WO2024175697A1 - Moyens et procédés de modulation du comportement - Google Patents

Moyens et procédés de modulation du comportement Download PDF

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Publication number
WO2024175697A1
WO2024175697A1 PCT/EP2024/054484 EP2024054484W WO2024175697A1 WO 2024175697 A1 WO2024175697 A1 WO 2024175697A1 EP 2024054484 W EP2024054484 W EP 2024054484W WO 2024175697 A1 WO2024175697 A1 WO 2024175697A1
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Prior art keywords
sugar
animal
oligosaccharide preparation
catalyst
fraction
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PCT/EP2024/054484
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English (en)
Inventor
Elkin AMAYA
Estel CANET-MARTINEZ
John Michael Geremia
Daniel Beat JOSS
Estefania Perez Calvo
Wen REN
Jerome SCHMEISSER
Ghislain Schyns
Clement SESTER
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DSM IP Assets BV
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DSM IP Assets BV
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Priority to JP2025544704A priority Critical patent/JP2026506852A/ja
Priority to EP24705697.1A priority patent/EP4669126A1/fr
Priority to CN202480013815.5A priority patent/CN120731013A/zh
Priority to KR1020257030885A priority patent/KR20250153809A/ko
Publication of WO2024175697A1 publication Critical patent/WO2024175697A1/fr
Priority to MX2025009882A priority patent/MX2025009882A/es
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]

Definitions

  • the present invention relates to a method for modulating behavior of an animal, improving animal welfare, and for increasing production of gamma-aminobutyric acid, kynurenine and/or serotonin improving animal performance, using an oligosaccharide preparation.
  • Secondary metabolites are molecules produced by an organism, and exert their function in interactions of the producing organism with its environment. Some well known secondary metabolites are caffeine, taxol, nicotine, artemisinin, morphine, botulinum toxin etc. Notably, many secondary metabolites may serve several purposes. For instance, caffeine is believed to be produced by plants as a pesticide, while humans consume caffeine to increase vigilance. Other secondary metabolites produced by one organism may act as a neurotransmitter in another organism. Neurotransmitters are signaling molecules which allow communication among cells of an organism. Receptors for neurotransmitters may be present e.g. on muscle cells or neurons. Gamma-aminobutyric acid (GABA; CAS-No.
  • GABA Gamma-aminobutyric acid
  • GABA slows or blocks specific nerve signals in the brain, thereby modulating levels of anxiety, fear and/or stress as perceivable by humans and animals. Accordingly, imbalanced GABA activity in the body can lead to associated disorders, such as anxiety disorders.
  • Kynurenine is known as a neuromodulator of stress.
  • TRP time to gouphan
  • kynurenine/tryptophan ratio is linked to higher social disturbance profile. It is hypothesized that an increase of the kynurenine/tryptophan ratio is responsible for reducing social disturbance behavior, thus improving animal welfare.
  • Serotonin within the central nervous system cannot cross the blood/brain barrier, while tryptophan is capable of crossing the blood/brain barrier. Therefore, higher tryptophan in the gut means more tryptophan will cross the blood/brain barrier, where it can be transformed to central serotonin.
  • Serotonin is the precursor of melatonin. An increase in serotonin level will thus cause an increase in melatonin level. It is known that melatonin and its precursor serotonin can impact the production of insulin and glucagon. An increase in the melatonin concentration can enhance the level of insulin and glucagon in animal body. It is also known that increased levels of insulin and glucagon enhance the synthesis of fat.
  • Both insulin and melatonin are further involved in regulating circadian rhythm. Changes in the light cycle affect the levels of insulin and melatonin produced by an animal. The changed level of insulin and melatonin in the body of the animal in turn regulates the animal’s physiological response to the light cycle change. Poultry production in general, and broiler rearing process is now going to long light time, as much as 23 hours a day. This illumination regimen strongly impacts production performance such as faster fat gain but is detrimental to animal welfare.
  • Tail biting is an abnormal, pathological behavior frequently observed in weaner and grower-finisher in commercial pig production, and thus a major health and welfare problem due to the risk of infection, the pain and stress experienced by the bitten animal as well as the additional stress experienced by the entire animal group.
  • Previous studies have shown that pigs that perform tail biting have a different concentration of neurotransmitters than pigs that do not develop that aggressive behavior. It is increasingly being recognized that neurochemicals or neurotransmitters produced from either or both, an animal and/or its microbiome represent a common language for host-microbe communication.
  • microbial endocrinology plays a role in animal behavior through the microbiota-gut-brain axis (MGBA) (Lyte et al. 2019. Animal. 13(11):2689-2698). It is known that gut microbiota are capable of synthesizing neuroactive molecules that are structurally similarly to host-derived neurotransmitters including y- aminobutyric acid (GABA), indole-derivatives and catecholamines such as dopamine and norepinephrine. Many of these co-synthesized neurotransmitters are involved in regulation of the host mood, behavior and cognition.
  • GABA y- aminobutyric acid
  • catecholamines such as dopamine and norepinephrine
  • GABAergic system is widely distributed in the hippocampus of piglets and has an inhibitory action on the neurons in the hippocampus.
  • GABA is synthesized through the decarboxylation of glutamate, catalyzed by glutamic acid decarboxylase (GAD), and is widely distributed in the nervous system to reduce the activity of neurons by binding to GABA receptors.
  • GABA glutamic acid decarboxylase
  • Brain levels of GABA are low in mice and rats that exhibit aggressive behaviors, which have been interpreted as being concordant with the proposed inhibitory role of GABA in aggression (Clement et al. 1987. Pharmacol Biochem Behav. 26(1):83-8).
  • direct supplementation with GABA can regulate stress, which is reflected in reducing aggressive behaviors and improving growth performance.
  • GABA supplementation could improve the transport stress of growing-finishing pigs (Bi et al. 2020. J Anim Physiol Anim Nutr. 104: 590-596). Notably however, direct external supply of GABA yielded conflicting results with limited efficacy so far. Also, the half-life of GABA in the body is short (e.g. less than 17 min in mice), thus further reducing efficacy and efficiency of direct GABA administration.
  • this objective is achieved by providing a method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; iv) for improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or v) for reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; the method comprising the steps of 1) providing an oligosaccharide preparation produced by or producible by a) condensing at least a first sugar with at least a second sugar using a catalyst, where
  • the method according to the invention for iv) improving animal performance of animals facing stress in particular for improving average body weight gain and/or reducing feed conversion ratio; relates to the improvement of animal performance of animals facing stress, which animals facing stress received the oligosaccharide preparation, compared to animals facing stress not having received the oligosaccharide preparation.
  • the method according to the invention for v) reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress relates to the reduction of cortisol in plasma, saliva and/or hair of animals facing stress, which animals facing stress received the oligosaccharide preparation, compared to animals facing stress not having received the oligosaccharide preparation.
  • the invention relates to a method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or iv) for improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; the method comprising the steps of 1) providing an oligosaccharide preparation, comprising 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; or b) condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid; thus producing the oligosaccharide preparation, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of
  • condensation or “condensation” as used herein are to be interpreted as the steps necessary for allowing combination of feed sugars or starting sugars, e.g. simple sugars (monosaccharides), with one another to form larger sugars (disaccharides, oligo- or polysaccharides). For instance, two monosaccharides may be combined with one another to form a disaccharide. Analogously, one monosaccharide may be condensed with a disaccharide to form a trisaccharide etc. In the course of such a condensation reaction, water molecule is released.
  • condensation is initiated by applying heat to a mixture of feed sugar(s) in the presence of a catalyst.
  • the invention relates to a method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; iv) for improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or v) for reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; the method comprising the steps of 1) providing an oligosaccharide preparation produced by or producible by condensing at least a first sugar with at least a second sugar using a catalyst, wherein the at least first sugar 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,
  • DP degree of polymer
  • the invention relates to a method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; iv) for improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or v) for reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; the method comprising the steps of 1) providing an oligosaccharide preparation produced by or producible by condensing at least a first sugar with at least a second sugar 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
  • DP degree of polymerization
  • the invention relates to a method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; iv) for improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or v) for reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; the method comprising the steps of 1) providing an oligosaccharide preparation produced by or producible by condensing at least a first sugar with at least a second sugar using a catalyst, wherein the at least first sugar 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
  • 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.
  • 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 glucose, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably glucose.
  • the at least first sugar and the at least second sugar of b) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) are not the same sugar(s) as 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).
