Combination comprising bifidobacterium bifidum ha-132 and human milk oligosaccharides

Abstract

The invention relates to a composition or a kit of parts comprising a probiotic and at least one human milk oligosaccharide (HMO), wherein the probiotic is the Bifidobacterium bifidum (B. bifidum) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and the at least one HMO is selected from a fucosylated HMO, a neutral core HMO, and a sialylated HMO, or is any mixture thereof, for use in supporting or improving cognitive development, emotional and behavioural development, immune system development and anti-inflammatory support, gut barrier function, and / or gut motility, in a subject. The invention further relates to a nutritional composition comprising a probiotic and at least one HMO, wherein the probiotic is the B. bifidum HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and the at least one HMO is a sialyllactose.

Description

[0001] COMBINATION COMPRISING BIFIDOBACTERIUM BIFIDUM HA-132 AND HUMAN MILK

[0002] OLIGOSACCHARIDES

[0003] FIELD

[0004] The present invention relates to the field of prebiotics and probiotics. In particular, the present invention relates to a composition or a kit of parts comprising a probiotic and at least one human milk oligosaccharide (HMO), wherein the probiotic is the Bifidobacterium bifidum (B. bifidum) HA- 132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and the at least one HMO is selected from a fucosylated HMO, a neutral core HMO, and a sialylated HMO, or is any mixture thereof, for use in supporting or improving cognitive development, emotional and behavioural development, immune system development and antiinflammatory support, gut barrier function, and / or gut motility, in a subject. The invention further relates to a nutritional composition comprising a probiotic and at least one HMO, wherein the probiotic is the B. bifidum HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and the at least one HMO is a sialyllactose.

[0005] DEPOSITS

[0006] Bifidobacterium bifidum HA-132 (Lallemand Health Solutions Inc.) has been deposited at Collection Nationale de Cultures de Microorganismes (CNCM; 25 Rue du Docteur Roux F-75724 Paris Cedex 15, France), according to the Budapest Treaty on September 19, 2022, under the accession number CNCM I-5898.

[0007] BACKGROUND

[0008] The human gut microbiome is composed of bacteria, archaea, viruses, and eukaryotic microbes that reside inter alia in the gut. These microbes have tremendous potential to impact our physiology, both in health and in disease. The microbiota of the human intestine is a complex and very dynamic microbial ecosystem, which is considered to serve numerous important functions for its human host, including protection against pathogens, induction of immune regulatory functions, nutrient processing, and metabolic functions. These basic functions affect, directly or indirectly, most of our physiologic functions. Various gut bacteria, including certain Bifidobacterium spp., can feed on oligosaccharides such as fructo-oligosaccharides (FOS), galacto-oligosaccharides (GOS), and human milk oligosaccharides (HMOs), whereas these structures are indigestible by humans. Due to the ability to feed beneficial microorganisms and induce the growth or activity of these, certain oligosaccharides are referred to as prebiotics.

[0009] Human milk oligosaccharides (HMOs) are a heterogeneous mixture of soluble glycans found in human milk, with more than 200 different structures identified in human milk to date. They are the third most abundant solid component after lactose and lipids in human milk and are present with concentrations of 5-25 g / l. Certain HMOs are believed to be important for the development of the infant gut microbiota, in particular by favouring the predominance of bifidobacteria. This is viewed as beneficial for infants because some strains of Bifidobacterium species may have a positive effect on gut health. In addition, some species of bifidobacterial, such as certain strains of B. bifidum and B. longum, may have immunomodulatory properties and may provide a protective effect for suppressing certain pathogens.

[0010] Combinations of prebiotics and probiotics are referred to as synbiotics or synbiotic compositions. Such synbiotics comprising Bifidobacterium spp. and specific HMOs have been described, primarily in the context of supplements for infant formula or for promoting certain Bifidobacteria species in the gastrointestinal tract of infants, but also for preventing or treating disease in adults. Disclosures of various Bifidobacterium bifidum strains that have been tested in combination with different prebiotics such as HMOs are for example:

[0011] W02009 / 077352 describes prevention of opportunistic infections in immune-compromised infants or young children by combining various Bifidobacterium spp. with a fucosylated oligosaccharide.

[0012] WO2023 / 118510 discloses a combination of Bifidobacterium breve and Bifidobacterium bifidum with certain properties in combination with an HMO selected from 2'-fucosyllactose, 3- fucosyllactose, 3'-sialyllactose and 6'-sialyllactose.

[0013] WO2024 / 013393 describes the use of Bifidobacterium bifidum DSM 32403 or Bifidobacterium bifidum HA-132 in combination with fucosylated oligosaccharides to increase nicotinamide mononucleotide (NMN) and / or nicotinamide adenine dinucleotide (NAD+) in the gut.

[0014] OBJECTIVE

[0015] An objective of the present disclosure is to provide a synbiotic combination (kit of parts) or nutritional composition comprising a Bifidobacterium bifidum HA-132 and mixtures of HMOs for use in supporting or improving cognitive development, emotional and behavioural development, immune system development and anti-inflammatory support, and / or gut barrier function and / or gut motility, in a subject, by increasing the levels of gamma-aminobutyric acid (GABA) and / or purines, including guanosine, inosine, adenosine, and hypoxanthine, in the gut of the subject. It is also an object to provide a nutritional composition comprising a synbiotic combination, which is capable of increasing the levels of one or more of these molecules in the gut of a subject.

[0016] SUMMARY

[0017] The present disclosure relates to a synbiotic combination with Bifidobacterium bifidum HA-132 or closely related strains and at least one HMO, such as mixtures of HMOs, which is capable of increasing the levels of gamma-aminobutyric acid (GABA), the short chain fatty acid (SOFA) acetate and / or purines in the gut of a subject and thus be useful for supporting or improving cognitive development, emotional and behavioural development, immune system development and anti-inflammatory support, gut barrier function, and / or gut motility, in a subject.

[0018] As will be described in detail below, GABA has been shown to be beneficial for cognitive development, emotional and behavioral development, such as stress and anxiety reduction and gut health. Purines are beneficial for gut health, especially in strengthening mucosal barrier functions. The invention provides for in situ production of these molecules in the gut, thereby providing a prolonged and naturally controlled production. As such, the synbiotic combination according to the invention may be useful in supporting or improving cognitive, emotional and behavioural development, immune system development and anti-inflammatory support, gut barrier function, and / or gut motility.

[0019] A first aspect of the present invention relates to a kit of parts or a nutritional composition comprising a probiotic and at least one human milk oligosaccharide (HMO), wherein: i. the probiotic is the Bifidobacterium bifidum (B. bifidum) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and ii. the at least one HMO is selected from a fucosylated HMO, a neutral core HMO, and a sialylated HMO, or is any mixture thereof, for use in supporting or improving one or more of the following: a) cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, b) emotional and behavioural development, such as stress and anxiety reduction, c) a healthy immune system, including immune system development and anti-inflammatory support, and / or d) gut health, including a healthy gut barrier function and / or gut motility, in a subject.

[0020] A second aspect of the present invention relates to a nutritional composition comprising a probiotic and at least one first human milk oligosaccharide (HMO), wherein: a) the probiotic is the Bifidobacterium bifidum (B. bifidum) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and b) the at least one first HMO is a sialyllactose, such as 3’-SL or 6’-SL or a neutral core HMO, such as LNnT or LNT.

[0021] A third aspect of the present invention relates to a non-medical use of the nutritional composition according to the second aspect of the invention or embodiments thereof, wherein the non-medical use relates to supporting one or more of the following: i. cognitive development of one or more of motor skills, ability to learn, language skills, and spatial cognition ability, ii. development of social skills, iii. immune system development, and / or iv. gut health, gut barrier function and / or gut motility, in a healthy subject.

[0022] DETAILED DESCRIPTION

[0023] Bifidobacteria species has been demonstrated to predominate in the gut microbiota of breastfed infants, whereas the colonization of such species is less predominant in non-breastfed infants and infants delivered by C-section. Human milk oligosaccharides (HMO) are the primary substrate for Bifidobacterium bifidum. Several B. bifidum strains have been subject to pre-clinical and clinical testing and have shown various benefits, including Bifidobacterium bifidum HA-132 (see for example Saturio et al 2021 Microorganisms 9:2415; Ford et al 2020 Journal of the Academy of Nutrition and Dietetics 120(4):500-516.E10 and Brown et al 2022 Frontiers in Nutrition 9:1063121).

[0024] Definitions

[0025] A “kit of parts”, as used herein, has the same meaning as the term “combination” which is sometimes used. It indicates that each of the components - i.e., the probiotic and the at least one HMO, may be acquired as separate “parts” or compositions for later admixture before administration to the subject, or that each of the components may be administered separately to the subject for mixing in the gut.

[0026] A “nutritional composition”, as used herein, refers to any composition formulated from one or more separate ingredients which are suitable for human or animal consumption, such as an infant formula or a dietary supplement. It can contain sources of protein, lipids, vitamins, minerals and / or digestible carbohydrates and can be in dry, such as powdered, or liquid forms, preferably it is in a dry form. The composition can be designed to be the sole source of nutrition or a nutritional supplement. A synthetic nutritional composition is a composition where at least one of the ingredients is obtained by a biological process (e.g., enzymatic or fermentation) or chemical process. In the context of the present disclosure mothers milk is not considered to be a synthetic nutritional composition since it is not formulated from separate ingredients. In preferred embodiments the nutritional composition is a synthetic nutritional composition.

[0027] A “probiotic”, as used herein, refers to bacteria, which, when ingested in adequate amounts, provide a benefit to the host (human or animal) by replenishing or otherwise supplementing its natural gastrointestinal flora or by eliminating undesired bacteria in its gastrointestinal (Gl) tract or by executing beneficial metabolic activities along the Gl tract or by stimulating its immune system.

[0028] A “human milk oligosaccharide” or “HMO”, as used herein, refers to a complex carbohydrate found in human breast milk. The HMOs have a core structure comprising a lactose unit at the reducing end that can be elongated by one or more beta-N-acetyl-lactosaminyl and / or one or more beta- lacto-N-biosyl unit, and this core structure can be substituted by an alpha-L-fucopyranosyl and / or an alpha-N-acetyl-neuraminyl (sialyl) moiety. HMO structures are, e.g., disclosed by Xi Chen in Chapter 4 of Advances in Carbohydrate Chemistry and Biochemistry 2015 vol 72. In the context of the present disclosure, lactose (a disaccharide) is not regarded as an HMO species. HMOs can be non-acidic (or neutral) or acidic. Neutral HMOs are devoid of a sialyl residue and while acidic HMOs have at least one sialyl residue in their structure. The non-acidic (or neutral) HMOs can be fucosylated or non-fucosylated fucosylated (neutral-core) HMOs. The HMOs falling into these groups are described in further details in the section “Human Milk Oligosaccharides (HMOs)”.

[0029] “Gut” or “intestine”, are used interchangeably herein and refers to the portion of the gastrointestinal tract consisting of the small intestine and the large intestine. The “large intestine” {intestinum crassum) is the lower part of the gastrointestinal tract and is also referred to herein as “colon”.

[0030] A "subject", as referred to herein, may be a human or a mammal, or another domestic animal such as a pet (cats, dogs, rodents, rabbits, avian species, reptiles, etc.), livestock and performance animals (pigs, poultry, goat, sheep and cows) and working animals (horses, oxen, camels, donkeys and elephants) with a gut microbiome. In some embodiments, the subject is a healthy individual. In some embodiments, the subject is an infant. In some embodiments, the subject is a non-infant individual. In preferred embodiments, the subject is a healthy non-infant individual. Preferably, the subject is a human. The human may be an infant. As used herein, the term “infant” in the context of a human means a human of less than 3 years of age. The infant may be a pre-term infant, meaning that it is delivered before 37 weeks of pregnancy. The infant may be delivered by C-section, which means it has not been exposed to vaginas natural microbiota of the delivering woman. In a preferred embodiment, the human is a non-infant, such as a child, an adult or an elderly individual. As used herein in the context of a human, a “child” is a human of 3 to 12 years of age, an “adult” is a human of at least 18 years of age, and an "elderly individual" means a human of at least 60 years, preferably above 65 years, more preferably above 70 years of age. In another preferred embodiment, the human is a healthy individual.

[0031] The terms “treat” or “treatment” or “treating” as used herein refer to both treatment of an existing disease {e.g., a disease, condition or disorder as referred to herein) or prevention of a disease, i.e., prophylaxis. Maintenance and / or promotion of health in an individual not suffering from a disease, but who may be susceptible to the development of an unhealthy condition, is considered non-medical treatment in the context of the present invention.

[0032] An "effective amount" of a composition of the present disclosure means an amount that renders a desired health benefit / treatment outcome in the subject it is administered to when compared to a non-administered subject. An effective amount can be administered in one or more doses to achieve the desired treatment outcome.

