Anti-regurgitation formula milk powder

Abstract

The present invention is in the field of thickened formula milk powder for infants suffering from regurgitation. In particular, the infant formula is thickened with a combination of xanthan gum and locust bean gum and comprises a mixture of specific non-digestible oligosaccharides.

Description

Technical Field

[0001] The present invention belongs to the field of thickened formula milk for infants suffering from regurgitation. Background Art

[0002] Breast milk is the preferred method of infant feeding. When breastfeeding is not possible or the mother chooses not to breastfeed, commercially available infant formula suitable as a complete nutrition is available.

[0003] Functional gastrointestinal disorders (FGIDs) are very common in the first year of life. FGIDs are the most common reason for parents to seek pediatric consultation. Regurgitation is the most prevalent FGID, and it is estimated that up to 80% of infants globally are affected in the first two months of life (Czinn et al., 2013, Pediatric Drugs 15:19 - 27). Gastroesophageal reflux (GER) is the backward flow of gastric contents along the esophagus, sometimes even into or out of the mouth. At the lower end of the esophagus, the lower esophageal sphincter (LES) opens when swallowing food and then usually closes again to keep the gastric contents in place. When the LES does not work properly, with GER, the gastric contents (including hydrochloric acid) come into contact with the esophagus, throat, nasal cavity, lungs, and / or teeth, causing pain and damage. Over time, repeated exposure of these areas to acid leads to a gradual increase in damage and more serious complications. When reflux can be seen, the diagnostic term regurgitation is used. When regurgitation of gastric contents causes complications or leads to tissue damage or inflammation, it is called gastroesophageal reflux disease (GERD). GER and GERD, also hereinafter referred to as GER(D), are problems that are particularly prone to occur in infancy. In addition to the problems already mentioned above, GER(D) in infants can also cause infant dehydration, impaired growth, and / or growth retardation.

[0004] Antiregurgitation formula milk with thickeners to increase the viscosity of the formula milk in the bottle and in the stomach is available on the market. Typically used thickeners are starches and indigestible polysaccharides, such as locust bean gum. An example is Nutrilon A.R., which has complete whey protein and casein from milk and about 0.5 g / 100 ml of locust bean gum as a thickener.

[0005] However, subjects suffering from regurgitation usually also suffer from other functional gastrointestinal disorders (FGIDs), including lower intestinal problems that may be caused by microbial dysbiosis.

[0006] WO 2010 / 120172 discloses an anti-regurgitation formula milk powder that is partially fermented and contains locust bean gum as a thickener. The effect on viscosity in the bottle and under gastric conditions is improved. This allows for a reduction in the level of the thickener classified as fiber and allows for the presence of additional fiber, which belongs to the category of indigestible oligosaccharides such as galactooligosaccharides and fructooligosaccharides, and has a beneficial effect on the gut microbiota.

[0007] WO 2021 / 151978 discloses an anti-regurgitation formula milk powder containing locust bean gum and indigestible oligosaccharides, which has an improved effect on functional gastrointestinal disorders (especially FGID of the lower intestine) in infants and young children.

[0008] WO 2004 / 002240 discloses the use of hydrocolloids as prebiotics in the preparation of products for consumption. After consuming the said products, hydrocolloids have the advantage of reducing gas release when fermented by bacteria in the gastrointestinal tract. Compositions containing hydrocolloids and methods of using these compositions in treatment methods are also provided.

[0009] US2013 / 0189398 discloses an infant formula milk powder specifically designed to reduce the occurrence of regurgitation, colic, and constipation episodes in infants fed with infant formula milk powder. To address regurgitation, potato starch is the preferred thickener. The presence of indigestible oligosaccharides to produce a bifidogenic effect is disclosed. Disadvantageously, starch replaces (partially) lactose as a source of digestible carbohydrates, which is undesirable for infants because lactose is the main source of digestible carbohydrates in human milk and has a lower glycemic index. In addition, starch can be degraded by α-amylase, so its effectiveness as a thickener may decrease over time during feeding. Since starch is digestible, it does not have a beneficial effect on the microbiota in the large intestine. Summary of the Invention

[0010] The inventors have worked on solving the problem of a suitable thickener or combination of thickeners that provides an appropriate viscosity in infant formula milk powder to ensure an anti-regurgitation effect. After applying several criteria related to the technical characteristics of the product and the shear viscosity under conditions of digestion in the bottle and simulated infant stomach, and after testing many thickeners and their combinations, it was found that the combination of xanthan gum (XG) and locust bean gum (LBG) is excellent.

[0011] Compared to that typically found in currently commercially available anti-reflux formula milks (such as Nutrilon Anti-Reflux formula milk), the XG / LBG combination requires significantly less thickening dietary fiber to be present in the formula milk, but this composition unexpectedly provides a similar high viscosity and optimal product characteristics with advantages during gastric digestion. This allows for the additional addition of additional non-thickening fibers with prebiotic activity, such as indigestible oligosaccharides.

[0012] Subsequently, the effect of the combination of XG, LBG, and indigestible oligosaccharides on infant microbiota activity was tested. The inventors were surprised to find that the combination of XG and LBG, long-chain fructooligosaccharides, and short-chain oligosaccharides selected from galactooligosaccharides and fructooligosaccharides showed a beneficial synergistic effect on the function of the infant gut microbiota. The addition of a mixture of galactooligosaccharides and long-chain fructooligosaccharides or a mixture of short-chain oligosaccharides and long-chain fructooligosaccharides to the XG / LBG combination resulted in a decrease in the amount and relative amount of butyrate, propionate, and gas, and this decrease was higher than expected based on a single thickener or indigestible oligosaccharide. In addition, in the presence of short-chain galactooligosaccharides, the amount of lactic acid increased synergistically. Thus, the combination of these specific thickeners and specific indigestible oligosaccharides synergistically shifts the activity of the infant gut microbiota towards an activity more similar to that observed in breastfed infants. Detailed Description

[0013] Accordingly, the present invention relates to a nutritional composition suitable for providing nutrition to an infant or young child, preferably an infant, comprising lipids, digestible carbohydrates, proteins, a thickener, and indigestible oligosaccharides, wherein the thickener comprises a combination of xanthan gum and locust bean gum and the indigestible oligosaccharides comprise long-chain fructooligosaccharides and short-chain oligosaccharides selected from galactooligosaccharides and fructooligosaccharides. The nutritional composition of the present invention can be in the form of a ready-to-drink liquid or in the form of a powder which is a ready-to-drink liquid after reconstitution with water.

[0014] The present invention further relates to a nutritional composition comprising lipids, digestible carbohydrates, proteins, a thickener, and indigestible oligosaccharides, wherein the thickener comprises a combination of xanthan gum and locust bean gum and the indigestible oligosaccharides comprise long-chain fructooligosaccharides and short-chain oligosaccharides selected from galactooligosaccharides and fructooligosaccharides, for use in the treatment or prevention of reflux in an infant or young child, preferably for use in the treatment of reflux.

[0015] The present invention can also be expressed as a method for treating or preventing reflux in an infant or young child, preferably a method for use in the treatment of reflux, which comprises administering to the infant a nutritional composition comprising lipids, digestible carbohydrates, proteins, a thickener, and indigestible oligosaccharides, wherein the thickener comprises a combination of xanthan gum and locust bean gum and the indigestible oligosaccharides comprise long-chain fructooligosaccharides and short-chain oligosaccharides selected from galactooligosaccharides and fructooligosaccharides.

[0016] The present invention can also be expressed as the use of lipids, digestible carbohydrates, proteins, thickeners, and indigestible oligosaccharides in the preparation of a nutritional composition for the treatment or prevention of reflux in infants or toddlers, preferably for the treatment of reflux, wherein the thickener comprises a combination of xanthan gum and locust bean gum and the indigestible oligosaccharides comprise long-chain fructooligosaccharides and short-chain oligosaccharides selected from galactooligosaccharides and fructooligosaccharides.

[0017] The present invention also relates to a method for preparing a nutritional composition comprising lipids, digestible carbohydrates, proteins, thickeners, and indigestible oligosaccharides, wherein the thickener comprises a combination of xanthan gum and locust bean gum and the indigestible oligosaccharides comprise long-chain fructooligosaccharides and short-chain oligosaccharides selected from galactooligosaccharides and fructooligosaccharides, the method comprising

[0018] a. providing a liquid nutritional composition comprising proteins, digestible carbohydrates, and lipids,

[0019] b. drying the liquid obtained in step a) into a powder, preferably by spray drying,

[0020] c. dry-mixing xanthan gum and locust bean gum and optionally waxy starch with the powder obtained in step b), and

[0021] d. adding indigestible oligosaccharides in step a) and / or step c).

[0022] Thickener

[0023] The present invention relates to the combination of a combination of xanthan gum and locust bean gum as a thickener for formula milk powder and a specific mixture of indigestible oligosaccharides. The inventors have specifically demonstrated that such a combination has a synergistic improvement effect on the microbiota activity of infants, making it closer to that of breastfed infants.

[0024] The choice of thickener was made after extensive testing of thickeners, their combinations, and at different levels and concentrations, taking into account user experience, regulatory requirements, viscosity characteristics in the bottle, and viscosity characteristics under infant gastric digestion conditions. It has been found that xanthan gum is a thickener that is suitable for combination with locust bean gum for the compositions according to the present invention, especially infant formula. Optionally, waxy starch is also included in the thickener combination. The inventors have specifically demonstrated that such a combination provides excellent viscosity, thereby reducing and preventing reflux or regurgitation. Combining XG and LBG allows them to be applied at a favorable low concentration, which still provides the required increased viscosity, and further allows for the presence of additional dietary fiber to ensure the required prebiotic effect.

[0025] Xanthan gum

[0026] Xanthan gum, also known as xanthan in the context of the present invention, is a branched polysaccharide used as a food additive with the code E415. It is produced by the bacterium Xanthomonas campestris. It consists of a combination of four subunits: glucose, mannose, glucuronic acid, and pyruvic acid. Xanthan gum is commercially available, such as Grinsted xanthan gum from DuPont Danisco.

[0027] Preferably, the total amount of xanthan gum is 0.02 to 0.12 g / 100 ml of the ready-to-drink liquid nutritional composition, more preferably 0.03 to 0.09 g / 100 ml, more preferably 0.03 to 0.07 g / 100 ml, and even more preferably 0.04 to 0.05 g / 100 ml of the ready-to-drink liquid nutritional composition. Preferably, the total amount of xanthan gum is between 0.03 and 0.18 g / 100 kcal, more preferably 0.04 to 0.13 g / 100 kcal, more preferably 0.04 to 0.10 g / 100 kcal, and even more preferably 0.06 to 0.07 g / 100 kcal. Preferably, the total amount of xanthan gum is 0.14 to 0.86 g / 100 g of the dry weight of the nutritional composition, more preferably 0.21 to 0.64 g / 100 g of the dry weight, more preferably 0.21 to 0.50 g / 100 g of the dry weight, and even more preferably 0.29 to 0.36 g / 100 g of the dry weight of the nutritional composition. If there is too much xanthan gum, the gastric viscosity will be too high. If the amount is too low, a reduced effect on GER can be expected.

