Mixture of human milk oligosaccharides and bifidomacterium longum infantis

Abstract

The present invention provides a combination of Bifidobacterium longum subsp. infantis, lacto- N-fucopentaose I (LNFP-I) and a HMO mixture for use in therapy by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

Description

[0001] MIXTURE OF HMOS AND B. INFANTIS

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture consisting of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosy I lactose (3FL) and / or lacto-N-neotetraose (LNnT). The present invention also relates to uses thereof, for example in promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4- guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child.

[0004] BACKGROUND TO THE INVENTION

[0005] Human milk oligosaccharides (HMOs) have become the subject of much interest in recent years due to their roles in numerous biological processes occurring in the human organism. Mammalian milk contains at least 130 of these complex oligosaccharides (Urashima et al, Milk Oligosaccharides, Nova Biomedical Books, New York, 2011 , ISBN: 978-1-61122-831-1).

[0006] Infancy, especially the first weeks, 3 months, 6 months or 12 months of life is a critical period for the establishment of a balanced gut microbiota. It is known that the modulation of the gut microbiota during infancy and early childhood can prospectively have a significant influence in the future health status of the body. For example, the gut microbiota can have an influence on the development of a strong immune system later in life, as well as normal growth, and even on the development of obesity later in life.

[0007] Bifidobacterium longum subsp. infantis (B. infantis) has been demonstrated to predominate in the gut microbiota of breastfed infants and to benefit the host by accelerating maturation of the immune response, balancing the immune system to suppress inflammation, improving intestinal barrier function, and increasing short-chain fatty acid (SCFA) production. Reduced abundance of Bifidobacterium species in infants and young children has been correlated to chronic diseases, including asthma and obesity, as well as to lower vaccine response. Researchers have postulated that loss of Bifidobacterium species in the infant gut in populations of developed countries is linked to increased incidence of allergic and autoimmune diseases. Due to the loss of Bifidobacterium species in the infant gut and low breast-feeding rates, there is a need to provide infants with both HMOs and HMO-utilizing bacteria such as B. longum subsp. infantis to support a healthy microbiome for long-term health.

[0008] Thus, there is a need for compositions to improve the health or prevent and / or treat disease in infants and young children. SUMMARY OF THE INVENTION

[0009] The present inventors have surprisingly found that the combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture can synergistically increase production of beneficial metabolites, such as uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine, in the gastrointestinal tract of infants and young children. These findings support the use of this combination in improving the health or preventing and / or treating disease in infants and young children.

[0010] Accordingly, in a first aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in therapy by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0011] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3- methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT), wherein the infant or young child was born by C-section and / or preterm, and / or small for gestational age (SGA) and / or with a low, very low, or extremely low weight at birth (LWB, VLBW, or ELBW) and / or experienced intra-uterine growth retardation (IUGR), and / or was hospitalized the first days following birth and / or took antibiotic the first days following birth and / or was sick at birth, and / or has impaired microbiota and / or dysbiosis of microbiota.

[0012] In a further aspect, the invention provides use of a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture to promote the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0013] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in improving metabolic health or treating and / or preventing a metabolic disorder in an infant or young child, for example by promoting the production of one or more of uridine, adenine, guanine, AMP, cAMP, and / or acetyl-carnitine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyl lactose (2FL), difucosy I lactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0014] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in improving cardiovascular health or treating and / or preventing a cardiovascular disease in an infant or young child, for example by promoting the production of one or more of uridine, cAMP, and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0015] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in improving neurological health, promoting brain growth and / or development, or treating and / or preventing a neurological disorder in an infant or young child, for example by promoting the production of one or more of uridine, guanine, cAMP, 4-guanidinobutyric acid, acetyl-carnitine and / or 3- methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0016] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in improving mental health or treating and / or preventing a mental disorder in an infant or young child, for example by promoting the production of one or more of guanine, 4-guanidinobutyric acid, and / or acetyl- carnitine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT). In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in improving respiratory health or treating and / or preventing a respiratory disease in an infant or young child, for example by promoting the production of cAMP and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosy I lactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0017] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in improving gastrointestinal health or treating and / or preventing gastrointestinal inflammation in an infant or young child, for example by promoting the production of one or more of adenine, cAMP and / or 4-guanidinobutyric acid in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyl lactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyl lactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0018] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in modulating / improving the immune response in an infant or young child, for example by promoting the production of cAMP and / or p-hydroxyphenyllactic acid in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3- fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0019] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in preventing and / or treating an infection in an infant or young child, for example by promoting the production of one or more of cAMP, p-hydroxyphenyllactic acid, and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0020] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in preventing and / or treating apnea in an infant or young child, for example by promoting the production of 3- methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0021] In a further aspect, the invention provides a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in improving musculoskeletal health, treating and / or preventing a musculoskeletal disorder, or promoting bone and / or muscle growth and / or development in an infant or young child, for example by promoting the production of uridine and / or AMP in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0022] In a further aspect, the invention provides a method of promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4- guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, the method comprising administering the infant or young child a combination of Bifidobacterium longum subsp. infantis, lacto-N- fucopentaose I (LNFP-I) and a HMO mixture, wherein the HMO mixture consists of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0023] The combination may be administered in any suitable manner. Suitably, the combination is administered separately, simultaneously or sequentially. In some embodiments, the combination is administered simultaneously. Suitably, the combination is provided in the form of a nutritional composition. In some embodiments, the nutritional composition is selected from an infant formula, a starter infant formula, a follow-on or follow-up formula, a baby food, an infant cereal composition, a growing-up-milk, a fortifier such as a human milk fortifier, or a supplement.

[0024] In a further aspect, the invention provides a nutritional composition comprising Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT). In some embodiments, the nutritional composition is selected from an infant formula, a starter infant formula, a follow-on or follow-up formula, a baby food, an infant cereal composition, a growing-up-milk, a fortifier such as a human milk fortifier, or a supplement. The combination or composition may comprise any suitable Bifidobacterium longum subsp. infantis in any suitable amounts. Suitably, the Bifidobacterium longum subsp. infantis is Bifidobacterium longum subsp. infantis LMG 11588 or a strain having an Average Nucleotide Identity (ANI) of at least 95%, preferably at least 99%, more preferably at least 99.9% to Bifidobacterium longum subsp. infantis LMG 11588. Suitably, the combination or composition comprises from 103to 1012cfu of Bifidobacterium longum subsp. infantis, preferably from 107to 1012cfu of Bifidobacterium longum subsp. infantis, more preferably from 108to 101° cfu of Bifidobacterium longum subsp. infantis per g of combination or composition on a dry weight basis.

[0025] The combination or composition may comprise LNFP-I in any suitable amounts. Suitably, the combination or composition comprises LNFP-I in a total amount of from 25 mg / L to 4000 mg / L of the combination or composition or from 0.02 g / 100 g to 3.75 g / 100 g of the combination or composition on a dry weight basis. In some embodiments, the combination or composition comprises LNFP-I in a total amount of from 50 mg / L to 2500 mg / L, preferably from 60 mg / L to 2000 mg / L, more preferably from 80 mg / L to 1500 mg / L of the combination or composition. In some embodiments, the combination or composition comprises LNFP-I in a total amount offrom 0.04 g / 100 g to 2 g / 100 g, preferably from 0.05 g / 100 g to 1 .6 g / 100 g, more preferably from 0.06 g / 100 g to 1.2 g / 100 g, most preferably from 0.07 g / 100 g to 0.8 g / 100 g of the combination or composition on a dry weight basis.

[0026] The combination or composition may comprise any suitable HMO mixture in any suitable amounts. Suitably, the total HMO concentration is from 0.5 to 10 g / L, preferably from 1 to 7.5 g / L. In some embodiments, the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and 3-fucosyllactose (3FL). In some embodiments, the HMO mixture consists essentially of: i. 16 wt% to 69 wt% of 2FL, preferably 22 wt% to 59 wt%; ii. 9 wt% to 24 wt% of LNT, preferably 12 wt% to 21 wt%; iii. 2 wt% to 10 wt% of DFL, preferably 3 wt% to 8 wt%; iv. 8 wt% to 26 wt% of 6SL and 3SL combined, preferably 11 wt% to 22 wt%; and v. 1810 wt% to 50 wt% of 3FL, preferably 11 wt% to 43 wt%. In other embodiments, the HMO mixture consists of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'- sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT). In some embodiments, the HMO mixture consists essentially of: i. 20 wt% to 60 wt% of 2FL, preferably 22 wt% to 55 wt%; ii. 4 wt% to 30 wt% of LNT, preferably 6 wt% to 20 wt%; iii. 1 wt% to 12 wt % of DFL, preferably 3 wt% to 8 wt%; iv. 7 wt% to 23 wt% of 6SL and 3SL combined, preferably 8 wt% to 22 wt%; v. 10 wt% to 50 wt% of 3FL, preferably 13 wt% to 46 wt%; and vi. 3 wt% to 25 wt% of LNnT, preferably 5 wt% to 20 wt%. In other embodiments, the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and lacto-N-neotetraose (LNnT). In some embodiments, the HMO mixture consists essentially of: i. 34 wt% to 85 wt% of 2FL, preferably 40 wt% to 71 wt% of 2FL; ii. 10 wt% to 40 wt% of LNT, preferably 12 wt% to 26 wt% of LNT ; iii. 4 wt% to 14 wt% of DFL, preferably 5 wt% to 10 wt% of DFL; iv. 9 wt% to 31 wt% of 6SL and 3SL combined, preferably 10 wt% to 28 wt% of 6SL and 3SL combined; and v. 6 wt% to 30 wt% of LNnT, preferably 7 wt% to 23 wt% of LNnT.

[0027] The infant or young child may be at risk and / or in need thereof. In some embodiments, the infant or young child was born by C-section and / or preterm, and / or small for gestational age (SGA) and / or with a low, very low, or extremely low weight at birth (LWB, VLBW, or ELBW) and / or experienced intra-uterine growth retardation (IUGR), and / or was hospitalized the first days following birth and / or took antibiotic the first days following birth and / or was sick at birth, and / or has impaired microbiota and / or dysbiosis of microbiota.

[0028] BRIEF DESCRIPTION OF THE FIGURES

[0029] Figure 1 : (A) Synergy size for compounds with a background of 49% 2’FL, 14% 3FL, 7% DFL, 6% 3’SL, 8% 6’SL, and 16% LNT dosed at total HMO concentration of 2.66 g / L. (B) Synergy size for compounds with a background of 38% 2’FL, 14% 3FL, 5% DFL, 4% 3’SL, 7% 6’SL, 13% LNT, and 19% LNnT dosed at total HMO concentration of 2.66 g / L Synergy size was calculated by first subtracting the blank value form all other values and then calculating the ratio of the blanked synbiotic condition concentrate and the summed blanked prebiotic condition and blanked probiotic condition. Synergy was defined as ratios > 1.

[0030] DETAILED DESCRIPTION OF THE INVENTION

[0031] Definitions

[0032] As used herein, the following terms have the following meanings.

[0033] The term "subject" may refer to an infant, young child, child, an infant small for gestational age (SGA) or a preterm infant.

[0034] The term "infant" means a child under the age of 12 months.

[0035] The expression "young child" means a child aged between one and three years, also called toddler.

[0036] The term “child” means a child aged between three and twelve years. Preferably, the term “child” means a child aged between three and six years. A "preterm" or "premature" subject means an infant or young child who was not born at term. Generally it refers to an infant or young child born prior 36 weeks of gestation.

[0037] By the expression "small for gestational age" or "SGA" it is referred to an infant or young child who is smaller in size than normal for their gestational age at birth, most commonly defined as a weight below the 10th percentile for the gestational age. In some embodiments, SGA may be associated with intrauterine growth restriction (IUGR), which refers to a condition in which a foetus is unable to achieve its potential size.

[0038] By the expression “low birth weight”, it should be understood as any body weight under 2500g at birth.

[0039] The expression "nutritional composition" means a composition which nourishes a subject. This nutritional composition is usually to be taken orally or intravenously. It may include a lipid or fat source, a carbohydrate source and / or a protein source. In a particular embodiment the nutritional composition is a ready-to-drink composition such as a ready-to-drink formula.

[0040] In a particular embodiment, the nutritional composition of the present invention is a "synthetic nutritional composition". The expression "synthetic nutritional composition" means a mixture obtained by chemical and / or biological means, which can be chemically identical to the mixture naturally occurring in mammalian milks (i.e. the synthetic nutritional composition is not breast milk).

[0041] The expression "infant formula" as used herein refers to a foodstuff intended for particular nutritional use by infants during the first months of life and satisfying by itself the nutritional requirements of this category of person (Article 2(c) of the European Commission Directive 91 / 321 / EEC 2006 / 141 / EC of 22 December 2006 on infant formulae and follow-on formulae). It also refers to a nutritional composition intended for infants and as defined in Codex Alimentarius (Codex STAN 72-1981) and Infant Specialities (incl. Food for Special Medical Purpose).

[0042] The expression "infant formula" encompasses both "starter infant formula" and "follow-up formula" or "follow-on formula".

[0043] A "follow-up formula" or "follow-on formula" is given from the 6th month onwards and includes “growing-up milk”. It constitutes the principal liquid element in the progressively diversified diet of this category of person.

[0044] The expression “growing-up milk” (or “GUM”) refers to a milk-based drink generally with added vitamins and minerals, that is intended for young children or children. The expression "baby food" means a foodstuff intended for particular nutritional use by infants or young children during the first years of life.

[0045] The expression "infant cereal composition" means a foodstuff intended for particular nutritional use by infants or young children during the first years of life.

[0046] The term "fortifier" refers to liquid or solid nutritional compositions suitable for mixing with breast milk or infant formula.

[0047] The expression “weaning period” means the period during which the mother's milk is substituted by other food in the diet of an infant or young child.

[0048] The "mother's milk" should be understood as the breast milk or the colostrum of the mother.

[0049] An “oligosaccharide” is a saccharide polymer containing a small number (typically three to ten) of simple sugars (monosaccharides).

[0050] The term "HMO" or "HMOs" refers to human milk oligosaccharide(s). These carbohydrates are resistant to enzymatic hydrolysis by digestive enzymes (e.g. pancreatic and / or brush border), indicating that they may display functions not directly related to their caloric value. It has especially been illustrated that they play a vital role in the early development of infants and young children, such as the maturation of the immune system. Many different kinds of HMOs are found in the human milk. Each individual oligosaccharide is based on a combination of glucose, galactose, sialic acid (N- acetylneuraminic acid), fucose and / or N- acetylglucosamine with many and varied linkages between them, thus accounting for the enormous number of different oligosaccharides in human milk - over 130 such structures have been identified so far. Almost all of them have a lactose moiety at their reducing end while sialic acid and / or fucose (when present) occupy terminal positions at the non-reducing ends. The HMOs can be acidic (e.g. charged sialic acid containing oligosaccharide) or neutral (e.g. fucosylated oligosaccharide). Some examples of HMOs are the fucosylated oligosaccharides, the N-acetylated oligosaccharides and / or the sialylated oligosaccharides.

[0051] A "fucosylated oligosaccharide" is an oligosaccharide having a fucose residue. It has a neutral nature. Some examples are LNFP-I (lacto-N-fucopentaose I), 2’-FL (2' fucosyllactose), 3-FL (3-fucosyllactose).

[0052] The expressions “fucosylated oligosaccharides comprising an alpha-1, 2-fucosyl- epitope” and “2-fucosylated oligosaccharides” encompass fucosylated oligosaccharides with a certain homology of form since they contain an alpha-1 , 2'-fucosyl-epitope, therefore a certain homology of function can be expected. The expression “N-acetylated oligosaccharide(s)” encompasses both “N-acetyl- lactosamine” and “oligosaccharide(s) containing N-acetyl-lactosamine”. They are neutral oligosaccharides having an N-acetyl-lactosamine residue. Suitable examples are LNT (lacto- N-tetraose), para-lacto-N-neohexaose (para-LNnH), LNnT (lacto-N-neotetraose) and any combinations thereof. Other examples are lacto-N-hexaose, lacto-N-neohexaose, para- lacto- N-hexaose, para-lacto-N-neohexaose, lacto-N-octaose, lacto-N- neooctaose, iso- lacto-N- octaose, para- lacto-N-octaose and lacto-N-decaose.

[0053] A "sialylated oligosaccharide" is a charged sialic acid containing oligosaccharide, i.e. an oligosaccharide having a sialic acid residue. It has an acidic nature. Some examples are 3’- SL (3’-sialyllactose) and 6’-SL (6’-sialyllactose). The expressions "sialylated oligosaccharide" and "sialyllactose (SL)" can be used interchangeably. The trisaccharide sialyllactose consists of lactose at the reducing terminus and one sialic acid residue at the non-reducing end via an alpha-2,3 binding or alpha-2,6 binding, resulting in 3'-SL and 6'-SL, respectively.

[0054] A "precursor of HMO" is a key compound that intervenes in the manufacture of HMO, such as sialic acid and / or fucose.

[0055] The term “GOS” as used herein means “Galacto-oligosaccharide". Galacto-oligosaccharides (GOS) as used herein typically consist of p-linked galactose moieties with galactose or glucose at the reducing end. Such GOS contains p-(1— >2), p-(1— >3), p-(1— >4), or p-(1— >6) linked galactose moieties and may have a degree of polymerization (DP) of 3-8 galactose units. The term GOS is therefore preferably referred to as oligosaccharide(s) comprising at least three galactose units, more preferably as oligosaccharide(s) comprising at least four galactose units, preferably having a degree of polymerization (DP) of 3-8 galactose units.

[0056] The nutritional composition of the present invention can be in solid form (e.g. powder) or in liquid form. The amount of the various ingredients (e.g. the oligosaccharides) can be expressed in g / 100g of composition on a dry weight basis when it is in a solid form, e.g. a powder, or as a concentration in g / L of the composition when it refers to a liquid form (this latter also encompasses liquid composition that may be obtained from a powder after reconstitution in a liquid such as milk, water, e.g. a reconstituted infant formula or a follow- on / follow-up formula or a growing-up milk or an infant cereal product or any other formulation designed for infant nutrition).

[0057] The expressions “infants / young children fed exclusively with human breast milk”, “infants or young children exclusively breast fed”, “exclusive breast fed infants or young children’ and “breast-fed infants / young children” can be used interchangeably. They refer to infants or young children fed with a great majority (i.e. at least 90%, or at least 95%, or at least 99%) or all (100%) of nutrients and / or energy originating from human breast milk.