  • the at least first sugar and the at least second sugar of b) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) are both glucose.
  • the at least first sugar of b) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) is glucose; and the at least second sugar of b) (condensing at least a first sugar with at least a 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.
  • the at least first sugar and the at least second sugar of 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 condensed with at least a third sugar selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from lactose.
  • the at least first sugar and the at least second sugar of b) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) is condensed with at least a third sugar selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from lactose.
  • a method according to the 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 lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose.
  • At least one of the sugars of a) is/are glucose; wherein said glucose is used at an amount from 50-99% (e.g. wt%) of all sugars that are being used in the condensation step.
  • the at least second sugar is used at an amount from 1-50% (e.g. wt%) of all sugars that are being used in the condensation step.
  • the at least third sugar is used at an amount from 1-50% (e.g. wt%) of all sugars that are being used in the condensation step.
  • a) is performed by condensing at least a first sugar with at least a second sugar and at least a third sugar using a catalyst, wherein the at least first sugar, the at least second sugar and the at least third sugar are used at a ratio of 50-99 : 1-50 : 1-50, preferably wherein at least one of the at least first sugar, the at least second sugar and the at least third sugar is glucose.
  • At least one of the sugars of b) is/are glucose; wherein said glucose is used at an amount from 50-99% (e.g. wt%) of all sugars that are being used in the condensation step.
  • the at least second sugar is used at an amount from 1-50% (e.g. wt%) of all sugars that are being used in the condensation step.
  • the at least third sugar is used at an amount from 1-50% (e.g. wt%) of all sugars that are being used in the condensation step.
  • b) is performed by condensing at least a first sugar with at least a second sugar and at least a third sugar using a catalyst, wherein the catalyst is methylphosphonic acid, wherein the at least first sugar, the at least second sugar and the at least third sugar are used at a ratio of 50- 99 : 1-50 : 1-50, preferably wherein at least one of the at least first sugar, the at least second sugar and the at least third sugar is glucose.
  • the invention relates to a method for increasing 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 producible by 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; or by b) condensing at least a first sugar with at least a second sugar 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 of more than 2; and contacting the microbial community with the oligosaccharide preparation.
  • DP degree of polymerization
  • the invention relates to a method for increasing 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 sugar with at least a second sugar using a catalyst, wherein the at least first sugar is N- acetylglucosamine; or b) condensing at least a first sugar with at least a second sugar 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 of more than 2; and contacting the microbial community with the oligosaccharide preparation.
  • DP degree of polymerization
  • a person skilled in the art will understand that the production of gamma-aminobutyric acid, kynurenine and/or serotonin (e.g. by a microbial community) will start upon contacting one or more cell(s) (e.g. of the microbial community) with the oligosaccharide preparation, and the amount(s) of gamma-aminobutyric acid, kynurenine and/or serotonin produced will increase with ongoing incubation or fermentation time.
  • the microbial community is an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably an animal intestinal microbial community, e.g.
  • pet animals dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, boar, zoo animals, horse, donkey, poultry, swine, ruminant, chicken, cow, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, crayfish, crab, oyster, mussel, clam, trout, tilapia, salmon, carp, catfish, tuna, preferably selected from the group consisting of poultry such as chicken, swine, and ruminant such as cow.
  • the present invention relates to a method for producing an oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) increasing a level of gamma-aminobutyric acid, kynurenine and/or serotonin in the body of the animal, in particular in the gastrointestinal tract and/or in the blood of the animal; and/or for iv) reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; the method comprising the step of: 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; or b) condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid; thus producing the oligos
  • the invention relates to a method for producing an oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) increasing a level of gamma-aminobutyric acid, kynurenine and/or serotonin in the body of the animal, in particular in the gastrointestinal tract and/or in the blood of the animal; and/or for iv) reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; the method comprising the step of: condensing at least a first sugar with at least a second sugar using a catalyst, wherein the at least first sugar is N-acetylglucosamine, and wherein the catalyst is methylphosphonic acid; thus producing the oligosaccharide preparation, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of
  • the invention relates to an oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; for iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or for v) reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; wherein the oligosaccharide preparation is produced or producible by 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; or by b) condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid; and wherein the oligosaccharide preparation is produced or
  • the invention relates to an oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; for iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or for v) reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; wherein the oligosaccharide preparation is produced or producible by condensing at least a first sugar with at least a second sugar using a catalyst, wherein the at least first sugar 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
  • DP degree of polymerization
  • 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.
  • 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 glucose, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably glucose.
  • the at least first sugar and the at least second sugar of b) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) are not the same sugar(s) as 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).
  • the at least first sugar and the at least second sugar of b) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) are both glucose.
  • the at least first sugar of b) (condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid) is glucose; and the at least second sugar of b) (condensing at least a first sugar with at least a 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.
  • the at least first sugar and the at least second sugar of 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 condensed with at least a third sugar selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from lactose.
  • the at least first sugar and the at least second sugar of b) are condensed with at least a third sugar selected from glucose, N- acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from lactose.
  • 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 lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose.
  • the invention relates to an oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; for iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or for v) reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; wherein the oligosaccharide preparation comprises at least two distinct 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
  • the invention relates to an oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; for iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or for v) reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress; wherein the oligosaccharide preparation comprises at least two distinct monosaccharide units selected from glucose, N-acetylglucosamine, and at least a third distinct 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
  • the oligosaccharide preparation according to the invention comprises at least 10 wt%, (11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50 wt% etc.) of the at least first fraction; preferably at least 20 wt%.
  • the oligosaccharide preparation according to the invention is comprised in a nutritional composition at an inclusion rate of at least 50 ppm (e.g. at least 50, 70, 100, 150, 200, 300, 400, 500 ppm).
  • 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 etc. fractions) each having a distinct DP.
  • the oligosaccharide preparation according to the invention comprises not more than three distinct monosaccharide units.
  • the oligosaccharide preparation according to the 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 by 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% anhydro-subunit containing oligosaccharides by relative abundance.
  • the oligosaccharide preparation according to the invention has a weight averaged molecular mass from 300 to 5000 g/mol (e.g. from about 2000 to 2800 g/mol, 2100 to 2700 g/mol, 2200 to 2600 g/mol, 2300 to 2500 g/mol, or 2320 to 2420 g/mol), 500 to 5000 g/mol, 700 to 5000 g/mol, 500 to 2000 g/mol, 700 to 2000 g/mol, 700 to 1500 g/mol, 300 to 1500 g/mol, 300 to 2000 g/mol, 300 to 700 g/mol, 400 to 1300 g/mol, 400 to 1200 g/mol, 400 to 1100 g/mol, 500 to 1300 g/mol, 500 to 1200 g/mol, 500 to 1100 g/mol, 600 to 1300 g/mol, 600 to 1200 g/mol, or 600 to 1100 g/mol); and/or wherein the oligo
  • the invention relates to a use of an oligosaccharide preparation according to the invention and as referred to herein, for i) modulating behavior of an animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; for iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; for v) increasing production of gamma-aminobutyric acid, kynurenine and/or serotonin by a microbial community; and/or for vi) reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress.
  • the invention relates to a use of an oligosaccharide preparation according to the invention and as referred to herein, for i) modulating behavior of a healthy animal, in particular for reducing aggressive behavior; for ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders of a healthy animal; for iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio of a healthy animal; for iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio of a healthy animal facing stress; for v) increasing production of gamma-aminobutyric acid, kynurenine and/or serotonin by a microbial community; and/or for vi) reducing cortisol concentration in plasma, saliva and/or hair of animals facing stress.
  • the invention relates to an oligosaccharide preparation for use in treatment, amelioration, prevention and/or prophylaxis of one or more disorders associated with an imbalanced level of gamma-aminobutyric acid, kynurenine and/or cortisol, wherein the one or more disorders are selected from the group of anxiety, stress, fear disorders, systemic inflammation, and local inflammation, wherein the oligosaccharide preparation is produced or producible by 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; or by b) condensing at least a first sugar with at least a second sugar 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
  • DP degree of poly
  • the invention relates to an oligosaccharide preparation for use in treatment, amelioration, prevention and/or prophylaxis of one or more disorders associated with an imbalanced level of gamma-aminobutyric acid, kynurenine and/or cortisol, wherein the one or more disorders are selected from the group of anxiety, stress, fear disorders, systemic inflammation, and local inflammation, in an animal suffering from or at risk at suffering from one or more disorders associated with an imbalanced level of gamma-aminobutyric acid and/or kynurenine, wherein the oligosaccharide preparation is produced or producible by 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; or by b) condensing at least a first sugar with at least a second sugar using a catalyst, wherein the catalyst is methylphosphonic acid; and wherein the oligosaccharide preparation is
  • Animals suffering from one or more disorders associated with an imbalanced level of gamma- aminobutyric acid and/or kynurenine may be animals (e.g. swine) e.g. being subjected to a situation prone to cause social unease for said animals, e.g. mixing of individuals amongst groups of animals. Analogously, e.g. animals about to being subjected to such situation(s) may be considered to be at risk at suffering from one or more disorders associated with an imbalanced level of gamma-aminobutyric acid and/or kynurenine.