[0028] Locust bean gum

[0029] In the context of the present invention, locust bean gum is also known as carob gum, ceratonia gum, carob bean gum, carobin, and is a galactomannan used as a food additive with the code E410. Locust bean gum can be isolated from the seeds of the carob tree. It consists of a linear main chain of D-mannopyranose units and side-branch units of D-galactopyranose, with an average of one D-galactopyranose unit branch per four D-mannopyranose units.

[0030] Preferably, locust bean gum is cold-soluble. In the temperature range of 10°C to 45°C, the solubility of cold-soluble locust bean gum in an aqueous medium is more than 60%. Cold-soluble locust bean gum differs from natural locust bean gum in that it has a lower average molecular weight. Locust bean gum is commercially available, such as Grindsted LBG or cold-soluble LBG from Danisco–DuPont.

[0031] Preferably, the total amount of locust bean gum is 0.02 to 0.45 g / 100 ml of the ready-to-drink liquid nutritional composition, more preferably 0.03 to 0.30 g / 100 ml. Preferably, the total amount of locust bean gum is 0.03 to 0.67 g / 100 kcal, more preferably 0.04 to 0.45 g / 100 kcal. Preferably, the total amount of locust bean gum is 0.14 to 3.21 g / 100 g of the dry weight of the nutritional composition, more preferably 0.21 to 2.14 g of the dry weight of the nutritional composition. If there is too much locust bean gum, the viscosity in the bottle will be too high. If the amount is too low, a reduced effect on GER can be expected.

[0032] Combination of xanthan gum and locust bean gum

[0033] It has been found that the combination of XG and LBG is superior to XG or LBG alone. Preferably, the combination of XG and LBG is present in the nutritional composition at the following levels: 0.05 to 0.50 g / 100 ml, preferably 0.06 to 0.40 g / 100 ml, even more preferably 0.07 to 0.30 g / 100 ml. Preferably, the combination of XG and LBG is present in the nutritional composition at the following levels: 0.07 to 0.75 g / 100 kcal, preferably 0.09 to 0.60 g / 100 kcal, even more preferably 0.10 to 0.45 g / 100 kcal. Preferably, the combination of XG and LBG is present in the nutritional composition at the following levels: 0.36 to 3.57 g / 100 g of dry weight, preferably 0.43 to 2.86 g / 100 g of dry weight, even more preferably 0.50 g / 100 g to 2.14 g / 100 g of dry weight. Preferably, the wt / wt ratio of XG and LBG is 0.1 to 6, preferably 0.5 to 5.

[0034] It has been found that the combination of XG and LBG has two optimal concentrations and ratios. The viscosity under conditions simulating infant digestion is most similar to the reference formula of Nutrilon Anti-Reflux formula containing LBG and intact milk protein, while still being able to reduce the amount of thickener.

[0035] In the first embodiment, the nutritional composition comprises small amounts of XG and LBG in specific ratios. Preferably, xanthan gum is present in an amount of 0.03 to 0.07 g / 100 ml, preferably 0.03 to 0.06 g / 100 ml, and even more preferably 0.04 to 0.05 g / 100 ml of the ready-to-drink nutritional composition. Preferably, locust bean gum is present in an amount of 0.03 to 0.07 g / 100 ml, more preferably 0.03 to 0.06 g / 100 ml, and even more preferably 0.04 to 0.05 g / 100 ml of the ready-to-drink nutritional composition. Preferably, the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, and even more preferably 0.9 to 1.1. Preferably, xanthan gum is present in an amount of 0.03 to 0.07 g / 100 ml, preferably 0.03 to 0.06 g / 100 ml, and even more preferably 0.04 to 0.05 g / 100 ml of the ready-to-drink nutritional composition, and locust bean gum is present in an amount of 0.03 to 0.07 g / 100 ml, more preferably 0.03 to 0.06 g / 100 ml, and even more preferably 0.04 to 0.05 g / 100 ml of the ready-to-drink nutritional composition, and the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, and even more preferably 0.9 to 1.1.

[0036] Alternatively, according to this embodiment, xanthan gum is present in an amount of 0.04 to 0.10 g / 100 kcal, preferably 0.04 to 0.09 g / 100 kcal, and even more preferably 0.06 to 0.07 g / 100 kcal. Preferably, locust bean gum is present in an amount of 0.04 to 0.10 g / 100 kcal, more preferably 0.04 to 0.09 g / 100 kcal, and even more preferably 0.06 to 0.07 g / 100 kcal. Preferably, the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, and even more preferably 0.9 to 1.1. Preferably, xanthan gum is present in an amount of 0.04 to 0.10 g / 100 kcal, preferably 0.04 to 0.09 g / 100 kcal, and even more preferably 0.06 to 0.07 g / 100 kcal, and locust bean gum is present in an amount of 0.04 to 0.10 g / 100 kcal, more preferably 0.04 to 0.09 g / 100 kcal, and even more preferably 0.06 to 0.07 g / 100 kcal, and the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, and even more preferably 0.9 to 1.1.

[0037] Alternatively, according to this embodiment, xanthan gum is present in an amount of: 0.21 to 0.50 g / 100 g dry weight of the nutritional composition, preferably 0.21 to 0.43 g / 100 g dry weight, even more preferably 0.29 to 0.36 g / 100 g dry weight. Preferably, locust bean gum is present in an amount of: 0.21 to 0.50 g / 100 g dry weight of the nutritional composition, more preferably 0.21 to 0.43 g / 100 g dry weight, even more preferably 0.29 to 0.36 g / 100 g dry weight of the nutritional composition. Preferably, the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, even more preferably 0.90 to 1.1. Preferably, xanthan gum is present in an amount of: 0.21 to 0.50 g / 100 g dry weight of the nutritional composition, preferably 0.21 to 0.43 g / 100 dry weight, even more preferably 0.29 to 0.36 g / 100 g dry weight, and locust bean gum is present in an amount of: 0.21 to 0.50 g / 100 g dry weight of the nutritional composition, more preferably 0.21 to 0.43 g / 100 g dry weight, even more preferably 0.29 to 0.36 g / 100 g dry weight of the nutritional composition, and the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, even more preferably 0.90 to 1.1.

[0038] At lower pH and digestion conditions, using the combination of XG and LBG at such low concentrations has been sufficient to achieve the desired high viscosity in the stomach. The ratio of these thickeners is approximately 1 and seems to have a synergistic effect on the thickening or viscosity-increasing properties, especially at low pH. Although the viscosity in the bottle is slightly lower than that of the reference formula milk powder Aptamil Anti-Reflux, the viscosity is much higher than that of the unthickened nutritional composition and is comparable to or even slightly higher than that of the commercial formula milk powder (Allernova AR+) with a thickener and deeply hydrolyzed milk protein.

[0039] For this embodiment, a further improvement to increase the viscosity in the bottle to a level comparable to that of Nutrilon Anti - Reflux formula would be to further add waxy starch, also known as amylopectin. It has been found that the presence of waxy starch causes an increase in viscosity in the bottle, but not under gastric conditions. Preferably, the total amount of waxy starch is 0.4 to 1.5 g / 100 ml of the ready - to - drink nutritional composition, more preferably 0.4 to 0.8 g / 100 ml. If there is too much waxy starch, the viscosity will be too high and it replaces more desirable carbohydrate sources such as lactose. Starch is not suitable as a digestible carbohydrate source in infant formula because it has a high glycemic index. If the amount is too low, the desired further improvement to increase the viscosity in the bottle will not be achieved. In one embodiment, preferably, xanthan gum is present in the following amounts: 0.03 to 0.07 g / 100 ml of the ready - to - drink nutritional composition, more preferably 0.03 to 0.06 g / 100 ml, even more preferably 0.04 to 0.05 g / 100 ml of the ready - to - drink nutritional composition, and locust bean gum is present in the following amounts: 0.03 to 0.07 g / 100 ml of the ready - to - drink nutritional composition, preferably 0.03 to 0.06 g / 100 ml, even more preferably 0.04 to 0.05 g / 100 ml of the ready - to - drink nutritional composition, and waxy starch is present at a level of 0.4 to 1.5 g / 100 ml of the ready - to - drink nutritional composition, more preferably 0.4 to 0.8 g / 100 ml of the ready - to - drink nutritional composition, and the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, even more preferably 0.9 to 1.1.

[0040] Alternatively, according to this embodiment, xanthan gum is present in the following amounts: 0.04 to 0.10 g / 100 kcal, preferably 0.04 to 0.09 g / 100 kcal, even more preferably 0.06 to 0.07 g / 100 kcal, and locust bean gum is present in the following amounts: 0.04 to 0.10 g / 100 kcal, more preferably 0.04 to 0.09 g / 100 kcal, even more preferably 0.06 to 0.07 g / 100 kcal, and waxy starch is present at a level of 0.6 to 2.2 g / 100 kcal, more preferably 0.6 to 1.2 g / 100 kcal, and the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, even more preferably 0.9 to 1.1.

[0041] Alternatively, according to this embodiment, xanthan gum is present in an amount of 0.21 to 0.50 g / 100 g dry weight of the nutritional composition, preferably 0.21 to 0.43 g / 100 dry weight, even more preferably 0.29 to 0.36 g / 100 g dry weight, and locust bean gum is present in an amount of 0.21 to 0.10 g / 100 g dry weight of the nutritional composition, more preferably 0.21 to 0.43 g / 100 g dry weight, even more preferably 0.29 to 0.36 g / 100 g dry weight of the nutritional composition, and waxy starch is present at a level of 2.9 to 10.7 g / 100 g dry weight of the nutritional composition, more preferably 2.9 to 5.7 g / 100 g dry weight of the nutritional composition, and the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, even more preferably 0.90 to 1.1.

[0042] In a second embodiment, the nutritional composition contains a low amount of XG and a high amount of LBG, resulting in a specific ratio different from the first embodiment. Preferably, the sum of xanthan gum and locust bean gum is present in an amount of 0.13 to 0.40 g / 100 ml of the ready-to-drink nutritional composition, preferably 0.20 to 0.40 g / 100 ml. Preferably, xanthan gum is present in an amount of 0.03 to 0.07 g / 100 ml, more preferably 0.03 to 0.06 g / 100 ml of the ready-to-drink nutritional composition. Preferably, locust bean gum is present in an amount of 0.10 to 0.35 g / 100 ml of the ready-to-drink nutritional composition, more preferably 0.20 to 0.30 g / 100 ml. Preferably, the wt / wt ratio of xanthan gum to locust bean gum is 1.5 to 6.0, more preferably 3.0 to 5.0. Preferably, xanthan gum is present in an amount of 0.03 to 0.07 g / 100 ml of the ready-to-drink nutritional composition, more preferably 0.03 to 0.06 g / 100 ml, locust bean gum is present at a level of 0.10 to 0.35 g / 100 ml of the ready-to-drink nutritional composition, more preferably 0.20 to 0.30 g / 100 ml, and the wt / wt ratio of xanthan gum to locust bean gum is 1.5 to 6.0, more preferably 3.0 to 5.0.

