CN112384079A

CN112384079A - Nutritional composition for treating diarrhea, preparation and treatment method thereof

Info

Publication number: CN112384079A
Application number: CN201880095363.4A
Authority: CN
Inventors: C·T·克尼平; B·斯塔尔
Original assignee: Nutricia NV
Current assignee: Nutricia NV
Priority date: 2018-07-12
Filing date: 2018-07-12
Publication date: 2021-02-19
Also published as: EP3820308A1; AU2018431641A1; WO2020013684A1

Abstract

The present invention relates to a composition comprising fucosylated human lacto-oligosaccharides, preferably 2' -fucosyllactose, for use in the treatment of virus-induced diarrhea. The composition may be an infant formula. Also claimed is the use of fucosylated human lacto-oligosaccharides for the preparation of a nutritional composition and for the treatment of virus-induced diarrhea in a human subject.

Description

Nutritional composition for treating diarrhea, preparation and treatment method thereof

Technical Field

The present invention relates to infant nutrition containing non-digestible oligosaccharides, in particular for use in the treatment of virus induced diarrhoea.

Background

Viral infections are common in people of all ages, but appear to be concentrated on infants and children. Many viral infections cause fever and physical pain or discomfort, but not so severe that most children infected with the virus will improve without treatment. Especially during infancy, parents have difficulty in determining whether their infants are potentially seriously infected and require immediate medical care.

Rotavirus (RV) is the major pathogen of acute infantile diarrhea, and if left untreated, can lead to dehydration or more serious complications and even death. Since the pathogenesis of RV in humans is not yet clear, RV vaccines have not been applied globally and regulation by nutritional intervention with bioactive ingredients is of interest. Recent studies have shown a possible role for HMOS in the prevention of RV-associated diarrhea (Laucirica et al J Nutr 2017; 147: 1709-.

Human milk is a preferred food for infants. Human milk provides several bioactive factors that benefit the relatively immature immune system of early-life newborns. These components are divided into two distinct groups according to their protective effect or their ability to promote maturation. In this sense, Human Milk Oligosaccharides (HMOS) are believed to play a role in both protection and maturation. However, breast-feeding infants are not always possible or desirable. In this case, infant formula or follow-on formula (follow on formula) is a good choice. In order to mimic the beneficial effects of human milk as closely as possible, these formulations should have an optimal composition.

The 162 HMOS structures found to date and described in Urshima et al (Trends in diabetes and diabetes technology, 2018, 30; 172, SE51-SE65) show a lactose, polylactosamine or lacto-N-disaccharide core, which can be further conjugated to fucose or sialic acid or both. This structure confers a high degree of chemical variability and protection from digestion. Thus, most HMOS is neither absorbed nor metabolized in the proximal intestine and reaches the distal intestine undigested, exerting a prebiotic effect on certain bacterial populations, strengthening the intestinal barrier, and preventing infection by intestinal pathogens. HMOS is present in relatively high proportions (5-20g/L) in human breast milk. The most abundant HMOS is 2 '-fucosyllactose (2' -FL), which accounts for about 20% of the total oligosaccharides in human breast milk. In vitro and in vivo studies of 2' -FL demonstrated immunomodulatory effects including promotion of anti-inflammatory activity (He et al Gut 2016; 65: 33-46) and inhibition of pathogen colonization (Ruiz-Palacios et al J Biol Chem 2003; 278: 14112-20).

WO 2016/139333 relates to nutritional compositions comprising fucosylated oligosaccharides, such as 2' -fucosyllactose, and specific N-acetylated oligosaccharides, such as lacto-N-neotetraose (LNnT), for use against gastrointestinal infections and inflammations in infants or young children.