[0058] The expression “conventional nutritional composition” refers to standard synthetic nutritional compositions such as infant formula, follow-up milks or growing-up milks already found in the market.

[0059] The terms “microbial”, “microflora” and “microbiota” can be used interchangeably.

[0060] The expressions “microbiota in the gut”, “microbiota of the gut”, “gut microbiota” and “intestinal microbiota” can be used interchangeably.

[0061] By the expressions “preventing” or “prevention”, it is meant avoiding that a physical state, a condition or their consequences occurs and / or decreasing its incidence (i.e. reduction of the frequency). Prevention also encompasses delay or prevention of the onset of the symptoms of the disease, disorder or condition. Prevention may be absolute (such that no disease occurs) or may be effective only in some individuals or for a limited amount of time.

[0062] By the expressions “treating” or “treatment”, it is meant a decrease of the duration and / or of the severity of a physical state, a condition or their consequences (e.g. a decrease or elimination of symptoms of the condition). Treatment also encompasses to reduce, alleviate or eliminate one or more symptoms associated with the disease, disorder or condition which is being treated and / or to slow down, reduce or block the progression of the disease, disorder or condition which is being treated.

[0063] The improvement of health, prevention and / or the treatment of a physical state, a condition or their consequences can occur during the treatment (i.e. during the administration of the composition of the present invention, either immediately after the start of the administration or some time after, e.g. some days or weeks after the start). But it can also encompass the improvement of health, prevention and / or the treatment later in life. The term “later in life” encompasses the effect after the termination of the intervention or treatment. The effect “later in life” can be from 1 week to several months, or even years, for example from 2 to 4 weeks, from 2 to 6 weeks, from 2 to 8 weeks, from 1 to 6 months or from 2 to 12 months. Suitably, the effect “later in life” can be from 12 months to 12 years, such as from 2 years to 10 years, or from 4 years to 5 years, after the termination of the intervention or treatment. Suitably, the effect “later in life” lasts until the subject is at least 5 years of age, such as at least 10 years of age, at least 20 years of age or at least 30 years of age.

[0064] The term “prebiotic” means non-digestible carbohydrates that beneficially affect the host by selectively stimulating the growth and / or the activity of healthy bacteria such as bifidobacteria in the colon of humans (Gibson GR, Roberfroid MB. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr. 1995;125:1401-12).

[0065] The term “probiotic” means microbial cell preparations or components of microbial cells with a beneficial effect on the health or well-being of the host. (Salminen S, Ouwehand A. Benno Y. et al. “Probiotics: how should they be defined” Trends Food Sci. Technol. 1999:10 107-10). The microbial cells are generally bacteria or yeasts.

[0066] The term “synbiotic” may refer to a component that contains both probiotics and prebiotics, or a live microbe and a substrate that is selectively utilized by the co-administered live microbe (see e.g. Swanson, K.S., et al., 2020. Nature Reviews Gastroenterology & Hepatology, 17(11), pp.687-701).

[0067] The term “cfu” should be understood as colony-forming unit.

[0068] All percentages are by weight unless otherwise stated.

[0069] All weights expressed in g per 100g of composition are dry weight unless otherwise stated.

[0070] The term “SCFA” means short chain fatty acid(s). The expression “increasing SCFA production” means that the amount of systemic and / or colonic SCFA, is higher in an individual fed with the nutritional composition according to the present invention in comparison with a standard. The SCFA production may be measured by techniques known by the skilled person such as by Gas-Liquid Chromatography.

[0071] In the present context, the term “gastrointestinal tract” includes the mouth, pharynx, oesophagus, stomach, small intestine, large intestine, rectum and anus. The term “intestine” includes the small intestine, the large intestine and rectum.

[0072] In addition, in the context of the invention, the terms "comprising" or "comprises" do not exclude other possible elements. The composition of the present invention, including the many embodiments described herein, can comprise, consist of, or consist essentially of the essential elements and limitations of the invention described herein, as well as any additional or optional ingredients, components, or limitations described herein or otherwise depending on the needs.

[0073] Any reference to prior art documents in this specification is not to be considered an admission that such prior art is widely known or forms part of the common general knowledge in the field.

[0074] The invention will now be described in further details. It is noted that the various aspects, features, examples and embodiments described in the present application may be compatible and / or combined together any combination thereof. Composition and combination

[0075] The invention provides a composition comprising or consisting of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture consisting of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0076] The invention also provides a combination of Bifidobacterium longum subsp. infantis, lacto-N- fucopentaose I (LNFP-I) and a HMO mixture consisting of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0077] The combination may be administered separately, simultaneously or sequentially. In preferred embodiments, the combination is administered simultaneously. In some embodiments, the combination is administered in the form of a composition, for example any composition described herein.

[0078] Bifidobacterium longum subsp. infantis

[0079] The composition or combination of the invention comprises Bifidobacterium longum subsp. infantis.

[0080] Bifidobacterium longum is a bacterium of the Bifidobacterium genus which is present in the human gastrointestinal tract. In 2002, three previously distinct species of Bifidobacterium, B. infantis, B. longum, and B. suis, were unified into a single species named B. longum with the biotypes infantis, longum, and suis, respectively (Sakata, S., et al., 2002. International journal of systematic and evolutionary microbiology, 52(6), pp.1945-1951).

[0081] Any suitable Bifidobacterium longum subsp. infantis strain may be used in the present invention. Such strains will be well-known to the skilled person. Suitable strains include Bifidobacterium longum subsp. infantis LMG 11588 (also known as Bifidobacterium longum subsp. infantis NCC3039 or Bifidobacterium longum subsp. infantis ATCC 17930) and Bifidobacterium longum subsp. infantis ATCC 15697 (also known as Bifidobacterium longum subsp. infantis NCC 3078).

[0082] The Bifidobacterium longum subsp. infantis may be a strain having at least 95%, at least 96%, at least 97%, at least 98%, at least 99% (suitably, at least 99.9%) ANI to Bifidobacterium longum subsp. infantis strain known to the skilled person.

[0083] Suitably, the Bifidobacterium longum subsp. infantis has at least 95%, at least 96%, at least 97%, at least 98%, at least 99% (suitably, at least 99.1 %, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%) AN I to Bifidobacterium longum subsp. infantis LMG 11588 (also known as Bifidobacterium longum subsp. infantis NCC3039 or Bifidobacterium longum subsp. infantis ATCC 17930). Preferably, the Bifidobacterium longum subsp. infantis has at least 99.9% ANI to Bifidobacterium longum subsp. infantis LMG 11588.

[0084] An example of a microorganism genome that has at least 99.9% ANI with B. longum subsp. infantis LMG 11588 can be found in PATRIC (https: / / www.patricbrc.org), genome ID 1678.111. Hence, suitably the B. longum subsp. infantis having the PATRIC genome ID 1678.111 may be used in the present invention.

[0085] Bifidobacterium longum subsp. infantis LMG 11588 is sold by the Belgian Coordinated Collections of Microorganisms (BCCM) under the LMG accession number LMG 11588.

[0086] Bifidobacterium longum subsp. infantis ATCC 15697 is sold by the American Type Culture Collection (ATCC) under the accession number ATCC 15697.

[0087] The composition or combination according to the invention may contain from 103to 1012cfu of Bifidobacterium longum subsp. infantis, more preferably from 107to 1012cfu such as from 108to 1O10cfu of Bifidobacterium longum subsp. infantis per g of composition or combination on a dry weight basis. Suitably, the Bifidobacterium longum subsp. infantis is administered to the subject in an amount of at least about 106cfu / day, at least about 107cfu / day, or at least about 108cfu / day. Suitably, the Bifidobacterium longum subsp. infantis is administered to the subject in an amount of about 1012cfu / day or less, about 1011cfu / day or less, or about 1O10cfu / day or less.

[0088] In one embodiment, the Bifidobacterium longum subsp. infantis is viable.

[0089] Bifidobacterium animalis subsp. lactis

[0090] In some embodiments, the composition or combination of the invention comprises Bifidobacterium longum subsp. lactis.

[0091] Any suitable Bifidobacterium animalis subsp. lactis (B. lactis) strain may be used in the present invention. Such strains will be well-known to the skilled person. Suitable strains include Bifidobacterium animalis subsp. lactis CNCM I-3446.

[0092] The Bifidobacterium animalis subsp. lactis may be a strain having at least 95%, at least 96%, at least 97%, at least 98%, at least 99% (suitably, at least 99.9%) ANI to a Bifidobacterium animalis subsp. lactis strain known to the skilled person. Suitably, the Bifidobacterium animalis subsp. lactis has at least 95%, at least 96%, at least 97%, at least 98%, at least 99% (suitably, at least 99.1 %, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%) AN I to Bifidobacterium animalis subsp. lactis CNCM I-3446. Preferably, the Bifidobacterium animalis subsp. lactis has at least 99.9% ANI to Bifidobacterium animalis subsp. lactis CNCM I-3446.

[0093] Bifidobacterium lactis CNCM 1-3446 was deposited with the Collection Nationale de Cultures de Microorganismes (CNCM), Institut Pasteur (INSTITUT PASTEUR, 25 RUE DU DOCTEUR ROUX, F-75724 PARIS CEDEX 15, FRANCE) by NESTEC S.A. (NESTEC S.A., AVENUE NESTLE 55, CH-1800 VEVEY) according to the Budapest Treaty on 7thJune 2005 receiving the deposit number CNCM 1-3446.

[0094] The composition or combination according to the invention may contain from 103to 1012cfu of Bifidobacterium animalis subsp. lactis, more preferably from 107to 1012cfu such as from 108to 101° cfu of Bifidobacterium animalis subsp. lactis per g of composition or combination on a dry weight basis. Suitably, the Bifidobacterium animalis subsp. lactis is administered to the subject in an amount of at least about 106cfu / day, at least about 107cfu / day, or at least about 108cfu / day. Suitably, the Bifidobacterium animalis subsp. lactis is administered to the subject in an amount of about 1012cfu / day or less, about 1011cfu / day or less, or about 101° cfu / day or less.

[0095] The composition or combination according to the invention may contain from 103to 1012cfu of Bifidobacteria (e.g. a combination of Bifidobacterium longum subsp. infantis and Bifidobacterium animalis subsp. lactis) more preferably from 107to 1012cfu such as from 108to 101° cfu of Bifidobacteria (e.g. of a combination of Bifidobacterium longum subsp. infantis and Bifidobacterium animalis subsp. lactis) per g of composition or combination on a dry weight basis. Suitably, the Bifidobacteria (e.g. a combination of Bifidobacterium longum subsp. infantis and Bifidobacterium animalis subsp. lactis) are administered to the subject in an amount of at least about 106cfu / day, at least about 107cfu / day, or at least about 108cfu / day. Suitably, the Bifidobacteria (e.g. a combination of Bifidobacterium longum subsp. infantis and Bifidobacterium animalis subsp. lactis) are administered to the subject in an amount of about 1012cfu / day or less, about 1011cfu / day or less, or about 101° cfu / day or less.

[0096] In one embodiment, the Bifidobacterium animalis subsp. lactis is viable.

[0097] Lacto-N-fucopentaose I (LNFP-I)

[0098] The composition or combination of the invention comprises lacto-N-fucopentaose I (LNFP-I). In some embodiments, LNFP-I is present in a total amount of from 10 mg / L to 5000 mg / L of the composition or combination according to the invention or of from 0.01 g / 100 g to 4 g / 100 g of the nutritional composition or combination according to the invention.

[0099] In some embodiments, LNFP-I is present in a total amount of from 25 mg / L to 4000 mg / L of the composition or combination according to the invention or of from 0.02 g / 100 g to 3.75 g / 100 g of the nutritional composition or combination according to the invention. Suitably, LNFP-I is present in a total amount of from 50 mg / L to 2500 mg / L, for example from 60 mg / L to 2000 mg / L, for example from 80 mg / L to 1500 mg / L, for example from 100 mg / L to 1000 mg / L, for example from 200 mg / L to 800 mg / L of the composition or combination according to the invention. Suitably, LNFP-I is present in a total amount of from 0.04 g / 100 g to 2 g / 100 g, for example from 0.05 g / 100 g to 1.6 g / 100 g, for example from 0.06 to 1.2 g / 100g, for example from 0.07 g / 100 g to 0.8 g / 100 g, for example from 0.1 g / 100g to 0.7 g / 100g of the composition or combination (dry weight).

[0100] In one embodiment, the composition or combination comprises from 0.015 wt.% to 3.8 wt.%, preferably from 0.08 wt.% to 1.2 wt.%, of lacto-N-fucopentaose I (LNFP-I) of the total wt.% of the composition or combination.

[0101] LNFP-I may be isolated by chromatography or filtration technology from a natural source such as animal milks. Suitably, the animal milk as used herein may be cow, sheep, goat, camel or buffalo milk. Preferably, the animal milk is cow’s milk. Preferably, the LNFP-I may be produced by biotechnological means using specific fucosyltransferases and / or fucosidases either through the use of enzyme-based fermentation technology (recombinant or natural enzymes) or microbial fermentation technology. In the latter case, microbes may either express their natural enzymes and substrates or may be engineered to produce respective substrates and enzymes. Single microbial cultures and / or mixed cultures may be used. Fucosylated oligosaccharide formation can be initiated by acceptor substrates starting from any degree of polymerization (DP), from DP = 1 onwards. Suitable techniques for producing LNFP-I are known in the art (see, for example, Hu et al., Carbohydr Polym, 2022, 297: 120017 and Derya et al., J Biotechnol., 2020, 318: 31-38). Alternatively, LNFP-I may be produced by chemical synthesis from lactose as initial acceptor substrate building an LNT backbone and free fucose as final donor substrate or by starting from LNT for example, produced by biotechnology or chemical synthesis, and using fucose.

[0102] Fucosylated oligosaccharides are also available for example from DSM of the Netherlands (for example, GlyCare™ 3FL 9001 and GlyCare™ LNFP-I / 2FL 8001) or from Elicityl of France (for example, 2FL, 3FL, DFL, LNFP-I). HMO mixture

[0103] The composition or combination of the invention comprises a HMO mixture consisting of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0104] In some embodiments, the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and 3- fucosyllactose (3FL).

[0105] In some embodiments, the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and lacto-N- neotetraose (LNnT).

[0106] In some embodiments, the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT).

[0107] In one embodiment, the HMO mixture comprises 2FL in an amount of from 16 wt% to 85 wt%. Suitably, the HMO mixture may comprise 2FL in an amount of from 31 wt% to 82 wt%, preferably from 41wt% to 70 wt%. Suitably, the HMO mixture may comprise 2FL in an amount of from 16 wt% to 69 wt%, preferably from 22 wt% to 59 wt%. Suitably, the HMO mixture may comprise 2FL in an amount of from 34 wt% to 85 wt%, preferably from 40 wt% to 71 wt%. Suitably, the HMO mixture may comprise 2FL in an amount of from 20 wt% to 60 wt%, preferably from 22 wt% to 55 wt%.

[0108] In one embodiment, the HMO mixture comprises LNT in an amount of from 4 wt% to 40 wt%. Suitably, the HMO mixture may comprise LNT in an amount of from 10 wt% to 27 wt%, preferably from 14 wt% to 23 wt%. Suitably, the HMO mixture may comprise LNT in an amount of from 9 wt% to 24 wt%, preferably 12 wt% to 21 wt%. Suitably, the HMO mixture may comprise LNT in an amount of from 10 wt% to 40 wt%, preferably 12 wt% to 26 wt%. Suitably, the HMO mixture may comprise LNT in an amount of from 4 wt% to 30 wt%, preferably 6 wt% to 20 wt%.

[0109] In one embodiment, the HMO mixture comprises DFL in an amount of from 1 wt% to 14 wt%. Suitably, the HMO mixture may comprise DFL in an amount of from 4 wt% to 11 wt%, preferably from 6 wt% to 10 wt%. Suitably, the HMO mixture may comprise DFL in an amount of from 2 wt% to 10 wt%, preferably from 3 wt% to 8 wt%. Suitably, the HMO mixture may comprise DFL in an amount of from 4 wt% to 14 wt %, preferably from 5 wt% to 10 wt%. Suitably, the HMO mixture may comprise DFL in an amount of from 1 wt% to 12 wt %, preferably from 2 wt% to 8 wt%.

[0110] In one embodiment, the HMO mixture comprises 6SL and 3SL combined in an amount of from

[0111] 7 wt% to 34 wt%. Suitably, the HMO mixture may comprise 6SL and 3SL combined in an amount of from 9 wt% to 34 wt%, preferably from 11 wt% to 29 wt%. Suitably, the HMO mixture may comprise 6SL and 3SL combined in an amount of from 8 wt% to 26 wt%, preferably from 11 wt% to 22 wt%. Suitably, the HMO mixture may comprise 6SL and 3SL combined in an amount of from 9 wt% to 31 wt%, preferably from 10 wt% to 28 wt%. Suitably, the HMO mixture may comprise 6SL and 3SL combined in an amount of from 7 wt% to 23 wt%, preferably from

[0112] 8 wt% to 22 wt%.

[0113] In one embodiment, the HMO mixture comprises 3FL in an amount of from 10 wt% to 50 wt%. Suitably, the HMO mixture may comprise 3FL in an amount of from 10 wt% to 50 wt%, preferably from 11 wt% to 43 wt%. Suitably, the HMO mixture may comprise 3FL in an amount of from 10 wt% to 50 wt%, preferably from 13 wt% to 46 wt%.

[0114] In one embodiment, the HMO mixture comprises LNnT in an amount of from 6 wt% to 30 wt%. Suitably, the HMO mixture may comprise LNnT in an amount of from 6 wt% to 30 wt%, preferably from 7 wt% to 22 wt%. Suitably, the HMO mixture may comprise LNnT in an amount of from 3 wt% to 25 wt%, preferably from 5 wt% to 20 wt%.

[0115] In some embodiments, the HMO mixture consists or consists essentially of: i. 16 wt% to 69 wt% of 2FL; ii. 9 wt% to 24 wt% of LNT ; iii. 2 wt% to 10 wt% of DFL; iv. 8 wt% to 26 wt% of 6SL and 3SL combined; and v. 10 wt% to 50 wt% of 3FL.

[0116] In some preferred embodiments, the HMO mixture consists or consists essentially of: i. 22 wt% to 59 wt% of 2FL; ii. 12 wt% to 21 wt% of LNT ; iii. 3 wt% to 8 wt% of DFL; iv. 11 wt% to 22 wt% of 6SL and 3SL combined; and v. 11 wt% to 43 wt% of 3FL.