[0033] “Enteral administration” means any conventional form for delivery of a composition to a subject that causes the deposition of the composition in the gastrointestinal tract (including the stomach). Methods of enteral administration include feeding through a naso-gastric tube or jejunum tube, oral, direct delivery to the gut, sublingual and rectal.

[0034] "Oral administration" means any conventional form of delivery of a composition to a subject, such as an infant ora non-infant, through the mouth. Accordingly, oral administration is a form of enteral administration.

[0035] Metabolites

[0036] Metabolites produced in the gut are small molecules generated as a result of the metabolic activities of the gut microbiota and host cells. These gut-derived metabolites include short-chain fatty acids (SCFAs), bile acids, vitamins, amino acids, and gases such as hydrogen and methane. They play crucial roles in maintaining gut health, regulating immune responses, influencing the gut-brain axis, and contributing to the overall metabolic homeostasis of the host. The gut metabolites are influenced by the nutrients consumed. Prebiotics and probiotics can for example enhance the growth of beneficial bacteria, leading to increased production of health-promoting metabolites.

[0037] Any variation in metabolites (i.e. , increase or decrease) effected or induced by the administration of the nutritional composition described herein is assessed relatively to the same subject prior to the administration of the composition described herein (i.e., the “non-administered subject(s)”). Alternatively, the comparative parameters may also constitute two cohorts of individuals, one cohort receiving a composition described herein (i.e., the “administered subjects”) and one or more additional cohorts receiving a placebo, such as a maltodextrin or lactose or alternatively the same mixture of HMOs without the probiotic (i.e., the “non-administered subjects”). The term “cohort” in this respect is understood as groupings of individuals with common traits, such as age, social and health factors. The size of the cohorts needed for comparative studies depends on the statistical variation observed within a cohort.

[0038] The present disclosure relates to a synbiotic combination with Bifidobacterium bifidum HA-132 or closely related strains and mixtures of HMOs, wherein the combination is capable of increasing the levels of gamma-aminobutyric acid (GABA), acetate, one or more short chain fatty acids (SCFAs), and / or purines, including guanosine, inosine, adenosine, and hypoxanthine, in the gut of a subject upon administration. The combination is a synbiotic combination or kit of parts of the present disclosure. The increase of the metabolites GABA and / or purines is preferably assessed by measuring the specific metabolite(s) in the gut prior to the administration of the composition of the present disclosure. As an alternative to measuring the metabolite(s) in the gut of a subject, the ability of a subject to produce the metabolite(s) following administration of the composition of the present disclosure can be accessed by fermenting a stool sample of said subject in the presence of a desired synbiotic composition, as described in the Examples. The increased levels of metabolite(s) upon administration of the synbiotic composition of the present disclosure can either be a direct increase due to the production of said metabolite(s) by Bifidobacterium bifidum HA-132 (or by closely related strains), or an indirect increase via the production of said metabolite(s) by the microbiota present in the gut of the subject. Such indirect increase of the metabolite(s) can be promoted or induced by metabolite(s) produced by Bifidobacterium bifidum HA-132 (or by closely related strains).

[0039] The gut-brain axis is the interplay between the brain and the gut microbiome and plays an important role in a wide range of physiological and psychological processes, including digestion, immune function, mood, and cognition (see for example Strandwitz 2018 Brain Research 1693:128-133). The effect of the microbiota of infants and the HMO composition in mother’s milk on cognitive abilities, such as motor skills, learning, language skills, spatial cognition ability, and anxiety reduction was studied by Cho et al. (2023 Front. Nutr. 10:1216327), which clearly indicates different benefits of certain HMOs and certain bacteria, for example in the modulation of or increase in the production of certain neurotransmitters.

[0040] Gamma-Aminobutyric Acid (GABA) is an important neurotransmitter that plays a crucial role in brain development and function. It is involved in regulating neuronal activity and is believed to have a calming effect on the brain. Studies have suggested that GABA may have a number of health benefits for children, including improving cognitive function, reducing anxiety and stress, and promoting healthy sleep patterns. GABA has also been shown to have a positive impact on certain developmental disorders, such as autism and attention deficit hyperactivity disorder (ADHD).

[0041] GABA can also be found in certain fermented foods, such as kimchi, yogurt, and kefir, and may be produced by some probiotic lactobacilli strains. GABA has also been shown to have potential benefits for gut health. For example, GABA may support gut barrier function and tight junctions and thereby contribute to ameliorating the effects caused by pathogens on the gut barrier (see for example Kaur et al. 2023 Microbial Cell Factories 22:256). Additionally, GABA may help to regulate gut motility, which could help to alleviate symptoms of constipation or diarrhea (Kerr and Ong, 2015 GABA Outside the CNS, GABA and Gut Motility, pp 29-44). Using a composition of the present disclosure to support the gut motility in a healthy individual not suffering from constipation or diarrhea is a non-medical use.

[0042] Furthermore, GABA may help to reduce inflammation in the gut, e.g. by reducing production of proinflammatory cytokines, such as TNF-a, IL6 and IL8, through downregulation of p38 mitogen- activated protein kinase (MAPK), which could be beneficial in treating or reducing symptoms for individuals with inflammatory bowel disease (IBD) or other gut-inflammatory conditions (see for example Aggarwal et al. 2018 J. Neurogastroenterol. Motil. 24(3): 422-430).

[0043] Immune system development refers to the process by which the immune system matures and becomes capable of defending the body against pathogens. The immune system consists of innate immunity (the body's first line of defence, including physical barriers, including the gut barrier, phagocytes, and natural killer cells) and adaptive immunity which involves highly specialized responses mediated by T cells and B cells. Using a composition of the present disclosure to support the development of the immune system in a healthy individual is a nonmedical use.

[0044] GABA plays a role in developing the immune system via the GABA receptors that are expressed on various immune cells, including T cells, B cells, macrophages, and dendritic cells. Activation of these receptors can influence the activity and behaviour of these immune cells and thereby influence the development of the immune system. The ability of GABA to modulate cytokine responses can influence B-cell activity and thereby affect immunoglobulin levels. GABA may therefore support the balancing of immunoglobulins of the different classes IgG, IgA, IgM, IgE, and IgD, thereby supporting maintenance of immunoglobulin homeostasis. In particular high levels of IgE can be associated with development of allergies.

[0045] Gut barrier function refers to the ability of the gut to prevent the leakage of harmful substances, such as toxins, bacteria, and undigested food particles, from the gut lumen into the bloodstream. The gut barrier is composed of several different components, including the gut epithelium, which is the layer of cells that lines the gut, as well as the mucus layer, which acts as a physical barrier to prevent direct contact between the gut epithelium and the gut lumen. The gut barrier also includes the tight junctions between gut epithelial cells, which help to regulate the passage of substances across the gut epithelium. When the gut barrier is compromised, harmful substances can leak into the bloodstream, and potentially trigger inflammation and immune responses. Several factors can contribute to the breakdown of the gut barrier, including poor diet, stress, infections, and medications such as nonsteroidal anti-inflammatory drugs (NSAIDs). Securing a strong gut barrier function in a healthy individual is therefore expected to protect this individual from developing unhealthy conditions, e.g., due to a poor diet. As described above GABA seems to have a positive effect on the gut barrier function.

[0046] The mucosal barrier is part of the gut barrier function and is a layer of mucus that lines the gastrointestinal tract and protects the body from harmful microorganisms and substances. The mucosal barrier function is not fully developed at birth, and the mucosal barrier continues to mature during the first year of life. In infants, this barrier is thinner and more permeable than in adults, which can make them more susceptible to infections and food allergies. The intestinal mucosa requires high levels of nucleotides for energy procurement, proliferation, and innate immunity. It has been shown that purine supplementation significantly improved intestinal epithelial cell wound healing and mucosal barrier functions.

[0047] Purines, such as guanosine, inosine, adenosine and hypoxanthine, may be produced by the gut microbiota, in particular E. coli K12 has been shown to be a source of purines in the gut (see for example Lee et al., 2018 J. Biol. Chem. 293:6039-6051 and Lee et al. 2020 iScience 101226). Promoting production of purines in the gut may therefore be beneficial for gut development, in particular in an infant, and can support or improve gut barrier functions in a subject.

[0048] In particular the purine adenosine also plays a role in the adenosine / ATP balance which affect immune responses via their interaction with adenosine receptors. Adenosine primarily have antiinflammatory effects and positively affects the differentiation of regulatory T-cells (Faas et al 2017 Molecular Aspects of Medicine 55: 9-19). Adenosine is therefore involved in immune system development.

[0049] SCFAs, of which butyrate, propionate, and acetate are the most predominant in the human gut, can have several health benefits, including potential benefits for gut immune health. SCFAs have been shown to have anti-inflammatory effects in the gut, which can help to promote a healthy immune response and reduce the risk and / or symptoms of inflammatory conditions in the gut, such as irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), Crohn’s disease, and / or ulcerative colitis. Additionally, SCFAs can help to support the growth and function of gut immune cells, such as T cells and regulatory T cells, which play a key role in maintaining a healthy immune system. Furthermore, SCFAs have been shown to help maintaining the gut barrier integrity, which can help to prevent harmful substances from entering the bloodstream and then trigger an immune response. Maintaining the gut barrier integrity can help to reduce the risk of allergies and chronic inflammation and other health issues. Examples of allergies where the risk may be reduced are in particular food allergies and potentially also drug allergies.

[0050] Acetate produced by bacteria acts in vivo to promote the defence functions of host epithelial cells. Acetate has been shown to promote intestinal antibody immunoglobulin (lg)A responses in the gut via the G-protein coupled receptor GPR43. Furthermore, acetate produced by B. infantis becomes a carbon source that stimulates growth and function of butyrate-producing microbes, such as Faecalibacterium prausnitzii.

[0051] In conclusion, GABA is therefore believed to be beneficial for cognitive development. In addition, both GABA, acetate and purines are beneficial for gut health, especially for the gut barrier and mucosal barrier functions. Furthermore, GABA and adenosine are beneficial in supporting the development of the immune system. Thus, both GABA and purines may contribute to preventing or treating various diseases or conditions of the gut and brain. An advantage of the present invention is that the kit of parts or composition disclosed herein provides a way to produce GABA and purines in situ in the gut. As such, these molecules do not have to be ingested and pass through the acidic environment of the stomach. Without being bound to any theory, the probiotic of the combination according to the invention would become a member of the microbiota community, and as such, it would be able to stay longer in the gut while providing a prolonged and naturally controlled production of GABA and purines as compared to orally administered GABA and / or purines if they can even survive the passage of the acidic environment in the stomach. Moreover, the combination of a prebiotic and a probiotic is easy to deliver, known to be safe to consume, and well accepted by the end-consumers or health care practitioners. Furthermore, the costs of such delivery are reasonable.

[0052] Use of the synbiotic combination or composition

[0053] The present disclosure relates to one or more methods of providing a subject with a health benefit by administering a synbiotic composition as disclosed herein. In particular the use, such as nonmedical use, of a composition or a kits of parts comprising a probiotic which is Bifidobacterium bifidum (B. bifidum) HA-132 (or closely related strains) and at least one HMO to achieve a health benefit is disclosed herein. Furthermore, an effective amount of the synbiotic composition disclosed herein can be used in a method for treating a condition, disease, or disorder.

[0054] The first aspect of the present disclosure relates to a kit of parts or a nutritional composition comprising a probiotic and at least one human milk oligosaccharide (HMO), such as a first HMO, wherein: i. the probiotic is the Bifidobacterium bifidum (B. bifidum) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and ii. the at least one HMO is selected from a fucosylated HMO, a neutral core HMO, and a sialylated HMO, or is any mixture thereof, for use in supporting or improving one or more of the following: a) cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, b) emotional and behavioural development, such as stress and anxiety reduction, c) a healthy immune system, including immune system development and anti-inflammatory support, and / or d) gut health, including a healthy gut barrier function and / or gut motility, in a subject.

[0055] In some embodiments, the subject is an infant, child, adult or elderly individual.

[0056] The kit of parts as disclosed herein may be used in the preparation of a nutritional composition or as a nutritional composition as such. The kit of parts or combination or composition can for example be used in an infant formula or dietary supplement. The kit of parts or combination or composition can also be used directly as a dietary supplement without addition of further active ingredients. Further details of nutritional compositions are described in the corresponding section below.

[0057] The constitutive elements of the nutritional composition or kit of parts in the first aspect are described in more detail in the sections below. It is understood that these elements can be integrated in the above aspect.

[0058] In some embodiments, the composition or the kit of parts (i.e, combination of the parts i) and ii)) disclosed herein increases gamma-aminobutyric acid (GABA), guanosine, inosine, adenosine, and / or hypoxanthine formation in the gut.