  • the oligosaccharide preparation for use in treatment, amelioration, prevention and/or prophylaxis is an oligosaccharide preparation according to the invention and as described herein.
  • the invention relates to an oligosaccharide preparation for use in treatment, amelioration, prevention and/or prophylaxis of one or more disorders associated with an imbalanced level of gamma-aminobutyric acid, kynurenine and/or cortisol, wherein the one or more disorders are selected from the group of anxiety, stress, fear disorders, systemic inflammation, and local inflammation, wherein the oligosaccharide preparation is comprised in a nutritional composition at an inclusion rate of at least 50 ppm (e.g. at least 50, 70, 100, 150, 200, 300, 400, 500 ppm).
  • ppm e.g. at least 50, 70, 100, 150, 200, 300, 400, 500 ppm
  • the animal referred to herein is selected from the group consisting of pet animals, dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, boar, zoo animals, horse, donkey, poultry, swine, ruminant, chicken, cow, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, crayfish, crab, oyster, mussel, clam, trout, tilapia, salmon, carp, catfish, tuna; preferably, the animal is selected from the group consisting of poultry such as chicken, swine, and ruminant such as cow; preferably swine.
  • systemic inflammation is the result of release of pro-inflammatory cytokines from immune-related cells and the chronic activation of the innate immune system. It contributes to the development of chronical disease conditions in animals.
  • the method according to the invention helps to reduce systemic inflammation of the animal.
  • the health of the host animal can be improved by way of decreasing local inflammation of the animal.
  • Local inflammation occurs within the area affected by the harmful stimulus.
  • Acute local inflammation develops within minutes or hours following a harmful stimulus, has a short duration, and primarily involves the innate immune system.
  • the method according to the invention helps to reduce local inflammation of the animal.
  • the health of the host animal can be improved by way of reducing the light regimen/duration into the daily circadian rhythm of the animal.
  • the circadian rhythms associated with light have important effects on the growth of production animals.
  • one way for increasing the growth rate and meat production is by prolongation of the illumination.
  • the illumination on poultry is extended to 23 hours a day, leaving the poultry under darkness for only one hour a day.
  • such a method may increase productivity, it has negative impacts on the health as well as the welfare of the animal.
  • the means, methods and uses according to the present invention help to increase the amount of melatonin and its precursor serotonin and thus restore the level of melatonin in animals which are subjected to prolonged illumination. Since artificially prolonged photoperiod leads to abnormal behavior such as aggressive interactions (tail biting, feather pecking, mobility/motility issues etc.) in poultry, restoring of melatonin level in such animals helps to improve the welfare of the animals.
  • the inventors of the present application have discovered that by compensating melatonin production by applying the means, methods and uses according to the invention, a stronger serotonergic flux is going into more melatonin and thus a reduction of the illumination regimen and a better animal welfare can be achieved.
  • the means and methods and uses of the present invention are inter alia applicable to production animals in general and may be provided to any suitable animal.
  • the animal is monogastric. It is generally understood that a monogastric animal has a singlechambered stomach.
  • the animal is a ruminant. It is generally understood that a ruminant has a multi-chambered stomach.
  • the animal is a ruminant in the pre-ruminant phase. Examples of such ruminants in the pre-ruminant phase include nursery calves.
  • the animal is a poultry (e.g. chicken, turkey), seafood (e.g. shrimp), sheep, cow, cattle, buffalo, bison, pig (e.g. nursery pig, grower/finisher pig), cat, dog, rabbit, goat, guinea pig, donkey, camel, horse, pigeon, ferret, gerbil, hamster, mouse, rat, bird, or human.
  • poultry e.g. chicken, turkey
  • seafood e.g. shrimp
  • sheep cow, cattle, buffalo, bison
  • pig e.g. nursery pig, grower/finisher pig
  • cat e.g. nursery pig, grower/finisher pig
  • cat dog, rabbit, goat, guinea pig, donkey, camel, horse, pigeon, ferret, gerbil, hamster, mouse, rat, bird, or human.
  • the animal is livestock. In some embodiments, the animal is a companion animal. In some embodiments, the animal is poultry. Examples of poultry include chicken, duck, turkey, goose, quail, or Cornish game hen. In one variation, the animal is a chicken. In some embodiments, the poultry is a layer hen, a broiler chicken, or a turkey.
  • the animal is a mammal, including, for example, a cow, a pig, a goat, a sheep, a deer, a bison, a rabbit, an, a llama, a mule, a horse, a reindeer, a water buffalo, a yak, a guinea pig, a rat, a mouse, an alpaca, a dog, or a cat.
  • the animal is a cow.
  • the animal is a pig.
  • the animal is a sow.
  • the invention is further characterized by the following items: [056] Item 1 : Method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; the method comprising the steps of providing an oligosaccharide preparation produced or producible by 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; or by b) condensing at least a first sugar with at least a second sugar using a catalyst; wherein the catalyst is methylphosphonic acid; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (
  • Item 2 Method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; the method comprising the steps of providing an oligosaccharide preparation produced or producible by 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; 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 of more than 2; feeding the oligosaccharide preparation to the animal; and increasing a
  • Item 3 Method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; the method comprising the steps of providing an oligosaccharide preparation produced or producible by b) condensing at least a first sugar with at least a second sugar 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 of more than 2; feeding the oligosaccharide preparation to the animal; and increasing a level of gamma-a
  • Item 4 Method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; the method comprising the steps of providing an oligosaccharide preparation, comprising 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; or comprising b) condensing at least a first sugar with at least a second sugar 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
  • Item 5 Method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; the method comprising the steps of providing an oligosaccharide preparation, comprising 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; 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 of more than 2; feeding the oligosaccharide preparation to the animal; and increasing a level of
  • Item 6 Method for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; iii) improving animal performance, in particular for improving average body weight gain and/or reducing feed conversion ratio; and/or iv) improving animal performance of animals facing stress, in particular for improving average body weight gain and/or reducing feed conversion ratio; the method comprising the steps of providing an oligosaccharide preparation, comprising b) condensing at least a first sugar with at least a second sugar 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 of more than 2; feeding the oligosaccharide preparation to the animal; and increasing a level of gamma-amino
  • DP
  • Item 7 Method for increasing production of gamma-aminobutyric acid, kynurenine and/or serotonin by a microbial community (e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • a microbial community e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • the method comprising the steps of providing an oligosaccharide preparation produced or producible by 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; or by b) condensing at least a first sugar with at least a second sugar using a catalyst; wherein the catalyst is methylphosphonic acid;
  • Item 8 Method for increasing production of gamma-aminobutyric acid, kynurenine and/or serotonin by a microbial community (e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • a microbial community e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • the method comprising the steps of providing an oligosaccharide preparation produced or producible by 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; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1 , at least a second
  • Item 9 Method for increasing production of gamma-aminobutyric acid, kynurenine and/or serotonin by a microbial community (e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • a microbial community e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • the method comprising the steps of providing an oligosaccharide preparation produced or producible by b) condensing at least a first sugar with at least a second sugar 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,
  • Item 10 Method for increasing production of gamma-aminobutyric acid, kynurenine and/or serotonin by a microbial community (e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • a microbial community e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • the method comprising the steps of providing an oligosaccharide preparation, comprising 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; or comprising b) condensing at least a first sugar with at least a second sugar using a catalyst; wherein the catalyst is methylphosphonic acid;
  • Item 11 Method for increasing production of gamma-aminobutyric acid, kynurenine and/or serotonin by a microbial community (e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • a microbial community e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • oligosaccharide preparation comprising 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; wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1 , at least a second fraction having
  • Item 12 Method for increasing production of gamma-aminobutyric acid, kynurenine and/or serotonin by a microbial community (e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • a microbial community e.g. an intestinal microbial community (i.e. as comprised in or obtainable from an intestinal sample, e.g. from caecum, ileum), preferably from an animal intestinal microbial community, e.g.