[0117] In some embodiments, the HMO mixture consists or consists essentially of: i. 34 wt% to 85 wt% of 2FL; ii. 10 wt% to 40 wt% of LNT ; iii. 4 wt% to 14 wt% of DFL; iv. 9 wt% to 31 wt% of 6SL and 3SL combined; and v. 6 wt% to 30 wt% of LNnT.

[0118] In some preferred embodiments, the HMO mixture consists or consists essentially of: i. 40 wt% to 71 wt% of 2FL; ii. 12 wt% to 26 wt% of LNT ; iii. 5 wt% to 10 wt% of DFL; and iv. 10 wt% to 28 wt% of 6SL and 3SL combined; and v. 7 wt% to 23 wt% of LNnT.

[0119] In some embodiments, the HMO mixture consists or consists essentially of: i. 20 wt% to 60 wt% of 2FL; ii. 4 wt% to 30 wt% of LNT ; iii. 1 wt% to 12 wt % of DFL; iv. 7 wt% to 23 wt% of 6SL and 3SL combined; v. 10 wt% to 50 wt% of 3FL; and vi. 3 wt% to 25 wt% of LNnT.

[0120] In some preferred embodiments, the HMO mixture consists or consists essentially of: i. 22 wt% to 55 wt% of 2FL; ii. 6 wt% to 20 wt% of LNT ; iii. 2 wt% to 8 wt % of DFL; iv. 8 wt% to 22 wt% of 6SL and 3SL combined; v. 13 wt% to 46 wt% of 3FL and vi. 5 wt% to 20 wt% of LNnT.

[0121] In the context of the present invention, the LNFP-I and the HMO mixture combined may be collectively referred to as a HMO mixture consisting of LNFP-I, 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT). This terminology may be used interchangeably.

[0122] For example, the composition of the invention may comprise or consist of Bifidobacterium longum subsp. infantis and a HMO mixture consisting of lacto-N-fucopentaose I (LNFP-I), 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT). For example, the combination of the invention may be Bifidobacterium longum subsp. infantis and a HMO mixture consisting of lacto-N-fucopentaose I (LNFP-I), 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'- sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT

[0123] In some embodiments, the LNFP-I and HMO mixture combined (i.e. the HMO mixture consisting of LNFP-I, 2FL, DFL, LNT, 6SL, and 3SL, and optionally 3FL and / or LNnT) consists or consists essentially of: i. 20 wt% to 46 wt% of 2FL; ii. 11 wt% to 17 wt% of LNT ; iii. 2 wt% to 7 wt% of DFL; iv. 9 wt% to 21 wt% of 6SL and 3SL combined; v. 9 wt% to 34 wt% of 3FL; and vi. 5 wt% to 32 wt% of LNFP-I.

[0124] In some preferred embodiments, the LNFP-I and HMO mixture combined (i.e. the HMO mixture consisting of LNFP-I, 2FL, DFL, LNT, 6SL, and 3SL, and optionally 3FL and / or LNnT) consists or consists essentially of: i. 22 wt% to 42 wt% of 2 FL; ii. 12 wt% to 15 wt% of LNT ; iii. 3 wt% to 6 wt% of DFL; iv. 9 wt% to 19 wt% of 6SL and 3SL combined; v. 11 wt% to 32 wt% of 3FL; and vi. 10 wt% to 19 wt% of LNFP-I.

[0125] In some embodiments, the LNFP-I and HMO mixture combined (i.e. the HMO mixture consisting of LNFP-I, 2FL, DFL, LNT, 6SL, and 3SL, and optionally 3FL and / or LNnT) consists or consists essentially of: i. 27 wt% to 41 wt% of 2FL; ii. 8 wt% to 15 wt% of LNT ; iii. 4 wt% to 6 wt% of DFL; iv. 8 wt% to 18 wt% of 6SL and 3SL combined; v. 13 wt% to 21 wt% of LNnT ; and vi. 7 wt% to 33 wt% of LNFP-I. In some preferred embodiments, the LNFP-I and HMO mixture combined (i.e. the HMO mixture consisting of LNFP-I, 2FL, DFL, LNT, 6SL, and 3SL, and optionally 3FL and / or LNnT) consists or consists essentially of: i. 32 wt% to 39 wt% of 2FL; ii. 10 wt% to 14 wt% of LNT ; iii. 4 wt% to 6 wt% of DFL; iv. 7 wt% to 15 wt% of 6SL and 3SL combined; v. 16 wt% to 20 wt% of LNnT ; and vi. 11 wt% to 23 wt% of LNFP-I .

[0126] In some embodiments, the LNFP-I and HMO mixture combined (i.e. the HMO mixture consisting of LNFP-I, 2FL, DFL, LNT, 6SL, and 3SL, and optionally 3FL and / or LNnT) consists or consists essentially of: i. 29 wt% to 40 wt% of 2FL; ii. 8 wt% to 13 wt% of LNT ; iii. 3 wt% to 11 wt % of DFL; iv. 3 wt% to 15 wt% of 6SL and 3SL combined; v. 11 wt% to 35 wt% of 3FL; vi. 1 wt% to 18 wt% of LNnT; and vii. 2 wt% to 24 wt% of LNFP-I.

[0127] In some preferred embodiments, the LNFP-I and HMO mixture combined (i.e. the HMO mixture consisting of LNFP-I, 2FL, DFL, LNT, 6SL, and 3SL, and optionally 3FL and / or LNnT) consists or consists essentially of: i. 32 wt% to 39 wt% of 2 FL; ii. 9 wt% to 12 wt% of LNT ; iii. 3 wt% to 11 wt % of DFL; iv. 4 wt% to 15 wt% of 6SL and 3SL combined; v. 12 wt% to 35 wt% of 3FL; vi. 1 wt% to 17 wt% of LNnT; and vii. 4 wt% to 14 wt% of LNFP-I.

[0128] When the composition or combination is in liquid form, the total HMO concentration is typically in the range from 0.5 to 10 g / L, preferably in the range from 1 to 7.5 g / L. Specific examples of the concentration level of total HMO, when the composition or combination is in liquid form, include 1 to 5 g / L, 1 to 4 g / L, 2 to 5 g / L, 1 to 3 g / L or 2 to 4 g / L. When the composition or combination is in solid form, the total HMO concentration is typically in the range from 0.35 to 7 wt% (g total HMO / 100 g dry composition), preferably in the range from 0.35 to 5 wt%. Specific examples of the concentration level of total HMO, when the composition or combination is in dry form, include 0.5 to 3.5 wt% (g total HMO per 100 g dry composition), 0.5 to 2.5 wt%, 1 to 3.5 wt%, 0.5 to 2 wt% or 1 to 2.5 wt%.

[0129] Therapeutic use

[0130] The present inventors have surprisingly found that the combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture can synergistically increase production of beneficial metabolites, such as uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract, e.g. the intestine, of a subject.

[0131] As described below, each of uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine are beneficial metabolites associated with improving health, treating and / or preventing disease, and / or promoting growth and / or development, etc.

[0132] In a first aspect, the invention provides the composition or combination of the invention for use in therapy in a subject, for example by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p- hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of the subject.

[0133] In a further aspect, the invention provides the composition or combination of the invention for use in improving metabolic health, cardiovascular health, neurological health, mental health, respiratory health, gastrointestinal health musculoskeletal health, and / or immune health in a subject, for example by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p- hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of the subject.

[0134] In a further aspect, the invention provides the composition or combination of the invention for use in promoting brain, bone, or muscle growth and / or development in a subject, for example by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl- carnitine and 3-methylxanthine in the gastrointestinal tract of the subject. In a further aspect, the invention provides the composition or combination of the invention for use in treating and / or preventing a metabolic disorder, a cardiovascular disease, a neurological disorder, a mental disorder, a respiratory disease, gastrointestinal inflammation, a musculoskeletal disorder, an immune disorder, an infection or apnea in a subject, for example by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of the subject.

[0135] In a further aspect, the invention provides the composition or combination of the invention for use in promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetylcarnitine and 3-methylxanthine in the gastrointestinal tract of a subject, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT), wherein the subject is at risk and / or in need.

[0136] In a further aspect, the invention provides the use of a composition or combination according to the invention for the manufacture of a medicament for a subject, for example by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3- methylxanthine in the gastrointestinal tract of the subject.

[0137] In a further aspect, the invention provides the use of a composition or combination according to the invention for the manufacture of a medicament for improving metabolic health, cardiovascular health, neurological health, mental health, respiratory health, gastrointestinal health musculoskeletal health, and / or immune health in a subject, for example by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3- methylxanthine in the gastrointestinal tract of the subject.

[0138] In a further aspect, the invention provides the use of a composition or combination according to the invention for the manufacture of a medicament for promoting brain, bone, or muscle growth and / or development in a subject, for example by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4- guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of the subject.

[0139] In a further aspect, the invention provides the use of a composition or combination according to the invention for the manufacture of a medicament for treating and / or preventing a metabolic disorder, a cardiovascular disease, a neurological disorder, a mental disorder, a respiratory disease, gastrointestinal inflammation, a musculoskeletal disorder, an immune disorder, an infection or apnea in a subject, for example by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4- guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of the subject.

[0140] In a further aspect, the invention provides a method of improving metabolic health, cardiovascular health, neurological health, mental health, respiratory health, gastrointestinal health, musculoskeletal health and / or immune health in a subject, the method comprising administering to the subject a composition or combination according to the invention, for example thereby promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of the subject.

[0141] In a further aspect, the invention provides a method of promoting brain, bone, or muscle growth and / or development in a subject, the method comprising administering to the subject a composition or combination according to the invention, for example thereby promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3- methylxanthine in the gastrointestinal tract of the subject.

[0142] In a further aspect, the invention provides a method of treating and / or preventing a metabolic disorder, a cardiovascular disease, a neurological disorder, a mental disorder, a respiratory disease, gastrointestinal inflammation, a musculoskeletal disorder, an immune disorder, an infection or apnea in a subject, the method comprising administering to the subject a composition or combination according to the invention, for example thereby promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3- methylxanthine in the gastrointestinal tract of the subject.

[0143] In some embodiments, the composition or combination of the invention promotes the production of the one or more beneficial metabolites in the gastrointestinal tract of a subject. In preferred embodiments, the composition or combination of the invention promotes the production of the one or more beneficial metabolites in the intestine of a subject. Uridine

[0144] In some embodiments, the composition or combination of the invention promotes the production of uridine in the gastrointestinal tract of a subject, for example when the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), 3'-sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT).

[0145] Uridine is an essential nucleotide with various health benefits. Supplementation of uridine has shown positive effects on cardiovascular health, such as preventing myocardial injury in rat models of acute ischemia and ischemia / reperfusion (Sci Rep. 2021 Aug 20; 11 (1 ): 16999), as well as protecting against diabetes-induced damage to cardiac mitochondria (Int J Mol Sci. 2022 Sep 13;23(18): 10633). Uridine also exhibits neuroprotective properties, demonstrating protective effects in a rat model of Parkinson's disease (Int J Mol Sci. 2024 Jul 6;25(13):7441). Additionally, uridine holds potential as a future therapeutic for osteoarthritis, as it has shown promising results in the treatment of OA in vivo (Cell Cycle. 2022 Jan;21(1):33-48). Finally, uridine supplementation could be beneficial by attenuating high fat diet-induced obesity and nonalcoholic fatty liver disease, as shown in a mice model (Food Funct. 2021 Mar 1 ;12(4):1829-1840).

[0146] In a first aspect, the present invention provides the composition or combination of the invention for use in therapy in a subject by promoting the production of uridine in the gastrointestinal tract of the subject.

[0147] In a further aspect, the invention provides the composition or combination of the invention for use in improving metabolic health, cardiovascular health, and / or neurological health in a subject by promoting the production of uridine in the gastrointestinal tract of the subject.

[0148] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a metabolic disorder, a cardiovascular disease, and / or a neurological disorder in a subject by promoting the production of uridine in the gastrointestinal tract of the subject.

[0149] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting brain growth and / or development in a subject by promoting the production of uridine in the gastrointestinal tract of the subject.

[0150] In a further aspect, the invention provides the use of a composition or combination according to the invention for use as a neuroprotectant in a subject by promoting the production of uridine in the gastrointestinal tract of the subject. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in improving musculoskeletal health or treating and / or preventing a musculoskeletal disorder in a subject by promoting the production of uridine in the gastrointestinal tract of the subject.

[0151] Adenine

[0152] In some embodiments, the composition or combination of the invention promotes the production of adenine in the gastrointestinal tract of a subject, for example when the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), 3'-sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT).

[0153] Adenine is an essential nucleotide, as it is involved in DNA and RNA synthesis as well as energy metabolism. Dietary adenine supplementation has been shown to inhibit fatty liver development in rats (Sci Rep. 2020 Dec 17; 10(1):22110) as well as preventing the decrease in cellular immunity in a ground-based microgravity analog (J Appl Physiol (1985). 2002 Jul;93(1): 161-6). Additionally, Fukada et al demonstrated that adenine could inhibit TNF-a signaling in intestinal epithelial cells and reduce mucosal inflammation in a mouse model of colitis (J Agric Food Chem. 2016 Jun 1 ;64(21):4227-34). Finally, adenine may also present some anti-cancer potency, as it can inhibits the growth of colon cancer cells (Evid Based Complement Alternat Med. 2019 Sep 12;2019:9151070 and Int J Med Sci. 2020 Feb 24;17(5):678-684).

[0154] In a first aspect, the present invention provides the composition or combination of the invention for use in therapy in a subject by promoting the production of adenine in the gastrointestinal tract of the subject.

[0155] In a further aspect, the invention provides the composition or combination of the invention for use in improving metabolic health and / or gastrointestinal health in a subject by promoting the production of adenine in the gastrointestinal tract of the subject.

[0156] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a metabolic disorder and / or gastrointestinal inflammation in a subject by promoting the production of adenine in the gastrointestinal tract of the subject.

[0157] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing intestinal inflammation in a subject by promoting the production of adenine in the gastrointestinal tract of the subject. Guanine

[0158] In some embodiments, the composition or combination of the invention promotes the production of guanine in the gastrointestinal tract of a subject, for example when the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), 3'-sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT).

[0159] Guanine is an essential nucleotide involved in DNA and RNA synthesis. Its derivatives, such as guanosine, hold potential health benefits in various areas, notably in mood disorders and treatment for depression (Pharmacol Biochem Behav. 2022 Jul;218:173422 and Front Psychiatry. 2021 Aug 4;12:701408). Guanosine is a purine nucleoside with important functions in cell metabolism and a protective role in response to degenerative diseases or injury. Evidence from rodent and cell models show a number of neurotrophic and neuroprotective effects of guanosine preventing deleterious consequences of seizures, spinal cord injury, pain, mood disorders and aging-related diseases, such as ischemia, Parkinson's and Alzheimer's diseases (Aging Dis. 2016 Oct 1 ;7(5):657-679).

[0160] In a first aspect, the present invention provides the composition or combination of the invention for use in therapy in a subject by promoting the production of guanine in the gastrointestinal tract of the subject.

[0161] In a further aspect, the invention provides the composition or combination of the invention for use in improving metabolic health, neurological health, and / or mental health in a subject by promoting the production of guanine in the gastrointestinal tract of the subject.

[0162] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a metabolic disorder, a neurological disorder, and / or a mental disorder in a subject by promoting the production of guanine in the gastrointestinal tract of the subject.

[0163] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting brain growth and / or development in a subject by promoting the production of guanine in the gastrointestinal tract of the subject.

[0164] In a further aspect, the invention provides the use of a composition or combination according to the invention for use as a neuroprotectant in a subject by promoting the production of guanine in the gastrointestinal tract of the subject. 1 Adenosine monophosphate (AMP)

[0165] In some embodiments, the composition or combination of the invention promotes the production of AMP in the gastrointestinal tract of a subject, for example when the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), 3'-sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT).

[0166] Adenosine monophosphate (AMP), an essential nucleotide, has been linked to increased weight gain, overall growth, and improved gut microbiota composition in formula-fed infants based on findings from several randomized clinical trials (Pediatrics. 2010 Oct; 126(4) :e946- 53 and Am J Clin Nutr. 2008 Jun;87(6):1785-92). Additionally, studies on mice models suggest that this metabolite may play a role in regulating food intake and energy expenditure (Food Nutr Res. 2022 Jun 30;66).

[0167] In a first aspect, the present invention provides the composition or combination of the invention for use in therapy in a subject by promoting the production of AMP in the gastrointestinal tract of the subject.

[0168] In a further aspect, the invention provides the composition or combination of the invention for use in improving metabolic health in a subject by promoting the production of AMP in the gastrointestinal tract of the subject.

[0169] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a metabolic disorder in a subject by promoting the production of AMP in the gastrointestinal tract of the subject.

[0170] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting bone and / or muscle growth and / or development in a subject (e.g. a formula-fed infant) by promoting the production of AMP in the gastrointestinal tract of the subject.

[0171] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting and / or maintaining a healthy gut microbiota in a subject (e.g. a formula-fed infant) by promoting the production of AMP in the gastrointestinal tract of the subject.

[0172] Cyclic adenosine monophosphate (cAMP)

[0173] In some embodiments, the composition or combination of the invention promotes the production of cAMP in the gastrointestinal tract of a subject, for example when the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), 3'-sialyllactose (3SL), and 3-fucosyllactose (3FL) or 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT).

[0174] Adenosine 3',5'-cyclic monophosphate (cAMP) is a nucleotide that serves as a crucial second messenger in multiple signal transduction pathways. It plays a regulatory role in diverse cellular functions, such as cell growth, differentiation, gene transcription, and protein expression (Mol Med Rep. 2016 May; 13(5):3715-23). Therefore, it is actively being investigated as a potential therapeutic target in numerous pathological conditions, including cardiovascular, neurological, respiratory (Pulm Pharmacol Ther. 2013 Feb;26(1):112-20 and Am J Physiol Lung Cell Mol Physiol. 2011 Mar;300(3):L453-61) and metabolic disorders. In preterm infants, levels of cAMP in blood sample has been associated with increased TLR- mediated production of acute-phase and anti-inflammatory cytokines in an in vitro model by Strunk et al (Pediatr Res. 2020 Nov;88(5):717-725).