[0059] A healthy immune system is characterized by its ability to protect the body from harmful pathogens, such as viruses and bacteria, while also recognizing and ignoring harmless substances, such as food and pollen. A healthy immune system is able to distinguish between self and non-self, and it responds appropriately to foreign invaders while leaving the body's own cells and tissues unharmed. Additionally, a healthy immune system is able to adapt and remember previous encounters with pathogens, allowing for a quicker and more effective response upon subsequent exposure. One component of obtaining a healthy immune system is to secure an optimal development of the immune system during infancy and childhood, which, as described in the above, is supported by GABA, acetate, and adenosine.

[0060] In embodiments, the reduced risk of inflammation can for example be assessed by a decreased production of pro-inflammatory cytokines like TNF-a, IL6 and IL8 in the gut mucosa, leading to improved anti-inflammatory support as well as maintaining balanced levels of immunoglobulins.

[0061] Furthermore, the levels of immunoglobulins can also be measured to determine if the immune system is balanced and therefore likely to be healthy. Balanced levels of immunoglobulins are preferably in the following ranges IgG: 700-1600 mg / dL, IgA: 70-400 mg / dL, IgM: 40-230 mg / dL and IgE: less than 100 ILI / mL.

[0062] In some embodiments, the improved anti-inflammatory support leads to a reduced risk of developing allergies as well as to reduced symptoms of irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), Crohn’s disease, and / or ulcerative colitis.

[0063] In some embodiments, the reduced risk of developing allergies is useful in preventing development of allergies such as food allergy and drug allergy.

[0064] A fourth aspect of the present disclosure relates to a non-medical use of a nutritional composition or a kit of parts comprising a probiotic and at least one human milk oligosaccharide (HMO), wherein: a) the probiotic is the Bifidobacterium bifidum (B. bifidum) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and b) the at least one HMO is selected from a fucosylated HMO, a neutral core HMO, and a sialylated HMO, or is any mixture thereof, wherein the use relates to supporting one or more of the following: i. cognitive development of one or more of motor skills, ability to learn, language skills, and spatial cognition ability, ii. development of social skills, iii. immune system development, and / or iv. gut barrier function and / or gut motility, in a subject.

[0065] The embodiments described herein for the 1staspect of the present disclosure, relating to the probiotic and to the at least one HMO, respectively, apply mutatis mutandis to the 4thaspect of the present disclosure.

[0066] For the non-medical uses, the subject is preferably a healthy subject, preferably a healthy infant or child.

[0067] In the context of the non-medical uses, it is not intended to treat any diseases that affect the health of the gut, in particular the gut barrier function and gut motility.

[0068] Reducing the risk of developing allergies, as well as reducing risk of inflammation are considered non-medical uses in the context of the present disclosure in so far that the subject is free of allergies and inflammatory conditions at the time when the subject is provided with / uses a kit of parts or a nutritional composition of the present disclosure.

[0069] Supporting cognitive development is considered a non-medical use in the context of the present disclosure in so far that the subject is free from diseases that affect cognitive abilities at the time when the subject is provided with / uses a kit of parts or a nutritional composition of the present disclosure.

[0070] Embodiments of the use (in particular the non-medical use) of the present disclosure may for example be the use of the nutritional composition or a kit of parts of the present disclosure to support or improve the immune system, such as in the case of the immune system development, by: a) increasing GABA, acetate and / or adenosine in the gut compared to its measured level prior to administration of the nutritional composition or the kit of parts; and / or b) decreasing pro-inflammatory cytokines selected from the group consisting of TNF-a, IL6 and IL8 compared to their measured levels prior to administration of the nutritional composition or the kit of parts; and / or c) maintaining balanced levels of immunoglobulins. Gut health refers to the overall health and function of the digestive system, including the stomach, small intestine, large intestine, and colon. A healthy gut is characterized by a balanced and diverse microbiome, the ability to absorb nutrients, regular bowel movements and a strong gut barrier function. In addition, a healthy gut is mostly free of bloating, excessive gas production, abdominal pain, and diarrhea. Using a composition of the present disclosure to support or maintain a healthy gut in a healthy individual not suffering from any diseases in the gut is a nonmedical use.

[0071] The gut barrier function refers to the ability of the gut lining to act as a physical and functional barrier between the digestive system and the rest of the body. The strength of the tight junctions is another part of the gut barrier function. The tightness of the junctions can be assessed by measuring the permeability of the gut lining to certain molecules. This can be done by administering a solution containing a small molecule, such as lactulose or mannitol, and then measuring the concentration of the molecule in the urine. If the gut lining is permeable, more of the molecule will be able to pass through the gut lining and into the bloodstream, resulting in higher levels of the molecule in the urine.

[0072] In some embodiments, the gut barrier function is improved or supported by an in situ increase in GABA and / or acetate levels in the gut.

[0073] In the context of the present disclosure, the expression “gut motility” refers to the stretching and contractions of the muscles in the gastrointestinal (Gl) tract, also known as bowel movements. These synchronized movements are essential for the movement of food along the digestive tract while ensuring the absorption of important nutrients.

[0074] Embodiments of the use (in particular the non-medical use) of the present disclosure may for example be the use of the nutritional composition or a kit of parts of the present disclosure, to support or improve gut barrier function and / or gut motility by: a) an in situ increase of purine formation in the gut, such as an increase in guanosine, inosine, adenosine, and / or hypoxanthine compared to their measured levels prior to administration of the nutritional composition or the kit of parts, b) an in situ increase of GABA in the gut compared to its measured level prior to administration of the nutritional composition or the kit of parts, c) an in situ increase of acetate in the gut compared to its measured level prior to administration of the nutritional composition or the kit of parts.

[0075] Cognitive development refers to the progressive growth and development of a person's thinking, reasoning, problem-solving, and decision-making abilities. This process starts from infancy and continues throughout adulthood.

[0076] From birth to about 2 years old, infants learn about the world through their senses and motor actions. Motor actions or motor skills relate to the coordination of various muscles and body parts to achieve a specific goal. Examples of motor skills include walking, running, jumping, throwing, and catching. Development of spatial recognition starts in infancy and continues into adolescence and is connected to the motor skills. Spatial recognition relates to the ability to perceive, analyse, and understand spatial relationships between objects and the environment. This includes the ability to recognize and remember the location of objects in space, to mentally manipulate objects in space, and to navigate through the environment.

[0077] From about 2 to 7 years old, children develop symbolic thinking, language, and the ability to understand others' perspectives. Language development refers to the process by which children acquire the ability to understand and use language. This process begins in infancy and continues through childhood and adolescence. Language development involves several key milestones, including learning to recognize and differentiate between speech sounds, developing a vocabulary of words, learning grammar and syntax, and developing the ability to use language for communication.

[0078] The ability to learn is developed early in life and continues to develop throughout the lifespan. Infants are born with the ability to learn and are constantly learning about their environment through their senses and experiences. As they grow, their ability to learn becomes more sophisticated and complex, allowing them to acquire new knowledge and skills. Learning can be defined as the process of acquiring new knowledge, skills, behaviours, or attitudes through experience, study, or instruction. It involves a change in behaviour or mental processes that results from experience or practice. Learning can occur through a variety of methods, including observation, trial and error, feedback, and instruction. In addition, there are a number of psychological tests for evaluating cognitive functions in infants and young children such as Agpar score (newborns), Bayley Scales of Infant and Toddler Development (BSID), Cognitive Assessment of Young Children (CAYC).

[0079] Emotional development begins in infancy, with the development of basic emotions such as happiness, sadness, and fear. As children grow, they develop more complex emotions such as empathy, jealousy, and pride, and they learn to regulate their emotions in response to social cues and expectations. Stress and anxiety can have a significant impact on emotional development, as they can interfere with a person's ability to regulate their emotions and form healthy relationships with others. Negative stress, also known as distress, is a type of stress that is characterized by a feeling of overwhelm, anxiety, or helplessness in response to a perceived threat or challenge. Anxiety is characterized by excessive worry, fear, or apprehension. Stress and anxiety reduction can for example be assessed by monitoring a decrease in for example cortisol and alpha-amylase levels in the saliva of the subject before and after the administration of the composition or kit of parts of the present disclosure. In addition, there are a number of psychological tests for anxiety and stress such as various self-reporting questionnaires including, Beck Anxiety Inventory, State-Trait Anxiety Inventory or Perceived Stress Scale. Behavioural development involves the development of social skills and appropriate behaviours in different contexts. This includes the development of social skills such as communication, cooperation, and conflict resolution, as well as the ability to follow rules and norms of behaviour. The use of a composition or kit of parts of the present disclosure in the development of social skills in a healthy individual is a non-medical use.

[0080] Embodiments of the use (in particular the non-medical use) of the present disclosure may for example be the use of the nutritional composition or a kit of parts of the present disclosure to support or improve cognitive, emotional and behavioral development by increasing GABA in the gut and / or brain compared to its measured level prior to administration of the nutritional composition or the kit of parts. In particular, the use may be for supporting the cognitive development of one or more of motor skills, ability to learn, language skills, and spatial cognition ability in a subject. Preferably, in a subject that is an infant, child or adolescent. In some embodiments, the probiotic and the at least one HMO are to be administered separately to the subject (as a kit of parts).

[0081] For example, the probiotic can be administered 1 to 5 hours before the HMO(s), to allow the probiotic to start propagating in the gut prior to administering the HMO(s).

[0082] The proper dosage of the composition or kit of parts of the present disclosure may be determined, at least in part, based upon factors such as immune status, body weight and age. In some cases, the dosage of the HMO(s) will be similar to that found for the specific HMOs in human breast milk.

[0083] The required amount of the at least one HMO or a mixture of HMOs would generally be in the range from about 0.5 g to about 25 g per day, in certain embodiments from about 1 g to about 20 g per day, in certain embodiments from about 2 g to about 15 g per day, from about 3 g to about 10 g per day, in certain embodiments from about 1 g to about 10 g per day.

[0084] Appropriate probiotic dose regimes can be determined based on the present disclosure and / or on factors known to a person of ordinary skill in the art. The dosage of the probiotic within the nutritional composition is expressed as colony forming units (CFU) / day. As used herein, CFU means the number of viable cells (i.e., probiotic cells which are able to multiply via binary fission under controlled conditions).

[0085] In some embodiments, the probiotic dose can be higher than T105and lower than T1012CFU / day.

[0086] In some embodiments, the probiotic dose can be at least T105, T106, 2- 106, 3- 106, 4- 106, 5- 106, 6-106, 7-106, 8-106, 9-106, 1 -107, 2-107, 3-107, 4-107, 5-107, 6-107, 7-107, 8-107, 9-107,1 -108, 2-108, 3-108, 4-108, 5- 108, 6- 108, 7- 108, 8- 108, 9- 108, 1 ■ 109, 2- 109, 3- 109, 4- 109, 5- 109, 6- 109, 7- 109, 8- 109, 9-109, T1010, 2-1O10, 3-1O10, 4-1O10, 5-1O10, 6-1O10, 7-1O10, 8-1O10, 9-1O10, T1011, 2-1011, 3 1011,4-1011, 5-1011, 6-1011, 7-1011, 8-1011, 9-1011, or at least 1 -1012, CFU / day. It is understood that an amount / dose pr day is a dose equivalent to the indicated amount pr day, meaning that the daily amount / dose can be combined to a dose every 2nd, 3rd, 4thor 5thday or to a weekly dose, alternatively a daily amount / dose can be split into multiple dosages and administered two, three or four times a day.

[0087] The probiotic

[0088] The probiotic according to the first aspect of the invention is the Bifidobacterium bifidum (B. bifidum) HA-132 strain, which has been deposited at Collection Nationale de Cultures de Microorganismes (CNCM) under the accession number CNCM I-5898, as described under ‘Deposit’ above.

[0089] The probiotic may also be a closely related strain having at least 97% average nucleotide identity (ANI) with CNCM I-3426, such as 97.1 %, such as 97.2%, such as 97.3%, such as 97.4%, such as 97.5%, such as 97.6%, such as 97.7%, such as 97.8%, such as 97.9%, such as 98.0%, such as 98.1 %, such as 98.2%, such as 98.3%, such as 98.4%, such as 98.5%, such as 98.6%, such as 98.7%, such as 98.8%, such as 98.9%, such as 99.0%, such as 99.1 %, such as 99.2%, such as 99.3%, such as 99.4%, such as 99.5%, such as 99.6%, such as 99.7%, such as 99.8%, and such as 99.9%, ANI.

[0090] The probiotic may also be a closely related strain having at least 98% average nucleotide identity (ANI) with CNCM I-3426, such as 98.1 %, such as 98.2%, such as 98.3%, such as 98.4%, such as 98.5%, such as 98.6%, such as 98.7%, such as 98.8%, such as 98.9%, such as 99.0%, such as 99.1 %, such as 99.2%, such as 99.3%, such as 99.4%, such as 99.5%, such as 99.6%, such as 99.7%, such as 99.8%, and such as 99.9%, ANI.