  • the method comprising the steps of providing an oligosaccharide preparation, comprising b) condensing at least a first sugar with at least a second sugar 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
  • Item 13 Method for producing an oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; and/or iii) increasing a level of gamma-aminobutyric acid, kynurenine and/or serotonin in the body of the animal, in particular in the gastrointestinal tract and/or in the blood of the animal; the method comprising the steps of 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; or b) condensing at least a first sugar with at least a second sugar using a catalyst; wherein the catalyst is methylphosphonic acid; thus producing the oligosaccharide preparation, wherein the oligosaccharide preparation comprises at least a first fraction having a degree of polymerization (DP) of 1
  • DP
  • Item 14 Method for producing an oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; and/or iii) increasing a level of gamma-aminobutyric acid, kynurenine and/or serotonin in the body of the animal, in particular in the gastrointestinal tract and/or in the blood of the animal; the method comprising the step of 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; thus 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 of more than 2.
  • DP degree of polymerization
  • Item 15 Method for producing an oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; and/or iii) increasing a level of gamma-aminobutyric acid, kynurenine and/or serotonin in the body of the animal, in particular in the gastrointestinal tract and/or in the blood of the animal; the method comprising the step of b) condensing at least a first sugar with at least a second sugar using a catalyst; wherein the catalyst is methylphosphonic acid; thus 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 of more than 2.
  • DP degree of polymerization
  • 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.
  • Item 17 The method according to any 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.
  • Item 18 The method according to any 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 selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably lactose.
  • a third sugar selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably lactose.
  • Item 19 The method according to any 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 selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably lactose.
  • a third sugar selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably lactose.
  • Item 20 The method according to any 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.
  • Item 21 The method according to any one of items 1-15, 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.
  • Item 22 The method according to any one of items 1-15, 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.
  • Item 23 The method according to any one of items 1-15, 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.
  • Item 24 The method according to any one of items 1-15, 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.
  • Item 25 The method according to any one of items 1-15, 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.
  • Item 26 The method according to any one of items 1-15, 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.
  • Item 27 The method according to any one of items 1-15, 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.
  • Item 28 The method according to any one of items 1-15, 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.
  • Item 29 The method according to any one of items 1-15, 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.
  • Item 30 The method according to any one of items 1-15, 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.
  • Item 31 The method according to any one of items 1-15, 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.
  • Item 32 The method according to any one of items 1-15, 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.
  • Item 33 The method according to any one of items 1-15, 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.
  • Item 34 The method according to any one of items 1-15, 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.
  • Item 35 The method according to any one of items 1-15, 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.
  • Item 36 The method according to any one of items 1-15, 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.
  • Item 37 The method according to any one of items 1-15, 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.
  • Item 38 The method according to any one of items 1-15, 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.
  • Item 39 The method according to any one of items 1-15, wherein the at least first sugar of step b) is glucose and wherein the at least second sugar of step b) is arabinose.
  • Item 40 The method according to any one of items 1-15, wherein the at least first sugar of step b) is glucose and wherein the at least second sugar of step b) is lactose.
  • Item 41 The method according to any one of items 1-15, wherein the at least first sugar of step b) is glucose and wherein the at least second sugar of step b) is N-acetylglucosamine.
  • Item 42 The method according to any one of items 1-15, wherein the at least first sugar of step b) is glucose and wherein the at least second sugar of step b) is xylose.
  • Item 43 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 arabinose.
  • Item 44 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 sucrose; and wherein the at least third sugar of step b) is arabinose.
  • Item 45 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 xylose; and wherein the at least third sugar of step b) is arabinose.
  • Item 46 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 arabinose; and wherein the at least third sugar of step b) is lactose.
  • Item 47 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 lactose.
  • Item 48 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 N-acetylglucosamine; and wherein the at least third sugar of step b) is lactose.
  • Item 49 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 sucrose; and wherein the at least third sugar of step b) is lactose.
  • Item 50 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 xylose; and wherein the at least third sugar of step b) is lactose.
  • 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.
  • Item 52 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 N-acetylglucosamine; and wherein the at least third sugar of step b) is mannose.
  • Item 53 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 sucrose; and wherein the at least third sugar of step b) is mannose.
  • Item 54 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 xylose; and wherein the at least third sugar of step b) is mannose.
  • Item 55 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 arabinose; and wherein the at least third sugar of step b) is N-acetylglucosamine.
  • Item 56 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 N-acetylglucosamine.
  • Item 57 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 N-acetylglucosamine.
  • Item 58 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 sucrose; and wherein the at least third sugar of step b) is N-acetylglucosamine.
  • Item 59 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 sucrose; and wherein the at least third sugar of step b) is N-acetylglucosamine.
  • Item 60 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 sucrose; and wherein the at least third sugar of step b) is N-acetylglucosamine.
  • Item 61 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 xylose; and wherein the at least third sugar of step b) is N-acetylglucosamine.
  • Item 62 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 xylose.
  • Item 63 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 sucrose; and wherein the at least third sugar of step b) is xylose.
  • Item 64 An oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; and/or iii) increasing a level of gamma-aminobutyric acid, kynurenine and/or serotonin in the body of the animal, in particular in the gastrointestinal tract and/or in the blood of the animal; wherein the oligosaccharide preparation is produced or producible by 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; or b) condensing at least a first sugar with at least a second sugar 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
  • DP
  • Item 65 The 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.
  • Item 66 The 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.
  • Item 67 The oligosaccharide preparation according to any one of items 64-66, wherein the at least first sugar of any one of step a) or b) and the at least second sugar of any one of step a) or b) are condensed with at least a third sugar selected from glucose, N-acetylglucosamine, lactose, mannose, glucosamine, fructose, sucrose, xylose, and arabinose; preferably selected from lactose.
  • Item 68 An oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; and/or iii) increasing a level of gamma-aminobutyric acid, kynurenine and/or serotonin in the body of the animal, in particular in the gastrointestinal tract and/or in the blood of the animal; wherein the oligosaccharide preparation comprises at least two distinct 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
  • Item 69 An oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; and/or iii) increasing a level of gamma-aminobutyric acid, kynurenine and/or serotonin in the body of the animal, in particular in the gastrointestinal tract and/or in the blood of the animal; wherein the oligosaccharide preparation comprises at least two distinct monosaccharide units selected from glucose, N-acetylglucosamine, and at least a third distinct 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
  • Item 70 The oligosaccharide preparation according to any one of items 64-69, wherein the oligosaccharide composition comprises at least 10 wt%, (11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50 wt% etc.) of the at least first fraction; preferably at least 20 wt%.
  • Item 72 The oligosaccharide preparation according to any one of items 64-71 , wherein the oligosaccharide preparation is comprised in a nutritional composition at an inclusion rate of at least 50 ppm (e.g. at least 50, 70, 100, 150, 200, 300, 400, 500 ppm).
  • Item 73 An oligosaccharide preparation for i) modulating behavior of an animal, in particular for reducing aggressive behavior; ii) improving animal welfare, in particular for reducing anxiety, stress and/or fear disorders; and/or iii) increasing a level of gamma-aminobutyric acid, kynurenine and/or serotonin in the body of the animal, in particular in the gastrointestinal tract and/or in the blood of the animal; wherein the oligosaccharide preparation is produced by or producible by the method according to any one of items 13-63.
  • Item 74 The oligosaccharide preparation according to any one of items 64-73, wherein the animal is selected from the group consisting of pet animals, dog, cat, canary, guinea pig, hamster, rabbit, mouse, rat, deer, boar, zoo animals, horse, donkey, poultry, swine, ruminant, chicken, cow, sheep, goat, pig, piglet, turkey, aquaculture, fish, shrimp, prawn, crayfish, crab, oyster, mussel, clam, trout, tilapia, salmon, carp, catfish, tuna, preferably, the animal is selected from the group consisting of poultry such as chicken, swine, and ruminant such as cow.
  • the animal is selected from the group consisting of poultry such as chicken, swine, and ruminant such as cow.
  • Item 75 The oligosaccharide preparation according to any one of items 64-74, 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 etc. fractions) each having a distinct DP.
  • 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 etc. fractions) each having a distinct DP.
  • Item 76 The oligosaccharide preparation according to any one of items 64-75, wherein the oligosaccharide preparation comprises not more than three distinct monosaccharide units.