Summary of The Invention

The inventors found that rotavirus-induced diarrhea can be ameliorated by the presence of 2' -fucosyllactose in a neonatal rat model. In particular an improvement in diarrhea was observed in terms of severity and duration of diarrhea. In animal models involving lactating rats, early nutritional intervention by administration of 2' -fucosyllactose was found to significantly reduce the maximum severity of diarrhea by 15% and the average severity of diarrhea by 22%. Furthermore, the duration of diarrhea was significantly reduced by 44% after administration of 2' -fucosyllactose. Furthermore, when stool weight was analyzed, an increase in stool weight after viral infection was observed, whereas the increase in stool weight was statistically significantly reduced after 2' -fucosyllactose administration.

Thus, the inventors found that supplementation with 2' -FL during RV infection induced a significant protective effect, significantly reducing the severity and duration of diarrhea and stool weight.

Detailed Description

Accordingly, the present invention relates to a method for the treatment of virus induced diarrhea in a human subject by administering a nutritional composition comprising fucosylated human milk oligosaccharides, preferably 2' -fucosyllactose.

In other words, the present invention relates to a nutritional composition comprising fucosylated human lacto-oligosaccharides, preferably 2' -fucosyllactose, for use in the treatment of virus-induced diarrhea in a human subject.

In a preferred embodiment, the virus-induced diarrhea is rotavirus-induced diarrhea.

For certain jurisdictions, the present invention may also be said to be the use of fucosylated human lactooligosaccharides, preferably 2' -fucosyllactose, for the preparation of a nutritional composition for the treatment of virus-induced diarrhea in a human subject.

The amount of fucosylated HMO, preferably 2' -fucosyllactose, is preferably an effective amount for treating and/or reducing the severity of virus-induced diarrhea and/or shortening the duration of virus-induced diarrhea in a human subject.

Fucosylated human milk oligosaccharides and 2' -fucosyllactose

Prebiotics are typically indigestible carbohydrate compounds, such as non-digestible oligosaccharides (NDO). These compounds pass through the first part of the gastrointestinal tract, without being substantially digested. In the intestine, these compounds are fermented by the microorganisms to release short-chain fatty acids and the like which are absorbed into the human body.

NDO are derived from a wide variety of sources, including human breast milk. These oligosaccharides are commonly referred to as Human Milk Oligosaccharides (HMOS). Typical NDO used in infant food are GOS and FOS.

Human milk is the first choice food for infants, also known as the gold standard. The levels of oligosaccharides contained in human milk are particularly high, on the order of 10 to 15g/L, which usually far exceeds the NDO levels in livestock's milk. Furthermore, HMOS is structurally different compared to NDO in livestock's milk. Human NDO are very complex, consisting of many heterogeneous groups of different compounds containing different sugar compositions. Large scale synthesis is complicated due to their complex and polymorphic structures. Therefore, it has not been technically and economically feasible to prepare compositions, such as infant formulas, containing the same NDO composition as human milk. In the method or use according to the invention, fucosylated non-digestible human milk oligosaccharides are used.

Fucosyllactose (FL) is a fucosylated non-digestible oligosaccharide present in human milk. It is not present in cow milk. It consists of three monosaccharide units (fucose, galactose and glucose) linked together. Lactose is a galactose unit linked to a glucose unit by a β 1,4 linkage. The fucose unit is linked to the galactose unit of the lactose molecule by an α 1,2 linkage (2 '-fucosyllactose, 2' -FL) or to the glucose unit of lactose by an α 1,3 linkage (3-fucosyllactose, 3-FL). 2' FL is the most abundant NDO in human milk. The HMOS used in the present invention is 2' -FL.

2' -FL, (β -L-Fuc- (1 → 2) - β -D-Gal- (1 → 4) -D-Glc) is commercially available from, for example, Sigma-Aldrich. Alternatively, it may be isolated from human milk, for example as described in Andersson & Donald,1981, J chromatography.211: 170-.

Preferably, the composition of the invention comprises 1mg to 3g 2 '-FL/100 ml, more preferably 10mg to 2g, even more preferably 20mg to 100mg 2' -FL/100 ml. The composition of the invention preferably comprises 0.007 to 20 wt.% 2' -FL, more preferably 0.07 to 10 wt.%, even more preferably 0.15 to 1 wt.%, based on dry weight. Lower amounts of fucosyllactose are less effective in ameliorating virus-induced diarrhea, while too high amounts result in unnecessarily high costs of the product.