[0091] The probiotic may also be a closely related strain having at least 99% average nucleotide identity (ANI) with CNCM I-3426, such as 99.1 %, such as 99.2%, such as 99.3%, such as 99.4%, such as 99.5%, such as 99.6%, such as 99.7%, such as 99.8%, and such as 99.9%, ANI.

[0092] Average nucleotide identity (ANI) is an in-silico method which can be used to determine genetic relatedness among bacterial strains. It is based on pairwise comparisons of all sequences shared by two strains and can be calculated using publicly available tools, such as OrthoANI with usearch (Yoon etal., 2017, Antonie van Leeuwenhoek, 110:1281-1286); ANI Calculator, JSpecies (Richter and Rossello-Mora, 2009, Proc Natl Acad Sci USA, 106:19126-19131); and JSpeciesWS (Richter et al., 2016, Bioinformatics, 32:929-931).

[0093] In some embodiments, the composition or the kit of parts of the present disclosure can further comprise at least one additional probiotic microorganism. In some other embodiments, the at least one additional probiotic microorganism is added to the first part of the kit which comprises B. bifidum HA-132 or a closely related strain. In some further embodiments, the at least one additional probiotic is added as a third part of the kit. The composition or the kit of part can comprise, as the at least one additional probiotic microorganism, a probiotic bacteria species from the Lactobacillus genus, Bacillus genus, Bifidobacteria genus, Enterococcus genus, Pediococcus genus and / or Streptococcus genus.

[0094] Lactobacillus species include, without any limitation, L. acidophilus, L. brevis, L. bulgaricus, L. casei, L. crispatus, L. delbrueckii, L. fermentum, L. gasseri, L. helveticus, L. lactis, L. plantarum, L. reuteri, L. rhamnosus, L. salivarius or L. paracasei. While it is not intended that the present disclosure be limited to any particular species of Lactobacillus, exemplary species and strains of Lactobacillus for the present disclosure includes, but are not limited to, the following well-known strains: L. helveticus / L. acidophilus HA-122 (sold by Lallemand Health Solutions ("LHS")), L. acidophilus R0418 (LHS), L. brevis HA-112 (LHS), L. casei HA-108 (LHS), L. casei R0215 (LHS), L. casei 431 (Chr. Hansen), L. delbrueckii bulgaricus HA-137 (LHS), L. fermentum HA-179 (LHS), L. helveticus HA-128 (LHS), L. helveticus HA-501 (LHS), L. helveticus R0052 (LHS), L. helveticus Lafti L10 (LHS), L. paracasei HA-196 (LHS), L. paracasei HA-274 (LHS), L. paracasei Lafti L26 (LHS), L. plantarum R0403 (LHS), L. plantarum R0202 (LHS), L. plantarum R1012 (LHS), L. plantarum TIFN101, L. reuteri HA-188 (LHS), L. reuteri (DSM12246, dsm-firmenich), L. rhamnosus HA-114 (LHS), L. rhamnosus HA-500 (LHS), L. rhamnosus R0011 (LHS), L. rhamnosus R0049 (LHS), L. rhamnosus R0343 (LHS), L. rhamnosus R1039 (LHS), L. salivarius HA-118 (LHS), L. salivarius R0078 (LHS), L. bulgaricus R0440 (LHS), L. lactis R1087 (LHS), L. rhamnosus GG (ATCC 53103) and / or L. rhamnosus LGG (DSM 33156, Chr. Hansen).

[0095] Bacillus species include, without any limitation, B. coagulans or B. subtilis. While it is not intended that the present disclosure be limited to any particular species of Bacillus, exemplary species and strains of Bacillus for the present disclosure includes, but are not limited to, the following well- known strains: B. subtilis R0179 (LHS) and / or B. subtilis HA-124 (LHS).

[0096] Bifidobacteria species include, without any limitation, B. bifidum, B. breve, B. infantis, B. lactis or B. longum. While it is not intended that the present disclosure be limited to any particular species of Bifidobacteria, exemplary species and strains of Bifidobacteria for the present disclosure includes, but are not limited to, the following well-known strains: B. bifidum R0071 (CNCM I-3426, LHS), B. breve R0070 (LHS), B. breve HA-129 (LHS), B. breve M-63 (NITE BP-02623, Morinaga), Bifidobacterium breve M-16V (LMG 23729, Morinaga), B. infantis R0033 (CNCM I-3424, LHS), B. infantis HA-116 (LHS), 8. lactis LAFTI® B94 (LHS), 8. lactis HA-194 (LHS), 8. lactis BB- 12(DSM 15954, Chr. Hansen), 8. longum R0175 (CNCM I-3470, LHS), 8. longum HA-135 (LHS), B. longum BB536 (ATCC BAA-999, Morinaga), B. longum DSM 32946, B. Infantis BB-02 (DSM 33361 , Istilos, Chr. Hansen), 8. Infantis LMG11588 (ATCC 17930), 8. infantis EVC001 (PTA- 125180), B. infantis Bi-26 (ATCC SD6720), 8. infantis M-63 (NITE BP-02623), B. infantis (ATCC 15697), B. infantis (NCTC 11817), B. infantis (NCTC 13219), B. infantis BT1 (KCTC 11859BP), B. infantis UBBI-01 , B. infantis IN-F29, B. infantis TPY 12-1 , B. infantis 1888B and / or B. infantis EK3. Enterococcus species include, without any limitation, E. faecium. While it is not intended that the present disclosure be limited to any particular species of Enterococcus, exemplary species and strains of Enterococcus for the present disclosure includes, but are not limited to, the following well-known strains: E. faecium R0026 (LHS) and / or E. faecium HA-127 (LHS).

[0097] Pediococcus species include, without any limitation, P. acidilactici. While it is not intended that the present disclosure be limited to any particular species of Pediococcus, exemplary species and strains of Pediococcus for the present disclosure includes, but are not limited to, the following well-known strain P. acidilactici HA-524 (LHS).

[0098] Streptococcus species include, without any limitation, S. thermophilus. While it is not intended that the present disclosure be limited to any particular species of Streptococcus, exemplary species and strains of Streptococcus for the present disclosure includes, but are not limited to, the following well-known strains: S. thermophilus R1018 (LHS), S. thermophilus R0083 (LHS) and / or S. thermophilus HA-110 (LHS).

[0099] In a preferred embodiment, the composition or the kit of parts of the present disclosure can use, as the at least one additional probiotic microorganism, one or more of the following probiotic species: B. bifidum R0033 (CNCM I-3424), B. bifidum R0071 (CNCM I-3426), B. breve HA-129 (CNCM I-5897), B. lactis LAFTI® B94 (CBS 118529), L. helveticus R0052 (CNCM 1-1722), L. rhamnosus HA-114 (LMG S-24117, CNCM I-5765), L rhamnosus R0011 (CNCM 1-1720), L. rhamnosus GG (ATCC 53103), B. Infantis LMG11588 (ATCC 17930), 8. infantis EVC001 (PTA- 125180), B. infantis Bi-26 (ATCC SD6720), 8. infantis M-63 (NITE BP-02623), B. infantis (ATCC 15697), B. infantis (NCTC 11817), B. infantis (NCTC 13219), B. infantis BT1 (KCTC 11859BP), B. infantis UBBI-01, B. infantis IN-F29, B. infantis TPY 12-1, B. infantis 1888B and B. infantis EK3.

[0100] In some embodiment, the probiotic bacteria of the present disclosure (8. bifidum HA-132 and / or potential added probiotic bacteria) can be provided in the form of viable cells, or in an inactivated form of non-viable cells (i.e., killed cultures). In a preferred embodiment, 8. bifidum HA-132 is provided in the form of viable cells. In some other embodiment, B. bifidum HA-132 is provided in the form of viable cells, and potential additional probiotic bacteria are provided in the form of viable cells and / or in an inactivated form of viable cells. As such, in some embodiments, the population of probiotic bacteria can be subjected to a thermal (heat or cold) treatment, a pH treatment, a radiation (sonication) treatment and / or a pressure treatment to kill the viable cells of the population of probiotic bacteria.

[0101] In a further embodiment, the probiotic bacteria of the present disclosure (B. bifidum HA-132 and / or potential additional probiotic bacteria) can be provided in encapsulated form to protect them from moisture during storage as well as from the stomach acidic conditions. The probiotic encapsulation processes are well known in the art, and can therefore be easily adapted by the one with skills in the art. Human Milk Oligosaccharide (HMO)

[0102] The at least one HMO according to the first aspect of the invention is selected from a fucosylated HMO, a neutral core HMO, and a sialylated HMO, or is any mixture thereof.

[0103] In some embodiments, the fucosylated HMO is selected from the group consisting of 2’- fucosyllactose (2’FL), 3-fucosyllactose (3-FL), difucosyllactose (DFL), lacto-N-fucopentaose I (LNFP-I), lacto-N-fucopentaose II (LNFP-II), lacto-N-fucopentaose III (LNFP-III), lacto-N- fucopentaose V (LNFP-V), lacto-N-difucohexaose I (LNDFH-I), lacto-N-difucohexaose II (LNDFH- II), lacto-N-difucohexaose III (LNDFH-I II), fucosyl-lacto-N-hexaose II (FLNH-II), , fucosyl-lacto-N- hexaose I (FLNH-I), fucosyl-para-lacto-N-hexaose I (FpLNH-l), fucosyl-para-lacto-N-neohexaose II (F-pLNnH II), and fucosyl-lacto-N-neohexaose (FLNnH).

[0104] In some embodiments, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of at least one HMO selected from 2’-FL, 3-FL and / or DFL, or a mixture thereof, such as 2’-FL and DFL or 2’-FL and 3-FL.

[0105] In some embodiments, the neutral core HMO is selected from the group consisting of lacto-N- triose II (LNT-II), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-neohexaose (LNnH), para-lacto-N-neohexaose (pLNnH), para-lacto-N-hexaose (pLNH), and lacto-N-hexaose (LNH).

[0106] In some embodiments, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of at least one HMO selected from LNT or LNnT.

[0107] In some embodiments, the sialylated HMO is selected from the group consisting of 3’-sialyllactose (3’-SL), 6’-sialyllactose (6’-SL), 3-fucosyl-3’-sialyllactose (FSL), 3’-O-sialyllacto-N-tetraose a (LST a), fucosyl-LST a (FLST a), 6’-O-sialyllacto-N-tetraose b (LST b), fucosyl-LST b (FLST b), 6’-O- sialyllacto-N-neotetraose (LST c), fucosyl-LST c (FLST c), 3’-O-sialyllacto-N-neotetraose (LST d), fucosyl-LST d (FLST d), sialyl-lacto-N-hexaose (SLNH), sialyl-lacto-N-neohexaose I (SLNH-I), sialyl-lacto-N-neohexaose II (SLNH-II), and disialyl-lacto-N-tetraose (DSLNT).

[0108] In some embodiments, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of at least one HMO selected from 3’-SL and / or 6’-SL, or a mixture thereof.

[0109] In preferred embodiments, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of at least one HMO, such as at least two HMOs, selected from a fucosylated HMO and / or a sialylated HMO, such as at least one HMO, such as at least two HMOs, selected from 2’-fucosyllactose (2’-FL), 3-fucosyllactose (3-FL), difucosyllactose (DFL), 3’-sialyllactose (3’-SL), and 6’-sialyllactose (6’-SL), or any mixture thereof.

[0110] In some embodiments, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL and 3’-SL. In some embodiment, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL and 6’-SL.

[0111] In some embodiments, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL and 3’-SL.

[0112] In some embodiment, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL and 6’-SL.

[0113] In some embodiments, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3’-SL and 6’-SL.

[0114] In some embodiment, the at least one HMO in the kits of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL, 3’-SL and 6’-SL.

[0115] In some embodiment, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3-FL, and 3’-SL.

[0116] In some embodiment, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3-FL, and 6’-SL.

[0117] In some embodiment, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3-FL, 3’-SL and 6’-SL.

[0118] In further embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises a fucosylated HMO and / or a sialylated HMO in combination with a neutral core HMO, such as lacto- / V-tetraose (LNT) or lacto-ZV-neotetraose (LNnT), preferably LNnT.

[0119] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL and LNnT.

[0120] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL and LNT.

[0121] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 3’-SL and LNnT.

[0122] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 3’-SL and LNT.

[0123] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 6’-SL and LNnT.

[0124] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 6’-SL and LNT.

[0125] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3’-SL and LNnT. In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3’-SL and LNT.

[0126] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 6’-SL and LNnT. In some embodiments, the mixture of HMOs comprises or consists essentially of 2’-FL, LNnT, 3’-SL and 6’-SL.

[0127] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 6’-SL and LNT.

[0128] In some embodiments, the mixture of HMOs comprises or consists essentially of 2’-FL, LNT, 3’- SL and 6’-SL.

