; Biotin 0.1 mg; Choline 100 mg; Manganese 40 mg; Iron 100 mg; Copper 15 mg; Zinc 65 mg; Iodine 2 mg; Selenium 0.4 mg.

[0316]

[0317]

[0318] A porcine cecal slurry was prepared by mixing 3.8 g of cecal sample in 25 mL of PBS (phosphate-buffered saline: 137 mM sodium chloride, 2.7 mM potassium chloride, 10 mM sodium hydrogen phosphate, 1.8 mM potassium dihydrogen phosphate, pH 7.4). After large particles in the mixture settled, the unsettled supernatant was decanted into a fresh container. The supernatant was then centrifuged (500 rpm, 5 min). After centrifugation, the supernatant was recovered and mixed with 50% glycerol to a final glycerol concentration of 15% (v / v) to form a porcine cecal slurry.

[0319] To simulate the actual diversity of the porcine cecal microbiota, three cecal samples containing different microbial compositions were tested. As identified by metagenomics, the cecal samples differed in diversity as well as microbial composition and ratios. In detail, in terms of phyla, cecal sample 1 contained 48.0%, 33.3%, 15.4%, and 3.3% of Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteria, respectively. Cecal sample 2 contained 54.9%, 27.2%, 15.3%, and 2.7% of Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteria, respectively. Cecal sample 3 contained 67.0%, 20.4%, 10.4%, and 2.3% of Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteria, respectively. In terms of genera, cecal sample 1 contained 15.1%, 8.5%, 14.6%, 5.2%, 3.8%, 3.6%, 4.2%, 23.4%, 6.2%, 11.1% and 4.3% of Bacteroides, Blautia, Clostridium, Escherichia, Faecalibacterium, Lachnoclostridium, Phocaeicola, Prevotella, Roseburia, Streptococcus and Turicibacter, respectively. Cecal sample 2 contained 10.8%, 5.5%, 3.7%, 12.3%, 6.0%, 10.3%, 4.9%, 17.9%, 4.4%, 9.9% and 14.3% of Bacteroides, Blautia, Campylobacter, Clostridium, Escherichia, Lactobacillus, Limosilactobacillus, Prevotella, Roseburia, Sarcina and Streptococcus, respectively.Cecal sample 3 contained 6.6%, 5.9%, 4.4%, 1.8%, 2.7%, 41.9%, 18.1%, 2.0%, 10.5%, 4.4% and 1.8% of Bacteroides, Blautia, Clostridium, Faecalibacterium, Lachnoclostridium, Lactobacillus, Limosilactobacillus, Phocaeicola, Prevotella, Roseburia and Ruminococcus, respectively.

[0320] After mixing 50 μL of porcine cecal slurry with 80 μL of feed extract solution, 70 μL of glycan solution, and 800 μL of minimal medium solution in a single well of a 96-deep-well plate, the microbiota began to produce GABA. The glycan solution was varied by testing several different oligosaccharide preparations prepared as described in Example 1. As controls, a blank control was included, in which the glycan solution was replaced with minimal medium solution; and a glucose control was included, in which the glycan solution contained 1% (v / v) glucose instead of the oligosaccharide preparation described herein. The glucose control was included to obtain a basal GABA response from the microbiota, which served as a reference for fold-change calculations. Since glucose is largely absorbed before actually reaching the small intestine, the 1% glucose control appropriately simulated the in vivo situation. The 96-deep-well plate was sealed with a porous, breathable adhesive film and incubated anaerobically at 37°C for 24 hours. Thereafter, the 96-deep-well plate was centrifuged to pellet the microbiota and insoluble material, and the supernatant was used for further analysis, particularly for determining GABA and / or KYN production.

[0321] Example 4

[0322] GABA was measured by liquid chromatography tandem mass spectrometry (HPLC-MS / MS). The supernatant obtained from the in vitro fermentation of Example 3 was centrifuged. Then, 200 μL of the supernatant was transferred to a 96-well plate. The components contained in these supernatants were separated on an Acquity UPLC H-class system, which includes a degasser, a quaternary pump, a sample manager, and a thermostat column compartment equipped with a TQDetector (Waters). The chromatographic columns used were a UPLC BEH C18 guard column, 2.1 x 50 mm, 1.7 μM (Waters reference: 1866003975) and an Acquity UPLC BEH C18, 2.1 x 150 mm, 1.7 μM (Waters reference: 176001048). The eluents were: A 99.8% H2O + 0.2% formic acid; and B 99.8% ACN (LC-MS grade) + 0.2% formic acid. The flow rate was 0.3 mL / min and the column temperature was 40° C. The autosampler was set at 15° C. and the run time was 7.5 minutes. The gradient profile is shown in Table 3.