[0043] Alternatively, according to this embodiment, the sum of xanthan gum and locust bean gum is present in an amount of 0.19 to 0.60 g / 100 kcal, preferably 0.30 to 0.60 g / 100 kcal. Preferably, xanthan gum is present in an amount of 0.04 to 0.10 g / 100 kcal, preferably 0.04 to 0.09 g / 100 kcal. Preferably, locust bean gum is present in an amount of 0.15 to 0.52 g / 100 kcal, more preferably 0.30 to 0.45 g / 100 kcal. Preferably, the wt / wt ratio of xanthan gum to locust bean gum is 1.5 to 6.0, more preferably 3.0 to 5.0. Preferably, xanthan gum is present in an amount of 0.04 to 0.10 g / 100 kcal, preferably 0.04 to 0.09 g / 100 kcal, locust bean gum is present in an amount of 0.15 to 0.52 g / 100 kcal, more preferably 0.30 to 0.45 g / 100 kcal, and the wt / wt ratio of xanthan gum to locust bean gum is 1.5 to 6.0, more preferably 3.0 to 5.0.

[0044] Alternatively, according to this embodiment, the sum of xanthan gum and locust bean gum is present in an amount of 0.93 to 2.86 g / 100 g dry weight of the nutritional composition, preferably 1.43 to 2.86 g / 100 g dry weight. Preferably, xanthan gum is present in an amount of 0.21 to 0.50 g / 100 g dry weight of the nutritional composition, preferably 0.21 to 0.43 g / 100 g dry weight. Preferably, locust bean gum is present in an amount of 0.71 to 2.50 g / 100 g dry weight of the nutritional composition, more preferably 1.43 to 2.14 g / 100 g dry weight. Preferably, the wt / wt ratio of xanthan gum to locust bean gum is 1.5 to 6.0, more preferably 3.0 to 5.0. Preferably, xanthan gum is present in an amount of 0.21 to 0.50 g / 100 g dry weight of the nutritional composition, preferably 0.21 to 0.43 g / 100 dry weight, locust bean gum is present in an amount of 0.71 to 2.50 g / 100 g dry weight of the nutritional composition, more preferably 1.43 to 2.14 g / 100 g dry weight, and the wt / wt ratio of xanthan gum to locust bean gum is 1.5 to 6.0, more preferably 3.0 to 5.0.

[0045] Since in this embodiment, the viscosity in the bottle is high enough, there is no need to add waxy starch as an additional thickener. Therefore, in this second embodiment, the amount of waxy starch is low or there is no waxy starch. Preferably, the amount of waxy starch is less than 0.2 g / 100 ml of the ready-to-drink nutritional composition, more preferably less than 0.1 g / 100 ml. Alternatively, the amount of waxy starch is less than 0.3 g / 100 kcal, more preferably less than 0.15 g / 100 kcal. Alternatively, the amount of waxy starch is less than 1.4 g / 100 g dry weight of the nutritional composition, more preferably less than 0.7 g / 100 g dry weight.

[0046] Waxy starch

[0047] The composition according to the invention may also comprise waxy starch as another thickener, in particular pre-cooked or pre-gelatinized waxy starch, preferably pre-gelatinized waxy starch. Waxy starch is derived from, for example, maize or potato or rice varieties that typically produce starch containing at least 98% amylopectin. Thus, as described above, in the context of the present invention, waxy starch is also defined as amylopectin. Waxy starch may also be denoted as viscous starch or isamylopectin. Dissolved waxy starch or amylopectin has a lower tendency to retrograde during storage and cooling and is suitable as a thickener. Starch is a mixture of two homopolymers, amylose and amylopectin, composed of D-glucopyranose units that are interconnected via α-(1-4) bonds and α-(1-6) bonds responsible for forming the branches in the molecular structure. Amylose is slightly branched with short branches and has a molecular weight that can range between 10,000 and 1,000,000 daltons. The molecule consists of 600 to 1000 glucose molecules; amylopectin or isamylopectin is a branched molecule that forms long branches every 24 to 30 glucose units via α-(1-6) bonds. Waxy starch or amylopectin is commercially available, for example, waxy maize starch from Cargill. Waxy starch has a thickening effect in the bottle. Under gastric conditions, after exposure to α-amylase during digestion, waxy starch does not produce a high viscosity.

[0048] Indigestible oligosaccharides

[0049] The nutritional composition of the invention comprises indigestible oligosaccharides (NDO). NDO contribute to the improvement of the gut microbiota and contribute to the improvement of the gastrointestinal barrier, thus contributing to the prevention or treatment of gastrointestinal disorders such as constipation, diarrhoea, gastrointestinal inflammation or gastrointestinal infection. Since the total amount of fibre permitted in infant formula is limited, a relatively low concentration of thickener will advantageously allow the presence of a large amount of additional NDO. It has been found that the combination of XG, LBG and NDO shows a beneficial synergistic effect on the function of the infant gut microbiota. It has been found that the combination of XG, LBG and NDO synergistically directs the activity of the infant gut microbiota in a direction more similar to that observed in breastfed infants. In particular, a mixture of galacto-oligosaccharides and long-chain fructo-oligosaccharides or a mixture of short-chain fructo-oligosaccharides and long-chain fructo-oligosaccharides, in combination with XG and LBG, results in a synergistic reduction in the amount and relative amount of butyrate, propionate and gas. In addition, in the presence of short-chain galacto-oligosaccharides, the amount of lactic acid increases synergistically.

[0050] Indigestible oligosaccharides comprise a mixture of long-chain indigestible oligosaccharides and short-chain indigestible oligosaccharides. Long-chain oligosaccharides are long-chain fructooligosaccharides (IcFOS) having an average degree of polymerization (DP) higher than 10, typically in the range of 10 - 100, preferably 15 - 50, and most preferably higher than 20. A specific type of long-chain fructooligosaccharide is inulin. Long-chain fructooligosaccharides are commercially available, and an example is Raftiline HP (Orafti).

[0051] Short-chain indigestible oligosaccharides have an average degree of polymerization less than 10, preferably at most 8, and preferably in the range of 2 - 7. In one embodiment, the nutritional composition comprises galactooligosaccharides, preferably β-galactooligosaccharides, preferably trans-galactooligosaccharides. Galactooligosaccharides preferably have an average degree of polymerization in the range of 2 - 8, preferably 3 - 7, and thus are short-chain oligosaccharides in the context of the present invention. (Trans)galactooligosaccharides are available, for example, under the trade names Vivina LGOS (Friesland Campina Domo Ingredients, Netherlands), Bimuno (Clasado), Cup-oligo (Nissin Sugar), and Oligomate 55 (Yakult). In one embodiment, the nutritional composition comprises short-chain fructooligosaccharides. Short-chain fructooligosaccharides preferably have an average degree of polymerization in the range of 2 - 8, preferably 3 - 7, and thus are short-chain fructooligosaccharides in the context of the present invention. scFOS can be an inulin hydrolyzate having an average degree of polymerization within the above (sub)range; such scFOS products are commercially available, for example, as Raftilose P95 (Orafti) or from Cosucra. Alternatively, scFOS can be synthesized enzymatically from sucrose by fructosyltransferase. Short-chain indigestible oligosaccharides can also comprise a mixture of galactooligosaccharides and / or fructooligosaccharides (i.e., scGOS and / or scFOS). If the formula is based on free amino acids, the preferred NDOs are not milk-derived. Preferably, the indigestible oligosaccharides in such a formula are scFOS rather than scGOS.

[0052] Most preferably, the short-chain indigestible oligosaccharides and long-chain fructooligosaccharides are present in a short-chain to long-chain weight ratio within the range of 1:99 to 99:1, more preferably 1:1 to 99:1, more preferably 4:1 to 97:3, even more preferably 5:1 to 95:5, even more preferably 7:1 to 95:5, even more preferably 8:1 to 10:1, and most preferably about 9:1. Most preferably, the short-chain galactooligosaccharides and long-chain fructooligosaccharides are present in a short-chain to long-chain weight ratio within the range of 1:99 to 99:1, more preferably 1:1 to 99:1, more preferably 4:1 to 97:3, even more preferably 5:1 to 95:5, even more preferably 7:1 to 95:5, even more preferably 8:1 to 10:1, and most preferably about 9:1. Most preferably, the short-chain fructooligosaccharides and long-chain fructooligosaccharides are present in a short-chain to long-chain weight ratio within the range of 1:99 to 99:1, more preferably 1:1 to 99:1, more preferably 4:1 to 97:3, even more preferably 5:1 to 95:5, even more preferably 7:1 to 95:5, even more preferably 8:1 to 10:1, and most preferably about 9:1.

[0053] Based on the dry weight of the nutritional composition, the nutritional composition preferably contains 0.05 to 20 wt% of said indigestible oligosaccharides, more preferably 0.5 to 15 wt%, even more preferably 1 to 10 wt%, and most preferably 2 to 10 wt%. When in liquid form, the nutritional composition preferably contains 0.01 to 1.0 g of indigestible oligosaccharides, more preferably 0.25 to 0.8 g, and even more preferably contains at least 0.4 g / 100 ml of ready-to-drink nutritional composition, preferably 0.4 to 0.8 g, more preferably 0.6 to 0.8 g / 100 ml of ready-to-drink nutritional composition.

[0054] Protein component

[0055] The nutritional composition contains protein. The protein can be derived from ruminant milk, such as casein or whey protein from milk.

[0056] The present invention advantageously relates to a composition and its use, wherein the protein source preferably provides 7% to 20% of the total calories of the composition, more preferably the protein source provides 8% to 17% of the total calories, and even more preferably the protein source provides 9% to 15% of the total calories of the composition. The present invention advantageously relates to a composition and its use, wherein the protein source preferably provides 1.0 g to 3.5 g of protein / 100 ml, more preferably the protein source provides 1.2 to 3.0 g / 100 ml, and even more preferably the protein source provides 1.4 to 2.5 g / 100 ml of a ready-to-drink nutritional composition. The present invention advantageously relates to a composition and its use, wherein the protein source preferably provides 1.5 g to 5.2 g of protein / 100 kcal, more preferably the protein source provides 1.8 to 4.5 g / 100 kcal, and even more preferably the protein source provides 2.1 to 3.7 g / 100 kcal. Alternatively, in the composition and its use according to the present invention, the amount of the protein source is preferably between 10 and 20 wt% based on the dry weight of the total composition, preferably between 11 and 18 wt%, and even more preferably between 12 and 16 wt% based on the dry weight of the total composition.