[0129] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL and LNnT.

[0130] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL and LNT.

[0131] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, LNnT and LNT

[0132] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL, LNnT and LNT.

[0133] In some embodiment, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3-FL, and LNnT.

[0134] In some embodiment, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3-FL, and LNT.

[0135] In some embodiment, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3-FL, LNnT and LNT.

[0136] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL, LNnT, 3’-SL and 6’-SL.

[0137] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL, LNT, 3’-SL and 6’-SL.

[0138] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL, LNnT, LNT, 3’-SL and 6’-SL.

[0139] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of 2’-FL, 3-FL, LNT, 3’-SL and 6’-SL.

[0140] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3-FL, LNnT, 3’-SL and 6’-SL. In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, DFL, LNT, LNnT, 3’-SL and 6’-SL.

[0141] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of 2’-FL, 3-FL, LNT, LNnT, 3’-SL and 6’-SL.

[0142] In some embodiments, if a fucosylated HMO is present in the at least one HMO in in the kit of parts or composition disclosed herein, the total amount of fucosylated HMO constitutes between 60 to 100 wt% of the total mixture of HMOs, such as between 60 to 95 wt% of the total mixture of HMOs, or such as between 71 to 96 wt%, or such as between 75 to 98 wt%, of the total mixture of HMOs.

[0143] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of HMOs, which mixture of HMOs constitutes at least 95 wt% of the total amount of HMOs in the kit of parts or composition, wherein the mixture is selected from: a) 2’-FL and 3’-SL, in a ratio between 9.5-18:1 of 2’-FL:3’-SL, such as between 11-16:1 , or b) 2’-FL and LNnT, in a ratio between 8-2:1 of 2’-FL:LNnT, such as between 4-6.4:1 , or c) 2’-FL and LNT, in a ratio between 1-6:1 of 2’-FL:LNT, such as between 2-4:1 , or d) 2’-FL and 6’-SL, in a ratio between 4-10:1 of 2’-FL:6’-SL, such as between 5.5-8:1 , or e) 2’-FL and DFL, in a ratio between 4-:12 of 2’-FL:3-FL, such as between 6-10:1 , or f) 2’-FL and 3-FL, in a ratio between 1-5:1 of 2’-FL:3-FL, such as between 2-4:1 , or g) LNnT and 3’-SL, in a ratio between 0.5-2:1 of LNnT:3’SL, such as between 1-2:1 , or h) LNT and 6’-SL, in a ratio between 0.5-4: 1 of LNT:6’SL, such as between 1-3:1 , or i) 2’-FL, LNT and LNnT, in a ratio between 4-8:1-3:1 of 2’-FL:LNT:LNnT, such as between 5- 7:1 .5-2.5:1 , or j) 2’-FL, LNnT and DFL, in a ratio between 6-11 :0.5-3:1 of 2’-FL: LNnT: DFL, such as between 7-9: 1-2:1 , or k) 2’-FL, LNT and DFL, in a ratio between 6-11 :0.5-4:1 of 2’-FL:LNT:DFL, such as between 7- 9:1.5-3:1 , or l) 2’-FL, DFL and 3’-SL, in a ratio between 8-14:1-2:1 of 2’-FL:DFL:3’-SL, such as between 10- 12:1.5:1 , or m) 2’-FL, LNnT and 3’-SL, in a ratio between 10-18:1-5:1 of 2’-FL:LNnT:3’-SL, such as between 12-16:2-4:1 , or n) 2’-FL, 6’-SL and LNnT, in a ratio between 3-9:0.5-2:1 of 2’-FL:6’-SL:LNnT, such as between 4-8:1-1 .5:1 or o) 2’-FL, 6’-SL, LNnT and 3’-SL, in a ratio between 8-16:0.5-4: 0.5-4: 1 of 2’-FL:6’-SL:LNnT:3’- SL, such as between 10-14:1-3:1-3:1 , or p) 2’-FL, LNT and 3’-SL, in a ratio between 8-16:2-6:1 of 2’-FL:LNT:3’-SL, such as between 10- 14:3-5:1 , or q) 2’-FL, LNT and 6’-SL, in a ratio between 4-8:1-4:1 of 2’-FL:LNT:6’-SL, such as between 5-

[0144] 7:1-3:1 , or r) 2’-FL, LNT, 6’-SL and 3’-SL, in a ratio between 8-16:2-6:0.5-4:1 of 2’-FL:6’-SL:LNT:3’-SL, such as between 10-14:3-5:1-3:1 , or s) 2’-FL, LNT, LNnT, and DFL in a ratio between 4-12:1-4:0.5-3:1 of 2’-FL:LNT:LNnT:DFL, such as between 6-8: 1.5-3.5: 1-2:1 , or t) 2’-FL, DFL, LNT, 6’-SL and 3’-SL, in a ratio between 10-18:1-3:2-6:1-5:1 of 2’-FL:DFL:LNT:6’- SL:3’-SL, such as between 10-14:1-2:3-5:1-3:1 , or u) 2’-FL, DFL, LNnT, 6’-SL and 3’-SL, in a ratio between 8-16:1-3:0.5-4: 0.5-4:1 of 2’- FL:DFL:LNnT:6’-SL:3’-SL, such as between 10-14:1-2:3-5:3-5:1 , or v) 2’-FL, 3-FL, LNT, 6’-SL and 3’-SL, in a ratio between 10-18:2-6:2-6:1-5:1 of 2’-FL:3- FL:LNT:6’-SL:3’-SL, such as between 10-14:3-5:3-5:1-3:1 , or w) 2’-FL, DFL, LNT, LNnT, 6’-SL and 3’-SL, in a ratio between 10-16:0.5-4:2-6:1-5:1-5:1 of 2’-FL:DFL:LNT:LNnT:6’-SL:3’-SL, such as between 10-14:3-5:3-5:1-3:1-3:1 , or x) 2’-FL, 3-FL, LNT, LNnT, 6’-SL and 3’-SL, in a ratio between 10-16:2-6:2-6:1-5:1-5:1 of 2’- FL:3-FL:LNT:LNnT:6’-SL:3’-SL, such as between 10-14:3-5:3-5:1-3:1-3:1.

[0145] In further preferred embodiments, the mixture is a) or m) or u).

[0146] In further preferred embodiments wherein the mixture is a), the ratio of 2’-FL:3’-SL is 16:1. In more preferred embodiments, the ratio is 11.5:1.

[0147] In further preferred embodiments wherein the mixture is b), the ratio of 2’-FL:LNnT is 4:1. In more preferred embodiments, the ratio is 6.1 :1

[0148] In further preferred embodiments wherein the mixture is c), the ratio of 2’-FL:6’-SL is 8:1. In more preferred embodiments, the ratio is 5.8:1.

[0149] In further preferred embodiments wherein the mixture is d), the ratio of 2’-FL:DFL:3’-SL is 12:1.5:1.

[0150] In further preferred embodiments wherein the mixture is e), the ratio of 2’-FL:LNnT:3’-SL is 16:4:1. In more preferred embodiment, the ratio of 12:2:1

[0151] In further preferred embodiments wherein the mixture is f), the ratio of LNnT:3’SL is 2:1.

[0152] All ratios between specific HMOs given herein are by weight.

[0153] In some embodiments, the at least one HMO in the kit of parts or composition disclosed herein comprises or consists essentially of a mixture of HMOs, which mixture of HMOs constitutes at least 95 wt% of the total amount of HMOs in the nutritional composition, and the mixture is selected from: a) 80 wt% to 95 wt% of 2’-FL and 5 wt% to 20 wt% DFL, such as between 85 wt% to 90 wt% of 2’-FL and 10 wt% to 15 wt% DFL, or b) 65 wt% to 85 wt% of 2’-FL and 15 wt% to 35 wt% 3-FL, such as between 70 wt% to 80 wt% of 2’-FL and 20 wt% to 30 wt% 3-FL, or c) 65 wt% to 85 wt% of 2’-FL and 15 wt% to 35 wt% of LNT, such as between 70 wt% to 80 wt% of 2’-FL and 20 wt% to 30 wt% LNT, or d) 88 wt% to 98 wt% of 2’-FL and 2 wt% to 12 wt% 3’-SL, such as between 90 wt% to 95 wt% of 2’-FL and 5 wt% to 10 wt% 3’-SL, or e) 75 wt% to 95 wt% of 2’-FL and 5 wt% to 25 wt% 6’-SL, such as between 80 wt% to 90 wt% of 2’-FL and 10 wt% to 20 wt% 6’-SL, or f) 75 wt% to 95 wt% of 2’-FL and 5 wt% to 25 wt% LNnT, such as between 80 wt% to 90 wt% of 2’-FL and 10wt% to 20 wt% LNnT, or g) 55 wt% to 75 wt% LNnT and 30 wt% to 45 wt% 3’-SL, such as between 60 wt% to 70 wt% LNnT and 30 wt% to 40 wt% 3’-SL, or h) 55 wt% to 75 wt% LNT and 30 wt% to 45 wt% 6’-SL, such as between 60 wt% to 70 wt% LNT and 30 wt% to 40 wt% 6’-SL, or i) 75 wt% to 93 wt% of 2’-FL and 4 wt% to 15 wt% DFL and 3 wt% to 10 wt% 3’-SL, such as between 90 wt% to 95 wt% of 2’-FL and 2 wt% to 12 wt% DFL and 5 wt% to 10 wt% 3’-SL, or j) 60 wt% to 80 wt% of 2’-FL and 10 wt% to 30 wt% LNT and 5 wt% to 15 wt% 6’-SL, such as between 62 wt% to 72 wt% of 2’-FL and 18 wt% to 25 wt% LNT and 9 wt% to 13 wt% 6’-SL, or k) 60 wt% to 80 wt% of 2’-FL, 18 wt% to 28 wt% of LNT, and 3 wt% to 10 wt% of 3’-SL, such as between 65 wt% to 75 wt% of 2’-FL, 20 wt% to 23 wt% of LNT, and 4 wt% to 8 wt% of 3’-SL, or l) 55 wt% to 80 wt% of 2’-FL, 16 wt% to 24 wt% of LNT, 7 wt% to 14wt% of 6’SL and 3 wt% to 9 wt% 3’-SL, such as between, or 60 wt% to 70 wt% of 2’-FL, 18 wt% to 22 wt% of LNT, 9 wt% to 12 wt% of 6’SL and 4 wt% to 7 wt% 3’-SL, or m) 70 wt% to 85 wt% of 2’-FL and 10 wt% to 25 wt% LNnT and 3 wt% to 10 wt% 3’-SL, such as between 75 wt% to 80 wt% of 2’-FL and 12 wt% to 20 wt% LNnT and 4 wt% to 8 wt% 3’-SL or n) 65 wt% to 85 wt% of 2’-FL, 8 wt% to 16 wt% of LNnT, and 9 wt% to 17 wt% of 6’-SL, such as between 70 wt% to 80 wt% of 2’-FL, 10 wt% to 14 wt% of LNnT, and 11 wt% to 15 wt% of 6’- SL, or o) 60 wt% to 85 wt% of 2’-FL, 9 wt% to 15 wt% of 6’-SL, 8 wt% to 14wt% of LNnT and 3 wt% to 9 wt% 3’-SL, such as between 65 wt% to 75 wt% of 2’-FL, 10 wt% to 12 wt% of 6’-SL, 9 wt% to 12wt% of LNnT and 4 wt% to 8 wt% 3’-SL, or p) 60 wt% to 75 wt% of 2’-FL, 15 wt% to 25 wt% LNT and 5 wt% to 16 wt% of LNnT, such as between 65 wt% to 70 wt% of 2’-FL,18 wt% to 22 wt% LNT and 9 wt% to 13 wt% of LNnT, or q) 65 wt% to 85 wt% of 2’-FL, 5 wt% to 15 wt% DFL and 8 wt% to 16 wt% of LNnT, such as between 70 wt% to 80 wt% of 2’-FL, 7 wt% to 11 wt% DFL and 10 wt% to 14 wt% of LNnT, or r) 60 wt% to 80 wt% of 2’-FL, 5 wt% to 14 wt% DFL and 18 wt% to 26 wt% of LNT, such as between 65 wt% to 75 wt% of 2’-FL, 6 wt% to 11 wt% DFL and 20 wt% to 24 wt% of LNT, or s) 75 wt% to 95 wt% of 2’-FL, 5 wt% to 15 wt% of DFL, and 3 wt% to 10 wt% of 3’-SL, such as between 80 wt% to 90 wt% of 2’-FL, 8 wt% to 12 wt% of DFL, and 4 wt% to 8 wt% of 3’-SL, or t) 55 wt% to 70 wt% of 2’-FL, 5 wt% to 12 wt% of DFL, 15 wt% to 22 wt% of LNT, and 5 wt% to 15 wt% of LNnT, such as between 58 wt% to 65 wt% of 2’-FL, 6 wt% to 10 wt% of DFL, 18 wt% to 22 wt% LNT and 8 wt% to 12 wt% of LNnT, or u) 55 wt% to 75 wt% of 2’-FL, 5 wt% to 10 wt% DFL, 12 to 25 wt% of LNT, 5 wt% to 15 wt% 6’SL and 3 wt% to 8 wt% of 3’-SL, such as between 57 wt% to 62 wt% of 2’-FL, 6 wt% to 9 wt% DFL, 15 to 20 wt% of LNT, 8 wt% to 12 wt% 6’SL and 4 wt% to 6 wt% of 3’-SL, or v) 48 wt% to 60 wt% of 2’-FL, 12 wt% to 20 wt% 3-FL, 12 to 25 wt% of LNT, 5 wt% to 15 wt% 6’SL and 2 wt% to 8 wt% of 3’-SL, such as between 50 wt% to 60 wt% of 2’-FL, 14 wt% to 18 wt% 3-FL, 15 to 20 wt% of LNT, 7 wt% to 11 wt% 6’SL and 3 wt% to 5 wt% of 3’-SL, or w) 45 wt% to 65 wt% of 2’-FL, 4 wt% to 10 wt% DFL, 12 wt% to 21 wt% of LNT, 6 wt% to 14 wt% of LNnT, 5 wt% to 15 wt% 6’SL and 3 wt% to 8 wt% of 3’-SL, such as between 50 wt% to 60 wt% of 2’-FL, 5 wt% to 8 wt% DFL, 15 wt% to 20 wt% of LNT, 7 wt% to 10 wt% of LNnT, 7 wt% to 11 wt% 6’SL and 2 wt% to 6 wt% of 3’-SL, or x) 40 wt% to 60 wt% of 2’-FL, 10 wt% to 20 wt% 3-FL, 10 wt% to 20 wt% of LNT, 4 wt to 12 wt% of LNnT 4 wt% to 12 wt% 6’SL and 2 wt% to 8 wt% of 3’-SL, such as between 45 wt% to 55 wt% of 2’-FL, 12wt% to 18 wt% 3-FL, 12 wt% to 18 wt% of LNT, 6 wt to 10 wt% of LNnT, 6 wt% to 10 wt% 6’SL and 3 wt% to 6 wt% of 3’-SL.