  • Item 77 The oligosaccharide preparation according to any one of items 64-76, 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% anhydro-subunit containing oligosaccharides by 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% anhydro-subunit containing oligosaccharides by relative abundance.
  • Item 78 The oligosaccharide preparation according to any one of items 64-77, wherein the oligosaccharide preparation has a weight averaged molecular mass from 300 to 5000 g/mol (e.g. from about 2000 to 2800 g/mol, 2100 to 2700 g/mol, 2200 to 2600 g/mol, 2300 to 2500 g/mol, or 2320 to 2420 g/mol), 500 to 5000 g/mol, 700 to 5000 g/mol, 500 to 2000 g/mol, 700 to 2000 g/mol, 700 to 1500 g/mol, 300 to 1500 g/mol, 300 to 2000 g/mol, 300 to 700 g/mol, 400 to 1300 g/mol, 400 to 1200 g/mol, 400 to 1100 g/mol, 500 to 1300 g/mol, 500 to 1200 g/mol, 500 to 1100 g/mol, 600 to 1300 g/mol, 600 to 1200 g/mol, or 600 to
  • Item 79 Use of an oligosaccharide preparation according to any one of items 64-78 for i) modulating behavior of an animal (e.g. a healthy animal), in particular for reducing aggressive behavior; ii) improving animal welfare (e.g. of a healthy animal), in particular for reducing anxiety, stress and/or fear disorders; iii) improving animal performance (e.g. of a healthy animal), in particular for improving average body weight gain and/or reducing feed conversion ratio; iv) improving animal performance of animals (e.g.
  • Item 80 An oligosaccharide preparation for use in treatment, amelioration, prevention and/or prophylaxis of one or more disorders associated with an imbalanced level of gamma- aminobutyric acid and/or kynurenine, wherein the one or more disorders are selected from the group of anxiety, stress, fear disorders, systemic inflammation, and local inflammation, wherein the oligosaccharide preparation is produced or producible by 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; or b) condensing at least a first sugar with at least a second sugar 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 of more than 2.
  • DP
  • Item 81 An oligosaccharide preparation for use in treatment, amelioration, prevention and/or prophylaxis of one or more disorders associated with an imbalanced level of gamma- aminobutyric acid and/or kynurenine, wherein the one or more disorders are selected from the group of anxiety, stress, fear disorders, systemic inflammation, and local inflammation, in an animal suffering from or at risk at suffering from one or more disorders associated with an imbalanced level of gamma-aminobutyric acid and/or kynurenine, wherein the oligosaccharide preparation is produced or producible by 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; or b) condensing at least a first sugar with at least a second sugar using a catalyst; wherein the catalyst is methylphosphonic acid; and wherein the oligosaccharide preparation comprises at least a
  • Item 82 The oligosaccharide preparation for use in treatment, amelioration, prevention and/or prophylaxis according to item 80 or 81 , wherein the oligosaccharide preparation is the oligosaccharide preparation according to any one of items 64-78.
  • Item 83 The oligosaccharide preparation for use in treatment, amelioration, prevention and/or prophylaxis according to any one of items 80-82, wherein the oligosaccharide preparation is comprised in a nutritional composition at an inclusion rate of at least 50 ppm (e.g. at least 50, 70, 100, 150, 200, 300, 400, 500 ppm).
  • at least 50 ppm e.g. at least 50, 70, 100, 150, 200, 300, 400, 500 ppm.
  • the oligosaccharide preparation may be provided in the form of a powderous formulation comprising at least 20% (w/w) of the oligosaccharide preparation as referred to 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 pm (e.g. 100-500, 200-500, 200-300 pm); and optionally 0-25% (wt/wt) of water and/or an auxiliary substance; wherein the % are based on the total weight of the powderous formulation.
  • a silica-based adsorbate e.g. diatomaceous earth, amorphous precipitated silica
  • such a powderous formulation may comprise 30-70% (wt/wt) of the oligosaccharide preparation as referred to herein; 30-70% (wt/wt) of a silica based adsorbate (e.g. having an average particle size of at least 50 pm); and 0-21 % (wt/wt) of water; wherein the % are based on the total weight of the powderous formulation.
  • the oligosaccharide preparation is formulated analogous to the protocol as described in any one of Examples 22-26 and 33 of WO 2020/097458.
  • an “oligosaccharide preparation” may comprise one or more monosaccharide(s) or sugar(s), and/or anhydro-monosaccharide(s), and/or one or more compound molecule(s) such as di-, tri-, tetra-saccharides or higher polymerized saccharides, containing two or more monosaccharide subunits, which are linked by glycosidic bonds to form the oligosaccharide(s).
  • An “oligosaccharide” may also refer to a compound molecule containing two or more monosaccharide subunits, where at least one monosaccharide unit is replaced by an anhydro-subunit.
  • oligosaccharide may be optionally functionalized.
  • oligosaccharide encompasses all species of the oligosaccharide, wherein each of the monosaccharide subunits in the oligosaccharide is independently and optionally functionalized and/or replaced with its corresponding anhydro-monosaccharide subunit.
  • an “anhydro-subunit” may be a product of reversible thermal dehydration of a monosaccharide or monosaccharide subunit, or a sugar caramelization product.
  • an “anhydro-subunit” may be an anhydro-monosaccharide such as anhydro-glucose.
  • an “anhydro-subunit” may be linked with one or more regular monosaccharide subunits and/or with one or more anhydro-monosaccharide subunits via glycosidic linkage(s).
  • An oligosaccharide may be characterized to contain two or more monosaccharide subunits linked by glycosidic bonds.
  • a “gluco-oligosaccharide” may refer to a compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds.
  • a “gluco-oligosaccharide” may also refer to compound containing two or more glucose monosaccharide subunits linked by glycosidic bonds, wherein at least one monosaccharide subunit is replaced by an anhydro-glucose subunit.
  • a “galacto-oligosaccharide” may refer to a compound containing two or more galactose monosaccharide subunits linked by glycosidic bonds.
  • a “galacto-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 anhydro-galactose subunit.
  • a gluco- galactose-oligosaccharide may be a compound comprising one or more glucose monosaccharide subunits and one or more galactose monosaccharide subunits linked by glycosidic bonds.
  • a gluco-galactose oligosaccharide may also refer to a compound, wherein at least one of the monosaccharide subunits is replaced with its respective anhydro-monosaccharide subunit.
  • a gluco-galacto-xylo-oligosaccharide may refer to a compound produced by the condensation reaction of glucose, galactose, and xylose.
  • An oligosaccharide preparation comprising gluco- galacto-xylo-oligosaccharides may comprise gluco-galactose-oligosaccharides, gluco-xylo- oligosaccharides, galacto-xylo-oligosaccharides, gluco-galacto-xylo-oligosaccharides, and compounds containing one or more glucose monosaccharide subunits, one or more xylose monosaccharide subunits, and one or more galactose monosaccharide subunits linked by glycosidic bonds, as well as anhydro-monosaccharide subunit(s) thereof.
  • a “monosaccharide subunit” may refer to a monosaccharide monomer in an oligosaccharide.
  • oligosaccharide having a degree of polymerization (DP) of 1 the oligosaccharide may be referred to as a monosaccharide subunit or monosaccharide.
  • DP degree of polymerization
  • its monosaccharide subunits are linked via glycosidic bonds.
  • the term “regular monosaccharide” may refer to a monosaccharide that does not contain an anhydro-subunit.
  • the term “regular disaccharide” may refer to a disaccharide that does not contain an anhydro-subunit.
  • the term “regular subunit” may refer to a subunit that is not an anhydro-subunit.
  • a DP1 fraction i.e. a fraction having a DP of 1 comprising 10% anhydro-subunit containing oligosaccharides by relative abundance may refer to a plurality of DP1 oligosaccharides, wherein 10%, by number, of the DP1 oligosaccharides are anhydro-monosaccharides.
  • a distribution of the degree of polymerization of the oligosaccharide preparation may be determined by any suitable analytical method and instrumentation, including but not limited to end group method, osmotic pressure (osmometry), ultracentrifugation, viscosity measurements, light scattering method, size exclusion chromatography (SEC), SEC-MALLS, field flow fractionation (FFF), asymmetric flow field flow fractionation (A4F), high-performance liquid chromatography (HPLC), and mass spectrometry (MS).
  • the distribution of the degree of polymerization may be determined and/or detected by mass spectrometry, such as MALDI-MS, LC-MS, or GC-MS.