In one embodiment, the nutritional composition for said use according to the invention does not comprise lacto-N-neotetraose (LNnT). In one embodiment, the nutritional composition for use according to the invention does not comprise a human milk oligosaccharide other than 2' -FL.

Non-digestible oligosaccharides other than human milk oligosaccharides

The nutritional composition preferably further comprises non-digestible oligosaccharides other than HMOS. Non-digestible oligosaccharides other than HMOS can have an effect on stool consistency, in particular loosening of the stool, and this effect therefore leads to the obvious opposite result, i.e. diarrhea is not improved. However, when correcting the common effects provided by non-digestible oligosaccharides other than HMO, in particular when correcting the intrinsic effects of non-digestible oligosaccharides other than HMO on stool consistency, their addition was found to have an additional effect in significantly reducing the severity score and duration of diarrhea.

NDO is preferably not or only partially digested in the intestine by the action of acids or digestive enzymes present in the human upper gastrointestinal tract, particularly in the small intestine and stomach, and is fermented by the human intestinal microbiota. For example, sucrose, lactose, maltose and ordinary maltodextrins are considered digestible.

Preferably, the present composition comprises non-digestible oligosaccharides having a DP between 2 and 250, more preferably between 2 and 60. The non-digestible oligosaccharide is preferably at least one, more preferably at least two selected from the group consisting of: fructooligosaccharides, galactooligosaccharides, xylooligosaccharides, arabino-oligosaccharides, arabinogalacto-oligosaccharides, oligoglucosides, chitooligosaccharides, gluco-oligosaccharides, galacto-oligosaccharides, galactomannan-oligosaccharides and oligomannose. The group of fructooligosaccharides comprises inulin and the group of galactooligosaccharides comprises transgalactooligosaccharides or beta-galactooligosaccharides.

More preferably, the composition of the invention comprises Fructooligosaccharides (FOS) and/or Galactooligosaccharides (GOS), preferably the galactooligosaccharides comprise beta-and/or alpha-galactooligosaccharides. More preferably, the fructooligosaccharide is a long chain fructooligosaccharide (lcFOS). More preferably, the galactooligosaccharide is a short chain galactooligosaccharide and the beta-galactooligosaccharide is a short chain beta-galactooligosaccharide. Most preferably, the composition comprises long chain fructooligosaccharides and short chain galactooligosaccharides. The weight ratio of short-chain galacto-oligosaccharides to long-chain fructo-oligosaccharides is preferably 100:1 to 1:10, preferably 20:1 to 1:1, preferably about 9: 1.

The galactooligosaccharide is preferably a beta-galactooligosaccharide. In a particularly preferred embodiment, the composition of the invention comprises beta-galactooligosaccharides ([ galactose ] n-glucose; wherein n is an integer from 2 to 60, i.e. 2, 3, 4, 5, 6, …, 59, 60; preferably n is selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10) wherein the galactose units are mostly linked together by beta bonds. The beta-galactooligosaccharides are also known as Transgalactooligosaccharides (TOS). For example, β -galactooligosaccharides may be sold under the trade name Vivinal (TM) (Borculo Domo Ingredients, Netherlands). Another suitable source is Bi2Munno (Classado). Preferably, the galactooligosaccharides comprise beta-1, 3, beta-1, 4 and/or beta-1, 6 linkages. In a preferred embodiment, the galactooligosaccharide comprises at least 80% beta-1, 4 and beta-1, 6 linkages, based on the total linkages, more preferably at least 90%. In another preferred embodiment, the galactooligosaccharide comprises at least 50% beta-1, 3 linkages, based on the total linkages, more preferably at least 60% based on the total linkages.