[0154] The production of HMOs is well known. See for example Bych et al., 2019, Current Opinion in Biotechnology, 56:130-137, for a review on HMO production. In a preferred embodiment, a HMO, e.g., 2’-FL or 3’-SL, may be produced synthetically, meaning it is produced ex vivo chemically and / or biologically, e.g., by means of a chemical reaction, an enzymatic reaction or from recombinant cell cultures. See, for example, the methods described in W02012 / 127410, WO2010 / 115934, WO2010 / 115935, WO2013 / 139344 or WO2022 / 136337 or EP4239066.

[0155] Nutritional composition

[0156] In some embodiments, the probiotic and the at least one HMO are for use in a nutritional composition, such as an infant formula, growing up formula, a dietary supplement or a medical nutrition product. The kit of parts or nutritional composition of the present disclosure is particular suitable for infant formula and growing up formula. In one embodiment the nutritional composition may be in the form of a kit of parts with the probiotic in one compartment (also referring as a part) and the at least one HMO in another compartment and an instruction describing the best intake form. The parts may individually be in the form of a powder, a dry composition, a liquid or a gel.

[0157] In some embodiments, the probiotic and the at least one HMO are co-formulated into the same composition.

[0158] The co-formulated nutritional composition may be provided as a powder, a dry composition or a gel comprising the probiotic(s) and the at least one HMO. The formulation may be a premix suitable for mixing with other ingredients. The formulation may be a ready to use formulation, such as a unit dosage form, i.e., a capsule, tablet or sachet / stick pack or a formulation that needs to be dissolved in a liquid prior to use.

[0159] In some embodiments, the composition, such as the co-formulated composition, is a dry composition, such as a powder or granulate, which comprises less than 5 wt%, such as less than 3 wt%, and such as less than 1 wt%, water. A low water content is preferred to avoid that the probiotic assimilates the HMOs if they are co-formulated in the composition.

[0160] For dietary supplements the dry composition may be formulated into a tablet or capsule or sachet / stick pack or a gummy.

[0161] The nutritional composition can additionally contain or be mixed with sources of protein, lipids, vitamins, minerals and / or digestible carbohydrates. The composition can be designed to be the sole source of nutrition or a nutritional supplement.

[0162] Probiotics / bacteria can be dried in multiple ways, e.g., spray drying, freeze drying (lyophilization), drum drying or fluid bed drying. It is advantageous to have the bacteria in a dry inactive state, since this prevents them from consuming carbohydrates they may be formulated together with, such as lactose, HMOs or GOS, e.g. in an infant formula.

[0163] The probiotics / bacterial cells can be regenerated or revived (also termed reconstitution) by rehydration either in the stomach or gut of the consumer or by suspending the nutritional formulation in a water containing liquid prior to consumption. Preferably, the bacterial cells are viable after the reconstitution. The viability of the bacteria following reconstitution may be assessed by spreading them on an agar plate with suitable growth medium and counting the number of colonies formed after incubation for a predefined time (plate counting). Alternatively, FACS analysis may be used.

[0164] Synbiotic compositions

[0165] In addition to the kit of parts for use in providing health benefits, the present disclosure also relates to synbiotic compositions and / or combinations. In the second aspect, the invention relates to a nutritional composition comprising a probiotic and at least one first human milk oligosaccharide (HMO), wherein: a) the probiotic is the Bifidobacterium bifidum (B. bifidum) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and b) the at least one first HMO is a sialyllactose, such as 3’-SL or 6’-SL or a neutral core HMO, such as LNnT or LNT.

[0166] In some embodiments, the composition comprises at least one second HMO which is a fucosylated HMO.

[0167] In some embodiments, the composition comprises at least one first HMO which is a sialyllactose, such as 3’-SL or 6’-SL, and a second HMO selected from: i. a fucosylated HMO, and ii. a neutral core HMO.

[0168] In some embodiments, the composition comprises at least one first HMO which is a neutral core HMO, such as LNnT or LNT, and a second HMO selected from: i. a fucosylated HMO, and ii. sialyllactose, such as 3’-SL or 6’-SL.

[0169] Fucosylated HMOs and neutral core HMOs are described in more detail in the section entitled “Human Milk Oligosaccharides”. The combination / mixture of HMOs in this section apply mutatis mutandis to the second aspect of the present disclosure in so far that they comprise a sialyllactose.

[0170] In embodiments the nutritional composition comprises at least a probiotic and a mixture of human milk oligosaccharides (HMOs), wherein a) the probiotic is the Bifidobacterium bifidum (B. bifidum) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and b) the mixture of HMOs constitutes at least 95 wt%, such as at least 97 wt%, of the total amount of HMOs in the nutritional composition, wherein the mixture is selected from i. 2’-FL and 3’-SL, or ii. 2’-FL and 6’-SL, or iii. 2’-FL and LNnT, or iv. 2’-FL and LNT, or v. 2’-FL, LNnT and 3’-SL, or vi. 2’-FL, 6’-SL, LNnT and 3’-SL, or vii. 2’-FL, DFL, LNnT, 6’-SL and 3’-SL, or viii. 2’-FL, 3-FL, LNT, 6’-SL and 3’-SL, or ix. 2’-FL, DFL, LNT, LNnT, 6’-SL and 3’-SL, or x. 2’-FL, 3-FL, LNT, LNnT, 6’-SL and 3’-SL, or xi. 2’-FL, DFL, LNT, 6’-SL and 3’-SL.

[0171] In preferred embodiments the nutritional composition of the present disclosure comprises an HMO mixture selected from i) or iii) or vii) or viii) or ix) or x) or xi) or xii) above. Most preferably the nutritional composition of the present disclosure comprises an HMO mixture selected from i) or iii) above.

[0172] In additional embodiments, the at least one first HMO and the at least one second HMO constitute a mixture of HMOs, which mixture of HMOs constitutes at least 95 wt% of the total amount of HMOs in the composition, and the mixture is selected from: a) 2’-FL and 3’-SL, in a ratio between 9.5-18:1 of 2’-FL:3’-SL, such as between 11-16:1 , or b) 2’-FL and 6’-SL, in a ratio between 4-10:1 of 2’-FL:6’-SL, such as between 5.5-8:1 , or c) 2’-FL, DFL and 3’-SL, in a ratio between 8-14:1-2:1 of 2’-FL:DFL:3’-SL, such as between 10- 12:1.5:1 , or d) 2’-FL, LNnT and 3’-SL, in a ratio between 10-18:1-5:1 of 2’-FL:LNnT:3’-SL, such as between 12-16:2-4:1 , or e) LNnT and 3’-SL, in a ratio between 0.5-2: 1 of LNnT:3’-SL, such as between 1-2:1 , or f) 6’-SL and LNnT, in a ratio between 1-2:1 of 6’-SL:LNnT, such as between 1-1 .5:1 , or g) 2’-FL, DFL, LNT, 6’-SL and 3’-SL, in a ratio between 10-16:1-3:2-6:1-5:1 of 2’-FL:DFL:LNT:6’- SL:3’-SL, such as between 10-14:1-2:3-5:1-3:1 , or h) 2’-FL, DFL, LNnT, 6’-SL and 3’-SL, in a ratio between 8-16:1-3:0.5-4: 0.5-4:1 of 2’- FL:DFL:LNnT:6’-SL:3’-SL, such as between 10-14:1-2:3-5:3-5:1 , or i) 2’-FL, 3-FL, LNT, 6’-SL and 3’-SL, in a ratio between 10-16:2-6:2-6:1-5:1 of 2’-FL:3- FL:LNT:6’-SL:3’-SL, such as between 10-14:3-5:3-5:1-3:1 , or j) 2’-FL, DFL, LNT, LNnT, 6’-SL and 3’-SL, in a ratio between 10-16:0.5-4:2-6:1-5:1-5:1 of 2’- FL:DFL:LNT:LNnT:6’-SL:3’-SL, such as between 10-14:3-5:3-5:1-3:1-3:1 , or k) 2’-FL, 3-FL, LNT, LNnT, 6’-SL and 3’-SL, in a ratio between 10-16:2-6:2-6:1-5:1-5:1 of 2’- FL:3-FL:LNT:LNnT:6’-SL:3’-SL, such as between 10-14:3-5:3-5:1-3:1-3:1.

[0173] In a further embodiment, the nutritional composition comprises at least one additional probiotic. Preferably, the additional probiotic microorganism is a species selected from Bifidobacterium longum, Bifidobacterium lactis and / or Bifidobacterium infantis.

[0174] In further preferred embodiments, wherein the mixture is a), the ratio of 2’-FL:3’-SL is 16:1. In more preferred embodiments, the ratio is 11.5:1.

[0175] In further preferred embodiments, wherein the mixture is b), the ratio of 2’-FL:6’-SL is 8:1. In more preferred embodiments, the ratio is 5.8:1.

[0176] In further preferred embodiments, wherein the mixture is c), the ratio of 2’-FL:DFL:3’-SL is 12:1.5:1. In further preferred embodiments, wherein the mixture is d), the ratio of 2’-FL:LNnT:3’-SL is 16:4:1. In more preferred embodiments, the ratio is 12:2:1.

[0177] In further preferred embodiments, wherein the mixture is e), the ratio of LNnT:3’-SL is 2:1.

[0178] In further preferred embodiments, wherein the mixture is f), the ratio of 6’-SL:LNnT is 1 :1.

[0179] It is understood that the synbiotic compositions described in the above can be applied in medical as well as non-medical uses described herein on the same terms as any kit of parts described herein.

[0180] In a fifth aspect, the invention relates to a kit of parts comprising a probiotic and at least one first human milk oligosaccharide (HMO), wherein: a) the probiotic is the Bifidobacterium bifidum (B. bifidum) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and b) the at least one first HMO is a sialyllactose, such as 3’-SL or 6’-SL, or a neutral core HMO, such as LNnT or LNT.

[0181] The embodiments of the second aspect of the invention as defined herein apply mutatis mutandis to the fifth aspect.

[0182] EXAMPLES

[0183] Materials and methods

[0184] Materials:

[0185] Table 1: PD media recipe

[0186] The probiotic strains were the Bifidobacterium bifidum (B. bifidum) HA-132 (CNCM I-5898) from Lallemand Health Solutions (LHS). The dose used in all the experiments was T107CFU.