[0323] Table 3: LC flow rate parameters.

[0324] Time (min) Flow rate (ml / min) ％A 0 0.3 95.0 0.25 0.3 95.0 2.70 0.3 85.0 4.00 0.3 60.0 5.00 0.3 0.0 5.20 0.3 0.0 6.00 0.3 0.0 6.10 0.3 95.0 7.20 0.3 95.0

[0325] As a standard, GABA powder with a calibration range of 3 to 0.0003 μg / mL was used. The chromatographic injection volume was 2 μL, and the syringe was rinsed after each injection. The MS method used was multiple reaction monitoring (MRM) with positive electrospray ionization. For GABA, the parent fragment m / z was 103.72 and the daughter fragment m / z was 85.85. The dwell time was 0.025 s, the cone voltage was 10 V, and the collision voltage was 13 V.

[0326] Kynurenine (KYN) was quantified by ELISA kit according to the supplier's recommendations (FI-EM1862; Euromedex). Briefly, 50 μL of standard, blank, or sample was added to 50 μL of biotin-labeled antibody working solution and incubated at 37°C for 45 minutes. The plate was washed three times with wash buffer, and 100 μL of HRP-streptavidin conjugate was added to each well. The plate was incubated at 37°C for 30 minutes. The plate was then washed five times with wash buffer. 90 μL of TMB substrate was added to each well, followed by incubation at 37°C in the dark for 10-20 minutes. Then, 50 μL of stop solution was added to each well, the absorbance was measured at 450 nm, and correlated with a standard curve of known KYN concentrations.

[0327] Example 5

[0328] As described in Example 3, the synthetic oligosaccharide prepared product of preparation among the embodiment 1 is subjected to the in vitro fermentation of pig microbial group separately, and is analyzed as described in Example 4.Microbial group is contacted with any synthetic oligosaccharide prepared product and the impact in terms of GABA generation is calculated as the log2 times of the GABA that produces compared with the GABA concentration obtained from the microbial group that contacts with glucose, referring to Table 4.Though compared with glucose control (such as OP ID 1,6,14,17,21,23,24,38,41,46,59), some oligosaccharide prepared products do not affect or only slightly affect GABA and produce, but find that other oligosaccharide prepared products can cause the GABA microorganism that is significantly larger than glucose control and produces.Especially, find that the oligosaccharide prepared product that comprises N-acetylglucosamine increases GABA and produces.Similarly, find that the oligosaccharide prepared product that produces with methylphosphonic acid as catalyst increases microbial GABA output. In particular, oligosaccharide preparations made from the same feed sugars but using catalysts other than methylphosphonic acid were found to have no effect or much lower effect on GABA than their corresponding oligosaccharide preparations using methylphosphonic acid as a catalyst (e.g., OP ID 30 compared to 58; 48 compared to 23; 10 compared to 21; 9 compared to 39; 15 compared to 25 and 26).

[0329] Table 4: Log2-fold change in GABA production for individual oligosaccharide preparations compared to glucose control (FC / Glu).

[0330]

[0331]

[0332] Example 6

[0333] In order to prove the ability of the in vitro results of the aforementioned embodiments to be converted into in vivo effects, a 14-day feeding test was carried out in pigs. For this reason, five different treatments were tested, with 12 animals used for each treatment: as a negative control, basic pig feed was used. As a positive control, 30g GABA was added per ton of feed in the same basic pig feed. It has been described that this GABA is supplemented with 30g / MT to increase growth performance, improve stress-related parameters and reduce the incidence of bites (Li et al. 2015.Can.J.Anim.Sci.95:165-171; Bi et al. 2020.J Anim Physiol Anim Nutr.104:590-596). Three test treatments were prepared by spraying any one of the oligosaccharide preparations of OP ID 50, 43 and 57 as mash onto basic pig feed. The oligosaccharide preparation was provided in a syrup form at a concentration of 70 Brix and added to the feed at a level of 1000mg / kg of inclusion compound. For negative and positive controls, an equal volume of water was sprayed as mash on the basal pig feed instead of the syrup of any test treatment.

[0334] The composition of the basal pig feed is shown in Table 5 below. During the experimental period, feed was provided as a mash for ad libitum consumption. Phytase (Ronozme HiPhos) at 100 mg / kg feed was included in the basal diet as a premix using ground corn as a carrier.