Fructooligosaccharides are NDOs comprising beta-linked chains of fructose units with a DP or average DP between 2 and 250, more preferably between 2 and 100, even more preferably between 10 and 60. The fructooligosaccharides comprise inulin, fructan and/or mixed polyfructose. A particularly preferred fructooligosaccharide is inulin. Fructooligosaccharides suitable for use in the compositions are also commercially available, for example

(Orafti). Preferably, the average DP of the fructooligosaccharides is above 20.

In a preferred embodiment, the composition comprises a mixture of inulin and short chain fructooligosaccharides. In a preferred embodiment, the composition comprises a mixture of galacto-oligosaccharides and fructo-oligosaccharides, wherein the fructo-oligosaccharides are selected from short chain fructo-oligosaccharides and inulin, more preferably inulin. A mixture of at least two different non-digestible oligosaccharides advantageously stimulates beneficial bacteria of the intestinal microbiota to a greater extent. Preferably the weight ratio of the two different non-digestible oligosaccharides, preferably the mixture of galacto-oligosaccharides and fructo-oligosaccharides, is between 25 and 0.05, more preferably between 20 and 1. Galactooligosaccharides, preferably beta-galactooligosaccharides, are more capable of stimulating bifidobacteria. Preferably, the composition of the invention comprises galacto-oligosaccharides, preferably beta-galacto-oligosaccharides, with a Degree of Polymerization (DP) of 2 to 10, and/or fructo-oligosaccharides with a DP of 2 to 60.

In a preferred embodiment, the composition comprises a combination of fucosylated human lactooligosaccharides (preferably 2' -fucosyllactose), short chain galactooligosaccharides and long chain fructooligosaccharides. In a preferred embodiment, the weight ratio of the sum of short chain galacto-oligosaccharides and long chain fructo-oligosaccharides comprised in the nutritional composition to 2' -fucosyllactose is preferably from 20:1 to 1:100, preferably from 10:1 to 1:50, more preferably from 4:1 to 1:20, most preferably from 1:1 to 1: 15.

LC-PUFA

The composition may also comprise long chain polyunsaturated fatty acids (LC-PUFA). LC-PUFA are fatty acids wherein the acyl chain has a length of 20 to 24 carbon atoms (preferably 20 or 22 carbon atoms) and wherein the acyl chain comprises at least two unsaturated bonds between said carbon atoms in the acyl chain. More preferably, the present 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 DHA, EPA and/or ARA. Such LC-PUFAs have a further beneficial effect on improving the integrity of the intestinal barrier.

The preferred content of LC-PUFA in the composition of the invention is not more than 15 wt.%, preferably not more than 10 wt.%, even more preferably not more than 5 wt.% of the total fatty acids. Preferably, the present composition comprises at least 0.2 wt.% LC-PUFA, more preferably DHA, based on total fatty acids, preferably at least 0.25 wt.%, more preferably at least 0.35 wt.%, even more preferably at least 0.5 wt.%. The composition of the invention preferably comprises ARA and DHA, wherein the weight ratio ARA/DHA is preferably above 0.25, preferably above 0.5, more preferably from 0.75 to 2, even more preferably from 0.75 to 1.25. The weight ratio is preferably 20 or less, more preferably 0.5 to 5. The amount of DHA is preferably above 0.2 wt.%, more preferably above 0.3 wt.%, more preferably at least 0.35 wt.%, even more preferably 0.35-0.6 wt.% of the total fatty acids.

Composition comprising a metal oxide and a metal oxide

The present invention advantageously relates to a composition for a given use, wherein lipids provide 5 to 50% of the total calories, proteins provide 5 to 50% of the total calories, and carbohydrates provide 15 to 90% of the total calories. Preferably, in the composition of the invention, the lipid provides 35 to 50% of the total calories, the protein provides 7.5 to 12.5% of the total calories, and the carbohydrate provides 40 to 55% of the total calories. To calculate the percentage of total calories of the protein component, the total energy provided by the protein, peptide and amino acid needs to be considered.