[0057] Preferably, the protein, more preferably the milk protein, is hydrolyzed. The hydrolyzed protein will have improved digestibility and will further improve gastrointestinal comfort. Preferably, the nutritional composition contains hydrolyzed whey protein. Preferably, the protein source in the formula milk of the present invention is suitable for infants suffering from allergies, especially food allergies, more especially allergies to dietary proteins, and even more especially milk protein allergies. Such infants typically also have a high incidence of gastrointestinal reflux. Therefore, the protein source is preferably a deeply hydrolyzed protein, free amino acids, or a combination thereof.

[0058] The deeply hydrolyzed protein can be derived from milk protein, such as whey protein or casein. The deeply hydrolyzed protein is typically treated with an ultrafiltration step after hydrolysis with a proteolytic enzyme to remove potentially allergenic intact proteins and large peptides. Preferably, the deeply hydrolyzed protein is deeply hydrolyzed whey protein.

[0059] The deeply hydrolyzed protein, preferably the deeply hydrolyzed whey protein, has a degree of hydrolysis of 16% to 50%, more preferably 20% to 30%, and even more preferably 20% to 25% (of protein). The degree of hydrolysis is defined as the percentage of peptide bonds broken down by enzymatic hydrolysis, where 100% is the total potential peptide bonds present. Too low a degree of hydrolysis will result in an undesired high level of intact allergenic proteins or allergic peptides.

[0060] In contrast, partially hydrolyzed proteins typically have a degree of (protein) hydrolysis of 5% to 15%, which makes them suitable for inducing tolerance because, to some extent, the allergenic parts of the protein remain intact, but makes them unsuitable for infants or young children who already have allergies.

[0061] The peptide size and molecular weight distribution can be determined by conventional methods known to the person skilled in the art, such as HPLC or size exclusion chromatography (SEC), in particular high performance size exclusion chromatography. Examples thereof are described below: Saint-Sauveur et al., “Immunomodulating properties of a whey protein isolate, its enzymatic digest and peptide fractions [Immunomodulating properties of a whey protein isolate, its enzymatic digest and peptide fractions]”, Int. Dairy Journal [International Dairy Journal] (2008) Vol. 18 (3) pp. 260-270. Briefly, the total surface area of the chromatogram is integrated and divided into mass ranges expressed as a percentage of the total surface area. The mass ranges are calibrated using peptides / proteins of known molecular weight.

[0062] In the context of the present invention, the protein hydrolysate preferably comprises 55% to 85% of peptides with a molecular weight below 1000 Da, 15% to 45% of peptides with a molecular weight between 1000 and less than 5000 Da, and 0% to 1% of peptides or proteins with a molecular weight of 5000 Da and above, all based on the total protein of the hydrolysate. In a preferred embodiment, the protein hydrolysate, preferably a whey protein hydrolysate, preferably a deeply hydrolyzed whey protein, comprises peptides with the following size distribution: based on the total protein of the hydrolysate, 60%-90% with a size <1 kDa, 10%-40% with a size of 1 to <5 kDa, 0%-0.5% with a size of 5 to <10 kDa, and 0%-0.2% with a size >10 kDa.

[0063] In a preferred embodiment of the present invention, the protein hydrolysate, preferably a whey protein hydrolysate, preferably a deeply hydrolyzed whey protein, comprises less than 100 mg of peptides with a size greater than 5 kDa per gram of protein hydrolysate, preferably less than 50 mg / g of protein hydrolysate, more preferably less than 10 mg / g of protein hydrolysate. Preferably, the protein hydrolysate, preferably a whey protein hydrolysate, preferably a deeply hydrolyzed whey protein, comprises less than 30 mg of peptides with a size greater than 10 kDa per gram of protein hydrolysate, preferably less than 10 mg / g of protein hydrolysate, most preferably less than 5 mg / g of protein hydrolysate.

[0064] Preferably, the protein hydrolysate, preferably whey protein hydrolysate, preferably extensively hydrolyzed whey protein contains less than 0.8 micrograms, more preferably less than 0.2 micrograms of allergenic β-lactoglobulin per gram of protein. In the context of the present invention, the expression "allergenic β-lactoglobulin" refers to intact or immunogenic β-lactoglobulin, without considering extensively hydrolyzed β-lactoglobulin. Allergenic β-lactoglobulin can be determined by methods known in the art (e.g., ELISA).

[0065] For infants who are severely allergic such that extensively hydrolyzed proteins (e.g., extensively hydrolyzed whey protein) are still a problem as a protein source, or for infants suffering from multiple food allergies, it is advantageous that the protein source consists essentially of free amino acids. Preferably, the protein source contains all essential amino acids. The optimal amino acid profile for infant formula is known in the art and is present in amino acid-based infant formulas (e.g., Neocate). A preferred embodiment of the amino acid composition is given in Example 11.

[0066] Probiotics

[0067] Preferably, the nutritional composition contains lactic acid-producing bacteria selected from the group consisting of Bifidobacterium and / or Lactobacillus genera, particularly Bifidobacterium. Bifidobacterium is present in high amounts in the gut microbiota of breastfed infants. Adding one or more strains belonging to the genus Bifidobacterium to the nutritional composition will further improve the gut microbiota and its activity. More preferably, the nutritional composition contains Bifidobacterium breve. Strains of such Bifidobacterium breve species are commercially available or can be isolated from the microbiota of the infant. An example of a commercially available strain is Bifidobacterium breve M-16V from Morinaga. The amount of Bifidobacterium and / or Lactobacillus is preferably 10 4 to 10 11 cfu / g dry weight of the nutritional composition.

[0068] Nutritional composition

[0069] The nutritional composition according to the present invention can be used as an infant or older infant formula, nutritional therapy, food for special medical purposes, or nutritional supplement. The nutritional composition is preferably an oral composition. The nutritional composition is orally administered or intended to be orally administered to a subject in need, particularly children and infants, including toddlers, preferably children under 6 years of age, preferably typically infants or toddlers from 0 to 36 months of age, more preferably infants from 0 to 12 months of age, most preferably infants from 0 to 6 months of age.

[0070] Thus, in some embodiments, the nutritional composition is an infant formula, a follow-on formula or a toddler formula (also known as growing-up milk or toddler milk), preferably an infant formula or a follow-on formula, most preferably an infant formula. The terms "infant formula" and "follow-on formula" are well-defined and are uniformly controlled by international regulatory bodies. It recommends nutritional value and formula composition, which requires the prepared milk to contain no less than 60 kcal (250 kJ) and no more than 70 kcal (295 kJ) of energy per 100 ml. The EU, FDA and other regulatory bodies have set nutritional requirements based on this. This calorie density ensures the optimal ratio between water and calorie consumption.

[0071] Suitably, the nutritional composition is in powder form, which can preferably be reconstituted with water to form a liquid. When the nutritional composition is in liquid form, the preferred daily administration volume ranges from about 80 to 2500 ml per day, more preferably about 450 to 1000 ml.

[0072] The nutritional composition according to the present invention comprises lipids, preferably lipids suitable for infant nutrition as known in the art. The lipids of the nutritional composition preferably provide 2.8 to 7.0 g, more preferably 4.0 to 6.0 g / 100 kcal of the nutritional composition. When in liquid form, the nutritional composition preferably contains 1.9 to 4.7 g of lipids / 100 ml, more preferably 2.7 to 4.0 g / 100 ml. Based on dry weight, the nutritional composition of the present invention preferably contains 12.5 to 40 wt% lipids, more preferably 19 to 30 wt% lipids.

[0073] The nutritional composition according to the invention may further comprise long-chain polyunsaturated fatty acids (LC-PUFAs). LC-PUFAs are fatty acids in which the acyl chain has a length of 20 to 24 carbon atoms and in which the acyl chain contains at least two unsaturated bonds between the carbon atoms in the acyl chain. More preferably, the nutritional composition comprises at least one LC-PUFA selected from the group consisting of eicosapentaenoic acid (EPA, 20:5n3), docosahexaenoic acid (DHA, 22:6n3), arachidonic acid (ARA, 20:4n6) and docosapentaenoic acid (DPA, 22:5n3), preferably, the nutritional composition comprises at least DHA. Such LC-PUFAs have a further beneficial effect on reducing the risk of allergy. Based on the total fatty acids, the preferred content of LC-PUFAs in the nutritional composition does not exceed 15 wt%, preferably does not exceed 10 wt%, even more preferably does not exceed 5 wt%. Preferably, based on the total fatty acids, the nutritional composition comprises at least 0.2 wt%, preferably at least 0.25 wt%, more preferably at least 0.35 wt%, even more preferably at least 0.5 wt% LC-PUFA. Based on the total fatty acids, the amount of DHA is preferably at least 0.2 wt%, more preferably at least 0.3 wt%, more preferably at least 0.35 wt%, even more preferably 0.35 - 0.6 wt%.

[0074] The nutritional composition comprises digestible carbohydrates. Typically, digestible carbohydrates known in the art suitable for infant nutritional compositions are, for example, selected from digestible polysaccharides (such as starch, maltodextrin), digestible monosaccharides (such as glucose, fructose) and digestible disaccharides (such as lactose, sucrose). Particularly suitable are lactose and / or maltodextrin. Preferably, the nutritional composition comprises lactose. Lactose is the main digestible carbohydrate in milk and has a relatively low glycemic index. For those infants who require an amino acid-based formula, lactose is not desired as it is derived from milk. In such cases, preferably, the nutritional composition comprises maltodextrin. Maltodextrin consists of D-glucose units linked in chains of variable length. The glucose units are mainly linked by α(1→4) glycosidic bonds and typically consist of a mixture of chains 3 to 17 glucose units long. Maltodextrin is classified by DE (dextrose equivalent) and if maltodextrin is included as a digestible carbohydrate in the nutritional composition of the invention, the maltodextrin preferably has a DE of 3 to 47.

[0075] Minerals, vitamins, trace elements and other micronutrients are present as known in the art to comply with the directives regarding infant and follow-on formulae and foods for medical purposes intended for infants.

[0076] Since the nutritional composition according to the present invention contains a thickener, the shear viscosity of the reconstituted ready-to-drink nutritional composition or formula milk powder is higher than that of a standard infant formula without a thickener. The shear viscosity is measured at a shear rate of 10 s -1 at 37 °C as measured by a viscometer from Anton Paar. Preferably, the ready-to-drink nutritional composition has the following shear viscosity in a bottle: 20 to 100 mPa·s, more preferably 25 to 90 mPa·s, even more preferably 30 to 80 mPa·s. These are the viscosities observed in thickened anti-reflux formula milks that have a clinically proven effect on treating and preventing reflux.