[0335] Table 5: Basic pig feed. * Premixed vitamin and mineral content per kilogram of diet: Vitamin A 6510 IU; Vitamin E 75 mg; Vitamin K 2 mg; Vitamin D3 2007 IU; Vitamin B1 0.99 mg; Vitamin B2 5.02 mg; Vitamin B6 2.01 mg; Vitamin B12 0.03 mg; Pantothenic acid 17.5 mg; Folic acid 0.51 mg; Biotin 0.1 mg; Choline 100 mg; Manganese 40 mg; Iron 100 mg; Copper 15 mg; Zinc 65 mg; Iodine 2 mg; Selenium 0.4 mg.

[0336]

[0337] Animal technical parameters were monitored (mortality, weight gain, final body weight, FCR - feed conversion ratio). GABA and KYN were measured in plasma and fecal samples; cortisol (COR), adrenocorticotropic hormone (ACTH), serum neuropeptide Y (NPY) and lactate were measured in plasma samples.

[0338] Performance Statistical Analysis: For this study, the experimental unit was the pen, n = 4 pigs / pen. Data were analyzed by one-way ANOVA followed by Tukey's multiple comparison test. The effect of treatment was considered significant at p < 0.05. For analysis, JMP software (version 16.0) was used. Statistical Biological Analysis: For GABA and KYN and stress-related markers COR, ACTH, NPY, glucose / lactose, in this study, the experimental unit was the pig, n = 12 pigs / treatment. Data were analyzed by one-way ANOVA followed by Tukey's multiple comparison test. The effect of treatment was considered significant at p < 0.05. For analysis, JMP software (version 16.0) was used.

[0339] Compared to day 0, the fold changes in plasma GABA on day 12 were 1.07 and 1.09 for the positive control and OP ID 57 treatment, respectively; a slight increase of 1.12 for OP ID 50 treatment; and a significant increase of 1.73 for OP ID 43 treatment. The fold changes in plasma COR, a stress marker, were significantly reduced with OP ID 50 and 43 treatments (0.27 and 0.38, respectively), compared to the fold changes for the negative control, positive control, and OP ID 57 treatment (0.66, 0.70, and 0.77, respectively). Consistent with these findings, the fold changes in plasma NPY, a stress resistance marker, were significantly increased (0.93 and 1.60, respectively) with OP ID 50 and 43 treatments (compared to day 0), compared to the fold changes for the negative control, positive control, and OP ID 57 treatment (0.57, 0.51, and 0.50, respectively). Thus, the findings demonstrate the scalability of the in vitro results to in vivo effects. As a further consequence of the identified differences in GABA and stress-related markers, a lower incidence of aggressive behaviors (e.g., tail biting) as well as reduced stress, fear, and anxiety, as well as improved animal performance in terms of weight gain and FCR, could be observed.

[0340] Example 7

[0341] In another feeding trial, 288 pigs were used for a 44-day study, including a 28-day growth period and a 16-day fattening period. Six treatments were tested, with 48 animals used in each treatment. As a negative control, a basal diet feed was provided. The basal diet feed is shown in Table 6 below. The GABA control was prepared by supplementing 30mg GABA / kg feed in the basal diet feed. In four test treatments, the oligosaccharide preparation of OPID 54 was added to the basal diet feed at a rate of 50, 150, 250 or 500mg oligosaccharide preparations per kg feed. In order to compensate for the quality difference between the processed feed and other processed feeds comprising 500mg oligosaccharide preparations / kg feed, silicon dioxide was added. Specifically, 0.5 kg SiO2 / 1000 kg feed was added to the negative control treatment feed; 0.47 kg SiO2 / 1000 kg feed was added to the GABA control treatment feed in addition to 0.03 kg GABA per 1000 kg feed; and 0.45 kg, 0.35 kg, and 0.25 kg SiO2 per 1000 kg feed were added to treatment feeds containing 50 mg, 150 mg, and 250 mg of the oligosaccharide preparation per 1000 kg feed, respectively.

[0342] As in the previous examples, during the experimental period, feed was provided as a mash for ad libitum consumption. 100 mg / kg of phytase Ronozme HiPhos was included in a basal diet (2*150 g) in a premix, using ground corn as a carrier. The main feed raw materials were ground using a 4 mm sieve in a hammer mill and mixed with other ingredients (minerals, vitamins, and amino acids) in a 1500 L mixer. The polysaccharide product was provided in the form of a syrup sprayed on the mash feed. For the negative control feed, the same volume of water was sprayed.