The composition of the present invention preferably comprises at least one lipid selected from animal lipids (excluding human lipids) and vegetable lipids. Preferably, the composition of the present invention comprises a combination of vegetable lipids and at least one oil selected from the group consisting of fish oil, animal oil, algal oil, fungal oil and bacterial oil. The present composition comprising 2' -FL is not human milk.

The composition of the invention preferably comprises a protein. The protein component used in the nutritional formulation is preferably selected from the group consisting of non-human animal proteins (preferably milk proteins, preferably proteins from bovine milk), plant proteins (preferably soy protein and/or rice protein), free amino acids and mixtures thereof. The composition of the invention preferably contains casein, whey, hydrolysed casein and/or hydrolysed whey protein. Preferably, the protein comprises intact protein, more preferably intact bovine whey protein and/or intact bovine casein protein.

The present composition preferably comprises digestible carbohydrates. The present composition preferably comprises a digestible carbohydrate component wherein at least 35 wt.%, more preferably at least 50 wt.%, more preferably at least 75 wt.%, even more preferably at least 90 wt.%, most preferably at least 95 wt.% is lactose. The composition of the invention preferably comprises at least 25 grams lactose per 100 grams dry weight of the composition of the invention, preferably at least 40 grams lactose per 100 grams.

When the nutritional composition is a liquid, its caloric density is preferably from 0.1 to 2.5kcal/ml, even more preferably from 0.5 to 1.5kcal/ml, most preferably from 0.6 to 0.8 kcal/ml. The amount of nutritional composition administered per day is preferably from 50 to 2000ml, more preferably from 200 to 1500, most preferably from 400 to 1000 ml.

In one embodiment, the present invention relates to a supplement suitable for fortifying human milk to fortify human milk fortified with a standard human milk fortifier or to fortify a standard preterm formula. In the context of the present invention, the supplement does not contain all the macronutrients and micronutrients required by preterm infants to achieve growth similar to that of a fetus, as well as satisfactory functional development. The term "preterm birth" herein is synonymous with premature birth and refers to any human infant born prior to the 37 th week of gestation.

Thus, in one embodiment the nutritional composition according to the invention or for said use according to the invention comprises protein, fat and/or digestible carbohydrates and is selected from the group consisting of infant initial formula, infant follow-up formula, baby milk, preterm formula, post-discharge formula and human milk fortifier.

Applications of

According to the present invention, virus induced diarrhea is treated by administering a nutritional composition comprising fucosylated human lacto-oligosaccharides, preferably 2' -fucosyllactose. In a preferred embodiment, the diarrhea treated is caused by rotavirus, also known as rotavirus-induced diarrhea.

In a preferred embodiment, the treatment of viral-induced diarrhea is to reduce the severity of viral-induced diarrhea. In another preferred embodiment, the treatment of viral-induced diarrhea is to reduce the duration of viral-induced diarrhea. Preferably, the treatment of viral-induced diarrhea is to reduce the severity and shorten the duration of viral-induced diarrhea.

In a preferred embodiment, the fucosylated human lactooligosaccharide, preferably 2' -fucosyllactose, is administered or used in order to reduce the severity of virus-induced diarrhea. In another preferred embodiment, the fucosylated human lactooligosaccharide, preferably 2' -fucosyllactose, is administered or used in order to shorten the duration of virus-induced diarrhea. Preferably, the fucosylated human lactooligosaccharide, preferably 2' -fucosyllactose, is administered or used in order to reduce the severity and duration of the virus-induced diarrhea.

The nutritional composition for use according to the invention is for use in a human subject. Preferably, the human subject is an infant or a young child. Infants are human subjects of 0-12 months of age. Young children are human subjects of 12-36 months of age. In a preferred embodiment, the human subject is a preterm or prematurely born infant. In particular, this subgroup of infants may benefit from the administration of fucosylated human milk oligosaccharides, preferably 2 '-fucosyllactose, or fucosylated human milk oligosaccharides, preferably 2' -fucosyllactose, in combination with long chain fructooligosaccharides and short chain galactooligosaccharides, since unlike infants born at term, the organs constituting the intestine of preterm infants are fragile and immature and therefore require a reduction in the duration of diarrhea and/or a reduction in the severity of diarrhea as the treatment progresses.