[0077] Use

[0078] The composition according to the method or use of the present invention is preferably administered enterally, more preferably orally. The composition of the present invention can advantageously be used as a complete nutrition for infants. The nutritional composition is preferably for infants or toddlers, more preferably infants. Infants are defined as 0 - 12 months of age, and toddlers are defined as 12 - 36 months of age.

[0079] Preferably, the method or use of the present invention is for treating and / or preventing, preferably preventing reflux. In the context of the present invention, reflux, also known as gastroesophageal reflux (GER), is the backward flow of gastric contents up the esophagus and sometimes even into or out of the mouth. At the lower end of the esophagus, the lower esophageal sphincter (LES) opens when swallowing food and then normally closes again to keep the gastric contents in place. When the LES does not function properly, with reflux, the gastric contents (including hydrochloric acid) can come into contact with the esophagus, throat, nasal cavity, lungs, and / or teeth. When reflux can be seen, the diagnostic term regurgitation is used. Regurgitation is the most common functional gastrointestinal disorder (FGID), estimated to affect up to 30% of infants worldwide and 80% of infants under 2 months of age. When the regurgitation of gastric contents causes complications or leads to tissue damage or inflammation, it is called gastroesophageal reflux disease (GERD). Reflux and GERD are problems that are particularly common in infancy. Billead et al. 1990 EJCN [European Journal of Clinical Nutrition] 44, 577 - 583 showed that infants with GER have a slightly faster gastric emptying. This means that the viscosity under simulated gastric digestion conditions in the first 50 minutes of digestion is particularly important. After 70 min, the relevance of having a high viscosity decreases, and the stomach has mostly emptied. On the contrary, a higher viscosity after 70 to 120 min is less desirable because it means that the viscosity of the nutrients in the small intestine may be too high and may impair, for example, the bioaccessibility of micronutrients.

[0080] The combination of thickeners in the nutritional composition of the present invention results in viscosities in the bottle and viscosity kinetics under gastric digestion conditions that are very similar to those of a commercial formula proven to have anti-regurgitation properties (Bellaiche et al., 2022, JPGN [Journal of Pediatric Gastroenterology and Nutrition] 73:579-585), while having a lower amount of thickeners. Thus, it is reasonable that, according to the present invention, these formulae with a combination of xanthan gum, locust bean gum and optionally waxy starch will treat and prevent reflux / regurgitation.

[0081] The nutritional composition of the present invention is preferably suitable for providing nutrition to food allergy subjects. The nutritional composition of the present invention is preferably specifically intended for food allergy infants and / or food allergy toddlers, more preferably infants. Such allergic infants more often suffer from reflux and other gastrointestinal disorders. The nutritional composition of the present invention preferably comprises a deeply hydrolyzed whey protein and / or free amino acids, and is preferably used for treating food allergies, more preferably cow's milk protein allergy. The nutritional composition of the present invention preferably comprises a deeply hydrolyzed whey protein and / or free amino acids, and is preferably used for the dietary management of food allergies, more preferably the dietary management of cow's milk protein allergy.

[0082] Preferably, the nutritional composition of the present invention is used for treating and / or preventing, preferably preventing reflux in food allergy subjects. Preferably, the use for treating and / or preventing reflux is in infants or toddlers, preferably, the use for treating and / or preventing reflux is in infants or toddlers suffering from or at risk of allergy, preferably allergic infants or toddlers, preferably wherein the allergy is cow's milk protein allergy.

[0083] In one aspect, the present invention provides a beneficial effect on the gut microbiota of infants or toddlers. The use of a mixture of galacto-oligosaccharides and long-chain fructo-oligosaccharides or a mixture of short-chain fructo-oligosaccharides and long-chain fructo-oligosaccharides in combination with xanthan gum and locust bean gum results in a decrease in the amount and relative amount of butyrate and propionate and gas, and this decrease is higher than that expected based on a single thickener or indigestible oligosaccharides. In addition, in the presence of short-chain galacto-oligosaccharides, the amount of lactic acid increases synergistically. Thus, the combination of these specific thickeners and specific indigestible oligosaccharides synergistically shifts the activity of the infant gut microbiota towards an activity more similar to that observed in breastfed infants.

[0084] Accordingly, the present invention also relates to a nutritional composition according to the present invention as defined herein, which is used to regulate the activity of the gut microbiota of an infant or young child (preferably an infant) to be similar to the activity found in the microbiota of a healthy breastfed infant, wherein the regulation is at least one, preferably at least two, selected from the following, compared with the gut microbiota activity of the same group of infants or young children fed with a nutritional composition that does not contain the combination of xanthan gum and locust bean gum thickeners as defined in any of the preceding claims and indigestible oligosaccharides:

[0085] a. increasing the amount of acetate or the relative amount of acetate based on total SCFA,

[0086] b. increasing the amount of L-lactate,

[0087] c. reducing the gas production,

[0088] d. reducing the amount of propionate or the relative amount of propionate based on total SCFA, and

[0089] e. reducing the amount of butyrate or the relative amount of butyrate based on total SCFA.

[0090] In other words, the present invention relates to a method for regulating the activity of the gut microbiota of an infant or young child (preferably an infant), the method comprising administering a nutritional composition according to the present invention as defined herein and regulating the gut microbiota activity to be similar to the activity found in the microbiota of a healthy breastfed infant, wherein the regulation is at least one, preferably at least two, selected from the following, compared with the gut microbiota activity of the same group of infants or young children fed with a nutritional composition that does not contain the combination of xanthan gum and locust bean gum thickeners as defined in any of the preceding claims and indigestible oligosaccharides:

[0091] a. increasing the amount of acetate or the relative amount of acetate based on total SCFA,

[0092] b. increasing the amount of L-lactate,

[0093] c. reducing the gas production,

[0094] d. reducing the amount of propionate or the relative amount of propionate based on total SCFA, and

[0095] e. reducing the amount of butyrate or the relative amount of butyrate based on total SCFA.

[0096] The present invention also relates to a nutritional composition according to the present invention as defined herein, which is used to regulate the activity of the gut microbiota of an infant or young child (preferably an infant) to be similar to the activity found in the microbiota of a healthy breastfed infant, wherein the gut microbiota activity is measured by one or more of the following:

[0097] a. The amount of acetate or the relative amount of acetate based on total SCFA,

[0098] b. The amount of L - lactate,

[0099] c. The amount of gas produced,

[0100] d. The amount of propionate or the relative amount of propionate based on total SCFA, and

[0101] e. The amount of butyrate or the relative amount of butyrate based on total SCFA.

[0102] In other words, the present invention relates to a method for modulating the activity of the gut microbiota in an infant or toddler (preferably an infant), the method comprising administering a nutritional composition according to the invention as defined herein and modulating the gut microbiota activity to a level found in the microbiota of a healthy breastfed infant, wherein the gut microbiota activity is determined by one or more of the following:

[0103] 1. The amount of acetate or the relative amount of acetate based on total SCFA,

[0104] 2. The amount of L - lactate,

[0105] 3. The amount of gas produced,

[0106] 4. The amount of propionate or the relative amount of propionate based on total SCFA, and

[0107] 5. The amount of butyrate or the relative amount of butyrate based on total SCFA.

[0108] Examples

[0109] Example 1: Guar gum, used as the sole thickener in a formula milk powder without intact proteins, does not have a high enough viscosity under gastric conditions Table 1: Infant formula milk powder under infant gastric digestion conditions at 37

[0110] The formula milks tested were:

[0111] 1) Pepti Syneo IF, a commercial infant formula containing deeply hydrolyzed whey protein and no thickener.

[0112] 2) Nutrilon A.R.1, a commercial anti - regurgitation infant formula containing intact proteins (casein and whey protein) and 0.5 g LBG / 100 ml as a thickener. This formula containing intact proteins has been clinically proven to be effective against regurgitation and is a reference in terms of viscosity values.

[0113] 3) Pepti Syneo IF containing 0.5 g LBG / 100 ml. The LBG (Grindsted LBG 860, Danisco 10120192) was dry mixed into the powdered composition to obtain a final concentration of 0.5 g / 100 ml.

[0114] 4) Pepti Syneo IF containing 2 g / 100 ml of waxy maize starch (waxy maize starch (Hiform, Cargill, Haubourdin, France)).

[0115] 5) Allernova AR + a commercially available anti-regurgitation infant formula containing a mixture of deeply hydrolyzed casein, highly esterified and weakly esterified pectins, tapioca starch, and locust bean gum as a thickening agent.

[0116] According to the manufacturer's instructions, the powdered formula was reconstituted with tap water pre-warmed to 37 °C and the bottle was shaken by hand for 30 seconds. The reconstituted sample was kept in a water bath maintained at 37 °C.

[0117] Semi-dynamic gastric model

[0118] The gastric digestion of thickened formula was simulated in a bioreactor using a fermenter device (Dasgip, Eppendorf, Germany) allowing pH control, substrate pumping, and overhead stirring at 60 rpm in a 37 °C water bath. The in vitro digestion protocol was selected according to the INFOGEST consensus (slightly modified) (Minekus et al., 2014, Food Funct. 5, 1113 - 1114). Modifications included the simulation of continuous swallowing of unstimulated saliva (Havenaar et al., 2013, Int J Parma 457:327 - 332; Bourlieu et al., 2014, Crit Rev Food Sci Nutr 54:1427 - 1457; Davis et al., 2009, Psychoneuroendocrinology, 34:795 - 804). The model was scaled up 2:1 to simulate the dietary intake of 200 mL of formula by 6-month-old infants.

[0119] After reconstruction, 400 mL of infant formula was placed in a bioreactor. Initial sampling began after 10 min, and then the pH was balanced for 5 min to 6.2. Then, a 66 mL volume of simulated stimulated saliva (106 mM NaCl, 30 mM KCl, 2.0 mM CaCl2, 0.6 mg / mL α-amylase (SIGMA, A9857)) was introduced into the bioreactor. At the same time, continuous unstimulated saliva (106 mM NaCl, 30 mM KCl, 2.0 mM CaCl2, 0.38 mg / mL α-amylase (SIGMA, A9857)) and simulated gastric juice residue (106 mM NaCl, 30 mM KCl, 0.51 mM CaCl2, 0.05 mg / mL pepsin (SIGMA P6887), 0.125 mg / mL lipase (SIGMA, 80612)) were added to represent the formula entering the infant's stomach. The simulated gastric digestion period lasted 120 min, with continuous addition of simulated unstimulated saliva (1.5 mL) and simulated gastric juice (140 mL), and acidified to pH = 4.4 (using 0.5 M HCl). During simulated gastric digestion, the bioreactor contents were sampled regularly at 0, 10, 30, 50, 70, 90, and 120 min for rotational rheological viscosity measurement.