[0343] Table 6: Basal diet. *Premixed vitamin and mineral content per kilogram of diet: Vitamin A 6510 IU; Vitamin E 75 mg; Vitamin K 2 mg; Vitamin D3 2007 IU; Vitamin B1 0.99 mg; Vitamin B2 5.02 mg; Vitamin B6 2.01 mg; Vitamin B12 0.03 mg; Pantothenic acid 17.5 mg; Folic acid 0.51 mg; Biotin 0.1 mg; Choline 100 mg; Manganese 40 mg; Iron 100 mg; Copper 15 mg; Zinc 65 mg; Iodine 2 mg; Selenium 0.4 mg. **Premixed vitamin and mineral 3145 provided per kilogram of diet: Vitamin A: 15,000 IU; Vitamin E: 100 mg; Vitamin K: 20.0 mg; Vitamin C: 100 mg; Vitamin B1: 3 mg; Vitamin B2: 10 mg; Vitamin B6: 6 mg; Vitamin B12: 0.04 mg; Niacin: 60.0 mg; Pantothenic acid: 25.0 mg; Folic acid: 1.5 mg; Biotin 0.2 mg; Choline: 325 mg; Manganese: 60 mg; Iron: 200 mg; Copper: 140 mg; Zinc: 100 mg; Iodine: 2 mg; Selenium: 0.4 mg.

[0344]

[0345]

[0346] In order to study the effect of providing the oligosaccharide preparation according to the present invention to animals, particularly with regard to animal welfare, a mixing challenge was carried out. It is known that the agitated behavior after mixing individual pigs between groups is an animal welfare issue, resulting in skin damage on the body, decreased growth performance and increased aggressive behavior. At the beginning of the experiment, pigs were grouped by weight. On the 28th day, all pigs were weighed before mixing, and then one pig was selected from each fence and exchanged with pigs of similar weight from the same treatment group but from another fence. Performance data (weight, weight gain, FCR), social behavior and skin lesions were monitored. Metagenomic analysis was performed from fecal samples. GABA, serotonin (5-HT), Trp (tryptophan) and KYN were analyzed from plasma and fecal samples. COR, ACTH, NPY, chromogranin A (CgA) and haptoglobin (HPT) were analyzed from plasma samples. Statistics were performed according to the above examples.

[0347] Consistent with the previous examples, when the oligosaccharide preparation of the present invention was given to animals along with their feed, an increase in plasma GABA fold change was observed compared to the negative control and GABA control. Furthermore, when the oligosaccharide preparation of the present invention was given to animals along with their feed, a significant decrease in plasma cortisol fold change and a significant increase in plasma 5-HT fold change were observed compared to the negative control. Between days 0 and 28 of the experiment (i.e., prior to the admixture challenge), the average daily gain (ADG) of the test treatment group was 3.0% higher than that of the negative control: 862 g / d and 837 g / d, respectively. Admixture had a negative impact on ADG, as reflected by ADG values ​​of 769 g / d and 810 g / d for the negative control and test treatment groups on days 28-42, respectively. However, despite the stress imposed on the animals during admixture, the ADG of the test treatment group receiving the oligosaccharide preparation was 5.3% higher than that of the negative control. When comparing ADG on days 0-42, the test treatment group (845 g / d) was significantly (P=0.04) higher than the negative control group (814 g / d). Thus, it was found that feeding the oligosaccharide preparation according to the present invention generally improves the performance of animals in terms of ADG, especially when the animals are under stress. In the negative control group and the test treatment group, no significant effect was found on the average daily feed intake (ADFI) at 2004 g / day and 1991 g / day on days 0-28; 2577 g / day and 2605 g / day on days 28-42; and 2195 g / day and 2196 g / day on days 0-42, respectively. Thus, feed conversion ratio was reduced by 3.3% in the test treatment group (2.32) compared to the negative control (2.40) on days 0-28; and by 1.1% in the test treatment group (3.32) compared to the negative control (3.36) on days 28-42; and significantly (P=0.07) reduced by 3.5% in the test treatment group (2.60) compared to the negative control (2.70) on days 0-42. It was concluded that the animals in the negative control group did not convert the energy of the feed consumed during the stress period into animal growth, while the animals receiving the oligosaccharide preparation were less sensitive to stress and more successfully converted the energy from the feed into growth.