[0175] In a first aspect, the present invention provides the composition or combination of the invention for use in therapy in a subject by promoting the production of cAMP in the gastrointestinal tract of the subject.

[0176] In a further aspect, the invention provides the composition or combination of the invention for use in improving metabolic health, cardiovascular health, neurological health, and / or respiratory health in a subject by promoting the production of cAMP in the gastrointestinal tract of the subject.

[0177] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a metabolic disorder, a cardiovascular disease, a neurological disorder, and / or a respiratory disease in a subject by promoting the production of cAMP in the gastrointestinal tract of the subject.

[0178] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing an infection (e.g. a respiratory infection) in a subject by promoting the production of cAMP in the gastrointestinal tract of the subject. In some embodiments, the subject is a preterm infant or young child.

[0179] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in modulating / improving the immune response in a subject by promoting the production of cAMP in the gastrointestinal tract of the subject. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in increasing Toll-like Receptors- TLR-mediated production of acutephase and anti-inflammatory cytokines in an infant born prematurely, optionally wherein the composition or combination promotes the production of cAMP in the gastrointestinal tract of the infant born prematurely.

[0180] 4-guanidinobutyric acid

[0181] In some embodiments, the composition or combination of the invention promotes the production of 4-guanidinobutyric acid in the gastrointestinal tract of a subject, for example when the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N- tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and 3-fucosyllactose (3FL).

[0182] 4-Guanidinobutyric acid is one of the precursor of GABA, an essential neurotransmitter with known health benefits. Decreased GABA levels are linked to anxiety, stress, fear, seizure disorders, movement disorders, attention deficit hyperactivity disorder, panic disorder, and mood disorders such as depression (Mol Psychiatry. 2010 Nov 16;16(4):383-406). Additionally, Hwang et al demonstrated in a rat model, the potential of 4-guanidinobutyric acid against gastric lesions and as potential therapeutics for gastritis (Biomol Ther (Seoul). 2012 Mar;20(2):239-44).

[0183] In a first aspect, the present invention provides the composition or combination of the invention for use in therapy in a subject by promoting the production of 4-guanidinobutyric acid in the gastrointestinal tract of the subject.

[0184] In a further aspect, the invention provides the composition or combination of the invention for use in improving neurological health, mental health, and / or gastrointestinal health in a subject by promoting the production of 4-guanidinobutyric acid in the gastrointestinal tract of the subject.

[0185] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a neurological disorder, a mental disorder and / or gastrointestinal inflammation in a subject by promoting the production of 4- guanidinobutyric acid in the gastrointestinal tract of the subject.

[0186] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting brain growth and / or development in a subject by promoting the production of 4-guanidinobutyric acid in the gastrointestinal tract of the subject. In a further aspect, the invention provides the use of a composition or combination according to the invention for use as a neuroprotectant in a subject by promoting the production of 4- guanidinobutyric acid in the gastrointestinal tract of the subject. p-hydroxyphenyllactic acid

[0187] In some embodiments, the composition or combination of the invention promotes the production of p-hydroxyphenyllactic in the gastrointestinal tract of a subject, for example when the HMO mixture consists of 2'-fucosyl lactose (2FL), difucosy I lactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N- neotetraose (LNnT).

[0188] A recent study indicated that Bifidobacterium species, promoted by breastmilk, produce aromatic lactic acids (including p-hydroxyphenyllactic acid) in the gut of infants (Nat Microbiol. 2021 Nov;6(11): 1367-1382). The study suggests that p-hydroxyphenyllactic may have an impact on immune function in the early stages of life. P-hydroxyphenyllactic acid also demonstrated potential as pathogens inhibitor (Agric Food Chem. 2019 May 22;67(20):5820- 5826). P-hydroxyphenyllactic has been associated with innate immune (Zugasti et al., Nature Immunology, 2014, 15: 833-838) and antimicrobial (Pahalagedara et al., Metabolites, 2023, 13(2): 252) functions. An isomer of PLA, 3-(4-hydroxyphenyl)propionate (HO-PPA), is also associated with antiviral (Hooda et al., Viruses, 2022, 14: 1778) and anti-inflammatory (Wei et al., FASEB J, 2020, 34: 16117-16128) activities.

[0189] In a first aspect, the present invention provides the composition or combination of the invention for use in therapy in a subject by promoting the production of p-hydroxyphenyllactic acid in the gastrointestinal tract of the subject.

[0190] In a further aspect, the invention provides the composition or combination of the invention for use in improving immune health in a subject by promoting the production of p- hydroxyphenyllactic acid in the gastrointestinal tract of the subject.

[0191] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing an immune disorder and / or an infection in a subject by promoting the production of p-hydroxyphenyllactic acid in the gastrointestinal tract of the subject.

[0192] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in modulating / improving the immune response in a subject by promoting the production of p-hydroxyphenyllactic acid in the gastrointestinal tract of the subject. A cetyl-carnitin e

[0193] In some embodiments, the composition or combination of the invention promotes the production of acetyl-carnitine in the gastrointestinal tract of a subject, for example when the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and 3-fucosyllactose (3FL) or 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'- sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT).

[0194] The primary focus of current research on acetyl-carnitine revolves around its potential as a neuroprotective agent (Neurochem Res. 2017 Jun;42(6):1661-1675). In animal models with rat pups after brain injury, supplementation of L-acetyl-carnitine demonstrated neuroprotecting effects (Magn Reson Med. 2015 Dec;74(6): 1530-42). In addition, Nasca et al. provided evidence of acetyl-carnitine's potential as an antidepressant and biomarker for major depressive disorder (PNAS 2018 Aug 21 ;115(34): 8627-8632). Furthermore, in case reports involving children at risk of Type 1 diabetes, the administration of acetyl-L-carnitine (at a dosage of 50 mg / kg / day) and nicotinamide (at a dosage of 25 mg / kg / day) resulted in a delay in the development or even remission of type 1 diabetes in certain patients (J Pediatr Endocrinol Metab. 2013;26(3-4):347-55).

[0195] In a first aspect, the present invention provides the composition or combination of the invention for use in therapy in a subject by promoting the production of acetyl-carnitine in the gastrointestinal tract of the subject.

[0196] In a further aspect, the invention provides the composition or combination of the invention for use in improving metabolic health, neurological health, and / or mental health in a subject by promoting the production of acetyl-carnitine in the gastrointestinal tract of the subject.

[0197] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a metabolic disorder, a neurological disorder and / or a mental disorder in a subject by promoting the production of acetyl-carnitine the gastrointestinal tract of the subject.

[0198] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting brain growth and / or development in a subject by promoting the production of acetyl-carnitine in the gastrointestinal tract of the subject.

[0199] In a further aspect, the invention provides the use of a composition or combination according to the invention for use as a neuroprotectant in a subject by promoting the production of 4- acetyl-carnitine in the gastrointestinal tract of the subject. 3-methylxanthine

[0200] In some embodiments, the composition or combination of the invention promotes the production of 3-methylxanthine in the gastrointestinal tract of a subject, for example when the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and 3-fucosyllactose (3FL).

[0201] 3-methylxanthine (3-MX), a derivative of theophylline, is a member of the methylxanthine family. Methylxanthines have gained significant attention as potential therapeutic agents for various health benefits, including neurodegenerative diseases, cardiovascular and respiratory conditions (Mol Nutr Food Res. 2017 Jun;61(6) and Methylxanthines: Dietary Sources, Bioavailability, and Health Benefits. In Fruit and Vegetable Phytochemicals, E.M. Yahia (Ed.)). In premature infants, methylxanthines have been utilized as treatments for apnea (Paediatr Drugs. 2014 Apr; 16(2): 169-77 and J Paediatr Child Health. 2009 Oct; 45(10):587-92). Although 3-MX is considered less potent than its precursor, theophylline, it still holds potential as a beneficial metabolite, notably in cancer treatment (mSystems. 2024 Jul 23;9(7):e0130123).

[0202] In a first aspect, the present invention provides the composition or combination of the invention for use in therapy in a subject by promoting the production of 3-methylxanthine in the gastrointestinal tract of the subject.

[0203] In a further aspect, the invention provides the composition or combination of the invention for use in improving cardiovascular health, neurological health, and / or respiratory health in a subject by promoting the production of 3-methylxanthine in the gastrointestinal tract of the subject.

[0204] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a cardiovascular disease, a neurological disorder, a respiratory disease and / or a apnea in a subject by promoting the production of 3- methylxanthine the gastrointestinal tract of the subject. In some embodiments, the subject is a preterm infant or young child.

[0205] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing infection (e.g. respiratory infection) in a subject by promoting the production of 3-methylxanthine the gastrointestinal tract of the subject. Metabolic health

[0206] The invention provides the use of a composition or combination according to the invention for use in improving metabolic health or treating and / or preventing a metabolic disorder in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of one or more of uridine, adenine, guanine, AMP, cAMP, and / or acetyl-carnitine in the gastrointestinal tract of the subject (e.g. infant or young child).

[0207] Improving metabolic health may reduce the risk of developing a metabolic disorder. A metabolic disorder may refer to a disorder that negatively alters the body's processing and distribution of macronutrients, such as proteins, fats, and carbohydrates. Metabolic disorders may include metabolic syndrome, which is a complex condition characterized by insulin resistance, abdominal obesity, dyslipidemia, hypertension, and hyperglycemia (Reprod Toxicol. 2016 Oct 17;68:3-33).

[0208] In some embodiments, the composition or combination according to the invention improves metabolic health by promoting nucleic acid synthesis and / or energy metabolism, for example wherein the combination promotes the production of one or more of uridine, adenine, guanine, AMP, and / or cAMP in the gastrointestinal tract of the subject. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in improving metabolic health or treating and / or preventing a metabolic disorder in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of AMP in the gastrointestinal tract of the subject (e.g. infant or young child).

[0209] In some embodiments, the metabolic disorder is fatty liver disease. Dietary adenine supplementation has been shown to inhibit fatty liver development in rats (Sci Rep. 2020 Dec 17;10(1):22110). In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing metabolic fatty liver disease in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of adenine in the gastrointestinal tract of the subject (e.g. infant or young child).

[0210] In some embodiments, the metabolic disorder is type 1 diabetes. In case reports involving children at risk of Type 1 diabetes, the administration of acetyl-L-carnitine (at a dosage of 50 mg / kg / day) and nicotinamide (at a dosage of 25 mg / kg / day) resulted in a delay in the development or even remission of type 1 diabetes in certain patients. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing type 1 diabetes in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of acetyl-carnitine in the gastrointestinal tract of the subject (e.g. infant or young child).

[0211] Cardiovascular health

[0212] The invention provides the use of a composition or combination according to the invention for use in improving cardiovascular health or treating and / or preventing a cardiovascular disease in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of one or more of uridine, cAMP, and / or 3-methylxanthine in the gastrointestinal tract of the subject (e.g. infant or young child).

[0213] Improving cardiovascular health may reduce the risk of developing a cardiovascular disease. Cardiovascular disease may refer to diseases involving the heart or blood vessels, including diseases of the cardiac muscle and of the vascular system supplying the heart, brain, and other vital organs (Gaziano, T., et al. , 2006. Cardiovascular disease. Disease Control Priorities in Developing Countries. 2nd edition).

[0214] Supplementation of uridine has shown positive effects on cardiovascular health, such as preventing myocardial injury in rat models of acute ischemia and ischemia / reperfusion (Sci Rep. 2021 Aug 20; 11 (1): 16999), as well as protecting against diabetes-induced damage to cardiac mitochondria (Int J Mol Sci. 2022 Sep 13;23(18): 10633). In some embodiments, the composition or combination according to the invention improves cardiovascular health by preventing myocardial injury or protecting against diabetes-induced damage to cardiac mitochondria, optionally wherein the composition or combination promotes the production of uridine in the gastrointestinal tract of the subject.

[0215] Neurological health

[0216] The invention provides the use of a composition or combination according to the invention for use in improving neurological health or treating and / or preventing a neurological disorder in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of one or more of uridine, guanine, cAMP, 4-guanidinobutyric acid, acetyl-carnitine and / or 3-methylxanthine in the gastrointestinal tract of the subject (e.g. infant or young child).

[0217] Improving neurological health may reduce the risk of developing a neurological disorder. A neurological disorder may refer to a disorder of the nervous system (World Health Organization, 2006. Neurological disorders: public health challenges. World Health Organization). In some embodiments, the composition or combination of the invention acts as a neuroprotective agent, for example protecting against brain injury. Uridine exhibits neuroprotective properties, demonstrating protective effects in a rat model of Parkinson's disease (Int J Mol Sci. 2024 Jul 6;25(13):7441). Evidence from rodent and cell models also show a number of neurotrophic and neuroprotective effects of guanosine preventing (Aging Dis. 2016 Oct 1 ;7(5):657-679). The primary focus of current research on acetyl-carnitine revolves around its potential as a neuroprotective agent (Neurochem Res. 2017 Jun;42(6): 1661 -1675). In a further aspect, the invention provides the use of a composition or combination according to the invention for use as a neuroprotective agent in a subject, optionally wherein the composition or combination promotes the production of one or more of uridine, guanine, and / or acetyl-carnitine in the gastrointestinal tract of the subject.

[0218] In some embodiments, the neurological disorder is a neurodegenerative disease. In addition to uridine and guanine, methylxanthines have gained significant attention as potential therapeutic agents for various health benefits, including neurodegenerative diseases, (Mol Nutr Food Res. 2017 Jun;61(6)). In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a neurodegenerative disease in a subject, optionally wherein the composition or combination promotes the production of one or more of uridine, guanine, and / or 3-methylxanthine in the gastrointestinal tract of the subject.

[0219] In some embodiments, the composition or combination of the invention promotes brain growth and / or development. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting brain growth and / or development in a subject, optionally wherein the composition or combination promotes the production of one or more of uridine, guanine, cAMP, 4-guanidinobutyric acid, acetyl-carnitine and / or 3- methylxanthine in the gastrointestinal tract of the subject. As used herein, “promoting brain growth and / or strength” may refer to the support of normal brain growth and / or development, for example during childhood and adolescence. Supporting normal brain growth and / or development may result in normal brain anatomy and physiology. Suitable methods and parameters to determine brain growth and development will be known to the skilled person. For example, improved brain growth and / or development may improve neurofunctional and visual performance. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting cognitive development, visual acuity, and / or motor skill development in a subject, optionally wherein the composition or combination promotes the production of one or more of uridine, guanine, cAMP, 4- guanidinobutyric acid, acetyl-carnitine, and / or 3-methylxanthine in the gastrointestinal tract of the subject. Mental health

[0220] The invention provides the use of a composition or combination according to the invention for use in improving mental health or treating and / or preventing a mental disorder in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of one or more of guanine, 4-guanidinobutyric acid, and / or acetyl-carnitine in the gastrointestinal tract of the subject (e.g. infant or young child).

[0221] Improving mental health may reduce the risk of developing a mental disorder. A mental disorder may refer to a disease which affects cognition, emotion, and behavioural control (Hyman, S., et al., 2006. Mental disorders. Disease control priorities related to mental, neurological, developmental and substance abuse disorders, pp.1-20).

[0222] In some embodiments, the mental disorder is a mood disorder, such as a depressive disorder. Guanosine, a derivative of guanine holds potential health benefits in various areas, notably in mood disorders and treatment for depression (Pharmacol Biochem Behav. 2022 Jul;218:173422 and Front Psychiatry. 2021 Aug 4;12:701408). 4-Guanidinobutyric acid is one of the precursor of GABA, an essential neurotransmitter with known health benefits. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a mood disorder in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of guanine and / or 4-guanidinobutyric acid in the gastrointestinal tract of the subject (e.g. infant or young child).

[0223] Respiratory health

[0224] The invention provides the use of a composition or combination according to the invention for use in improving respiratory health or treating and / or preventing a respiratory disease in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of cAMP and / or 3-methylxanthine in the gastrointestinal tract of the subject (e.g. infant or young child).

[0225] Improving respiratory health may reduce the risk of developing a respiratory disease. A respiratory disease may refer to a disease which affects the respiratory tract and may include lung diseases (Ann Am Thorac Soc. 2014 Mar;11 (3):404-6).

[0226] In some embodiments, the respiratory disease is asthma or chronic obstructive pulmonary disease (COPD). cAMP is actively being investigated as a potential therapeutic target in numerous pathological conditions, including respiratory disease such as asthma and COPD (Pulm Pharmacol Ther. 2013 Feb;26(1):112-20). In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing asthma or COPD in a subject, optionally wherein the composition or combination promotes the production of cAMP in the gastrointestinal tract of the subject.

[0227] In some embodiments, the respiratory disease is a respiratory tract infection. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing asthma a respiratory tract infection in a subject, optionally wherein the composition or combination promotes the production of cAMP and / or 3-methylxanthine in the gastrointestinal tract of the subject.

[0228] Gastrointestinal health

[0229] The invention provides the use of a composition or combination according to the invention for use in improving gastrointestinal health or treating and / or preventing gastrointestinal inflammation in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of adenine and / or 4-guanidinobutyric acid in the gastrointestinal tract of the subject (e.g. infant or young child).

[0230] Improving gastrointestinal health may reduce the risk of developing a gastrointestinal disorder, such as gastrointestinal inflammation. A gastrointestinal disorder may refer to a disease which affects the gastrointestinal tract (Gut. 1999 Sep;45(Suppl 2): 1160— 1168). In some embodiments, the gastrointestinal disorder is gastrointestinal inflammation, for example intestinal inflammation or gastritis.

[0231] In some embodiments, the gastrointestinal disorder is intestinal inflammation (e.g. colitis). Fukada et al demonstrated that adenine could inhibit TNF-a signaling in intestinal epithelial cells and reduce mucosal inflammation in a mouse model of colitis (J Agric Food Chem. 2016 Jun 1 ;64(21):4227-34). In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing intestinal inflammation (e.g. colitis) in a subject, optionally wherein the composition or combination promotes the production of adenine in the gastrointestinal tract of the subject.

[0232] In some embodiments, the gastrointestinal disorder is stomach inflammation (e.g. gastritis). Hwang et al demonstrated in a rat model, the potential of 4-guanidinobutyric acid against gastric lesions and as potential therapeutics for gastritis (Biomol Ther (Seoul). 2012 Mar;20(2):239-44). In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing stomach inflammation (e.g. gastritis) in a subject, optionally wherein the composition or combination promotes the production of 4-guanidinobutyric acid in the gastrointestinal tract of the subject. Musculoskeletal health

[0233] The invention provides the use of a composition or combination according to the invention for use in improving musculoskeletal health or treating and / or preventing a musculoskeletal disorder in a subject (e.g. an infant or young child), for example by promoting the production of uridine in the gastrointestinal tract of the subject (e.g. infant or young child). Uridine has shown promising results in the treatment of osteoarthritis in vivo (Cell Cycle. 2022 Jan;21 (1):33-48).