  • the distribution of the degree of polymerization may be determined and/or detected by SEC, such as gel permeation chromatography (GPC).
  • the distribution of the degree of polymerization may be determined and/or detected by HPLC, FFF, or A4F.
  • the degree of polymerization of the oligosaccharide preparation may be determined based on its molecular weight and molecular weight distribution (for a more detailed description see WO 2020/097458).
  • An oligosaccharide preparation may comprise n distinct fractions, wherein each fraction has a distinct DP, and wherein n is any integer, e.g. 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10 etc.
  • each of the n fractions of oligosaccharides of the oligosaccharide preparation as described herein independently comprises an anhydro-subunit level.
  • the DP1 fraction comprises 10% anhydro-subunit containing oligosaccharides by relative abundance
  • the DP2 fraction comprises 15% anhydro-subunit containing oligosaccharides by relative abundance.
  • DP1 , DP2, and DP3 fraction each comprises 5%, 10%, and 2% anhydro-subunit containing oligosaccharides by relative abundance, respectively.
  • two or more fractions of oligosaccharides may comprise similar level of anhydro-subunit containing oligosaccharides.
  • the DP1 and DP3 fraction each comprises about 5 % anhydro-subunit containing oligosaccharides by relative abundance.
  • the level of anhydro-subunits may be determined by any suitable analytical methods, such as nuclear magnetic resonance (NMR) spectroscopy, mass spectrometry, HPLC, FFF, A4F, or any combination thereof.
  • the level of anhydro-subunits is determined, at least in part, by mass spectrometry such as MALDI-MS.
  • the level of anhydro-subunits may be determined, at least in part, by NMR.
  • the level of anhydro-subunits may be determined, at least in part, by HPLC.
  • the level of anhydro-subunits may be determined by MALDI-MS, as illustrated in more detail in WO 2020/097458.
  • the oligosaccharide preparation described herein comprise a variety of glycosidic linkages.
  • the type and distribution of the glycosidic linkages may depend on the source and manufacturing method of the oligosaccharide preparation.
  • the type and distribution of various glycosidic linkages may be determined and/or detected by any suitable methods known in the art such as NMR.
  • the glycosidic linkages are determined and/or detected by proton NMR, carbon NMR, 2D NMR such as 2D JRES, HSQC, HMBC, DOSY, COSY, ECOSY, TOCSY, NOESY, or ROESY, or any combination thereof.
  • the glycosidic linkages are determined and/or detected, at least in part, by proton NMR. In some embodiments, the glycosidic linkages are determined and/or detected, at least in part, by carbon NMR. In some embodiments, the glycosidic linkages are determined and/or detected, at least in part, by 2D HSQC NMR.
  • an oligosaccharide preparation may comprise one or more a-(1 ,2) glycosidic linkages, a-(1 ,3) glycosidic linkages, a-(1 ,4) glycosidic linkages, a-(1 ,6) glycosidic linkages,
  • the oligosaccharide preparations may have a glycosidic bond type distribution of about from 0 to 60 mol%, 5 to 55 mol%, 5 to 50 mol%, 5 to 45 mol%, 5 to 40 mol%, 5 to 35 mol%, 5 to 30 mol%, 5 to 25 mol%, 10 to 60 mol%, 10 to 55 mol%, 10 to 50 mol%, 10 to 45 mol%, 10 to 40 mol%, 10 to 35 mol%, 15 to 60 mol%, 15 to 55 mol%, 15 to 50 mol%, 15 to 45 mol%, 15 to 40 mol%, 15 to 35 mol%, 20 to 60 mol%, 20 to 55 mol%, 20 to 50 mol%, 20 to 45 mol%, 20 to 40 mol%, 20 to 35 mol%, 25 to 60 mol%, 25 to 55 mol%, 25 to 50 mol%, 25 to 45 mol%, 25 to 40 mol%, or 25 to 35 mol%,
  • the molecular weight and molecular weight distribution of the oligosaccharide preparation may be determined by any suitable analytical means and instrumentation, such as end group method, osmotic pressure (osmometry), ultracentrifugation, viscosity measurements, light scattering method, SEC, SEC-MALLS, FFF, A4F, HPLC, and mass spectrometry.
  • the molecular weight and molecular weight distribution are determined by mass spectrometry, such as MALDI-MS, LC-MS, or GC-MS.
  • the molecular weight and molecular weight distribution are determined by size exclusion chromatography (SEC), such as gel permeation chromatography (GPC).
  • SEC size exclusion chromatography
  • GPC gel permeation chromatography
  • the molecular weight and molecular weight distribution are determined by HPLC.
  • the molecular weight and molecular weight distribution are determined by MALDI- MS.
  • the oligosaccharide preparation has a weight averaged molecular mass from 300 to 5000 g/mol (e.g. from about 2000 to 2800 g/mol, 2100 to 2700 g/mol, 2200 to 2600 g/mol, 2300 to 2500 g/mol, or 2320 to 2420 g/mol), 500 to 5000 g/mol, 700 to 5000 g/mol, 500 to 2000 g/mol, 700 to 2000 g/mol, 700 to 1500 g/mol, 300 to 1500 g/mol, 300 to 2000 g/mol, 300 to 700 g/mol, 400 to 1300 g/mol, 400 to 1200 g/mol, 400 to 1100 g/mol, 500 to 1300 g/mol, 500 to 1200 g/mol, 500 to 1100 g/mol, 600 to 1300 g/mol, 600 to 1200 g/mol, or 600 to 1100 g/mol); and/or wherein the oligosaccharide preparation
  • the species of oligosaccharides present in an oligosaccharide preparation referred to herein may depend on the type of the feed sugar(s).
  • the oligosaccharide preparations comprise a gluco-oligosaccharide when the feed sugars comprise glucose.
  • the oligosaccharide preparations comprise a fructose-oligosaccharide when the feed sugars comprise fructose.
  • the oligosaccharide preparations comprise gluco-fructo-oligosaccharides when the feed sugars comprise fructose and glucose.
  • the oligosaccharide preparations comprise one or more species of monosaccharide subunits.
  • the oligosaccharide preparation may comprise oligosaccharides with 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, or more different species of monosaccharides subunits.
  • An oligosaccharide preparation may be manufactured by heating an agueous composition comprising one or more feed sugar(s) 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-guinolinesulfonic acid hydrate; a-hydroxy-2- pyridinemethanesulfonic acid; (P)-camphor-IO-sulfonic acid; butylphosphonic acid; diphenylphosphinic acid; hexylphosphonic acid; methylphosphonic acid; phenylphosphinic acid; phenylphosphonic acid; tert-butylphosphonic acid; SS)-VAPOL hydrogenphosphate; 6- guinolinesulfonic acid, 3-(1-pyridinio)-1 -propa
  • the polymerization of the feed sugars is achieved by a step-growth polymerization. In some embodiments, the polymerization of the feed sugars is achieved by polycondensation.
  • Feed Sugar The one or more feed sugars used in the methods of manufacturing oligosaccharide preparations described herein may comprise one or more types of sugars.
  • the one or more feed sugars comprise monosaccharides, disaccharides, trisaccharides, tetrasaccharides, or any mixtures thereof.
  • Feed sugar(s) may be e.g. one or more selected from glucose, lactose, galactose, mannose, glucosamine, N-acetylglucosamine, arabinose, xylose, sucrose, and fructose.
  • Feed sugars were selected from glucose, lactose, galactose, mannose, glucosamine, N-acetylglucosamine, arabinose, xylose, sucrose, and fructose.
  • Catalysts were selected from sulfuric acid, acetic acid, propionic acid, butanoic acid, L- glutamic acid, L-lysine, phosphoric acid, (+)-camphor-10-sulfonic acid, methylphosphonic acid, L- cysteic acid, hydrochloric acid, citric acid, lactic acid, 3-(1-pyridinio)-1 -propanesulfonate, 2- pyridinesulfonic acid, 3-pyridinesulfonic acid, a-hydroxy-2-pyridinemethanesulfonic acid, (P)- camphor-10-sulfonic acid, butylphosphonic acid, diphenylphosphinic acid, hexylphosphonic acid, phenylphosphinic acid, phenylphosphonic acid, tert-butylphosphonic acid, 2-(2- pyridinyl)ethanesulfonic acid, 3-(2-pyridyl)-5,6-diphenyl-1 ,2,4
  • Feed sugars of OP IDs 1-61 as listed in the order of Table 1 below were used at 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, 95:5, 95:5, 95:5, 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 ,
  • Table 1 Feed sugars and catalysts used for preparing synthetic oligosaccharide preparations with the indicated oligosaccharide preparation identifier (OP ID).