[0120] Viscosity measurement

[0121] The rheological behavior of each sample was evaluated using a rheometer Anton Paar MCR101, whose parallel plate geometry (PP50) was positioned at a gap of 1 mm and heated at 37 °C. After pre-shearing for 10 s and waiting for another 10 s period, the shear was measured from 2 to 100 s -1 pre-shearing for 10 seconds and waiting for another 10 seconds period, the shear was measured from 2 to 100 s -1 . The viscosity values were given at a shear rate of 10 s -1 . Typically representing gastric shear, and expressed as the mean + / - SEM of 3 to 8 replicates.

[0122] The results are shown in Table 1. Compared to the anti-reflux formula milks (2 and 5), the extensively hydrolyzed formula milk without LBG (1) showed significantly lower viscosities. After reconstitution in the bottle, the extensively hydrolyzed formula milk supplemented with 0.5 g / 100 ml LBG (3) reached a viscosity level similar to that of the anti-reflux formula milks (2 and 5). However, after 30 min under gastric conditions, a significant decrease in the viscosity level was observed. Thus, the solution of adding LBG is not suitable for eHF formula milk. The interaction between LBG and intact proteins such as casein in the stomach results in sufficient gastric viscosity, but due to the lack of such proteins in the extensively hydrolyzed hypoallergenic formula milk, the observed gastric viscosity was too low. In addition, the amount of LBG used (0.5 g / 100 ml) is higher than the desired amount because this leaves insufficient space in the formula for the desired amount of prebiotics such as the scGOS / lcFOS mixture. However, when compared to the standard anti-reflux formula milk, a lower amount of LBG resulted in too low a viscosity in the bottle (data not shown). Using starch as the sole thickener (4) resulted in good viscosity in the bottle, but after 30 min under gastric conditions, the viscosity was similar to that of the formula milk without a thickener. The commercial AR formula milk containing a thickener mixture (cassava starch, LBG, HMP, and LMP; 5) showed a lower viscosity in the bottle but a higher viscosity after 50 to 70 min. Too high a viscosity is not desirable before entering the duodenum at the end of gastric digestion as this may affect mineral bioavailability.

[0123] Example 2: Select a combination of thickeners to prepare a formula milk powder containing extensively hydrolyzed whey protein to have sufficient viscosity and product technical characteristics after reconstitution into a ready-to-drink formula milk powder °C and 10 s -1 Viscosity at shear rate

[0124]

[0125] Example 3: Fermentation of thickeners and indigestible oligosaccharides scGOS / lcFOS Table 2: Conditions in fecal slurry fermentation

[0126] Several formulas of infant formula milk containing extensively hydrolyzed proteins were developed considering i) the maximum amount of total fiber (including indigestible thickeners) should be 0.8 g / 100 ml, ii) the prebiotic fiber combination (such as GOS / lcFOS or scFOS / lcFOS) should be present in an amount of at least 0.4 g / 100 ml (preferably higher), iii) the regulatory required amounts of protein, calories, and the caloric contributions of fat and protein and carbohydrates should be met, and iv) adding thickeners does not overly dilute the existing formula to ensure that the requirement for the minimum amount of DHA is met without the need to add additional DHA.

[0127] As a reference, the viscosity in the bottle and under gastric conditions should be comparable to that of Nutrilon Anti-reflux Follow-on Formula Stage 1 (containing 0.48 g LBG / 100 ml). After 70 to 90 min of digestion, the viscosity should not be too high either in the bottle or under gastric conditions, as this would affect drinking through the nipple and may respectively affect the bioavailability of (trace) nutrients. Other criteria are related to user experience and product stability, as explained in more detail.

[0128] Thickening agents tested

[0129] - Waxy maize starch (Hiform, Cargill, Aubourdan, France)

[0130] - Xanthan gum (Keltrol Grindsted 808MAS-SH clarified form CP Kelco or IFF)

[0131] - Locust bean gum (Grindsted LBG 860, Danisco, Valencia, Spain)

[0132] - β-Glucan (PromOat, Lantmannen, Kimstad, Sweden)

[0133] - Amidated low-methoxylated pectin (Amid CF 010-D, HERBSTREITH&FOX GmbH&Co, Neurenberg,

[0134] - Non-amidated low-methoxylated pectin (Classic CF 714, HERBSTREITH&FOX GmbH&Co, Neurenberg, Germany)

[0135] - Amidated high-methoxylated pectin (Amid CS 005, HERBSTREITH&FOX GmbH&Co, Neurenberg, Germany)

[0136] - Non-amidated high-methoxylated pectin (Classic CU 70, HERBSTREITH&FOX GmbH&Co, Neurenberg, Germany)

[0137] Over 40 different concentrations of single thickening agents and thickening agent combinations were tested. Each thickening agent at different concentrations was weighed and then dry-blended into the powdered hydrolyzed infant formula Pepti as described in Example 1. Reconstitution and viscosity measurements were carried out as described in Example 1.

[0138] Viscosity measurements were carried out using a rheometer (Anton Paar GmbH MCR302, Austria) equipped with a thermostatic bath at 37 °C. Viscosity values were given at a shear rate of 10 s -1 −1.

[0139] Results:

[0140] Single fibers:

[0141] According to Example 1, 0.5 g / l LBG as the sole thickener resulted in a satisfactory viscosity in the bottle. The product was stable and no large "fish eyes" were observed. "Fish eyes" are transparent and shiny particles, which, if too large, are not only visually unappealing to consumers but may also affect the passage of the formula milk through the nipple. When compared to Nutrilon Anti-Reflux Formula Stage 1, a lower concentration of LBG resulted in too low a viscosity in the bottle. However, as known from Example 1, when 0.5 g LBG / 100 ml was used in a formula milk with hydrolyzed protein or free amino acids, too low a viscosity was observed under gastric conditions.

[0142] It was found that using waxy starch alone as a thickener was not suitable because a large amount (2 g / 100 ml) was required to increase the viscosity in the bottle to a sufficiently high level, which imposed formulation limitations. The product was stable and no large "fish eyes" were observed. A concentration of 1.5 g / 100 ml waxy starch or lower resulted in a lower viscosity of the formula milk than Nutrilon Anti-Reflux Formula Stage 1. In addition, the viscosity under gastric conditions was not affected by waxy starch because α-amylase in saliva would degrade waxy starch, resulting in too low a viscosity under gastric conditions, as can be seen in Example 1.

[0143] Using β-glucan (0.4 g / 100 ml) as a thickener in a hydrolyzed protein formula milk resulted in highly unstable products with separation, and the measurable viscosity in the separated products was too low.

[0144] Using a single pectin of HM, HMA, LM or LMA pectin (0.3 g / 100 ml) resulted in too low a viscosity of the formula milk. These formula milks were unstable because the products separated. It was expected that increasing the pectin level would further destabilize the formula milk. In addition, large "fish eyes" were also observed. Relevantly, the thickener can be dry-blended into the base powder of infant formula milk and does not need to be processed in the wet phase of the manufacturing process, as this is not attractive for commercial production, resulting in low formulation flexibility and possible technical problems when spray-dried. Therefore, these pectins are less attractive.

[0145] As a single thickener, the level of xanthan gum is close to the permitted maximum (0.1 g / 100 ml), resulting in too low viscosity in the bottle for the formula milk powder compared to Aptamil Anti-Reflux 1.

[0146] Combination of two fibers:

[0147] The pectin mixture of HM(A) and LMA pectin (each 0.2 g / 100 ml) results in the formula milk powder being too thin in the bottle compared to Aptamil Anti-Reflux 1. The product is unstable. Similar to the single fiber, large "fish eyes" were observed.

[0148] The mixture of pectin and waxy starch shows a sufficiently high viscosity in the bottle only when using 2 g / 100 ml of starch, which brings formula limitations. Less waxy starch results in the formula milk powder being too thin. Generally, many test combinations in which the thickener is dry-blended into the powdered formula milk powder are unstable and / or show large "fish eyes".

[0149] The mixture of pectin and XG with the maximum concentration permitted by regulations is still too thin when compared to Aptamil Anti-Reflux 1.

[0150] LBG and waxy starch show a sufficient viscosity in the bottle, but since waxy starch does not contribute to the viscosity in the stomach, it can be expected that LBG will have the same low viscosity in a protein formula milk powder that is deeply hydrolyzed under gastric conditions.

[0151] The combination of LBG and XG shows a promising viscosity level in the bottle, but this depends on the concentration and ratio. If the weight ratio is close to 1, i.e., between 0.7 and 1.5, the total amount of the two thickeners should be between 0.05 - 0.2 g / 100 ml. When it is higher than 0.2 g / 100 ml, the formula milk powder is too thick. When the total amount of the two thickeners is lower than 0.05 g / 100 ml, the viscosity is much lower than that of Aptamil Anti-Reflux 1.

[0152] For the combination of LBG and XG, where the amount of locust bean gum is higher than XG, i.e., the ratio is higher than 3, the LBG concentration is at least 0.15 g / 100 ml and at most 0.35 g / 100 ml, while the amount of xanthan gum should be at least 0.03 g / 100 ml and at most 0.07 g / 100 ml, good viscosity was observed.

[0153] Combination of three fibers:

[0154] If the weight ratio of XG and LBG is close to 1, i.e., between 0.7 and 1.5, and the total amount of XG and LBG is not higher than 0.2 g / 100 ml or lower than 0.05 g / 100 ml, then the combination of XG, LBG and waxy starch shows good results in terms of viscosity. In particular, in the range of 0.5 to 1.5 g / 100 ml XG+LBG, the additional presence of a relatively low amount of waxy starch (e.g., 0.4 to 1.5 g / 100 ml) is sufficient to further increase the viscosity to the level observed for the first stage of Nutrilon Anti-reflux formula milk powder.

[0155] The combination of pectin, XG and waxy starch shows good viscosity, but requires a very close amount of pectin (HMA or HM) to the maximum amount allowed by regulations. In addition, a higher concentration of XG (0.08 to 0.1 g / 100 ml) is required. Some of the combinations tested show large "fish eyes". In addition, large "fish eyes" were observed in some of the combinations of pectin, XG and LBG tested.

[0156] It is clear from these experiments that the combination of XG / LBG (with or without waxy starch) is the most promising from the perspective of in-bottle viscosity and other product technical characteristics.

[0157] Table 3: Formation of fermentation end products after 48 h of fermentation by the infant gut microbiota (in μmol / g of fiber)

[0158] Stool samples were collected from formula-fed infants (5.5 months old) and breastfed infants (3 months old). The infants had no gastrointestinal problems and had not used antibiotics for nearly a month. The stool samples were combined, homogenized, divided into smaller aliquots, and mixed with glycerol (10%) in an anaerobic chamber. Subsequently, the aliquots were stored at -80 °C.

[0159] Indigestible oligosaccharides were added at a concentration of 100 mg dietary fiber (DP≥2) per 6 ml of stool suspension, see the conditions in Table 2. The source of GOS was Vivinal GOS (Friesland Campina, Netherlands), and the source of lcFOS was Raftilin HP (Orafti). The sources of xanthan gum and locust bean gum were the same as in Examples 1 and 2.