[0234] Improving musculoskeletal health may reduce the risk of developing a musculoskeletal disorder. A musculoskeletal disorder may refer to injuries or pain in the human musculoskeletal system, including the joints, ligaments, muscles, nerves, tendons, and structures that support limbs, neck and back.

[0235] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a musculoskeletal disorder (e.g. bone disease) in a subject, optionally wherein the composition or combination promotes the production of uridine in the gastrointestinal tract of the subject. Metabolic bone disease is frequent in preterm and / or low birth weight infants and / or infants suffering from suboptimal intra-uterine nutrition and leads to an increased risk of bone fractures in these populations (Arch Dis Child Fetal Neonatal Ed 2002 86: F82-F85). Infants, children and adolescents suffering from growth retardation due to malnutrition and / or disease are also frequently affected by these conditions.

[0236] Immune response

[0237] The invention provides the use of a composition or combination according to the invention for use in improving immune health, treating and / or preventing an immune disorder, and / or modulating / improving the immune response in a subject (e.g. an infant or young child), for example by promoting the production of cAMP and / or p-hydroxyphenyllactic acid in the gastrointestinal tract of the subject (e.g. infant or young child).

[0238] Improving immune health may reduce the risk of developing an immune disorder, Suitably, modulating / improving the immune response comprises supporting or promoting protective immunity. Suitably, promoting protective immunity means one or more of the following: prevention of infection, anti-pathogen activity, improving or increasing the function of immune cells, limiting pathogen expansion, promoting pathogen clearance, restriction of pathogen dissemination, recovery from infection, reducing the risk of secondary infection, and / or limiting immune mediated pathology following infection. Suitably, promoting protective immunity means improving or increasing the function of immune cells. For example, promoting protective immunity can be defined by the three levels of immune defence against pathogens (i) mucosal barrier functions of the lung and gastrointestinal tract, (ii) the innate immune response (for example, macrophages with antimicrobial activity) and (iii) the adaptive immune response (including, for example, CD8 T cell activation, which increases anti-viral immunity in the lung).

[0239] Suitably, improving or increasing the function of immune cells can be an increase in total serum immunoglobulin concentration or an increase in cytokine secretion from immune cells or a combination thereof. The increase in cytokine secretion can be an increase in IL-6, IFN- gamma, I L12p70, or any combination thereof. The increase in total serum immunoglobulin concentration or increase in cytokine secretion can be an increase of about 5, 10, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1 ,000 percent or more (or any range or value between about 5 and about 1 ,000 percent) as compared to a corresponding subject who does not receive the composition or combination of the invention.

[0240] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting a healthy immune system in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of cAMP in the gastrointestinal tract of the subject (e.g. infant or young child).

[0241] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting a healthy immune system in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of p- hydroxyphenyllactic acid in the gastrointestinal tract of the subject (e.g. infant or young child).

[0242] Infection

[0243] The invention provides the use of a composition or combination according to the invention for use in preventing and / or treating an infection in a subject (e.g. an infant or young child), for example by promoting the production of one or more of cAMP, p-hydroxyphenyllactic acid, and / or 3-methylxanthine in the gastrointestinal tract of the subject (e.g. infant or young child). In some embodiments, the subject is a preterm infant or young child.

[0244] The infection may be a viral, bacterial or fungal infection. The infection may be a gastrointestinal infection or a respiratory tract infection. The gastrointestinal infection may be an intestinal infection or a stomach infection. Particular examples of bacterial pathogens are Clostridium difficile (e.g. Clostridium difficile toxin A / B), Campylobacter, Salmonella, Shigella, Citrobacter, and Escherichia coli (e.g. E. coli 0157). The respiratory tract infection may be an infection in the upper respiratory tract or in the lower respiratory tract. Particular examples of a bacterial pathogen are Mycobacterium tuberculosis or Strepcoccus pneumoniae.

[0245] The symptoms most often associated with the bacterial infection, and which may be reduced by the composition of the invention, are irritation in the lungs, congestion in the lungs, excessive mucus production, fever, cough, wheezing, breathlessness, abdominal cramps, diarrhoea or vomiting. Suitably, the symptoms are abdominal cramps, diarrhoea or vomiting.

[0246] Viral respiratory infections, such as respiratory syncytial virus (RSV), affect nearly 90% of children by the age of two (Karpinnen et al, Clin Microbiol Infect, 2016;22;208.e1-e6). Such viral respiratory infections in infants and young children often lead to bronchiolitis, an inflammatory bronchial reaction in infants and young children (Pickles et al, J Pathol, 2015;235;266-276). Severe RSV-induced bronchiolitis is a major cause of morbidity and mortality in infants globally (Nair et al, Lancet, 2010;375;9725;2545-1555).

[0247] In a preferred embodiment, the viral infection is a viral respiratory tract infection. The viral respiratory tract infection may be a viral infection in the upper respiratory tract or in the lower respiratory tract. In a typical embodiment of the invention, the viral respiratory tract infection is caused by respiratory syncytial virus (RSV).

[0248] The disease associated with the viral infection will typically be common cold, influenza (flu), bronchitis, bronchiolitis, pneumonia, sore throat (pharyngitis), sinusitis, non-allergic rhinitis, severe acute respiratory syndrome (SARS), viral croup, otitis media, meningitis or diarrhoea. Typically, when the viral infection is in the respiratory tract, the disease associated with the respiratory tract infection is common cold, influenza (flu), bronchitis, bronchiolitis, pneumonia, sore throat (pharyngitis), sinusitis, non-allergic rhinitis, severe acute respiratory syndrome (SARS), viral croup or otitis media. Most often, the disease associated with the viral respiratory tract infection is common cold, influenza (flu), bronchitis, bronchiolitis or pneumonia.

[0249] Accordingly, in a preferred embodiment of the invention the composition or combination of the invention is for use in treating and / or preventing a disease associated with a viral respiratory tract infection selected from the group consisting of common cold, influenza (flu), bronchitis, bronchiolitis and pneumonia. In a more preferred embodiment, the disease associated with the respiratory tract infection is selected from the group consisting of bronchiolitis and pneumonia, in particular RSV-induced bronchiolitis and / or pneumonia, i.e. bronchiolitis and / or pneumonia caused by RSV. In an even more preferred embodiment, the disease associated with the respiratory tract infection is bronchiolitis, in particular RSV-induced bronchiolitis.

[0250] The symptoms most often associated with the viral infection, and which may be reduced by the composition of the invention, are irritation in the lungs, congestion in the lungs, excessive mucus production, fever, cough, wheezing, breathlessness, abdominal cramps, diarrhoea or vomiting.

[0251] The above-mentioned infections may be caused by a variety of different viruses, including respiratory syncytial virus (RSV), parainfluenza virus (PIV), influenza virus such as influenza virus A (IVA) and / or influenza virus B (IVB), rhinovirus (RV), adenovirus (ADV), metapneumovirus (MPV), bocavirus (BoV), coronavirus (CoV), myxovirus, herpesvirus, enterovirus (EV), parachovirus (PeV) or a combination thereof.

[0252] The composition of the invention is useful for treating and / or preventing infections, in particular respiratory tract infection in a human of any age. Thus, the human to be treated with the composition of the invention may be selected from the group consisting of 0 to <1 year (infants), 1 to <3 years (young children) and 3 to <6 years (children), including 3 to <5 years (pre-schoolers).

[0253] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing a respiratory tract infection, in a subject, optionally wherein the composition or combination promotes the production of one or more of cAMP, p-hydroxyphenyllactic acid, and / or 3-methylxanthine in the gastrointestinal tract of the subject.

[0254] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in reducing the risk of contracting an infection (e.g. a respiratory tract infection) in a subject and / or for use in reducing the symptoms associated with an infection infection (e.g. a respiratory tract infection) in subject, optionally wherein the composition or combination promotes the production of one or more of cAMP, p-hydroxyphenyllactic acid, and / or 3-methylxanthine in the gastrointestinal tract of the subject.

[0255] Apnea

[0256] The invention provides the use of a composition or combination according to the invention for use in treating and / or preventing apnea in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of 3-methylxanthine in the gastrointestinal tract of the subject (e.g. infant or young child). In premature infants, methylxanthines have been utilized as treatments for apnea (Paediatr Drugs. 2014 Apr; 16(2): 169-77 and J Paediatr Child Health. 2009 Oct; 45(10):587-92). In some embodiments, the subject is a preterm infant or young child.

[0257] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing apnea in a subject (e.g. an infant or young child born prematurely), optionally wherein the composition or combination promotes the production of 3-methylxanthine in the gastrointestinal tract of the subject (e.g. infant or young child born prematurely).

[0258] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in treating and / or preventing apnea in an infant born prematurely, optionally wherein the composition or combination promotes the production of 3- methylxanthine in the gastrointestinal tract of the infant born prematurely.

[0259] Bone and / or muscle growth / development

[0260] The invention provides the use of a composition or combination according to the invention for use in promoting bone and / or muscle growth and / or development in a subject (e.g. an infant or young child), for example by promoting the production of AMP in the gastrointestinal tract of the subject (e.g. infant or young child). Adenosine monophosphate (AMP) has been linked to increased weight gain, overall growth in formula-fed infants based on findings from several randomized clinical trials (Pediatrics. 2010 Oct; 126(4) :e946-53 and Am J Clin Nutr. 2008 Jun; 87(6): 1785-92).

[0261] The composition or combination of the present invention may promote bone growth and / or strength. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting bone growth and / or strength in a subject, optionally wherein the composition or combination promotes the production of AMP in the gastrointestinal tract of the subject. As used herein, “promoting bone growth and / or strength” may refer to the support of normal bone growth and / or strength, for example during childhood and adolescence. Supporting normal bone growth and / or strength may result in normal bone anatomy and physiology. Suitable methods and parameters to determine bone growth and bone strength will be known to the skilled person (see e.g. Donnelly, E., 2011. Clinical Orthopaedics and Related Research, 469(8), pp.2128-2138). Suitably, normal bone growth and / or strength may be determined using one or more bone parameter selected from: trabecular bone volume fraction (BV / TV), bone mineral density (BMD), bone mineral content

[0262] (BMC), cortical bone volume (Ct.BV), medio-lateral diameter, antero-posterior diameter, bone ultimate force (FMax), and bone stiffness. In some embodiments, normal bone growth and / or strength is determined using one or more bone parameter selected from: bone mineral density

[0263] (BMD), trabecular bone volume fraction (BV / TV), cortical bone volume (Ct.BV), and bone ultimate force (FMax). Suitable methods to determine these parameters will be available to the skilled person. The composition or combination of the present invention may promote muscle growth and / or strength. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting muscle growth and / or strength in a subject, optionally wherein the composition or combination promotes the production of AMP in the gastrointestinal tract of the subject. As used herein, “promoting muscle growth and / or strength” may refer to the support of normal muscle growth and / or strength, for example during childhood and adolescence. Supporting normal muscle growth and / or strength may result in normal muscle anatomy and physiology. Suitable methods and parameters to determine muscle growth and muscle strength will be known to the skilled person

[0264] The composition or combination of the present invention may promote catch-up growth e.g. in subjects with stunted growth and / or faltering growth. In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting catchup growth in a subject, optionally wherein the composition or combination promotes the production of AMP in the gastrointestinal tract of the subject. As used herein, “catch-up growth” may refer to height velocity above the limits of normal for age for at least 1 year after a transient period of growth inhibition and may be complete or incomplete (see e.g. Wit, J.M. and Boersma, B., 2002. Journal of Pediatric Endocrinology and Metabolism, 15, pp.1229-1242. Suitable method and parameters to determine catch-up growth will be known to the skilled person. Suitably, catch-up growth may be determined using height velocity or height standard deviation score (see e.g. Frongillo, E.A., Leroy, J.L. and Lapping, K., 2019. Advances in Nutrition, 10(3), pp.372-379 and Desmond, C. and Casale, D., 2017. PloS one, 12(12), p.e0189135). In some embodiments, the catch-up growth is determined in absolute terms of linear growth (i.e. the height deficit from the healthy reference population mean is reduced). In some embodiments, the catch-up growth is determined in relative terms of linear growth (i.e. the height-for-age z-score is improved and / or passes the -2SD or -1SD cut-off points).

[0265] Improved gut microbiota composition

[0266] The composition or combination of the present invention may support the development and / or maintenance of a healthy gut microbiota. Adenosine monophosphate (AMP) has been linked to improved gut microbiota composition in formula-fed infants based on findings from several randomized clinical trials (Pediatrics. 2010 Oct; 126(4) :e946-53 and Am J Clin Nutr. 2008 Jun; 87(6): 1785-92).

[0267] In a further aspect, the invention provides the use of a composition or combination according to the invention for use in promoting and / or maintaining a healthy gut microbiota in a subject (e.g. an infant or young child), optionally wherein the composition or combination promotes the production of AMP in the gastrointestinal tract of the subject (e.g. infant or young child). Non-therapeutic use

[0268] In a further aspect, the invention provides use of the composition or combination of the invention to promote the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of a subject.

[0269] In a further aspect, the invention provides a method of promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4- guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of a subject, the method comprising administering to the subject the composition or combination of the invention.

[0270] The use or method of the present invention may be referred to as a non-therapeutic use or method. In some embodiments, the subject is a healthy subject.

[0271] Subject

[0272] In some embodiments, the subject is a mammal, such as a human.

[0273] In some embodiments, the composition according to the invention is for use in infants, young children or children.

[0274] In one embodiment, the subject is an infant. In one embodiment, the subject is a young child. In one embodiment, the subject is a child.

[0275] The composition or combination according to the invention is for use in infants or young children. It is particularly adapted for infants under 6 months of age.

[0276] The composition or combination may be particularly useful for preterm infants and / or low or very low birth weight infants. In another particularly interesting embodiment, the composition or combination is used in infants delivered via Caesarean section. Antibiotic administration is a recommended medical practice for C-section birth in order to prevent infection. Such interventions are potent disruptors of microbial communities (the mother’s or the child’s) and C-section born infants may have a delayed and less optimal colonization of the large intestinal tract.

[0277] The infants, young children or children may be born term or preterm. In a particular embodiment, the composition or combination of the invention is for use in infants, young children or children that were born preterm. Preterm infants may be at increased risk of poor nutrient utilization, impaired lean body mass growth, fat accumulation in the visceral area and metabolic disease later in life. Thus, in one embodiment the composition or combination of the invention is for use in preterm infants.

[0278] In one embodiment, the subject is an infant or a young child that was born small for gestational age or low birth weight.

[0279] Infants or young children with low birth weight may or may not be preterm, and similarly, infants or young children who are small for gestational age may or may not be preterm.

[0280] The composition or combination of the present invention may also be used in an infant or a young child that was born by C-section or that was vaginally delivered.

[0281] All infants and young children can benefit from the invention as all of them are or can be, at a certain age, susceptible to acquiring an unbalanced intestinal / gut microbiota.

[0282] In some advantageous embodiments of the invention, the composition or combination is for use infants or young children having a fragile or unbalanced microbiota or dysbiosis of microbiota, such as preterm infants, infants born by Caesarean-section, infants born small for gestational age or with low birth weight, hospitalized infants / young children, infants / young children treated or having been treated by antibiotics, infants / young children sick at birth including necrotizing enterocolitis (NEC), and / or infants / young children suffering or having suffered from gut infection and / or gut inflammation.

[0283] It is indeed foreseen that the composition or combination of the invention may be even more beneficial to infants born with possibly impaired gut microbiota or fragile infants / young children (such as prematurely born infants and / or infants born by C-section). It is also foreseen that the composition or combination of the invention may be even more beneficial to infants / young children exhibiting intestinal disorders (such as diarrhea, infections or colic), especially after birth, for example, during the first 4 weeks after birth.

[0284] In embodiments of the invention, the infants born prematurely or born by caesarean section or born small for gestational age or with low birth weight, or exhibiting unbalanced or abnormal gut microbiota or suffering or having suffered from gut infection and / or gut inflammation, are targeted by the composition or combination of the present invention, and especially when the infants are 0-6 months of age. Without being bound by the theory, it is believed that younger infants benefit even more from the composition or combination of the invention, especially when the infants have (or are at risk of having) an unbalanced intestinal microbiota and / or have a fragile health condition (as exemplified by the conditions cited above).

[0285] The composition or combination can be administered (or given or fed) at an age and for a period that depends on the needs. In one embodiment, the infants or young children are 0-36 months of age, such as 0-12 months or 0-6 months of age. It is foreseen that the composition or combination of the invention may be even more beneficial to infants just after birth (0-4 weeks or 0-8 weeks) as their intestinal tract may be more fragile.

[0286] The mammal to be treated is preferably a human being, but the mammal may also be nonhuman mammal, such as a non-human mammal selected from the group consisting of pig, cow, horse, dog, cat, goat, sheep and rabbit.

[0287] In other embodiments, the subject is a juvenile animal, preferably wherein the animal is a pet.

[0288] A pet may be a mammal such as dogs or cats, or rodents such as mice, rats, and guinea pigs, rabbits, etc. In some embodiments, the pet is a small dog breed.

[0289] The term “juvenile” may refer to an individual that has not yet reached adulthood.

[0290] In some embodiments the composition or combination according to the invention can be for use before and / or during the weaning period.

[0291] In some preferred embodiments the composition or combination according to the invention is for use in a subject at risk and / or in need. The subject at risk and / or in need may be bottle-fed and / or formula-fed. The subject at risk and / or in need may be born by C-section and / or preterm, and / or small for gestational age (SGA) and / or with a low, very low, or extremely low weight at birth (LWB, VLBW, or ELBW) and / or experienced intra-uterine growth retardation (IUGR), and / or was hospitalized the first days following birth and / or took antibiotic the first days following birth and / or was sick at birth including necrotizing enterocolitis (NEC), and / or has impaired microbiota and / or dysbiosis of microbiota.