  • the amount of catalyst present in the oligosaccharide preparations after production was quantified by anion chromatography.
  • the analysis was performed on a Thermo-Fisher ICS-3000 ion chromatography system equipped with EG-3000 eluent generator, AS autosampler and CD- 3000 conductivity detector. Separation was achieved on an AS19 column (4.0 mm x 250 mm) protected with an AG19 precolumn (4.0 mm x 50 mm).
  • the mobile phase was a KOH gradient (0-55 mM) produced by an eluent generator (EGCII EluGen® cartridge).
  • the chromatographic parameters were: 1.0 mL/min flow rate, 25 pL injection volume, and 30 °C column temperature.
  • the columns used were a PL aquagel-OH guard column, 7.5 x 50 mm, 5 pM (Agilent Part No: PL1149-1530) and a PL aquagel-OH, 7.5 x 300 mm, 5 pM (Agilent Part No: PL1120-6520), whereas the used solvent was 200 mM aqueous sodium nitrate at a flow-rate of 0.9 mL/min. and a column temperature of 40 °C.
  • glucose for the quantification of total sugar content
  • pullulan standards for the size distribution
  • Oligosaccharide preparations of OP IDs 50, 43, 57, and 54 were found to have a DP1 content (DP: degree of polymerization; DP1 : degree of polymerization of 1) of 29.5 wt%, 27.3 wt%, 11.6 wt%, and 50.5 wt%, respectively; and to have a DP2+ content (oligosaccharides having a degree of polymerization of 2 or more) of 42.2 wt%, 39.2 wt%, 45.0 wt% and 22.1 wt%, respectively.
  • DP1 content DP: degree of polymerization
  • DP1 degree of polymerization of 1
  • DP2+ content oligosaccharides having a degree of polymerization of 2 or more
  • OP ID 54 Upon additional analysis of OP ID 54 via liquid chromatography with evaporative light scattering detector coupled to high-resolution mass spectrometry (LC-ELSD-HRMS) a DP1 content of 69.22 Area% was confirmed, a DP2 content of 24.64 Area% and a DP3+ content of 6.14 Area% were found. Further, the Mn (number averaged molecular mass) and Mw (weight averaged molecular mass) were determined. OP IDs 50, 43, 57, and 54 had an Mn of 477, 574, 710 and 690 g/mol, respectively; and an Mw of 578, 779, 1059, and 1008 g/mol, respectively.
  • refractive indices were determined at 78.9, 79.6, 77.0 and 73.6 Brix for the oligosaccharide preparations of OP IDs 50, 43, 57, and 54, respectively.
  • the minimal media 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 Phosphate dibasic, 1500 mg/L Sodium Phosphate dibasic, 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 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 in solution, 4 mg/L Iron Sulfate in solution, 1 mg/L Co
  • the glycan solutions were prepared to a concentration of 6 brix of each of the individual oligosaccharide preparations to be tested, diluted with distilled water. To this end, the oligosaccharide preparations OP ID 1-61 as prepared in Example 1 were used.
  • the feed extract solution was prepared by mixing ground pig feed (as detailed in Table 2) with 20 mL of distilled water, followed by incubating at 125 °C for 15 min. Thereafter, the mixture was pelleted by centrifugation The supernatant was incubated at 121 °C for 15 min. Thereafter, sterile distilled water was added to a final volume of 20 mL.
  • Table 2 Pig feed composition. *Premix vitamin mineral 3144 provided per kilogram of diet: Vitamin A 6510 I.U.; 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; Mn: 40 mg; Fe: 100 mg; Cu: 15 mg; Zn: 65 mg; I: 2 mg; Se: 0.4 mg.
  • Swine cecal slurry was prepared by mixing 3.8 g of caecal sample in 25 mL PBS (phosphate-buffered saline: 137 mM sodium chloride, 2.7 mM potassium chloride, 10 mM disodium hydrogenphosphate, 1.8 mM potassium dihydrogen phosphate, pH 7.4). After sedimentation of large particles within this mixture, the non-sedimented supernatant was decanted into a fresh vessel. Thereafter, this supernatant was subjected to centrifugation (500 rpm, 5 min). After centrifugation, the supernatant was recovered and mixed with 50% glycerol to a final glycerol concentration of 15% (v/v), thus forming the swine cecal slurry.
  • PBS phosphate-buffered saline: 137 mM sodium chloride, 2.7 mM potassium chloride, 10 mM disodium hydrogenphosphate, 1.8 mM potassium dihydrogen phosphate, pH 7.
  • caecal sample 1 contained 48.0%, 33.3%, 15.4% and 3.3% of Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteriae, respectively.
  • Caecal sample 2 contained 54.9%, 27.2%, 15.3% and 2.7% of Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteriae, respectively.
  • Caecal sample 3 contained 67.0%, 20.4%, 10.4%, and 2.3% of Firmicutes, Bacteroidetes, Proteobacteria, and Actinobacteriae, respectively.
  • caecal 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.
  • Caecal sample 2 contained 10.8%. 5.5%. 3.7%. 12.3%. 6.0%. 10.3%. 4.9%.
  • Bacteroides Blautia. Campylobacter, Clostridium, Escherichia, Lactobacillus, Umosilactobacillus, Prevotella, Roseburia, Sarcina, and Streptococcus, respectively.
  • Caecal 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, Umosilactobacillus, Phocaeicola, Prevotella, Roseburia, and Ruminococcus, respectively.
  • the glucose control was included to obtain a basal GABA response from the microbiome as a reference for fold changes calculations. Since glucose is mostly absorbed before actually reaching the caecum in the small intestine, the 1 % glucose control appropriately mimicked the in vivo situation.
  • the 96-deep well plates were sealed with a porous, breathable adhesive film, and incubated anaerobically for 24 h at 37 °C. Thereafter, the 96-deep well plates were centrifuged for pelleting the microbiota and non-dissolved matter, and the supernatants were used for further analysis, in particular for determination of production of GABA and/or KYN.
  • GABA was measured by liquid chromatography with tandem mass spectrometry (HPLC- MS/MS).
  • the supernatants obtained from the in vitro fermentations of Example 3 were subjected to centrifugation. Then, 200 pL of supernatant was transferred to 96-well plates. Components comprised in these supernatants were separated on an Acquity LIPLC H-class system comprising degasser, quaternary pump, sample manager and thermostat column compartment equipped with TQDetector (Waters).
  • the columns used were a LIPLC BEH C18 guard column, 2.1 x 50 mm, 1.7 pM (Waters reference: 1866003975) and an Acquity LIPLC BEH C18, 2.1 x 150 mm, 1.7 pM (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.
  • Flow rate was 0.3 mL/min, column temperature was 40 °C.
  • the autosampler was set to 15 °C, run time was 7.5 minutes. Gradient profile is shown in Table 3.
  • GABA powder was used at a calibration range from 3 to 0.0003 pg/mL.
  • the chromatographic injection volume was 2 pL, the syringe was washed after each injection.
  • the MS method used was a multiple reaction monitoring (MRM) with a positive electrospray ionization.
  • MRM multiple reaction monitoring
  • 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
  • the collision voltage was 13 V.
  • Kynurenine (KYN) quantification was performed by ELISA kit following the supplier’s recommendation (FI-EM1862; Euromedex). Briefly, 50 pL of standard, blank or samples were added to 50 pL of Biotin-labeled Antibody Working Solution and incubated for 45 min at 37 °C. The plate was washed three times with Wash Buffer, and 100 pL of HRP-streptavidin conjugate was added into each well. The plate was incubated for 30 minutes at 37 °C. The plate was then washed five times with Wash Buffer. 90 pL of TMB substrate was added to each well followed by an incubation at 37 °C in dark for 10-20 minutes. Then, 50 pL of stop solution was added to each well and the absorbance was measured at 450 nm and correlated to a standard curve of known KYN concentrations.
  • Example 5 The synthetic oligosaccharide preparations produced in Example 1 were individually subjected to swine microbiota in in vitro fermentations as described in Example 3, and analyzed as outlined in Example 4. The effect of contacting the microbiota with any one of the synthetic oligosaccharide preparations in terms of GABA production was calculated as Iog2 fold change of produced GABA compared to the GABA concentration obtained from the microbiota contacted with the glucose control, see Table 4. While some oligosaccharide preparations did not or only marginally affect GABA production as compared to the glucose control (e.g.