[0160] Table 4: L-lactate production at t = 24

[0161]

[0162] In the experiment, the fecal pool was thawed in a 37 °C water bath for 20 minutes. Subsequently, the fecal pool was placed in an anaerobic chamber. The feces were mixed with the fermentation medium at a ratio of 1:5 in a falcon tube. A sample of this fecal suspension was taken at t = 0, and 6 ml of this suspension was added to a sterile falcon tube together with the target substrate and mixed well. Then, 6 ml of the feces / substrate suspension was placed in a dialysis tube, and the air in the voids was removed. The dialysis tube was placed in a 100 ml Scott bottle containing 100 ml of dialysis medium. The Scott bottle was sealed and incubated at 37 °C. Samples of the dialysis medium (dialysate) and the fecal suspension (lumen) were collected at t = 24 and t = 48 hours to measure SCFA, D- and L-lactate, and gas volume.

[0163] The fermentation medium (Mc Bain and MacFarlane) contained buffered peptone water 3.0 g / l, yeast extract 2.5 g / l, tryptone 3.0 g / l, L-cysteine-HCl 0.4 g / l, bile salts 0.05 g / l, K2HPO4·3H2O 2.6 g / l, NaHCO3 0.2 g / l, NaCl 4.5 g / l, MgSO4·7H2O 0.5 g / l, CaCl2·2H2O 0.3 g / l, FeSO4·7H2O 0.005 g / l. The components were added one by one to 800 ml of water, the pH was adjusted to 5.5 ± 0.1 with K2HPO4 or NaHCO3, and the volume was made up to 1 liter. The medium was sterilized at 121 °C for 15 minutes and placed in the anaerobic chamber for at least 16 hours before use.

[0164] The dialysis medium contained K2HPO4·3H2O 2.6 g / l, NaHCO3 0.2 g / l, NaCl 4.5 g / l, MgSO4·7H2O 0.5 g / l, CaCl2·2H2O 0.3 g / l, FeSO4·7H2O 0.005 g / l. The pH was adjusted to 5.5 ± 0.1 with K2HPO4 or NaHCO3. Due to the formation of sediment, the medium was not sterilized. The medium was placed in the anaerobic chamber for at least 16 hours before use.

[0165] The pH was measured by directly immersing a 423 pH-electrode (Mettler Toledo, Columbus, Ohio, USA) connected to a Handy-lab pH meter (Schott Glas, Mainz, Germany) into the sample.

[0166] The gas volume was determined by a device that measures pressure and volume. Before measurement, the bottles were shaken well.

[0167] Quantitative determination of SCFAs acetic acid, propionic acid, n-butyric acid, isobutyric acid, n-valeric acid, and isovaleric acid was performed using a Shimadzu-GC2025 gas chromatograph and a flame ionization detector. Hydrogen was used as the mobile phase. The level of SCFAs was determined using 2-ethylbutyric acid as the internal standard. A calibration curve was constructed based on the peak area, and the concentration in the sample was calculated.

[0168] Lactate was enzymatically determined using an L-lactate detection kit containing D- and L-lactate dehydrogenases (Boehringer Mannheim, Mannheim, Germany). First, the samples were centrifuged at 13,000 rpm for 10 min at 4 °C, then the supernatants were heated at 100 °C for 10 min to inactivate all enzymes, and then the samples were centrifuged at 13,000 rpm for 10 min.

[0169] Results:

[0170] The levels of isobutyric acid, valeric acid, and isovaleric acid were below the detection limit. Lactate was formed at t = 24 h in the mixture containing scG / lcF, mainly L-lactate. Interestingly and advantageously, the amount of lactate in the scG / lcF / XG / LBG mixture was much higher than could be expected based on the fermentation characteristics of the individual components. Since lactate is an intermediate metabolite and is subsequently fermented by other bacteria, a synergistic excess of lactate was only observed at the earliest sampling time points. However, this indicates an increased activity of lactate-producing bacteria.

[0171] When xanthan gum or locust bean gum was the sole fiber, the content of butyric acid and especially propionic acid was high. Thus, the SCFA profile showed a lower % of acetic acid, and higher % of butyric acid and propionic acid. Interestingly, when the combination of scGOS / lcFOS / xanthan gum / locust bean gum was tested, the formation of butyric acid, especially propionic acid, was lower in amount and percentage than could be expected based on the fermentation characteristics of the individual components. Advantageously, the amount of gas formed was also lower than could be expected.

[0172] These effects occurred at t = 24 and 48 h. Table 3 shows SCFA production and gas production after 48 h, which represents the colonic transit time of the infant.

[0173] Example 4: Fermentation of thickeners and indigestible oligosaccharides scFOS / lcFOS

[0174]

[0175]

[0176] Repeat the same experiment, but use scGOS / lcFOS / LBG / XG mixtures of 0.8208 / 0.0912 / 0.044 / 0.044 and 0.6372 / 0.0708 / 0.235 / 0.057 respectively, and compare with XG alone, LBG alone or scGOS / lcFOS alone.

[0177] Similarly, for the two mixtures containing 4 fibers, the amount and relative amount of butyrate were lower than expected based on the results obtained with XG alone, LBG alone or scGOS / lcFOS alone. Additionally, the amounts of acetate and L-lactate (data shown in Table 4) increased and were much higher than expected based on the results obtained with XG alone, LBG alone or scGOS / lcFOS alone.

[0178] Table 5: Conditions in fecal slurry fermentation

[0179]

[0180] This indicates that the scG / lcF / XG / LBG mixture synergistically shifts the microbiota activity, lactate and SCFA formation towards a situation more similar to that observed in breastfed infants. It is well known that in healthy infants breastfed before weaning, the microbiota produces high levels of acetic acid and L-lactic acid and low levels of butyric acid and propionic acid compared to infants fed standard formula milk, or compared to infants with intestinal microbiota dysbiosis, or compared to allergic infants (Wopereis et al. Pediatr Allergy Immunol [Pediatric Allergy and Immunology] 2014:25:428 - 438). Compared to traditional prebiotics such as scGOS and lcFOS, thickeners such as fiber XG and LBG are considered to be less acidic and produce more propionic acid and butyric acid, but the presence of scGOS and lcFOS may unexpectedly overcome these effects.

[0181] Table 6: Formation of fermentation end products after 48 h of fermentation by the infant gut microbiota (average value, in μmol / g of fiber)

[0182] Conduct the fermentation experiment of fecal samples in the manner described in Example 3.

[0183] Add indigestible oligosaccharides at a concentration of 100 mg dietary fiber (DP≥2) per 6 ml fecal suspension. All conditions are in the following table. The source of scFOS is Raftilose P95 (Orafti), and the source of lcFOS is Raftilin HP (Orafti). The sources of xanthan gum and locust bean gum are the same as in Examples 1 and 2.

[0184] Example 5: Viscosity under gastric conditions. Select the optimal XG / LBG thickener composition

[0185]

[0186] Results:

[0187] The levels of isobutyric acid, valeric acid, and isovaleric acid were below the detection limit. Only a small amount of lactic acid was formed. In the mixture containing scFOS / lcFOS, the level of total SCFA (the sum of acetic acid, propionic acid, and butyric acid) was the highest.

[0188] When xanthan gum or locust bean gum was the sole fiber, the level of butyric acid, but especially propionic acid, was high. Thus, the SCFA profile showed a lower % of acetic acid, as well as higher % of butyric acid and higher % of propionic acid. Interestingly, when the combination of scFOS / lcFOS / XG / LBG was used, the formation of butyric acid and propionic acid was inhibited, and the amounts and percentages produced were much lower than expected based on the fermentation characteristics of the individual components. Interestingly and advantageously, the amount of gas formed was also lower than could be expected. This was observed at t = 24 and 48 h. Table 6 shows the effect at 48 h, which represents the colonic transit time of the infant. The gas production of the scF / lcF mixture was high, while that of XG or LBG was low, but in the scF / lcF / LBG / XG mixture, the gas production was lower than expected based on the individual components.

[0189] Table 7: Different infant formula milk powders for simulated gastric digestion at 37

[0190]

[0191]

[0192] This indicates that the mixture of scF / lcF / XG / LBG synergistically shifts the microbiota activity and SCFA formation towards a situation more similar to that observed in breastfed infants.

[0193] The same experiment was repeated, but using mixtures of scFOS / lcFOS / LBG / XG of 0.8208 / 0.0912 / 0.044 / 0.044 and 0.6372 / 0.0708 / 0.235 / 0.057.

[0194] Similarly, for both mixtures of mixtures containing thickeners and indigestible oligosaccharides, the amount and relative amount of butyrate were lower than expected based on the results obtained using individual XG, individual LBG, or individual scFOS / lcFOS. In addition, the gas production was lower than expected based on the results obtained using individual XG, individual LBG, or individual scGFOS / lcFOS.

[0195] Example 6: Role of thickeners in amino acid-based formula milk powder

[0196] Several combinations of XG and LBG were tested with the aim of determining the optimal XG / LBG thickener concentration required to make the gastric viscosity kinetics similar to that of the reference formula milk powder, Aptamil Anti-reflux Formula Stage 1.

[0197] According to the method of Example 1, the initial viscosities of Pepti Syneo thickened with different concentrations of LBG, waxy starch, XG or LBG-XG combinations were measured over time in the formula milk powder and during in vitro gastric digestion. Since waxy starch does not contribute to viscosity under gastric conditions (see Example 1), it was not further tested in the combinations under gastric conditions.

[0198] Subsequently, an XG / LBG concentration matrix was designed to cover the XG and LBG concentrations to be tested. Xanthan gum was tested at several concentrations (0.02, 0.04, 0.05, 0.06 and 0.12 g / 100 ml). Waxy starch: 2 g / 100 ml. XG / LBG 0.02 / 0.48; 0.10 / 0.05; 0.035 / 0.02; 0.035 / 0.19; 0.034 / 0.35; 0.043 / 0.02; 0.043 / 0.19; 0.043 / 0.36; 0.043 / 0.043; 0.06 / 0.02; 0.06 / 0.19; 0.06 / 0.36; and 0.05 / 0.46 g / 100 ml. A multiple linear regression model was developed to simulate the gastric viscosity over time. Using this model, a multi-factor equation solver found several optimal values of XG and LBG to thicken the formula milk powder under gastric conditions as close as possible to Aptamil Anti-reflux Formula Stage 1. The gastric viscosity predictions based on the XG / LBG concentrations showed different behaviors in the undigested / digested states when the ratio was close to 1 / 1 or deviated from 1 / 1. This finding was translated into 2 models, one for cases where the XG / LBG ratio was close to 1:1 and one for cases where the ratio deviated from 1 / 1. The optimal ratios were tested in a dynamic gastrointestinal model simulating infant gastric digestion.