[0187] Preparations of the following human milk oligosaccharides (HMOs) were used: 2’-Fucosyllactose (2’-FL; GLYCARE® 2FL 9000, dsm-firmenich), 3-fucosyllactose (3-FL; GLYCARE®3FL 9001 , dsm-firmenich), lacto- / V-neotetraose (LNnT; GLYCARE® LNnT 9000, dsm- firmenich), DFL (GLYCARE® 2’FL / DFL 8001 , dsm-firmenich), lacto- / V-tetraose (LNT; GLYCARE® LNT 8001 , dsm-firmenich), 3’-silyllactose (3’-SL; GLYCARE® 3’SL 9001 , dsm-firmenich), and 6’- silyllactose (6’-SL; GLYCARE® 6’SL 9001 , dsm-firmenich). .

[0188] In Examples 1 and 2 the oligosaccharides were dosed at 10g / L.

[0189] In Examples 4 and 5 the oligosaccharides were dosed in g / L according to Table 2 below.

[0190] Table 2: Oligosaccharide amounts (g / L) used in Examples 5 to 7

[0191] All oligosaccharides were prepared as 20% (w / v) stocks in H2O filtered with a Nalgene syringe with a 0.2 pm filter (Thermo cat#723-2520). After individual stocks were made, mixtures of human milk oligosaccharides were combined into master stocks to test combinations.

[0192] Singapore infant fecal samples were sourced by Sequential Skin, Ltd (Singapore) and stored as 10% w / v in PBS:glycerol from a collection kit (Norgen Biotek Corp).

[0193] Experimental description:

[0194] Sample preparation

[0195] The probiotic strains were streaked on MRS media plates (Anaerobe Systems, cat#AS-6429) and grown anaerobically in a Coy chamber (Coy Laboratory Products) at 37°C for two days until single colonies were observed. Colonies were picked and inoculated into 10 mL of PD media (Anaerobe Systems cat#5255) with 1 % (v / v) lactose and grown for six to eight hours until an ODeoo between 0.2-0.5.

[0196] At the same time, 10% fecal slurry infant sample from two to three donors was diluted 1 :10 into PD media with 0.25% lactose (v / v). All samples were grown anaerobically at 37°C.

[0197] Anaerobic chamber protocol (used in Examples 1 and 2)

[0198] After eight hours, the fecal slurry sample was diluted 1 :2 into fresh PD media (no sugar added). Samples were then prepped in triplicate replicates in 98 deep-well plates (Axygen, cat#P-2ML- SQ-CS) consisting of: probiotic diluted 1 :20 (5% v / v final), HMO or GOS (diluted 1 :10 to a final 1% v / v), and the remaining volume was filled with fecal slurry culture to 1 mL. Several wells were left empty as contamination controls. The plate was then sealed with a plate cover (Sigma-Aldrich, cat Z380059) and placed in an incubator / shaker (Benchmark Scientific) in the anaerobic chamber for 24 hours at 37°C. The samples were tested with two technical replicates. After 24 hours the experiment was ended and samples were spun down at 3000 rpm for 10 minutes at 4°C. 200 pL of supernatant was removed from each well, added to a 0.2 pm filter plate (PALL cat#8582), and spun down at 1000 rpm for 5 minutes. The collected filtered supernatants were sent for untargeted LC-MS using Hydrophilic interaction chromatography (HILIC) covering polar metabolites including organic acids, nucleic acids, amino acids, hydrophilic vitamins, phospholipids etc. and reverse phase LC-MS covering more non-polar metabolites, and lipids including mono-, di-, tri-acylglycerides, fatty acids, etc.

[0199] Biolector protocol (used in Examples 4 and 5)

[0200] After eight hours, the fecal slurry sample from three donors (not the same as used above) was diluted 1 :2 into fresh PD media (no sugar added). Samples were then prepped in triplicate replicates in 48 deep-well FlowerPlates (Beckman Coulter Life Sciences, cat#M2P-MTP-48- BOH1) consisting of: probiotic diluted 1 :20 (5% v / v final), HMO or GOS (diluted 1 :10 to final concentrations (see table 5), and the remaining volume was filled with fecal slurry culture to 1 mL. Several wells were left empty as contamination controls. The plate was then sealed with a plate cover (Sigma-Aldrich, cat Z380059), covered with an anaerobic gassing lid (Beckman Coulter Life Sciences, Cat# E-AN-300), and placed in BioLector XT Microbioreactor (Beckman Coulter Life Sciences, cat# M2P-G-BLXT). Samples were fermented anaerobically for 24 hours at37°C. The samples were tested with three technical replicates.

[0201] EXAMPLE 1

[0202] Testing composition with B. bifidum HA-132 and selected HMOs in mixed microbial communities to assess the production of metabolites of interest to the gut-brain axis

[0203] The purpose of this experiment was to test the effects of incubating infant fecal samples with different combinations containing the probiotic strain B. bifidum HA-132 and different human milk oligosaccharides (HMOs). Interesting metabolites with potential positive effect on the gut-brain axis were analysed following untargeted LC-MS, and GABA was identified as a metabolite which when increased in situ in the gut following administration of the compositions disclosed herein could lead to improved cognitive function, reduced stress and anxiety and positive immunomodulatory effects and positive effects on gut motility.

[0204] The amount of GABA produced in infant fecal samples incubated with different combinations of B. bifidum HA-132 and different oligosaccharides is shown in Table 3.

[0205] Table 3: Gamma-aminobutyric acid (GABA) production in mixed microbial communities incubated with test ingredients as specified.

[0206] *T107CFU

[0207] ’Change probiotic compared to no probiotic

[0208] In the present example the ability of the B. bifidum HA-132 to produce GABA was not assessed. From Table 3 it can be seen that B. bifidum HA-132, when combined with either 2’-FL or 3’-SL (Mix1 and Mix2, respectively), increased the formation of GABA by between about 130% and 400% compared to measured GABA production using the individual HMOs 2’-FL and 3’-SL (Ref1 and Ref2, respectively). The data shows the unique effect of the combination of the specific B. bifidum HA-132 strain with the selected oligosaccharides.

[0209] EXAMPLE 2

[0210] Testing composition with B. bifidum HA-132 and selected HMOs in mixed microbial communities to assess effect on production of metabolites of interest for strengthening the mucosal barrier function in the gut

[0211] The purpose of this experiment was to test the effects of incubating infant fecal samples with different combinations containing the probiotic strain B. bifidum HA-132 and different human milk oligosaccharides (HMOs). Interesting metabolites with potential positive effect on the mucosal barrier in the gut were analysed following untargeted LC-MS, and purine salvage pathway metabolites, which, when increased in situ in the gut following administration of the compositions disclosed herein, could lead to improved development of the infant gut including reduced risk of inflammation and reduced risk of developing allergies by strengthening the gut barrier function.

[0212] The amounts of purines, including guanosine, inosine, adenosine, and hypoxanthine, produced in infant fecal samples incubated with different combinations of B. bifidum HA-132 and different oligosaccharides is shown in Table 4.

[0213] Table 4: Purine production in mixed microbial communities incubated with test ingredients as specified. The results are based on the average of two donors with two technical replicates. *Change probiotic compared to no probioticnThe values are not quantized and reflect the peak intensity

[0214] In the present example the ability of the B. bifidum HA-132 strain to produce purines was not assessed. From Table 4 it can be seen that the specific 2’-FL and B. bifidum HA-132 combination (Mix1) was capable of increasing the level of all the four purines that were tested. It is worth mentioning that the measured adenosine level obtained via this specific combination (Mix1) was increased by more than 4000%. The 3’-SL I B. bifidum HA-132 combination also showed positive effect on all the purine production (Mix2).

[0215] Since purines are known to have a positive effect on the mucosal barrier and tight junctions in the gut, this is an indication that the combination of B. bifidum HA-132 and fucosylated or sialylated oligosaccharides can have a positive effect on gut health and gut development in infants. The combinations may also have a positive impact on immunity in that they may prevent entry of immunogenic / harmful substances from the gut into the blood stream.

[0216] EXAMPLE 3

[0217] Effects of a synbiotic formulations in gut health, in cognitive, emotional and behavioural development and in overall health (immune system development, anti-inflammatory support, gut barrier function and gut motility) in infants up to 24 months of age.

[0218] Study design: randomized, double-blind, parallel, placebo-controlled trial

[0219] Phase: II

[0220] Sample size: 80 participants.

[0221] 1. 40 participants on the claimed synbiotic composition, namely “2 g per day of a 12:1 [2’-FL : 3’-SL] HMO mixture + 2.109CFU per day of B. bifidum HA-132”,

[0222] 2. 40 participants on the placebo (i.e. , base formula feeding).

[0223] Study length: 24 months.

[0224] Visits: Baseline (Birth), 1 month, 3 months, 6 months, 12 months, 18 months and 24 months

[0225] Biological samples collected:

[0226] Mother: stool, blood, saliva, vaginal swab, nasal cavity, breastmilk.

[0227] Infant: umbilical cord blood, stool, saliva, blood (finger or heel prick), urine, nasal cavity.

[0228] Population (main inclusion criteria):

[0229] Healthy pregnant mothers and their healthy newborn;

[0230] Singleton pregnancy;

[0231] Birth weight of 2500g or more;

[0232] Gestational age 37-42 weeks (i.e., term pregnancy);

[0233] - Absence of diagnosed genetic disease or malformations during pregnancy. Population (main exclusion criteria):

[0234] Obstetric or fetal complications known for current pregnancy such as preterm labor (premature rupture of membranes or preterm labor), chorioamnionitis, placenta previa, active vaginal bleeding, cervical cerclage, fetal distress or anomalies, corticosteroid therapy;

[0235] Risk factors for obstetric complications during current pregnancy such as gestational diabetes, type I / II diabetes and high blood pressure (systole >140 and diastole >90);

[0236] Chorioamnionitis, pre-eclampsia or hepatogestosis;

[0237] Suspected malformation or serious condition of the fetus / neonate.

[0238] Study Description:

[0239] It is hypothesized that the synbiotic formulation, consumed over a 12-month period, is able to increase the levels of short chain fatty acids (SCFAs) and lactic acid, of purines (i.e. , guanosine, adenosine, inosine, hypoxanthine), and of neurotransmitters such as gamma-aminobutyric acid (GABA), and / or dopamine in the gut of a subject, with beneficial effects for the subject’s health, when compared to a non-administered subject.

[0240] Study objectives:

[0241] Primary objective

[0242] To measure the levels of SCFAs and lactic acid in stools of infant participants, measured by: Change from baseline in stool SCFAs and lactic acid levels as measured by LC / MSMS.

[0243] Hypothesis’. The study participants who consume the synbiotic formulation have greater mean increase change in SCFA levels from baseline to 24 months of follow up than the study participants on the placebo.

[0244] Secondary objectives

[0245] Changes in inflammation and oxidative markers from blood and urine samples

[0246] Including, but not limited to, anti-inflammatory cytokines TGF-pi , IL12, IL22, IL37, and IL38, and pro-inflammatory cytokines TNF-a, TNF- , IL1 , IL6, IL8, IL11 , IL17, IL18, IFN-a, IFN-p and IFN-y, Malondialdehyde (MDA), Glutathione (GSH), 8-Hydroxydeoxyguanosine (8- OHdG), Tryptophan, Kynurenine, Total antioxidant capacity, C-reactive protein (hsCRP)

[0247] Hypothesis’. The study participants who consume synbiotic formulation have a reduced mean concentration in inflammatory and / or oxidative markers after 24 months of follow up than the study participants on the placebo.

[0248] Reduction in the incidence of colics

[0249] Comparison of the incidence of colics or inconsolable crying episodes between the intervention groups and placebo group.

[0250] Hypothesis: There is less incidence of colics or inconsolable crying episodes in the synbiotic groups than in the placebo group after 3 months of intervention. Prevention of infections or allergic diseases

[0251] Comparison of rate / occurrence of infections or allergic diseases diagnosed during intervention (cold, flu, eczema, atopic dermatitis, diarrhea or gastroenteritis, food allergies, inflammatory bowel diseases, etc.)

[0252] Hypothesis’. There is, on average, less number of days of infections and less occurrence of infections and / or allergic diseases and / or inflammatory bowel diseases diagnosed in the synbiotic group than in the placebo group after 24 months of follow up.

[0253] Metagenomics and metabolomics

[0254] Changes in microbiome profiles at baseline versus 3 months, 6 months, 12 months, 18 months and 24 months (composition, diversity, and metabolites of interest) o 16S rRNA gene sequencing and shotgun metagenomics o Strain specific qPCR assays (strain recovery in biological samples) o Targeted and untargeted metabolomics

[0255] Hypothesis’. There is, on average, greater presence of beneficial microbes and metabolites of interest, and / or support the establishment and development of the gut microbiome, in the synbiotic group compared to the placebo group after 24 months of follow up.