  • oligosaccharide preparations were found capable of eliciting a significantly larger microbial production of GABA than the glucose control.
  • oligosaccharide preparations comprising N-acetylglucosamine were found to increase GABA production.
  • oligosaccharide preparations produced with methylphosphonic acid as catalyst were found to increase microbial GABA production.
  • oligosaccharide preparations produced from the same feed sugars but with a catalyst other than methylphosphonic acid had no or a considerable lower effect on GABA than their corresponding oligosaccharide preparations with methylphosphonic acid as catalyst (compare e.g. OP IDs 30 to 58; 48 to 23; 10 to 21 ; 9 to 39; 15 to 25 and to 26).
  • Table 4 Log2 fold changes in GABA production compared to a glucose control (FC/Glu) by individual oligosaccharide preparations.
  • Three test treatments were prepared by spraying either one of the oligosaccharide preparations of OP IDs 50, 43, and 57 onto the basal pig feed as mash.
  • the oligosaccharide preparations were supplied in syrup form at a concentration of 70 brix, and added to the feed at an inclusion level of 1000 mg/kg.
  • the same volume of water was sprayed on the basal pig feeds as mash, instead of the syrup of any of the test treatments.
  • the composition of the basal pig feed is shown in T able 5 below. Feeds were given for ad libitum consumption as mash during the experimental period. Phytase (Ronozme HiPhos) at 100 mg/kg feed was included in premix form using ground corn as carrier in the basal diet.
  • Table 5 Basal pig feed. *Premix vitamin mineral 3144 provided per kilogram of diet: Vitamin A 6510 I.U.; Vitamin E: 75 mg.; Vitamin K: 2 mg; Vitamin D3: 2007 III; 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; Mn: 40 mg; Fe: 100 mg; Cu: 15 mg; Zn: 65 mg; I: 2 mg; Se: 0.4 mg.
  • Zootechnical parameters (mortality, weight gain, final weight, FCR - feed conversion ratio) were monitored. GABA and KYN were determined in plasma and feces samples; cortisol (COR), adrenocorticotrophic hormone (ACTH), serum neuropeptide Y (NPY), and lactate were determined in plasma samples.
  • COR cortisol
  • ACTH adrenocorticotrophic hormone
  • NPY serum neuropeptide Y
  • lactate lactate were determined in plasma samples.
  • silica was added to compensate for the mass difference between the treatment feeds comprising 500 mg of oligosaccharide preparation per kg of feed and the other treatment feeds.
  • 0.5 kg SiO2 per 1000 kg of feed was added to treatment feeds of the negative control; 0.47 kg SiO2 per 1000 kg of feed was added to treatment feeds of the GABA control in addition to the 0.03 kg of GABA per 1000 kg of feed; 0.45 kg, 0.35 kg and 0.25 kg of SiO2 per 1000 kg of feed were added to the treatment feeds comprising 50, 150, and 250 mg of oligosaccharide preparation per 1000 kg of feed, respectively.
  • feeds were given for ad libitum consumption as mash during the experimental period.
  • Phytase Ronozme HiPhos at 100 mg/kg feed was included in the premix forms using ground corn as carrier in the basal diet (2*150 g). Feed main raw materials were milled in a hammer mill using a 4mm sieve and mixed with the other ingredients (minerals, vitamins and amino acids) in 1500 L mixer. The glycan products were supplied in syrup form sprayed on the mash feed. The same volume of water was sprayed for negative control feeds.
  • Table 6 Basal diet feeds. *Premix vitamin mineral 3144 provided per kilogram of diet: Vitamin A 6510 I.U.; Vitamin E: 75 mg.; Vitamin K: 2 mg; Vitamin D3: 2007 III; 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; Mn: 40 mg; Fe: 100 mg; Cu: 15 mg; Zn: 65 mg; I: 2 mg; Se: 0.4 mg.
  • Vitamin A 15000 I.U.
  • 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
  • Choline 325 mg
  • Mn 60 mg
  • Fe 200 mg
  • Cu 140 mg
  • Zn 100 mg
  • I 2 mg
  • Se 0.4 mg.
  • GABA serotonin (5-HT), Trp (tryptophan) and KYN were analyzed from plasma and feces samples.
  • Example 8 [187] A further animal trial was performed using 144 growing pigs (72 female, 72 uncastrated male), [Large White x Landrace] x Pietrain, undocked tails. Two treatments were tested: Basal diet feed as shown in Table 7 was fed as negative control. Basal diet feed supplemented with 250 g of dry OP ID 54 per MT of feed was used as test treatment. Feed was provided for ad libitum consumption in pellet form.
  • the trial was performed for a total of 105 days with an initial seven-days adaptation period (d-7 to dO), followed by diet treatment start at dO. At d55 and d56, the animals were subjected to a mixing challenge analogous to Example 7. The trial was concluded at d98 with a final weight of approximately 100-105 kg.
  • Feeding of the oligosaccharide compositions of the invention was found to improve animal performance (e.g. in terms of daily weight gain, feed intake, feed-to-gain ratio) throughout the trial, and in particular during the stress period established by the mixing challenge. Notably, during the mixing challenge week (d51-d59), animals receiving the oligosaccharide compositions of the invention showed significant improvement of growth performance (final weight, weight gain, FCR). Details are shown in Tables 8-11 below.
  • Saliva samples were collected on d-7 (baseline), d58 (after mixing challenge) and d97 (end of trial). Hair samples were collected on d-7 and d98 (to detect potential accumulation of cortisol during the treatment period d-7 to d98). Saliva and hair samples were tested for cortisol content, as non-invasive methods to measure stress response. Notably, the cortisol concentration in saliva samples was significantly reduced by 24.9% and by 23.9% on d58 and d97, respectively, in animals of the test treatment group compared to animals of the control group. Similarly, the cortisol concentration in hair samples on d98 was reduced by 17.5% in animals of the test treatment group compared to animals of the control group.
  • Animals receiving oligosaccharide preparations of the invention thus showed a significantly decreased stress response after the mixing challenge, and showed lower chronic stress caused by the mixing challenge, as found by lower accumulation of the stress-biomarker cortisol in hair samples.
  • the amount of severe or medium-severe lesions on ears, body and/or tails was found reduced on animals receiving oligosaccharide preparations of the invention, thus indicating a reduction in aggressive behavior of these animals compared to animals receiving the control diet.
  • Vitamin and mineral premix provided per kg feed: Vitamin A (E 672) 5500 Ul; vitamin D3 (E 671) 1100 Ul; vitamin E (alfa tocopherol) 25 mg; vitamin B1 0.5 mg; vitamin B2 1.4 mg; vitamin B6 1 mg; vitamin B12 8 pg; vitamin K3 0.5 mg; calcium panthotenate 5.6 mg; nicotinic acid 8 mg; choline 120 mg; Fe (E 1) (from FeSO4'7H2O) 80 mg; I (E 2) (from Ca(l20s)2) 0.5 mg; Co (E 3) (from 2CoCC>3-3Co(OH)2 H2O) 0.4 mg; Cu (E 4) (from CUSO4 5H2O) 5 mg; Cu (E 4) (from aminoacids quelate) 5 mg; Mn (E 5) (from MnO) 40 mg; Zn (E 6) (from ZnO) 100 mg; Se (E 8) (from Na2
  • Table 8 Animal performance for days 0-29. "Control” animals were fed the negative control diet; “test” animals were fed the test treatment diet.
  • Table 9 Animal performance for days 29-64. "Control” animals were fed the negative control diet;
  • test animals were fed the test treatment diet.
  • Table 10 Animal performance for days 64-98. "Control” animals were fed the negative control diet; “test” animals were fed the test treatment diet.
  • Table 11 Animal performance for days 51-59. "Control” animals were fed the negative control diet; “test” animals were fed the test treatment diet.

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Abstract

La présente invention concerne un procédé pour moduler le comportement d'un animal, améliorer le bien-être de l'animal, et augmenter la production d'acide gamma-aminobutyrique, de kynurénine et/ou de sérotonine de manière à améliorer les performances de l'animal, à l'aide d'une préparation oligosaccharidique.
PCT/EP2024/054484 2023-02-23 2024-02-22 Moyens et procédés de modulation du comportement Ceased WO2024175697A1 (fr)

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CN202480013815.5A CN120731013A (zh) 2023-02-23 2024-02-22 用于调节行为的手段和方法
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