[0199] The two optimal concentrations determined by the regression model were 1) a wt / wt ratio of XG / LBG of 1:1, with each fiber present at 0.044 / 0.044 wt% in the reconstituted formula milk powder, and 2) a wt / wt ratio of XG / LBG of 1:4.15, with LBG present at a concentration of 0.235 wt% and XG present at a concentration of 0.057 wt% in the reconstituted formula milk powder.

[0200] Table 8: Different infant formula milk powders for simulated gastric digestion at 37 °C and 10 s -1 Viscosity at the shear rate.

[0201]

[0202] The innovative low-allergenic formula milks with XG / LBG of 0.044 / 0.044 and 0.057 / 0.235 show the best ability to mimic the rheological gastric behavior found in the standard anti-reflux formula milks with proven efficacy. The advantage of XG / LBG 0.044 / 0.044 is the low total fiber content, and the advantage of XG / LBG 0.057 / 0.235 is that its viscosity in the bottle (t = 0) is more comparable to that of the standard anti-reflux formula milk.

[0203] Example 7: Anti-regurgitation formula milk powder with hydrolyzed whey protein and thickeners

[0204] In addition to the efficacy of thickeners in deeply hydrolyzed formula milks, the role of thickeners in amino acid-based formula milks was also tested. Thickener was added to Neocate Syneo, an infant formula milk that contains free amino acids as the protein source and is sold for infants with severe cow's milk protein allergy.

[0205] Example 8: Thickened formula milk powder with hydrolyzed whey protein ℃ and 10 s -1 Viscosity at the shear rate.

[0206]

[0207] As can be seen from Table 8, in amino acid-based formula milks, the combination of thickeners works well under gastric conditions, comparable to the Nutrilon anti-reflux formula milk and far higher than products without thickeners.

[0208] Example 9: Anti-regurgitation formula milk powder for allergic infants containing extensively hydrolyzed whey protein and thickeners

[0209] A packaged powdered infant formula milk, after being reconstituted with water according to the instructions on the package, contains per 100 ml (13.7 g of powder in 100 ml of final volume):

[0210] - 66 kcal

[0211] - 1.5 g of protein (partially hydrolyzed whey protein)

[0212] - 7.2 g of digestible carbohydrates

[0213] - 3.4 g of fat (mainly vegetable oil, fish oil)

[0214] - 0.712 g of indigestible oligosaccharides scGOS / lcFOS with a 9:1 ratio (source Vivinal GOS and Raftiline HP)

[0215] - 0.044 g of xanthan gum

[0216] - 0.044 g of locust bean gum

[0217] Minerals, vitamins, trace elements and other micronutrients as per the instructions.

[0218] Example 10: Anti-regurgitation formula milk powder for allergic infants containing extensively hydrolyzed whey protein and thickeners

[0219] A packaged powdered infant formula which, when reconstituted with water according to the instructions on the package, contains per 100 ml (13.7 g of powder in 100 ml final volume):

[0220] - 66 kcal

[0221] - 1.5 g protein (whey protein and casein)

[0222] - 7.2 g digestible carbohydrates

[0223] - 3.4 g fat (vegetable oil, fish oil)

[0224] - 0.61 g of indigestible oligosaccharides scGOS / lcFOS in a 9:1 ratio (source Vivinal GOS and Raftiline HP)

[0225] - 0.057 g xanthan gum

[0226] - 0.235 g locust bean gum

[0227] Minerals, vitamins, trace elements and other micronutrients as per the instructions.

[0228] Example 11: Anti-regurgitation formula milk powder for allergic infants containing free amino acids and thickeners

[0229] A packaged powdered infant formula which, when reconstituted with water according to the instructions on the package, contains per 100 ml (13.46 g of powder in 100 ml final volume):

[0230] - 66 kcal

[0231] - 1.6 g protein (deeply hydrolyzed whey protein)

[0232] - 7.1 g digestible carbohydrates (mainly lactose)

[0233] - 3.4 g fat (vegetable oil, fish oil)

[0234] - 0.4 g prebiotic: scGOS / lcFOS in a 9:1 ratio (source Vivinal GOS and Raftiline HP)

[0235] - 0.057 g xanthan gum

[0236] - 0.235 g locust bean gum

[0237] Such as minerals, vitamins, trace elements and other micronutrients according to the instructions.

[0238] ​

[0239] A packaged powdered follow-on infant formula which, when reconstituted with water according to the instructions on the package, contains per 100 ml (14.44 g of powder in 100 ml of final volume):

[0240] - 68 kcal

[0241] - 1.6 g of protein (deeply hydrolyzed whey protein)

[0242] - 7.8 g of digestible carbohydrates (mainly lactose) and includes 1.0 g of waxy maize starch.

[0243] - 3.2 g of fat (vegetable oil, fish oil)

[0244] - 0.7 g of prebiotic: 9:1 ratio of scGOS / lcFOS (source Vivinal GOS and Raftiline HP)

[0245] - 0.044 g of xanthan gum

[0246] - 0.044 g of locust bean gum

[0247] Such as minerals, vitamins, trace elements and other micronutrients according to the instructions.

[0248] ​

[0249] A packaged infant formula which, when reconstituted with water according to the instructions on the package, contains per 100 ml (14.7 g of powder in 100 ml of final volume):

[0250] - 67 kcal

[0251] - 1.8 g of protein equivalent (free amino acids) L-alanine, L-arginine, L-aspartic acid, L-cystine, L-glutamine, glycine, L-histidine, L-isoleucine, L-lysine, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, L-carnitine

[0252] - 7.3 g of digestible carbohydrates (mainly dried glucose syrup) and includes 1.0 g of waxy maize starch.

[0253] - 3.4 g of fat (vegetable oil)

[0254] -0.7 g of prebiotic: scFOS / lcFOS in a 9:1 ratio (source: Raftilose P95 and Raftiline HP)

[0255] -0.044 g of xanthan gum

[0256] -0.044 g of locust bean gum

[0257] Minerals, vitamins, trace elements and other micronutrients as per the instructions.

Claims

1. A nutritional composition suitable for providing nutrition to infants or toddlers, preferably infants, comprising lipids, digestible carbohydrates, proteins, thickeners, and indigestible oligosaccharides, wherein the thickener comprises a combination of xanthan gum and locust bean gum and the indigestible oligosaccharides comprise long-chain fructooligosaccharides and short-chain oligosaccharides selected from galactooligosaccharides and fructooligosaccharides.

2. The nutritional composition according to claim 1, which comprises short-chain fructooligosaccharides.

3. The nutritional composition according to claim 1, which comprises short-chain galactooligosaccharides.

4. The nutritional composition according to any one of claims 1-3, which is in the form of a ready-to-drink liquid or a powder, the powder form being a ready-to-use liquid after reconstitution with water, wherein the xanthan gum is present in an amount of 0.03 to 0.07 g / 100 ml, preferably 0.03 to 0.06 g / 100 ml, even more preferably 0.04 to 0.05 g / 100 ml of the ready-to-drink nutritional composition, and the locust bean gum is present in an amount of 0.03 to 0.07 g / 100 ml, more preferably 0.03 to 0.06 g / 100 ml, even more preferably 0.04 to 0.05 g / 100 ml of the ready-to-drink nutritional composition, and the wt / wt ratio of xanthan gum to locust bean gum is 0.7 to 1.5, more preferably 0.8 to 1.2, even more preferably 0.90 to 1.

1.

5. The nutritional composition according to claim 4, which further comprises waxy starch as an additional thickener in an amount of preferably 0.4 to 1.5 g / 100 ml of the ready-to-drink nutritional composition, more preferably 0.4 to 0.8 g / 100 ml of the ready-to-drink nutritional composition.

6. The nutritional composition according to any one of claims 1-3, wherein the xanthan gum is present in an amount of 0.03 to 0.07 g / 100 ml of the ready-to-drink nutritional composition, more preferably 0.03 to 0.06 g / 100 ml, and the locust bean gum is present in an amount of 0.10 to 0.35 g / 100 ml of the ready-to-drink nutritional composition, more preferably 0.20 to 0.30 g / 100 ml, and the wt / wt ratio of xanthan gum to locust bean gum is 1.5 to 6.0, more preferably 3.0 to 5.

0.

7. The nutritional composition according to claim 6, wherein the amount of waxy starch is less than 0.2 g / 100 ml of the ready-to-drink nutritional composition.

8. The nutritional composition according to any one of claims 1-7, wherein the amount of the indigestible oligosaccharides is at least 0.4 g / 100 ml of the ready-to-drink nutritional composition, preferably 0.4 to 0.8 g / 100 ml, more preferably 0.6 to 0.8 g / 100 ml of the ready-to-drink nutritional composition.

9. The nutritional composition according to any one of claims 1-8, wherein the long-chain fructooligosaccharides have an average degree of polymerization higher than 10, preferably in the range of 15-50, most preferably higher than 20, and the short-chain indigestible oligosaccharides have an average degree of polymerization less than 10, preferably at most 8, preferably in the range of 2-7.

10. The nutritional composition according to any one of claims 1 - 9, wherein the short-chain indigestible oligosaccharides and the long-chain fructooligosaccharides are present in a short-chain to long-chain weight ratio within the range of 1:99 - 99:1, more preferably 1:1 - 99:1, more preferably 4:1 - 97:3, even more preferably 5:1 - 95:5, even more preferably 7:1 - 95:5, even more preferably 8:1 - 10:1, and most preferably approximately 9:

1.

11. The nutritional composition according to any one of claims 1-10, wherein the ready-to-drink liquid has a shear viscosity of 20 to 100 mPa·s at a shear rate of 37 °C and 10 s -1 and more preferably 25 to 90 mPa·s, even more preferably 30 to 80 mPa·s.

12. The nutritional composition according to any one of claims 1 - 11, wherein the protein comprises hydrolyzed protein and / or free amino acids.

13. The nutritional composition according to any one of claims 1 - 12, which is in powder form and is suitable for reconstitution with water into an instant formula milk powder.

14. The nutritional composition according to any one of claims 1 - 13, which is an infant formula or a follow-on formula.

15. The nutritional composition according to any one of the preceding claims, which is used for the treatment or prevention of reflux in infants or toddlers, preferably for use in the treatment of reflux.

16. The nutritional composition according to claim 15, wherein the infant or toddler has an allergy or is at risk of developing an allergy, preferably has an allergy.

17. The nutritional composition according to claim 16, wherein the allergy is cow's milk protein allergy.

18. A method for preparing the nutritional composition according to any one of the preceding claims, the method comprising a. providing a liquid nutritional composition comprising protein, digestible carbohydrates, and lipids, b. drying the liquid obtained in step a) into a powder, preferably by spray drying, c. dry mixing xanthan gum, locust bean gum, and optionally waxy starch with the powder obtained in step b), and d. adding indigestible oligosaccharides in step a) and / or step c).

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