[0292] In one embodiment the composition or combination of the invention is given to the subject as a supplementary composition to the mother's milk. In some embodiments the subject receives the mother's milk during at least the first 2 weeks, first 1 , 2, 4, or 6 months. In one embodiment the composition or combination of the invention is given to the subject after such period of mother's nutrition, or is given together with such period of mother's milk nutrition. In another embodiment the composition or combination is given to the subject as the sole or primary nutritional composition during at least one period of time, e.g. after the 1st, 2ndor 4thmonth of life, during at least 1 , 2, 4 or 6 months. In one embodiment the composition or combination of the invention is a complete nutritional composition (fulfilling all or most of the nutritional needs of the subject). In another embodiment the composition or combination of the invention is a supplement or a fortifier intended for example to supplement human milk or to supplement an infant formula or a follow- on formula. Nutritional composition

[0293] In some embodiments, the composition or combination of the invention is in the form of a nutritional composition.

[0294] The nutritional composition according to the invention can be for example an infant formula, a starter infant formula, a follow-on or follow-up formula, a growing-up milk, a baby food, an infant cereal composition, a fortifier such as a human milk fortifier, a supplement, a pet food, or a pet food supplement. In some particular embodiments, the composition of the invention is an infant formula, a fortifier or a supplement that may be intended for the first 4 or 6 months of age. In a preferred embodiment the nutritional composition of the invention is an infant formula.

[0295] In some embodiments, the nutritional composition contains only intact proteins as protein sources.

[0296] In some embodiments, the nutritional composition contains intact and hydrolysed proteins as protein sources.

[0297] In some other embodiments, the nutritional composition of the present invention is a fortifier. The fortifier can be a breast milk fortifier (e.g. a human milk fortifier) or a formula fortifier such as an infant formula fortifier or a follow-on / follow-up formula fortifier.

[0298] When the nutritional composition is a supplement, it can be provided in the form of unit doses. In such cases it is particularly useful to define the amount of oligosaccharides and probiotics in terms of daily dose to be administered to the infant, young child or child.

[0299] When the nutritional composition is a supplement, it may comprise the Bifidobacterium longum subsp. infantis, optionally the Bifidobacterium animalis subsp. lactis, LNFP-I and / or the HMO mixture as described herein, and no other additional nutrient on top of the excipients necessary to obtain a stable nutritional composition.

[0300] The nutritional composition of the present invention can be in solid (e.g. powder), liquid or gelatinous form. In a specific embodiment the nutritional composition is a supplement, wherein the supplement is in powder form and provided in a sachet, preferably a sachet with 0.1 to 20 g per sachet, for example 1 to 10 g per sachet, or in the form of a syrup, preferably a syrup with a total solid concentration of 5 to 75 g / 100 mL (5 to 75% (w / v)). When the supplement is in powder form, it may comprise a carrier. It is however preferred that the supplement is devoid of a carrier. When the supplement is in the form of a syrup, the components are preferably dissolved or suspended in water acidified with citrate. When the nutritional composition is a supplement, it can be free of protein.

[0301] In a particular embodiment the nutritional composition according to the invention is a hypoallergenic composition. In another particular embodiment the composition according to the invention is a hypoallergenic nutritional composition.

[0302] Other ingredients

[0303] The composition or combination according to the present invention may also comprise any other suitable ingredients.

[0304] Suitably, the composition or combination according to the present invention may also comprise other types of oligosaccharide(s), polysaccharides and / or a fiber(s) and / or a precursor(s) thereof. The other oligosaccharide and / or fiber and / or precursor thereof may be selected from the list comprising human milk oligosaccharides (HMOs), galacto-oligosaccharides (GOS), fructo-oligosaccharides (FOS), xylooligosaccharides (XOS), cello-oligosaccharides (COS), arabinoxylans, arabinans, xylans, inulin, polydextrose, beta-glucans, pectins and any combination thereof and any derived preparations thereof (e.g. partial hydrolysis). They may be in an amount between 0 and 10% by weight of composition. In a particular embodiment, the composition or combination can also contain at least one BMO (bovine milk derived oligosaccharide).

[0305] Additional HMOs which may be included in the composition or combination according to the present invention may be selected from the group consisting of lacto-N- fucopentaose (e.g. lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V), lacto-N- fucohexaose, lacto-N-difucohexaose I, fucosyllacto-N-hexaose, fucosyllacto-N-neohexaose, difucosyllacto-N-hexaose I, difucosyllacto-N-neohexaose II, para-lacto-N-neohexaose (para- LNnH), lacto-N-hexaose, lacto- N-neohexaose, para-lacto-N-hexaose, para-lacto-N- neohexaose, lacto-N-octaose, lacto-N-neooctaose, iso-lacto-N-octaose, para- lacto-N- octaose, lacto-N-decaose, and any combination thereof.

[0306] In some embodiments, the composition or combination according to the invention comprises at least one additional HMO.

[0307] In other embodiments, the composition or combination according to the present invention is devoid of any further HMOs. Thus, the LNFP-I or HMO mixture as described herein may be the only HMOs in the composition or combination of the invention.

[0308] In some embodiments, the composition or combination of the present invention does not comprise at least one further probiotic (or probiotic strain), such as at least one further probiotic bacterial strain. In other embodiments, the composition or combination of the present invention can further comprise at least one further probiotic (or probiotic strain), such as at least one further probiotic bacterial strain.

[0309] The probiotic microorganisms most commonly used are principally bacteria and yeasts of the following genera: Lactobacillus spp., Lacticaseibacillus spp, Limosilactobacillus spp, Streptococcus spp., Enterococcus spp., Bifidobacterium spp. and Saccharomyces spp.

[0310] In some particular embodiments, the probiotic is a probiotic bacterial strain. In some specific embodiments, it is particularly Bifidobacteria and / or Lactobacilli.

[0311] Suitable probiotic bacterial strains according to the present invention include B. longum CNCM 1-2618 (B. longum NCC2705), Bifidobacterium breve sold by Danisco under the trademark Bb- 03, Bifidobacterium breve sold by Morinaga under the trade mark M-16V, Bifidobacterium breve sold by Morinaga under the trade mark B-3, Bifidobacterium breve sold by sold by Yakult under the trade mark BBG-01 , Bifidobacterium breve sold by Institut Rosell (Lallemand) under the trademark R0070, Lactobacillus rhamnosus ATCC 53103 available from Valio Oy of Finland under the trademark LGG, Lactobacillus rhamnosus CGMCC 1.3724, Lactobacillus paracasei CNCM 1-2116, Lactobacillus johnsonii CNCM 1-1225, Streptococcus salivarius DSM 13084 sold by BLIS Technologies Limited of New Zealand under the designation KI2.

[0312] In some embodiments, the composition or combination of the invention comprises Lactobacillus paracasei CNCM 1-2116 or a strain having an Average Nucleotide Identity (ANI) of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1 %, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% to Lactobacillus paracasei CNCM 1-2116.

[0313] In some embodiments, the composition or combination of the invention comprises Lactobacillus rhamnosus CGMCC 1.3724 or a strain having an Average Nucleotide Identity (ANI) of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% to Lactobacillus rhamnosus CGMCC 1.3724.

[0314] The composition or combination according to the invention may contain from 10e3 to 10e12 cfu of the at least one (further) probiotic strain, more preferably from 10e7 to 10e12 cfu such as from 10e8 to 10e10 cfu of probiotic strain per g of composition or combination on a dry weight basis. In one embodiment, the probiotics are viable. In another embodiment, the probiotics are nonreplicating or inactivated. There may be both viable probiotics and inactivated probiotics in some other embodiments. Probiotic components and metabolites can also be added.

[0315] The composition according to the invention generally contains a protein source. The protein can be in an amount of from 1.6 to 3 g per 100 kcal. In some embodiments, especially when the composition is intended for premature infants, the protein amount can be between 2.4 and 4 g / 1 OOkcal or more than 3.6 g / 1 OOkcal (or 3.6 g / 1 OOkcal or more). In some other embodiments the protein amount can be below 2.0 g per 100 kcal, e.g. between 1.8 to 2 g / 100 kcal, or in an amount below 1.8 g per 100 kcal.

[0316] Protein sources based on whey, casein and mixtures thereof may be used as well as protein sources based on soy. As far as whey proteins are concerned, the protein source may be based on acid whey or sweet whey or mixtures thereof and may include alpha-lactalbumin and beta-lactoglobulin in any desired proportions.

[0317] In some advantageous embodiments the protein source is whey predominant (i.e. more than 50% of proteins are coming from whey proteins, such as 51 , 60% or 70%).

[0318] The proteins may be intact or hydrolysed or a mixture of intact and hydrolysed proteins. By the term “intact” is meant that the main part of the proteins are intact, i.e. the molecular structure is not altered, for example at least 80% of the proteins are not altered, such as at least 85% of the proteins are not altered, preferably at least 90% of the proteins are not altered, even more preferably at least 95% of the proteins are not altered, such as at least 98% of the proteins are not altered. In a particular embodiment, 100% of the proteins are not altered.

[0319] The term “hydrolysed” means in the context of the present invention a protein which has been hydrolysed or broken down into its component amino acids. The proteins may be either fully or partially hydrolysed. It may be desirable to supply partially hydrolysed proteins (degree of hydrolysis between 2 and 20%), for example for infants or young children believed to be at risk of developing cow’s milk allergy. If hydrolysed proteins are required, the hydrolysis process may be carried out as desired and as is known in the art. For example, whey protein hydrolysates may be prepared by enzymatically hydrolysing the whey fraction in one or more steps. If the whey fraction used as the starting material is substantially lactose free, it is found that the protein suffers much less lysine blockage during the hydrolysis process. This enables the extent of lysine blockage to be reduced from about 15% by weight of total lysine to less than about 10% (or to about 10% or less) by weight of lysine; for example about 7% by weight of lysine which greatly improves the nutritional quality of the protein source. In an embodiment of the invention at least 70% of the proteins are hydrolysed, preferably at least 80% of the proteins are hydrolysed, such as at least 85% of the proteins are hydrolysed, even more preferably at least 90% of the proteins are hydrolysed, such as at least 95% of the proteins are hydrolysed, particularly at least 98% of the proteins are hydrolysed. In a particular embodiment, 100% of the proteins are hydrolysed.

[0320] In one particular embodiment the proteins of the nutritional composition are hydrolyzed, fully hydrolyzed or partially hydrolyzed. The degree of hydrolysis (DH) of the protein can be between 8 and 40, or between 20 and 60 or between 20 and 80 or more than 10, 20, 40, 60, 80 or 90 (or 10, 20, 40, 60, 80 or 90 or more).

[0321] The protein component can alternatively be replaced by a mixture or synthetic amino acid, for example for preterm or low birth weight infants.

[0322] In a particular embodiment, the composition or combination according to the invention is a hypoallergenic composition. In another particular embodiment, the composition or combination according to the invention is a hypoallergenic nutritional composition.

[0323] The composition according to the present invention generally contains a carbohydrate source. This is particularly preferable in the case where the composition of the invention is an infant formula. In this case, any carbohydrate source conventionally found in infant formulae such as lactose, sucrose, saccharose, maltodextrin, starch and mixtures thereof may be used although one of the preferred sources of carbohydrates is lactose.

[0324] The composition according to the present invention generally contains a source of lipids. This is particularly relevant if the composition of the invention is an infant formula. In this case, the lipid source may be any lipid or fat which is suitable for use in infant formulae. Some suitable fat sources include palm oil, structured triglyceride oil, high oleic sunflower oil and high oleic safflower oil, medium-chain-triglyceride oil. The essential fatty acids linoleic and a-linolenic acid may also be added, as well small amounts of oils containing high quantities of preformed arachidonic acid and docosahexaenoic acid such as fish oils or microbial oils. The fat source may have a ratio of n-6 to n-3 fatty acids of about 5:1 to about 15:1 ; for example about 8:1 to about 10:1.

[0325] The composition of the invention may also contain all vitamins and minerals understood to be essential in the daily diet and in nutritionally significant amounts. Minimum requirements have been established for certain vitamins and minerals. Examples of minerals, vitamins and other nutrients optionally present in the composition of the invention include vitamin A, vitamin B1 , vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorous, iodine, iron, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine, and L-carnitine. Minerals are usually added in salt form. The presence and amounts of specific minerals and other vitamins will vary depending on the intended population.

[0326] If necessary, the composition of the invention may contain emulsifiers and stabilisers such as soy, lecithin, citric acid esters of mono- and di-glycerides, and the like.

[0327] The composition of the invention may also contain other substances which may have a beneficial effect such as nucleotides, nucleosides, and the like.

[0328] The composition of the invention may also contain carotenoid(s). In some particular embodiments of the invention, the composition of the invention does not comprise any carotenoid.

[0329] Manufacture of a composition

[0330] The composition or combination according to the invention may be prepared in any suitable manner. A composition will now be described by way of example.

[0331] For example, a formula such as an infant formula may be prepared by blending together the protein source, the carbohydrate source and the fat source in appropriate proportions. If used, the emulsifiers may be included at this point. The vitamins and minerals may be added at this point but they are usually added later to avoid thermal degradation. Any lipophilic vitamins, emulsifiers and the like may be dissolved into the fat source prior to blending. Water, preferably water which has been subjected to reverse osmosis, may then be mixed in to form a liquid mixture. The temperature of the water is conveniently in the range between about 50°C and about 80°C to aid dispersal of the ingredients. Commercially available liquefiers may be used to form the liquid mixture.

[0332] The oligosaccharide(s) may be added at this stage, especially if the final product is to have a liquid form. If the final product is to be a powder, they may likewise be added at this stage if desired.

[0333] The liquid mixture is then homogenised, for example in two stages.

[0334] The liquid mixture may then be thermally treated to reduce bacterial loads, by rapidly heating the liquid mixture to a temperature in the range between about 80°C and about 150°C for a duration between about 5 seconds and about 5 minutes, for example. This may be carried out by means of steam injection, an autoclave or a heat exchanger, for example a plate heat exchanger. Then, the liquid mixture may be cooled to between about 60°C and about 85°C for example by flash cooling. The liquid mixture may then be again homogenised, for example in two stages between about 10 MPa and about 30 MPa in the first stage and between about 2 MPa and about 10 MPa in the second stage. The homogenised mixture may then be further cooled to add any heat sensitive components, such as vitamins and minerals. The pH and solids content of the homogenised mixture are conveniently adjusted at this point.

[0335] If the final product is to be a powder, the homogenised mixture is transferred to a suitable drying apparatus such as a spray dryer or freeze dryer and converted to powder. The powder should have a moisture content of less than about 5% (or about 5% or less) by weight. The oligosaccharide(s) may also or alternatively be added at this stage by dry-mixing or by blending them in a syrup form of crystals, along with the probiotic strain(s), and the mixture is spray-dried or freeze-dried.

[0336] If a liquid composition is preferred, the homogenised mixture may be sterilised then aseptically filled into suitable containers or may be first filled into the containers and then retorted.

[0337] In another embodiment, the composition of the invention may be a supplement such as a paediatric supplement. The supplement may be in the form of tablets, capsules, pastilles or a liquid for example. The supplement may further contain protective hydrocolloids (such as gums, proteins, modified starches), binders, film forming agents, encapsulating agents / materials, wall / shell materials, matrix compounds, coatings, emulsifiers, surface active agents, solubilizing agents (oils, fats, waxes, lecithins etc.), adsorbents, carriers, fillers, cocompounds, dispersing agents, wetting agents, processing aids (solvents), flowing agents, taste masking agents, weighting agents, jellifying agents and gel forming agents. The supplement may also contain conventional pharmaceutical additives and adjuvants, excipients and diluents, including, but not limited to, water, gelatine of any origin, vegetable gums, lignin- sulfonate, talc, sugars, starch, gum arabic, vegetable oils, polyalkylene glycols, flavouring agents, preservatives, stabilizers, emulsifying agents, buffers, lubricants, colorants, wetting agents, fillers, and the like.

[0338] Further, the supplement may contain an organic or inorganic carrier material suitable for oral or parenteral administration as well as vitamins, minerals trace elements and other micronutrients in accordance with the recommendations of Government bodies such as the USRDA.

[0339] This disclosure is not limited by the exemplary methods and materials disclosed herein, and any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of this disclosure. Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, any nucleic acid sequences are written left to right in 5' to 3' orientation; amino acid sequences are written left to right in amino to carboxy orientation, respectively.

[0340] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limits of that range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within this disclosure. The upper and lower limits of these smaller ranges may independently be included or excluded in the range, and each range where either, neither or both limits are included in the smaller ranges is also encompassed within this disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in this disclosure.

[0341] It must be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

[0342] The terms "comprising", "comprises" and "comprised of" as used herein are synonymous with "including", "includes" or "containing", "contains", and are inclusive or open-ended and do not exclude additional, non-recited members, elements or method steps. The terms "comprising", "comprises" and "comprised of" also include the term "consisting of".

[0343] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of chemistry, biochemistry, molecular biology, microbiology and immunology, which are within the capabilities of a person of ordinary skill in the art. Such techniques are explained in the literature. See, for example, Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press; Ausubel, F.M. et al. (1995 and periodic supplements) Current Protocols in Molecular Biology, Ch. 9, 13 and 16, John Wiley & Sons; Roe, B., Crabtree, J. and Kahn, A. (1996) DNA Isolation and Sequencing: Essential Techniques, John Wiley & Sons; Polak, J.M. and McGee, J.O’D. (1990) In Situ Hybridization: Principles and Practice, Oxford University Press; Gait, M.J. (1984) Oligonucleotide Synthesis: A Practical Approach, IRL Press; and Lilley, D.M. and Dahlberg, J.E. (1992) Methods in Enzymology: DNA Structures Part A: Synthesis and Physical Analysis of DNA, Academic Press. Each of these general texts is herein incorporated by reference. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that such publications constitute prior art to the claims appended hereto.

[0344] The invention will now be further described by way of Examples, which are meant to serve to assist one of ordinary skill in the art in carrying out the invention and are not intended in any way to limit the scope of the invention.

[0345] EXAMPLES

[0346] Bifidobacterium infantis synergistically produces beneficial metabolites when combined with a blend of HMOs, including LNFP-I

[0347] An ex vivo fecal fermentation study was conducted using an ex-vivo human Gastrointestinal Tract replica model and microbiota colonic incubations as described in Front Microbiol. 2023 Apr 14;14:1131662.