Preferably, the human subject is administered a fucosylated human milk oligosaccharide, preferably 2 '-fucosyllactose, or a nutritional composition comprising a fucosylated human milk oligosaccharide, preferably 2' -fucosyllactose, daily. Furthermore, the administration of the nutritional composition comprising fucosylated human lactooligosaccharide, preferably 2' -fucosyllactose, or the use thereof, preferably starts before the occurrence of virus-induced diarrhea and lasts for a period of time during which the subject may be diagnosed with diarrhea.

In a preferred embodiment, the human subject is vaginally born, also known as naturally born.

In a preferred embodiment, the reduction in the severity of the virus-induced diarrhea and/or the reduction in the duration of the virus-induced diarrhea is accompanied by an improvement in the elimination of the virus or a reduction in the count of virus particles in the faeces in the human subject. The present inventors have found that the addition of 2' -fucosyllactose, short chain galacto-oligosaccharides and long chain fructo-oligosaccharides reduces shedding of viral particles from human subject faeces. This is particularly advantageous for preventing viral transmission or for preventing secondary rounds of viral diarrhoea through self-contamination by faeces after infection with a primary virus, such as rotavirus. Thus, in a preferred embodiment, the reduction in severity of the viral-induced diarrhea and/or the reduction in duration of the viral-induced diarrhea is accompanied by a reduction in viral elimination in the human subject.

The composition of the invention is preferably administered enterally, more preferably orally. The composition of the invention is preferably a nutritional formula, preferably an infant formula. The composition of the invention can advantageously be used as a complete nutrition for infants. The composition of the invention preferably comprises lipids, proteins and carbohydrates, and is preferably administered in liquid form. The present invention includes dry compositions (e.g., powders) accompanied by instructions for mixing the dry compositions (particularly nutritional compositions) with a suitable liquid (e.g., water).

Detailed Description

Example 1

Newborn rats were divided into 5 groups of 24 animals each: reference (REF) group, rotavirus infection (RV) group, and 3 rotavirus infection groups supplemented with the following respectively: a) mixture of scGOS and lcFOS (RV + GOS/FOS group); b)2 '-FL (RV + 2' -FL group); and c) scGOS/lcFOS and 2 '-FL (RV + GOS/FOS + 2' -FL group).

The same normalized volume/body weight (4.5 μ L/g/day) of all products was orally administered to lactating rats once daily from day two to day sixteenth of life (corresponding to a strict lactation period). The RV + GOS/FOS group was supplemented with 0.8g scGOS/lcFOS/100g body weight. The RV +2 '-FL group was supplemented with 0.2g of 2' -FL per 100g of body weight. The RV + GOS/FOS + 2' -FL group received the same dose of both products as given alone and maintained the administration volume (4.5. mu.L/g/day). The REF group and RV group were given the same volume of water.

RV (simian SA-11) was obtained as described previously (Perrez-Cano et al. Peditar Res 2007; 62: 658-63), and inoculated on day 5 of life (4X 10) in all experimental groups except the REF group⁸Tissue culture infectious dose 50[ TCID50]Rat), the REF group received the same volume of Phosphate Buffered Solution (PBS) under the same conditions.

Body weight was recorded daily throughout the study to assess weight gain. Half of each group of animals (n-12) were sacrificed on day 8 to analyze variables associated with peak diarrhea and the other half (n-12/group) were sacrificed on day 16 to analyze the effect of the supplement after diarrhea was resolved. In addition, the length of the nose-anus and tail is measured to determine the body/tail ratio, in terms of body weight/body length²(g/cm²) Calculated Body Mass Index (BMI) and^0.33length) × 1000 (g)^0.33/cm) calculated Lee index.