[0256] Gastrointestinal Health

[0257] Changes in gastrointestinal health and associated quality of life throughout intervention using the capture of frequency (mean daily number of stools) and consistency of bowel movement (Amsterdam infant stool scale)

[0258] Biomarkers of nutrients absorption (Arginine, Threonine, polyamines (putrescine, cadaverine) in feces)

[0259] Fecal inflammatory mediators: gastrointestinal function through the measurement of fecal inflammatory mediators

[0260] Fecal gut barrier function: gastrointestinal function through the measurement of Gl barrier function markers.

[0261] Hypothesis’. Participants in the synbiotic group report, on average, less gastrointestinal symptoms (e.g., constipation, diarrhea) and better nutrients absorption and a better barrier function than participants in the placebo group, after 24 months of follow up.

[0262] Cognition and mental health

[0263] Changes in cognitive status throughout intervention assessed by:

[0264] - Agpar score (newborns), Bayley Scales of Infant and Toddler Development (BSID), Cognitive Assessment of Young Children (CAYC)

[0265] Changes in mental health state including stress, sleep quality and anxiety assessed by: Mother’s daily / weekly diary Biological markers (e.g., salivary slgA, alpha-amylase levels, cortisol, melatonin, gut neurotransmitters (e.g., levels of GABA, dopamine, serotonin in feces)

[0266] Hypothesis: Participants in the synbiotic group have a better cognitive development than participants in the placebo group, after 24 months of follow up, when controlling fortheir baseline values.

[0267] Muscle Health and Growth Rate

[0268] Changes in muscle strength assessed by handgrip muscle strength (HGS) via dynamometer

[0269] - Antropometric measurements

[0270] Body composition (PEA POD)

[0271] Hypothesis: Participants in the synbiotic group have, on average, better muscle strength and a normal growth rate and body composition compared to participants in the placebo group, after 24 months of follow up.

[0272] Exploratory

[0273] - Infant fecal bacteria oligosaccharide consumption and infant fecal sialic acid concentrations

[0274] Hypothesis: Participants in the synbiotic group have an improved and selective consumption of HMO’s and higher levels of sialic acid concentration than participants in the placebo group, after 24 months of follow up, when controlling for their baseline values.

[0275] Safety objectives

[0276] To assess the safety and tolerability of a synbiotic formulation in the study population, measured by:

[0277] Growth rate and anthropometric measurements

[0278] Changes (outside normal variation) in clinical hematology markers (e.g., Complete Blood Count (CBC) including hemoglobin, hematocrit, red blood cell indices, platelet count, leukocyte count and differential elicited after baseline and throughout intervention

[0279] Changes (outside normal variation) in clinical chemistry markers (Comprehensive Metabolic Panel (CMP) including glucose, calcium, albumin, protein, sodium, potassium, bicarbonate, chloride, blood urea nitrogen, creatinine, alkaline phosphatase, alanine amino transferase, aspartate amino transferase and bilirubin) elicited after baseline and throughout intervention

[0280] - Adverse events (AEs) and Serious adverse events (SAEs) until study completion: Incidence of Adverse Events and treatments gastrointestinal symptoms and related symptoms (diarrhea, vomiting, constipation, colic, irritability) after synbiotic supplementation will be determined and reported. General health status of the infant such as occurrence of any illness, health care visits for sickness, fever, antibiotic and medication use and parental assessments of infant's overall health will also be documented. We do not expect a significant difference for AEs / SAEs or product tolerance between the synbiotic and placebo group.

[0281] Hypothesis: Participants in the synbiotic group have, on average, a normal growth rate and less values outside normal variation for markers associated with organ failure / tissue degeneration than participants in the placebo group.

[0282] EXAMPLE 4

[0283] Testing composition with B. bifidum HA-132 and selected HMO mixtures in mixed microbial communities to assess the production of GABA

[0284] The purpose of this experiment was to expand the knowledge obtained on individual HMOs in Example 1 to mixtures of 2’-FL with additional HMOs.

[0285] In addition, the dosage of the HMOs in the present example were reduced to levels closer to what is used in infant formula today. It should also be noted that the fecal sample donors are different than in Example 2, and therefore the microbial communities are different and the data in this example are therefore not directly comparable to the data in Example 2.

[0286] The amount of GABA produced in infant fecal samples incubated with different combinations of B. bifidum HA-132 and different oligosaccharides is shown in Table 5.

[0287] Table 5: Gamma-aminobutyric acid (GABA) production in mixed microbial communities incubated with test ingredients as specified. Oligosaccharide amounts are in g / L. Amounts of the individual HMOs in the mixtures are specified in Table 2 in the method section

[0288] *T107CFU

[0289] #% change in sample with probiotic compared to non-probiotic [(mix - ref) / ref]- 100%

[0290] From Table 5 it can be seen that B. bifidum HA-132, when combined with a HMO mixture of 2’- FL and 3’-SL or 2’-FL and LNnT (MixB and MixC, respectively), increased the formation of GABA as compared to the HMO mixture without the B. bifidum HA-132 (Ref-ll and Ref-Ill, respectively). In addition, the 2’-FL and 3’-SL HMO mixture in combination with B. bifidum HA-132 produced more GABA than the B. bifidum HA-132 strain combined with 2’-FL alone. EXAMPLE 5

[0291] Testing composition with B. bifidum HA-132 and selected HMO mixtures in mixed microbial communities to assess the effect on production of the short chain fatty acid (SCFA) acetate The purpose of this experiment was to test the effects of incubating infant fecal samples (mixed microbial communities) with compositions containing the probiotic strain Bifidobacterium bifidum HA-132 and different human milk oligosaccharide (HMO) mixtures on the production of acetate.

[0292] The amount of acetic acid (acetate) and lactic acid (lactate), produced in infant fecal samples incubated with B. bifidum HA-132 and / or different oligosaccharides is shown in Table 6. Table 6: Acetic acid production in mixed microbial communities incubated with test ingredients as specified. Oligosaccharide amounts are in g / L. Amounts of the individual HMOs in the mixtures are specified in Table 2 in the method section.

[0293] *% change in sample with probiotic compared to non-probiotic[(mix - ref) / ref]- 100%

[0294] *T107CFU As can be seen from Table 6, the combinations of B. bifidum HA-132 with 2’-FL alone (MixA) or a HMO mixture of 2’-FL and 3’-SL or 2’-FL and LNnT (MixB and MixC, respectively) had an increased production of acetate as compared to the HMO or HMO mixture without the B. bifidum HA-132. In addition, the mixtures of 2’-FL and either 3’-SL or LNnT in combination with B. bifidum HA-132 produced more acetate than the B. bifidum HA-132 strain combined with 2’-FL alone.

Claims

CLAIMS1 . A kit of parts or a nutritional composition comprising a probiotic and at least one human milk oligosaccharide (HMO), wherein: i. the probiotic is the Bifidobacterium bifidum (B. bifid urn) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and ii. the at least one HMO is selected from a fucosylated HMO, a neutral core HMO, and a sialylated HMO, or is any mixture thereof, for use in supporting or improving one or more of the following: a) cognitive development, such as motor skills, learning, language skills, and spatial cognition ability, b) emotional and behavioural development, such as stress and anxiety reduction, c) a healthy immune system, including immune system development and antiinflammatory support, and / or d) gut health, including a healthy gut barrier function and / or gut motility, in a subject.

2. The kit of parts or the nutritional composition for the use according to claim 1 , wherein the at least one HMO is selected from 2’-fucosyllactose (2’-FL), 3-fucosyllactose (3-FL), difucosyllactose (DFL), 3’-sialyllactose (3’-SL), and 6’-sialyllactose (6’-SL), or is any mixture thereof.

3. The kit of parts or the nutritional composition for the use according to claim 1 or 2, wherein the at least one HMO is a mixture of 2’-FL and 3’-SL.

4. The kit of parts or the nutritional composition for the use according to claim 2 or 3, further comprising a neutral core HMO, such as lacto-ZV-tetraose (LNT) or lacto-ZV-neotetraose (LNnT), preferably LNnT.

5. The kit of parts or the nutritional composition for the use according to any one of the preceding claims, wherein the at least one HMO comprises or consists essentially of a mixture of HMOs, which mixture of HMOs constitutes at least 95 wt% of the total amount of HMOs in the kit of parts, and wherein the mixture is selected from: a) 2’-FL and 3’-SL, or b) 2’-FL and LNnT, or c) 2’-FL, DFL and 3’-SL, or d) 2’-FL, LNnT and 3’-SL, or e) LNnT and 3’-SL.

6. The kit of parts or the nutritional composition for the use according to any one of the preceding claims, wherein the at least one HMO comprises or consists essentially of a mixture of HMOs,which mixture of HMOs constitutes at least 95 wt% of the total amount of HMOs in the kit of parts, and wherein the mixture is selected from: a) 2’-FL and 3’-SL, in a ratio between 9.5-18:1 of 2’-FL:3’-SL, such as between 11-16:1 , or b) 2’-FL and LNnT, in a ratio between 8-2:1 of 2’-FL:LNnT, such as between 4-6.4:1 , or c) 2’-FL, DFL and 3’-SL, in a ratio between 8-14:1-2:1 of 2’-FL:DFL:3’-SL, such as between 10-12:1.5:1 , or d) 2’-FL, LNnT and 3’-SL, in a ratio between 10-18:1-5:1 of 2’-FL:LNnT:3’-SL, such as between 12-16:2-4:1 , or e) LNnT and 3’-SL, in a ratio between 0.5-2: 1 of LNnT:3’SL, such as between 1-2:1 , preferably wherein the mixture is a), b) or e).

7. The kit of parts or the nutritional composition for the use according to any one of the preceding claims, wherein the probiotic and the at least one HMO are for use in a nutritional composition, such as an infant formula or a dietary supplement.

8. The kit of parts or the nutritional composition for the use according to any one of the preceding claims, wherein the probiotic and the at least one HMO are co-formulated into the same composition.

9. The kit of parts or the nutritional composition for the use according to claim 8, wherein the composition is a dry composition, such as a powder or granulate, which comprises less than 5 wt%, such as less than 3 wt%, and such as less than 1 wt% water.

10. The kit of parts or the nutritional composition for the use according to any one of the preceding claims, wherein the combination of the parts i) and ii) increases gamma-aminobutyric acid (GABA), guanosine, inosine, adenosine, and / or hypoxanthine formation in the gut.11 . The kit of parts or the nutritional composition for the use according to any one of the preceding claims, wherein the improved anti-inflammatory support leads to a reduced risk of developing allergies as well as to reduced symptoms of irritable bowel syndrome (IBS), inflammatory bowel disease (IBD), Crohn’s disease, and / or ulcerative colitis.

12. The kit of parts or the nutritional composition for the use according to any one of the preceding claims, wherein the subject is an infant, child, adult or elderly individual.

13. A nutritional composition comprising a probiotic and at least one first human milk oligosaccharide (HMO), wherein: a) the probiotic is the Bifidobacterium bifidum (B. bifid urn) HA-132 strain (CNCM I-5898) or a strain having at least 97% average nucleotide identity with CNCM I-5898, and b) the at least one first HMO is a sialyllactose, such as 3’-SL or 6’-SL or a neutral core HMO, such as LNnT or LNT.

14. The nutritional composition according to claim 13, wherein the composition comprises at least one second HMO, which is a fucosylated HMO.

15. The nutritional composition according to claim 13 or 14, wherein the mixture of HMOs constitutes at least 95 wt% of the total amount of HMOs in the nutritional composition, and wherein the mixture is selected from a) 2’-FL and 3’-SL, or b) 2’-FL and LNnT, or c) 2’-FL, DFL and 3’-SL, or d) 2’-FL, LNnT and 3’-SL, or e) LNnT and 3’-SL.

16. The nutritional composition according to any one of claims 13 to 15, wherein the at least one first HMO and the at least one second HMO constitute a mixture of HMOs, which mixture of HMOs constitutes at least 95 wt% of the total amount of HMOs in the composition, and wherein the mixture is selected from: a) 2’-FL and 3’-SL, in a ratio between 9.5-18:1 of 2’-FL:3’-SL, such as between 11-16:1 , or b) 2’-FL, DFL and 3’-SL, in a ratio between 8-14:1-2:1 of 2’-FL:DFL:3’-SL, such as between 10-12:1.5:1 , or c) 2’-FL, LNnT and 3’-SL, in a ratio between 10-18:1-5:1 of 2’-FL:LNnT:3’-SL, such as between 12-16:2-4:1 , or d) LNnT and 3’-SL, in a ratio between 0.5-2: 1 of LNnT:3’-SL, such as between 1-2:1 , or e) 6’-SL and LNnT, in a ratio between 1-2:1 of 6’-SL:LNnT, such as between 1-1 .5:1.

17. A non-medical use of the nutritional composition according to any one of claims 13 to 16, wherein the non-medical use relates to supporting one or more of the following: i. cognitive development of one or more of motor skills, ability to learn, language skills, and spatial cognition ability, ii. development of social skills, iii. immune system development, and / or iv. gut health, gut barrier function and / or gut motility, in a healthy subject.

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