[0348] Example 8

[0349] A further animal experiment was conducted using 144 growing pigs (72 females, 72 intact males) of the [Large White x Landrace] x Pietrain breed, with undocked tails. Two treatments were tested: a basal diet as shown in Table 7 was fed as a negative control. A basal diet supplemented with 250 g of dry OP ID54 per metric ton of feed was used as the test treatment. Feed was provided in pellet form for ad libitum consumption.

[0350] The experiment lasted for 105 days, with an initial 7-day adaptation period (d-7 to d0), followed by dietary treatment starting on d0. On d55 and d56, the animals were subjected to a mixed challenge similar to that in Example 7. The experiment ended on d98, with final body weights of approximately 100-105 kg.

[0351] It was found that throughout the trial, particularly during the stress period established by the mixed challenge, feeding the oligosaccharide composition of the present invention improved animal performance (e.g., in terms of daily weight gain, feed intake, feed to weight gain ratio, etc.). Notably, during the mixed challenge week (d51-d59), animals receiving the oligosaccharide composition of the present invention showed significant improvements in growth performance (final body weight, weight gain, FCR). See Tables 8-11 below for details.

[0352] Saliva samples were collected on d-7 (baseline), d58 (after the mixed challenge), and d97 (end of the experiment). Hair samples were collected on d-7 and d98 (to detect potential accumulation of cortisol during the treatment period d-7 to d98). As a non-invasive method for measuring stress response, saliva and hair samples were tested for cortisol content. Notably, the cortisol concentrations in saliva samples of the test treatment group were significantly reduced by 24.9% and 23.9% on d58 and d97, respectively, compared to the animals in the control group. Similarly, the cortisol concentrations in hair samples of the test treatment group were reduced by 17.5% on d98, compared to the animals in the control group. Thus, animals receiving the oligosaccharide preparation of the present invention exhibited a significantly reduced stress response after the mixed challenge and exhibited lower chronic stress caused by the mixed challenge, as found by the lower accumulation of the stress biomarker cortisol in the hair samples. Consistent with these findings, one and two days after the mixed challenge, animals that received the oligosaccharide preparation of the invention were found to have a reduced number of severe or moderately severe lesions on the ears, body and / or tail compared to animals that received the control diet, thus indicating a reduction in aggressive behavior in these animals.

[0353] Table 7: Basic diet feed. 1Vitamin and mineral premix provided per kg feed: Vitamin A (E 672) 5500 IU; Vitamin D3 (E 671) 1100 UI; Vitamin E (α-tocopherol) 25 mg; Vitamin B1 0.5 mg; Vitamin B2 1.4 mg; Vitamin B6 1 mg; Vitamin B12 8 μg; Vitamin K3 0.5 mg; Calcium pantothenate 5.6 mg; Nicotinic acid 8 mg; Choline 120 mg; Fe (E 1) (from FeSO4·7H2O) 80 mg; I (E 2) (from Ca(I2O3)2) 0.5 mg; Co (E 3) (from 2CoCO3·3CO(OH)2·H2O) 0.4 mg; Cu (E 4) (from CuSO4·5H2O) 5 mg; Cu (E 4) (from amino acid chelate) 5 mg; Mn (E 5) (from MnO) 40 mg; Zn (E6) (from ZnO) 100 mg; Se (E 8) (from Na2SeO3) 0.25 mg. 2 Noxyfeed is an ITPSA product from Barcelona, ​​Spain. It contains BHT + propyl gallate (56%) and citric acid (14%).

[0354]

[0355]

[0356] Table 8: Animal performance on days 0-29. "Control" animals were fed a negative control diet; "Test" animals were fed a test treatment diet.

[0357]

[0358] Table 9: Animal performance on days 29-64. "Control" animals were fed a negative control diet; "Test" animals were fed a test treatment diet.

[0359]

[0360]

[0361] Table 10: Animal performance on days 64-98. "Control" animals were fed a negative control diet; "Test" animals were fed a test treatment diet.

[0362]

[0363] Table 11: Animal performance on days 51-59. "Control" animals were fed a negative control diet; "Test" animals were fed a test treatment diet.

[0364]

Claims

1. For the following methods: i) regulating animal behavior, particularly to reduce aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; and / or v) reducing plasma, saliva and / or hair cortisol concentrations in animals exposed to stress; The method comprises the following steps: Provided is an oligosaccharide preparation, which is produced or capable of being produced by the following steps: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2; feeding the oligosaccharide preparation to the animal; as well as Increase the level of gamma-aminobutyric acid, kynurenine and / or serotonin in the animal's body, especially in the gastrointestinal tract and / or blood of the animal.