Stool sample collection and clinical indices

Stool sampling was performed daily by gently pressing and massaging the abdomen throughout the study (from day 4 to day 16 of life). Fecal samples were stored at-20 ℃ for analysis of RV shedding. Severity of diarrhea was expressed as stool weight and stool samples were scored from 1 to 4 (diarrhea index [ DI ]) according to color, texture and amount as follows: normal stool (1); soft yellow-green feces (2); completely loose yellowish green faeces (3); a large amount of watery feces (4). A score of 2 or more indicates diarrheal stools, while a score of < 2 indicates no diarrhea. The area of severity under the curve (S-AUC), consistent with the time that the animals showed diarrhea over the course of days 5-11, was calculated as the total value of severity. Maximum Severity (MS) was defined as the highest score for the period of diarrhea. The period of Diarrhea (DP) is calculated by counting the number of days the animals show a DI ≧ 2.

In the RV + GOS/FOS and RV + GOS/FOS + 2' -FL groups, DI, S-AUC, MS and DP were normalized (nDI, nS-AUC, nMS and nDP) while taking into account the baseline values before and after the period of diarrhea due to intrinsic fecal aspects of GOS/FOS supplementation, as described previously (Rigo-Adroller et al. Eur J Nutr 2017; 56: 1657-70).

Statistical analysis

Statistical analysis was performed using statistical software package for social sciences (SPSS v22.0) (IBM, Chicago, IL, USA). Data were tested for homogeneity of variance and normal distribution using the Levene and Shapiro-Wilk tests, respectively. When the data is homogeneous and behaving normally, a conventional one-way ANOVA test is performed, followed by post hoc Bonferroni. Otherwise, the nonparametric Kruskal-Wallis test is performed first, followed by the post-operative Mann-Whitney U (MWU) test. Finally, the chi-square test was used to compare the frequency of diarrhea occurrence. When p < 0.05, a significant difference was established.

Results

Growth of

RV infection did not produce any significant changes in growth, either during the peak diarrhea (day 8) or at the end of the study (day 16), as shown by the results for body weight, body/tail ratio, BMI and Lee index (table 1). At the end of the study (day 16, p < 0.05), the body weight of the group supplemented with scGOS/lcFOS was slightly higher, although none of these growth changes changed BMI, but some differences were observed in the body/tail length ratio and Lee index. At some time points, all supplements increased body/tail length ratio compared to REF or RV. Furthermore, the Lee index is only reduced during the peak of diarrhea compared to REF.

TABLE 1 variables relating to growth

Results are expressed as mean ± s.e.m. (n-12); p < 0.05 compared to REF (by MWU test); compared with the RV, the composite material has the advantages that,^#p < 0.05 (by MWU assay).

Clinical variables for assessing severity and duration of diarrhea

Rats vaccinated with RV showed diarrhea feces from day 6 to day 11 of life, with the highest score at day 8. As shown in previous studies using the SA11 RV infection model, GOS/FOS caused changes in fecal consistency, increasing the number of feces considered diarrhea (DI ≧ 2) before, during, and after the course of diarrhea (Rigo-Adroller et al Eur J Nutr 2017; 56: 1657-70). However, supplementation with GOS/FOS and GOS/FOS + 2' -FL showed a tendency to reduce the severity of diarrhea at the point of greatest impact. When normalized to the severity score value of the group supplemented with GOS/FOS, significant reductions were found on days 7, 8 and 9 (p < 0.05). Supplementation with 2' -FL did not induce this basal fecal consistency effect and showed a tendency to reduce the severity of diarrhea.

Other indicators of the severity and duration of diarrhea were also calculated (table 2). With respect to the maximum severity of diarrhea (MS), supplementation with 2 '-FL and GOS/FOS + 2' -FL resulted in a score reduction of up to 15% and 10%, respectively, (p < 0.05) compared to the RV group. Upon normalization of the index (nMS), a smaller score was observed for the GOS/FOS group, showing a 28-30% decrease (p < 0.05). Calculation of the area of severity under the curve (S-AUC) shows the average severity for each group. For the RV group, a clear trend of decreasing S-AUC by 22% was observed for the supplemental 2' -FL. Only after normalized area did nS-AUC decrease in animals supplemented with GOS/FOS and GOS/FOS + 2' -FL (45% and 58%, respectively, p < 0.05). Finally, an improvement was also found in reducing the Diarrheal Phase (DP), which was reduced by more than 50% (p < 0.05) due to the supplement.