[0348] Eight arms were used in the study. These consisted of:

[0349] 1. HMO blend I: 2’FL, 3FL, DiFL, 3’SL, 6’SL, LNT

[0350] 2. HMO blend II: 2’FL, 3FL, DiFL, 3’SL, 6’SL, LNT, LNnT

[0351] 3. HMO blend I + LNFP-I

[0352] 4. HMO blend II + LNFP-I

[0353] 5. HMO blend I + B. infantis LMG11588

[0354] 6. HMO blend II + B. infantis LMG11588

[0355] 7. HMO blend I + LNFP-I + B. infantis LMG11588

[0356] 8. HMO blend II + LNFP-I + B. infantis LMG11588

[0357] Each arm consisted of 6 bioreactors (n = 6) seeded with the faecal microbiota of 6 individual donors. The same fecal inoculum of the 6 donors was used for all arms. Donors were 3-month- old infants (± 2 weeks) exclusively fed with formulae without HMO / probiotics. The exclusion criteria were antibiotic use in 30 days before sample delivery for the study and previous NEC or gut surgery. Intestinal absorption and colonic incubation conditions were then conducted using ex-vivo SIFR protocols (cf. Front Microbiol. 2023 Apr 14; 14:1131662). B. infantis was inoculated at a dose of 5.0E+07 CFU / mL. The carbohydrate LNFP-I was dosed at a concentration of 0.30 g / L, while the HMO blend was dosed at a final concentration of 2.66g / L. Controls with no added carbohydrate source, and each individual carbohydrate source, were performed. Colonic incubations were carried out for 24-hours, with sampling at 0 and 24 hours for Metabolite analysis by LC-MS. LC-MS analysis was carried out using a Thermo Scientific Vanquish LC coupled to Thermo Q Exactive HF MS. An electrospray ionization interface was used as ionization source. Analysis was performed in negative and positive ionization mode. The LIPLC was performed using a slightly modified version of the protocol described by Doneanu et al. (Anal Chem, 2015, 87, 10283-10291). Peak areas were extracted using Compound Discoverer 3.1 (Thermo Scientific). In addition to the automatic compound extraction by Compound Discoverer 3.1 , a manual extraction of compounds included in an inhouse library was performed using Skyline 21.1 (MacCoss Lab Software). After data acquisition, the average value for each compound in each condition was calculated. These conditions were blank (no LNFP-I nor B. infantis), prebiotic (only LNFP-I), probiotic (only B. infantis), and synbiotic (LNFP-I and B. infantis). An HMO blend (either HMO blend I or blend II) without LNFP-I was present in all conditions as a background. Synergy size was calculated by first subtracting the blank value form all other values and then calculating the ratio of the blanked synbiotic condition concentrate and the summed blanked prebiotic condition and blanked probiotic condition. Synergy was defined as ratios > 1.

[0358] The combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) with a background of 49% 2’FL, 14% 3FL, 7% DFL, 6% 3’SL, 8% 6’SL, and 16% LNT dosed at total HMO concentration of 2.66 g / L synergistically increased production of cAMP, 4- guanidinobutyric acid, acetyl-carnitine and 3-methylxanthine (see Figure 1A). The combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) with a background of 38% 2’FL, 14% 3FL, 5% DFL, 4% 3’SL, 7% 6’SL, 13% LNT, and 19% LNnT dosed at total HMO concentration of 2.66 g / L synergistically increased production of uridine, adenine, guanine, AMP, cAMP, p-hydroxyphenyllactic acid, and acetyl-carnitine (see Figure 1 B).

[0359] EMBODIMENTS

[0360] Various preferred features and embodiments of the present invention will now be described with reference to the following numbered clauses:

[0361] 1. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in therapy by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p- hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'-fucosyl lactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosy I lactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0362] 2. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p- hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT), wherein the infant or young child was born by C-section and / or preterm, and / or small for gestational age (SGA) and / or with a low, very low, or extremely low weight at birth (LWB, VLBW, or ELBW) and / or experienced intra-uterine growth retardation (IUGR), and / or was hospitalized the first days following birth and / or took antibiotic the first days following birth and / or was sick at birth, and / or has impaired microbiota and / or dysbiosis of microbiota.

[0363] 3. Use of a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture to promote the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p- hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT), wherein the use is for example non-therapeutic.

[0364] 4. The combination for use according to clause 1 or 2, or the use according to clause 3, wherein the combination is administered separately, simultaneously or sequentially, preferably wherein the combination is administered simultaneously. 5. The combination for use according to any of clauses 1-2 or 4, or the use according to clause 3 or 4, wherein the combination is provided in the form of a nutritional composition, preferably wherein the nutritional composition is selected from an infant formula, a starter infant formula, a follow-on or follow-up formula, a baby food, an infant cereal composition, a growing-up-milk, a fortifier such as a human milk fortifier, or a supplement.

[0365] 6. The combination for use according to any of clauses 1-2 or 4-5, or the use according to any of clauses 3-5, wherein the Bifidobacterium longum subsp. infantis is Bifidobacterium longum subsp. infantis LMG 11588 or a strain having an Average Nucleotide Identity (AN I) of at least 95%, preferably at least 99%, more preferably at least 99.9% to Bifidobacterium longum subsp. infantis LMG 11588.

[0366] 7. The combination for use according to any of clauses 1-2 or 4-6, or the use according to any of clauses 3-6, wherein the combination comprises from 103to 1012cfu of Bifidobacterium longum subsp. infantis, preferably from 107to 1012cfu of Bifidobacterium longum subsp. infantis, more preferably from 108to 101° cfu of Bifidobacterium longum subsp. infantis per g of combination or composition on a dry weight basis.

[0367] 8. The combination for use according to any of clauses 1-2 or 4-7, or the use according to any of clauses 3-7, wherein the combination further comprises Bifidobacterium animalis subsp. lactis, optionally wherein the Bifidobacterium animalis subsp. lactis is Bifidobacterium animalis subsp. lactis CNCM I-3446 or a strain having an Average Nucleotide Identity (ANI) of at least 95%, preferably at least 99%, more preferably at least 99.9% to Bifidobacterium animalis subsp. lactis CNCM I-3446.

[0368] 9. The combination for use according to clause 8, or the use according to clause 8, wherein the combination comprises from 103to 1012cfu of Bifidobacterium longum subsp. lactis, preferably from 107to 1012cfu of Bifidobacterium longum subsp. lactis, more preferably from 108to 101° cfu of Bifidobacterium longum subsp. lactis per g of combination or composition on a dry weight basis.

[0369] 10. The combination for use according to any of clauses 1-2 or 4-9, or the use according to any of clauses 3-9, wherein the combination comprises LNFP-I in a total amount of from 25 mg / L to 4000 mg / L of the combination or composition or from 0.02 g / 100 g to 3.75 g / 100 g of the combination or composition on a dry weight basis.

[0370] 11. The combination for use according to any of clauses 1-2 or 4-10, or the use according to any of clauses 3-10, wherein the combination comprises LNFP-I in a total amount of from 50 mg / L to 2500 mg / L, preferably from 60 mg / L to 2000 mg / L, more preferably from 80 mg / L to 1500 mg / L of the combination or composition; or from 0.04 g / 100 g to 2 g / 100 g, preferably from 0.05 g / 100 g to 1.6 g / 100 g, more preferably from 0.06 g / 100 g to 1.2 g / 100 g, most preferably from 0.07 g / 100 g to 0.8 g / 100 g of the combination or composition on a dry weight basis.

[0371] 12. The combination for use according to any of clauses 1-2 or 4-11 , or the use according to any of clauses 3-11 , wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and 3-fucosyllactose (3FL), and / or wherein the HMO mixture consists essentially of: i. 16 wt% to 69 wt% of 2FL, preferably 22 wt% to 59 wt%; ii. 9 wt% to 24 wt% of LNT, preferably 12 wt% to 21 wt%; iii. 2 wt% to 10 wt% of DFL, preferably 3 wt% to 8 wt%; iv. 8 wt% to 26 wt% of 6SL and 3SL combined, preferably 11 wt% to 22 wt%; and v. 10 wt% to 50 wt% of 3FL, preferably 11 wt% to 43 wt%.

[0372] 13. The combination for use according to clause 12, or the use according to clause 12, wherein the LNFP-I and HMO mixture combined consists essentially of: i. 20 wt% to 46 wt% of 2FL, preferably 22 wt% to 42 wt% of 2FL; ii. 11 wt% to 17 wt% of LNT, preferably 12 wt% to 15 wt% of LNT; iii. 2 wt% to 7 wt% of DFL, preferably 3 wt% to 6 wt% of DFL; iv. 9 wt% to 21 wt% of 6SL and 3SL combined, preferably 9 wt% to 19 wt% of 6SL and 3SL combined; v. 9 wt% to 34 wt% of 3FL, preferably 11 wt% to 32 wt% of 3FL; and vi. 5 wt% to 32 wt% of LNFP-I, preferably 10 wt% to 19 wt% of LNFP-I.

[0373] 14. The combination for use according to any of clauses 12-13, or the use according to any of clauses 12-13, wherein the combination promotes the production of one or more of cAMP, 4- guanidinobutyric acid, acetyl-carnitine and 3-methylxanthine, preferably wherein the combination promotes the production of each of cAMP 4-guanidinobutyric acid, acetylcarnitine and 3-methylxanthine.

[0374] 15. The combination for use according to any of clauses 1 -2 or 4-11 , or the use according to any of clauses 3-11 , wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), 3- fucosyllactose (3FL), and lacto-N-neotetraose (LNnT), and / or wherein the HMO mixture consists essentially of: i. 20 wt% to 60 wt% of 2FL, preferably 22 wt% to 55 wt%; ii. 4 wt% to 30 wt% of LNT, preferably 6 wt% to 20 wt%; iii. 1 wt% to 12 wt % of DFL, preferably 3 wt% to 8 wt%; iv. 7 wt% to 23 wt% of 6SL and 3SL combined, preferably 8 wt% to 22 wt%; v. 10 wt% to 50 wt% of 3FL, preferably 13 wt% to 46 wt%; and vi. 3 wt% to 25 wt% of LNnT, preferably 5 wt% to 20 wt%

[0375] 16. The combination for use according to clause 15, or the use according to clause 15, wherein the LNFP-I and HMO mixture combined consists essentially of: i. 29 wt% to 40 wt% of 2FL, preferably 32 wt% to 39 wt% of 2FL; ii. 8 wt% to 13 wt% of LNT, preferably 9 wt% to 12 wt% of LNT; iii. 3 wt% to 11 wt % of DFL; iv. 3 wt% to 15 wt% of 6SL and 3SL combined, preferably 4 wt% to 15 wt% of 6SL and 3SL combined; v. 11 wt% to 35 wt% of 3FL, preferably 12 wt% to 35 wt% of 3FL; vi. 1 wt% to 18 wt% of LNnT, preferably 1 wt% to 17 wt% of LNnT; and vii. 2 wt% to 24 wt% of LNFP-I, preferably 4 wt% to 14 wt% of LNFP-I.

[0376] 17. The combination for use according to any of clauses 15-16, or the use according to any of clauses 15-16, wherein the combination promotes the production of one or more of uridine, adenine, guanine, AMP, cAMP, p-hydroxyphenyllactic acid and acetyl-carnitine, preferably wherein the combination promotes the production of each of uridine, adenine, guanine, AMP, cAMP, p-hydroxyphenyllactic acid and acetyl-carnitine.

[0377] 18. The combination for use according to any of clauses 1-2 or 4-11 , or the use according to any of clauses 3-11 , wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and lacto-N-neotetraose (LNnT), and / or wherein the HMO mixture consists essentially of: i. 34 wt% to 85 wt% of 2FL, preferably 40 wt% to 71 wt% of 2FL; ii. 10 wt% to 40 wt% of LNT, preferably 12 wt% to 26 wt% of LNT ; iii. 4 wt% to 14 wt% of DFL, preferably 5 wt% to 10 wt% of DFL; iv. 9 wt% to 31 wt% of 6SL and 3SL combined, preferably 10 wt% to 28 wt% of 6SL and 3SL combined; and v. 6 wt% to 30 wt% of LNnT, preferably 7 wt% to 23 wt% of LNnT.

[0378] 19. The combination for use according to clause 18, or the use according to clause 18, wherein the LNFP-I and HMO mixture combined consists essentially of: i. 27 wt% to 41 wt% of 2FL, preferably 32 wt% to 39 wt% of 2FL; ii. 8 wt% to 15 wt% of LNT, preferably 10 wt% to 14 wt% of LNT ; iii. 4 wt% to 6 wt% of DFL, preferably 4 wt% to 6 wt% of DFL; iv. 8 wt% to 18 wt% of 6SL and 3SL combined, preferably 7 wt% to 15 wt% of 6SL and 3SL combined; v. 13 wt% to 21 wt% of LNnT, preferably 16 wt% to 20 wt% of LNnT ; and vi. 7 wt% to 33 wt% of LNFP-I, preferably 11 wt% to 23 wt% of LNFP-I.

[0379] 20. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is improving metabolic health or treating and / or preventing a metabolic disorder in an infant or young child, optionally wherein the combination promotes the production of uridine, adenine, guanine, AMP, cAMP, and / or acetyl-carnitine in the gastrointestinal tract of the infant or young child.

[0380] 21. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is improving cardiovascular health or treating and / or preventing a cardiovascular disease in an infant or young child, optionally wherein the combination promotes the production of uridine, cAMP, and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child.

[0381] 22. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is improving neurological health, promoting brain growth and / or development, or treating and / or preventing a neurological disorder in an infant or young child, optionally wherein the combination promotes the production of uridine, guanine, cAMP, 4-guanidinobutyric acid, acetyl-carnitine and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child. 23. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is improving mental health or treating and / or preventing a mental disorder in an infant or young child, optionally wherein the combination promotes the production of guanine, 4- guanidinobutyric acid, and / or acetyl-carnitine in the gastrointestinal tract of the infant or young child.

[0382] 24. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is improving respiratory health or treating and / or preventing a respiratory disease in an infant or young child, optionally wherein the combination promotes the production of cAMP and / or 3- methylxanthine in the gastrointestinal tract of the infant or young child.

[0383] 25. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is improving gastrointestinal health or treating and / or preventing gastrointestinal inflammation in an infant or young child, optionally wherein the combination promotes the production of adenine and / or 4-guanidinobutyric acid in the gastrointestinal tract of the infant or young child.

[0384] 26. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is modulating / improving the immune response in an infant or young child, optionally wherein the combination promotes the production of cAMP and / or p-hydroxyphenyllactic acid in the gastrointestinal tract of the infant or young child.

[0385] 27. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is preventing and / or treating an infection in an infant or young child, optionally wherein the combination promotes the production of cAMP, p-hydroxyphenyllactic acid, and / or 3- methylxanthine in the gastrointestinal tract of the infant or young child.

[0386] 28. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is preventing and / or treating apnea in an infant or young child, optionally wherein the combination promotes the production of 3-methylxanthine in the gastrointestinal tract of the infant or young child.

[0387] 29. The combination for use according to any of clauses 1-2 or 4-19, wherein the therapy is improving musculoskeletal health, treating and / or preventing a musculoskeletal disorder, or promoting bone and / or muscle growth and / or development in an infant or young child, optionally wherein the combination promotes the production of uridine and / or AMP in the gastrointestinal tract of the infant or young child.

[0388] 30. The combination for use according to any of clauses 1 , 4-29, or use according to any of clauses 3-19, wherein the infant or young child was born by C-section and / or preterm, and / or small for gestational age (SGA) and / or with a low, very low, or extremely low weight at birth (LWB, VLBW, or ELBW) and / or experienced intra-uterine growth retardation (IUGR), and / or was hospitalized the first days following birth and / or took antibiotic the first days following birth and / or was sick at birth, and / or has impaired microbiota and / or dysbiosis of microbiota.

[0389] 31. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in improving metabolic health or treating and / or preventing a metabolic disorder in an infant or young child, for example by promoting the production of one or more of uridine, adenine, guanine, AMP, cAMP, and / or acetyl-carnitine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0390] 32. The combination for use according to clause 31 , wherein the combination improves metabolic health by promoting nucleic acid synthesis and / or energy metabolism, for example wherein the combination promotes the production of uridine, adenine, guanine, AMP, and / or cAMP in the gastrointestinal tract of the infant or young child; or wherein the metabolic disorder is fatty liver disease, for example wherein the combination promotes the production of adenine in the gastrointestinal tract of the infant or young child; or wherein the metabolic disorder is type 1 diabetes, for example wherein the combination promotes the production of acetylcarnitine in the gastrointestinal tract of the infant or young child.

[0391] 33. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in improving cardiovascular health or treating and / or preventing a cardiovascular disease in an infant or young child, for example by promoting the production of one or more of uridine, cAMP, and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0392] 34. The combination for use according to clause 33, wherein the combination improves cardiovascular health by preventing myocardial injury or protecting against diabetes-induced damage to cardiac mitochondria, for example wherein the combination promotes the production of uridine in the gastrointestinal tract of the infant or young child.

[0393] 35. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in improving neurological health, promoting brain growth and / or development, or treating and / or preventing a neurological disorder in an infant or young child, for example by promoting the production of one or more of uridine, guanine, cAMP, 4- guanidinobutyric acid, acetyl-carnitine and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosy I lactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0394] 36. The combination for use according to clause 35, wherein the combination improves neurological health by neuroprotection, for example wherein the combination promotes the production of uridine, guanine and / or acetyl-carnitine in the gastrointestinal tract of the infant or young child.

[0395] 37. The combination for use according to clause 35, wherein the combination promotes brain growth and / or development, for example wherein the combination promotes the production of AMP in the gastrointestinal tract of the infant or young child.

[0396] 38. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in improving mental health or treating and / or preventing a mental disorder in an infant or young child, for example by promoting the production of one or more of guanine, 4-guanidinobutyric acid, and / or acetyl-carnitine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosy I lactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0397] 39. The combination for use according to clause 38, wherein the mental disorder is a mood disorder, such as a depressive disorder, for example wherein the combination promotes the production of guanine and / or 4-guanidinobutyric acid in the gastrointestinal tract of the infant or young child.

[0398] 40. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in improving respiratory health or treating and / or preventing a respiratory disease in an infant or young child, for example by promoting the production of cAMP and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyl lactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyl lactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0399] 41. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in improving gastrointestinal health or treating and / or preventing gastrointestinal inflammation in an infant or young child, for example by promoting the production of adenine and / or 4-guanidinobutyric acid in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0400] 42. The combination for use according to clause 41 , wherein the gastrointestinal inflammation is intestinal inflammation, for example wherein the combination promotes the production of adenine in the gastrointestinal tract of the infant or young child; or wherein the gastrointestinal inflammation is gastritis, for example wherein the combination promotes the production of 4- guanidinobutyric acid in the gastrointestinal tract of the infant or young child.