2. Methods for increasing the production of gamma-aminobutyric acid, kynurenine, and / or serotonin by a microbial community; The method comprises the following steps: Provided is an oligosaccharide preparation, which is produced or capable of being produced by the following steps: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2; and contacting the microbial community with the oligosaccharide preparation.

3. A method for producing an oligosaccharide preparation for: i) regulating animal behavior, particularly to reduce aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia; iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in an animal, particularly in the gastrointestinal tract and / or blood of the animal; and / or iv) reducing plasma, saliva and / or hair cortisol concentrations in animals exposed to stress; The method comprises the following steps: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid; Thereby the oligosaccharide preparation is produced, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2 and at least a third fraction having a DP greater than 2.

4. The method according to any one of the preceding claims, wherein the at least second sugar of step a) is selected from the group consisting of glucose, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably glucose.

5. The method according to any one of the preceding claims, wherein the at least first sugar of step b) is glucose, and wherein the at least second sugar of step b) is selected from N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose.

6. The method according to any one of the preceding claims, wherein the at least first sugar of either step a) or b); and the at least second sugar of either step a) or b) are condensed with at least a third sugar selected from the group consisting of glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably lactose.

7. An oligosaccharide preparation for: i) regulating animal behavior, particularly to reduce aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia; iii) increasing the level of gamma-aminobutyric acid, kynurenine and / or serotonin in an animal, particularly in the gastrointestinal tract and / or blood of the animal; and / or iv) reducing plasma, saliva and / or hair cortisol concentrations in animals exposed to stress; The oligosaccharide preparation is produced or can be produced by the following steps: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid; And wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2.

8. The oligosaccharide preparation according to claim 7, wherein the at least second sugar of step a) is selected from the group consisting of glucose, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably glucose.

9. The oligosaccharide preparation according to claim 7 or 8, wherein the at least first sugar of step b) is glucose, and wherein the at least second sugar of step b) is selected from N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose.

10. The oligosaccharide preparation according to any one of claims 7 to 9, wherein the at least first saccharide of any one of steps a) or b); and the at least second saccharide of any one of steps a) or b) are condensed with at least a third saccharide selected from the group consisting of glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose and arabinose; preferably lactose.

11. The oligosaccharide preparation according to any one of claims 7 to 10, wherein the oligosaccharide composition comprises at least 10% by weight (11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, etc.) of the at least first fraction; preferably at least 20% by weight.

12. The oligosaccharide preparation according to any one of claims 7 to 11, wherein the oligosaccharide preparation is included in the nutritional composition at an inclusion rate of at least 50 ppm (e.g. at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm).

13. Use of the oligosaccharide preparation according to any one of claims 7 to 12 for: i) regulating animal behavior, particularly to reduce aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and / or phobia; iii) improving animal performance, in particular for improving average weight gain and / or reducing feed conversion ratio; iv) improving animal performance in animals exposed to stress, in particular for improving average weight gain and / or reducing feed conversion ratio; v) increasing the production of gamma-aminobutyric acid, kynurenine and / or serotonin by the microflora; and / or vi) Reducing Cortisol concentrations in the plasma, saliva and / or hair of animals exposed to stress.

14. An oligosaccharide preparation for use in treating, ameliorating, preventing and / or prophylaxis of one or more conditions associated with an imbalance in gamma-aminobutyric acid, kynurenine and / or cortisol levels; wherein the one or more conditions are selected from anxiety, stress, phobias, systemic inflammation and local inflammation; wherein the oligosaccharide preparation is produced or capable of being produced by the following steps: a) condensing at least a first saccharide with at least a second saccharide using a catalyst, wherein the at least first saccharide is N-acetylglucosamine; or b) condensing at least a first saccharide with at least a second saccharide using a catalyst; wherein the catalyst is methylphosphonic acid; And wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1, at least a second fraction having a DP of 2, and at least a third fraction having a DP greater than 2.

15. An oligosaccharide preparation for use in treating, ameliorating, preventing and / or prophylaxis of one or more conditions associated with an imbalance in gamma-aminobutyric acid, kynurenine and / or cortisol levels according to claim 14; wherein the one or more conditions are selected from anxiety, stress, phobias, systemic inflammation and local inflammation; wherein the oligosaccharide preparation is included in the nutritional composition at an inclusion rate of at least 50 ppm (e.g., at least 50 ppm, 70 ppm, 100 ppm, 150 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm).

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