TABLE 2 analysis of other variables related to severity, incidence and duration of diarrhea

Results for severity and duration variables are expressed as mean ± s.e.m. (n-12-24, depending on the number of stool samples obtained per day); compared with the RV, the composite material has the advantages that,^#p < 0.05 (by MWU test and Chi Square test)

Stool weight was also measured as an objective severity variable for the course of diarrhea. Animals receiving GOS/FOS and GOS/FOS + 2' -FL had higher fecal weights before RV inoculation (corresponding to days 2-5 of life) compared to the REF and RV groups, which could be related to previously reported direct changes in fecal consistency. Following infection with virus (corresponding to days 6-11 of life), RV groups showed an increase in stool weight (p < 0.05). The effect of oligosaccharide supplementation was observed on days 6-8, which reduced the increase in stool weight, despite the statistical difference (p < 0.05) only in supplementation with 2' -FL. The lower stool weights observed in the 2' -FL group were maintained until the end of the diarrhea period (days 9-11). At the end of the course of diarrhea (days 12-16 of late life in diarrhea), the fecal weight of RV and 2 '-FL was similar to that of REF, while supplementation with GOS/FOS and GOS/FOS + 2' -FL continued to show higher values due to nonpathogenic changes in fecal consistency.

Example 2: reducing the severity of or shortening rotavirus-induced diarrhea in infants Infant formula

The infant formula according to the invention comprises per 100ml (13.9 dry weight):

1.4g protein (whey and casein)

7.3g digestible carbohydrates (including lactose)

3.6g fat (vegetable fat, fish oil)

0.6g non-digestible oligosaccharides, wherein 60mg 2' -fucosyllactose and 480mg beta-galacto-oligosaccharides and 60mg fructo-oligosaccharides

According to the guidelines also contain: choline, inositol, taurine, minerals, trace elements and vitamins.

Claims

1. A nutritional composition comprising fucosylated human lacto-oligosaccharides, preferably 2' -fucosyllactose, for use in the treatment of virus-induced diarrhea in a human subject.

2. Nutritional composition for use according to claim 1, wherein the virus-induced diarrhea is rotavirus-induced diarrhea.

3. Nutritional composition for use according to claim 1 or 2, wherein the treatment is reducing the severity of virus-induced diarrhoea.

4. Nutritional composition for use according to any one of the preceding claims, wherein the treatment is a reduction in the duration of virus-induced diarrhea.

5. Nutritional composition for use according to any one of the preceding claims, wherein the nutritional composition further comprises at least one, more preferably at least two non-digestible oligosaccharides with a DP between 2 and 250, wherein the non-digestible oligosaccharides are selected from the group consisting of fructooligosaccharides, galactooligosaccharides, xylooligosaccharides, arabinooligosaccharides, arabinogalactooligosaccharides, glucooligosaccharides, chitooligosaccharides, glucomannooligosaccharides, galactomannooligosaccharides and oligomannose.

6. Nutritional composition for use according to claim 5, wherein the non-digestible oligosaccharides are selected from the group consisting of fructooligosaccharides and galactooligosaccharides.

7. Nutritional composition for use according to any one of the preceding claims, wherein the nutritional composition is an infant formula or a follow-on formula, preferably an infant formula.

8. Use of fucosylated human milk oligosaccharides, preferably 2' -fucosyllactose, for the preparation of a nutritional composition for the treatment of virus-induced diarrhea in a human subject.

9. A method for treating virus-induced diarrhea in a human subject by administering a nutritional composition comprising fucosylated human lacto-oligosaccharides, preferably 2' -fucosyllactose.