[0401] 43. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in modulating / improving the immune response in an infant or young child, for example by promoting the production of cAMP and / or p-hydroxyphenyllactic acid in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N- neotetraose (LNnT).

[0402] 44. The combination for use according to clause 43, wherein the combination modulates / improves the immune response by supporting or promoting protective immunity, for example wherein the combination promotes the production of cAMP and / or p- hydroxyphenyllactic acid in the gastrointestinal tract of the infant or young child.

[0403] 45. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in preventing and / or treating an infection in an infant or young child, for example by promoting the production of one or more of cAMP, p-hydroxyphenyllactic acid, and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyl lactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0404] 46. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in preventing and / or treating apnea in an infant or young child, for example by promoting the production of 3-methylxanthine in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0405] 47. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP- I) and a HMO mixture for use in improving musculoskeletal health, treating and / or preventing a musculoskeletal disorder, or promoting bone and / or muscle growth and / or development in an infant or young child, for example by promoting the production of uridine and / or AMP in the gastrointestinal tract of the infant or young child, wherein the HMO mixture consists of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosy I lactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0406] 48. The combination for use according to any of clauses 30-48, wherein the combination is defined according to any of clauses 4-19.

[0407] 49. The combination for use according to any of clauses 30-48, wherein the infant or young child was born by C-section and / or preterm, and / or small for gestational age (SGA) and / or with a low, very low, or extremely low weight at birth (LWB, VLBW, or ELBW) and / or experienced intra-uterine growth retardation (IUGR), and / or was hospitalized the first days following birth and / or took antibiotic the first days following birth and / or was sick at birth, and / or has impaired microbiota and / or dysbiosis of microbiota.

[0408] 50. A nutritional composition comprising Bifidobacterium longum subsp. infantis, lacto-N- fucopentaose I (LNFP-I) and a HMO mixture, wherein the HMO mixture consists of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

[0409] 51. The nutritional composition according to clause 50, wherein the nutritional composition is selected from an infant formula, a starter infant formula, a follow-on or follow-up formula, a baby food, an infant cereal composition, a growing-up-milk, a fortifier such as a human milk fortifier, or a supplement.

[0410] 52. The nutritional composition according to clause 50 or 51 , wherein the Bifidobacterium longum subsp. infantis is Bifidobacterium longum subsp. infantis LMG 11588 or a strain having an Average Nucleotide Identity (ANI) of at least 95%, preferably at least 99%, more preferably at least 99.9% to Bifidobacterium longum subsp. infantis LMG 11588.

[0411] 53. The nutritional composition according to any of clauses 50-52, wherein the composition comprises from 103to 1012cfu of Bifidobacterium longum subsp. infantis, preferably from 107to 1012cfu of Bifidobacterium longum subsp. infantis, more preferably from 108to 101° cfu of Bifidobacterium longum subsp. infantis per g of composition on a dry weight basis.

[0412] 54. The nutritional composition according to any of clauses 50-53, wherein the composition further comprises Bifidobacterium animalis subsp. lactis. 55. The nutritional composition according to clause 54, wherein the Bifidobacterium animalis subsp. lactis is Bifidobacterium animalis subsp. lactis CNCM 1-3446 or a strain having an Average Nucleotide Identity (ANI) of at least 95%, preferably at least 99%, more preferably at least 99.9% to Bifidobacterium animalis subsp. lactis CNCM 1-3446.

[0413] 56. The nutritional composition according to clrim 54 or 55, wherein the composition comprises from 103to 1012cfu of Bifidobacterium longum subsp. lactis, preferably from 107to 1012cfu of Bifidobacterium longum subsp. lactis, more preferably from 108to 101° cfu of Bifidobacterium longum subsp. lactis per g of composition on a dry weight basis.

[0414] 57. The nutritional composition according to any of clauses 50-56, wherein the composition comprises LNFP-I in a total amount of from 25 mg / L to 4000 mg / L of the composition or of from 0.02 g / 100 g to 3.75 g / 100 g of the composition on a dry weight basis.

[0415] 58. The nutritional composition according to any of clauses 50-57, wherein the composition comprises LNFP-I in a total amount of from 50 mg / L to 2500 mg / L, preferably from 60 mg / L to 2000 mg / L, more preferably from 80 mg / L to 1500 mg / L of the composition; or from 0.04 g / 100 g to 2 g / 100 g, preferably from 0.05 g / 100 g to 1 .6 g / 100 g, more preferably from 0.06 g / 100 g to 1.2 g / 100 g, most preferably from 0.07 g / 100 g to 0.8 g / 100 g of the composition on a dry weight basis.

[0416] 59. The nutritional composition according to any of clauses 50-58, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), 3'-sialyllactose (3SL), and 3-fucosyl lactose (3FL).

[0417] 60. The nutritional composition according to clause 59, wherein the HMO mixture consists essentially of: i. 16 wt% to 69 wt% of 2FL, preferably 22 wt% to 59 wt%; ii. 9 wt% to 24 wt% of LNT, preferably 12 wt% to 21 wt%; iii. 2 wt% to 10 wt% of DFL, preferably 3 wt% to 8 wt%; iv. 8 wt% to 26 wt% of 6SL and 3SL combined, preferably 11 wt% to 22 wt%; and v. 10 wt% to 50 wt% of 3FL, preferably 11 wt% to 43 wt%.

[0418] 61. The nutritional composition according to clause 59 or 60, wherein the LNFP-I and HMO mixture combined consists essentially of: i. 20 wt% to 46 wt% of 2FL, preferably 22 wt% to 42 wt% of 2FL; ii. 11 wt% to 17 wt% of LNT, preferably 12 wt% to 15 wt% of LNT; iii. 2 wt% to 7 wt% of DFL, preferably 3 wt% to 6 wt% of DFL; iv. 9 wt% to 21 wt% of 6SL and 3SL combined, preferably 9 wt% to 19 wt% of 6SL and 3SL combined; v. 9 wt% to 34 wt% of 3FL, preferably 11 wt% to 32 wt% of 3FL; and vi. 5 wt% to 32 wt% of LNFP-I, preferably 10 wt% to 19 wt% of LNFP-I.

[0419] 62. The nutritional composition according to any of clauses 50-58, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'- sialyllactose (6SL), 3'-sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT).

[0420] 63. The nutritional composition according to clause 62, wherein the HMO mixture consists essentially of: i. 20 wt% to 60 wt% of 2FL, preferably 22 wt% to 55 wt%; ii. 4 wt% to 30 wt% of LNT, preferably 6 wt% to 20 wt%; iii. 1 wt% to 12 wt % of DFL, preferably 3 wt% to 8 wt%; iv. 7 wt% to 23 wt% of 6SL and 3SL combined, preferably 8 wt% to 22 wt%; v. 10 wt% to 50 wt% of 3FL, preferably 13 wt% to 46 wt%; and vi. 3 wt% to 25 wt% of LNnT, preferably 5 wt% to 20 wt%

[0421] 64. The nutritional composition according to clause 62 or 63, wherein the LNFP-I and HMO mixture combined consists essentially of: i. 29 wt% to 40 wt% of 2FL, preferably 32 wt% to 39 wt% of 2FL; ii. 8 wt% to 13 wt% of LNT, preferably 9 wt% to 12 wt% of LNT; iii. 3 wt% to 11 wt % of DFL; iv. 3 wt% to 15 wt% of 6SL and 3SL combined, preferably 4 wt% to 15 wt% of 6SL and 3SL combined; v. 11 wt% to 35 wt% of 3FL, preferably 12 wt% to 35 wt% of 3FL; vi. 1 wt% to 18 wt% of LNnT, preferably 1 wt% to 17 wt% of LNnT; and vii. 2 wt% to 24 wt% of LNFP-I, preferably 4 wt% to 14 wt% of LNFP-I.

[0422] 65. The nutritional composition according to any of clauses 50-64, for use in therapy in an infant or young child.

[0423] 66. The nutritional composition according to any of clauses 50-64, for use in improving metabolic health, cardiovascular health, neurological health, mental health, respiratory health, gastrointestinal health, musculoskeletal health or immune health in an infant or young child.

[0424] 67. The nutritional composition according to any of clauses 50-64, for use in treating and / or preventing a metabolic disorder, a cardiovascular disease, a neurological disorder, a mental disorder, a respiratory disease, gastrointestinal inflammation, a musculoskeletal disorder, an immune disorder, an infection or apnea in an infant or young child.

[0425] 68. The nutritional composition according to any of clauses 50-64, for use in modulating / improving the immune response in an infant or young child.

[0426] 69. The nutritional composition according to any of clauses 50-64, for use in promoting brain, bone, or muscle growth and / or development in an infant or young child.

[0427] 70. Use of the nutritional composition according to any of clauses 50-64, to promote the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3- methylxanthine in the gastrointestinal tract of an infant or young child, wherein the use is for example non-therapeutic.

[0428] 71 . A method of promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, the method comprising administering the infant or young child a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture, wherein the HMO mixture consists of 2'-fucosy I lactose (2FL), difucosyllactose (DFL), lacto-N- tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3- fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT), wherein the use is for example non- therapeutic.

[0429] 72. The method according to clause 71 , wherein the combination is defined according to any of clauses 4-19.

[0430] All publications mentioned in the above specification are herein incorporated by reference. Various modifications and variations of the described methods and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in molecular biology or related fields are intended to be within the scope of the following claims.

[0431] (Original in Electronic Form)

[0432] (This sheet is not part of and does not count as a sheet of the international application)

[0433] (Original in Electronic Form)

[0434] (This sheet is not part of and does not count as a sheet of the international application) (Original in Electronic Form)

[0435] (This sheet is not part of and does not count as a sheet of the international application) (Original in Electronic Form)

[0436] (This sheet is not part of and does not count as a sheet of the international application)

[0437] FOR RECEIVING OFFICE USE ONLY

[0438] FOR INTERNATIONAL BUREAU USE ONLY

Claims

CLAIMS1. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in therapy by promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p- hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosy I lactose (3FL) and / or lacto-N-neotetraose (LNnT).

2. A combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture for use in promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p- hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT), wherein the infant or young child was born by C-section and / or preterm, and / or small for gestational age (SGA) and / or with a low, very low, or extremely low weight at birth (LWB, VLBW, or ELBW) and / or experienced intra-uterine growth retardation (IUGR), and / or was hospitalized the first days following birth and / or took antibiotic the first days following birth and / or was sick at birth, and / or has impaired microbiota and / or dysbiosis of microbiota.

3. Use of a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture to promote the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p- hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

4. The combination for use according to claim 1 or 2, or the use according to claim 3, wherein the combination is administered separately, simultaneously or sequentially, preferably wherein the combination is administered simultaneously.

5. The combination for use according to any of claims 1-2 or 4, or the use according to claim 3 or 4, wherein the combination is provided in the form of a nutritional composition, preferably wherein the nutritional composition is selected from an infant formula, a starter infant formula,72a follow-on or follow-up formula, a baby food, an infant cereal composition, a growing-up-milk, a fortifier such as a human milk fortifier, or a supplement.

6. The combination for use according to any of claims 1-2 or 4-5, or the use according to any of claims 3-5, wherein the Bifidobacterium longum subsp. infantis is Bifidobacterium longum subsp. infantis LMG 11588 or a strain having an Average Nucleotide Identity (AN I) of at least 95%, preferably at least 99%, more preferably at least 99.9% to Bifidobacterium longum subsp. infantis LMG 11588; and / or wherein the combination comprises from 103to 1012cfu of Bifidobacterium longum subsp. infantis, preferably from 107to 1012cfu of Bifidobacterium longum subsp. infantis, more preferably from 108to 101° cfu of Bifidobacterium longum subsp. infantis per g of combination or composition on a dry weight basis.

7. The combination for use according to any of claims 1-2 or 4-6, or the use according to any of claims 3-6, wherein the combination comprises LNFP-I in a total amount of from 25 mg / L to 4000 mg / L of the combination or composition or from 0.02 g / 100 g to 3.75 g / 100 g of the combination or composition on a dry weight basis.

8. The combination for use according to any of claims 1-2 or 4-7, or the use according to any of claims 3-7, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and 3- fucosyllactose (3FL), or wherein the HMO mixture consists essentially of: i. 16 wt% to 69 wt% of 2FL, preferably 22 wt% to 59 wt%; ii. 9 wt% to 24 wt% of LNT, preferably 12 wt% to 21 wt%; iii. 2 wt% to 10 wt% of DFL, preferably 3 wt% to 8 wt%; iv. 8 wt% to 26 wt% of 6SL and 3SL combined, preferably 11 wt% to 22 wt%; and v. 10 wt% to 50 wt% of 3FL, preferably 11 wt% to 43 wt%.

9. The combination for use according to any of claims 1-2 or 4-7, or the use according to any of claims 3-7, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), 3-fucosyllactose (3FL), and lacto-N-neotetraose (LNnT), or wherein the HMO mixture consists essentially of: i. 20 wt% to 60 wt% of 2FL, preferably 22 wt% to 55 wt%; ii. 4 wt% to 30 wt% of LNT, preferably 6 wt% to 20 wt%; iii. 1 wt% to 12 wt % of DFL, preferably 3 wt% to 8 wt%;73iv. 7 wt% to 23 wt% of 6SL and 3SL combined, preferably 8 wt% to 22 wt%; v. 10 wt% to 50 wt% of 3FL, preferably 13 wt% to 46 wt%; and vi. 3 wt% to 25 wt% of LNnT, preferably 5 wt% to 20 wt%.

10. The combination for use according to any of claims 1-2 or 4-7, or the use according to any of claims 3-7, wherein the HMO mixture consists of 2'-fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), 3'-sialyllactose (3SL), and lacto-N- neotetraose (LNnT), or wherein the HMO mixture consists essentially of: i. 34 wt% to 85 wt% of 2FL, preferably 40 wt% to 71 wt% of 2FL; ii. 10 wt% to 40 wt% of LNT, preferably 12 wt% to 26 wt% of LNT ; iii. 4 wt% to 14 wt% of DFL, preferably 5 wt% to 10 wt% of DFL; iv. 9 wt% to 31 wt% of 6SL and 3SL combined, preferably 10 wt% to 28 wt% of 6SL and 3SL combined; and v. 6 wt% to 30 wt% of LNnT, preferably 7 wt% to 23 wt% of LNnT.

11. The combination for use according to any of claims 1-2 or 4-10, wherein the therapy is:(a) improving metabolic health or treating and / or preventing a metabolic disorder in an infant or young child, optionally wherein the combination promotes the production of one or more of uridine, adenine, guanine, AMP, cAMP, and / or acetyl-carnitine in the gastrointestinal tract of the infant or young child; and / or(b) improving cardiovascular health or treating and / or preventing a cardiovascular disease in an infant or young child, optionally wherein the combination promotes the production of one or more of uridine, cAMP, and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child; and / or(c) improving neurological health, promoting brain growth and / or development, or treating and / or preventing a neurological disorder in an infant or young child, optionally wherein the combination promotes the production of one or more of uridine, guanine, cAMP, 4-guanidinobutyric acid, acetyl-carnitine and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child; and / or(d) improving mental health or treating and / or preventing a mental disorder in an infant or young child, optionally wherein the combination promotes the production of one or74more of guanine, 4-guanidinobutyric acid, and / or acetyl-carnitine in the gastrointestinal tract of the infant or young child; and / or(e) improving respiratory health or treating and / or preventing a respiratory disease, such as a respiratory tract infection, in an infant or young child, optionally wherein the combination promotes the production of cAMP and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child; and / or(f) improving gastrointestinal health or treating and / or preventing gastrointestinal inflammation in an infant or young child, optionally wherein the combination promotes the production of adenine and / or 4-guanidinobutyric acid in the gastrointestinal tract of the infant or young child; and / or(g) modulating the immune response in an infant or young child, optionally wherein the combination promotes the production of cAMP and / or p-hydroxyphenyllactic acid in the gastrointestinal tract of the infant or young child; and / or(h) preventing and / or treating an infection in an infant or young child, optionally wherein the combination promotes the production of one or more of cAMP, p- hydroxyphenyllactic acid, and / or 3-methylxanthine in the gastrointestinal tract of the infant or young child; and / or(i) preventing and / or treating apnea in an infant or young child, optionally wherein the combination promotes the production of 3-methylxanthine in the gastrointestinal tract of the infant or young child; and / or(j) improving musculoskeletal health, treating and / or preventing a musculoskeletal disorder, or promoting bone and / or muscle growth and / or development in an infant or young child, optionally wherein the combination promotes the production of uridine and / or AMP in the gastrointestinal tract of the infant or young child.

12. A nutritional composition comprising Bifidobacterium longum subsp. infantis, lacto-N- fucopentaose I (LNFP-I) and a HMO mixture, wherein the HMO mixture consists of 2'- fucosyllactose (2FL), difucosyllactose (DFL), lacto-N-tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3-fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

13. The nutritional composition according to claim 12, for use in therapy in an infant or young child, preferably for use in: (a) improving metabolic health, cardiovascular health, neurological health, mental health, respiratory health, gastrointestinal health, musculoskeletal health, or immune health in an infant or young child; (b) treating and / or preventing a metabolic disorder,75a cardiovascular disease, a neurological disorder, a mental disorder, a respiratory disease, gastrointestinal inflammation, a musculoskeletal disorder, an immune disorder, an infection or apnea in an infant or young child; (c) modulating the immune response in an infant or young child; or (d) promoting brain, bone, or muscle growth and / or development in an infant or young child.

14. Use of the nutritional composition according to claim 12, to promote the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4- guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child.

15. A method of promoting the production of one or more beneficial metabolites selected from uridine, adenine, guanine, AMP, cAMP, 4-guanidinobutyric acid, p-hydroxyphenyllactic acid, acetyl-carnitine and 3-methylxanthine in the gastrointestinal tract of an infant or young child, the method comprising administering the infant or young child a combination of Bifidobacterium longum subsp. infantis, lacto-N-fucopentaose I (LNFP-I) and a HMO mixture, wherein the HMO mixture consists of 2'-fucosy I lactose (2FL), difucosyllactose (DFL), lacto-N- tetraose (LNT), 6'-sialyllactose (6SL), and 3'-sialyllactose (3SL), and optionally 3- fucosyllactose (3FL) and / or lacto-N-neotetraose (LNnT).

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