US20090221486A1

US20090221486A1 - Method for treating and/or preventing infections in infants delivered via caesarean section

Info

Publication number: US20090221486A1
Application number: US12/091,050
Authority: US
Inventors: Joachim Schmitt; Bernd Stahl; Jan Knol
Original assignee: Nutricia NV
Current assignee: Nutricia NV
Priority date: 2005-10-21
Filing date: 2006-10-06
Publication date: 2009-09-03
Also published as: WO2007046699A3; ES2857732T3; CA2620821C; ES2451020T3; EP2638812A1; PT2407036E; SG10201503970XA; PL3326634T3; HUE028348T2; EP2984943A1; PT1940245E; DK2105055T3; WO2007046698A1; EP2140771A1; BRPI0617653B8; EP2407036B1; EP1940250B1; RU2424736C2; CN103816173B; RU2008119992A

Abstract

The present invention provides the use of a composition comprising uronic acid oligosaccharide for the manufacture of a composition for enteral administration to an infant delivered via caesarean section.

Description

FIELD OF THE INVENTION

The present invention relates to methods for feeding infants delivered via caesarean section.

BACKGROUND OF THE INVENTION

Human milk comprises non-digestible oligosaccharides which specifically stimulate the growth of lactic acid producing bacteria, such as species belonging to the genus Bifidobacterium and Lactobacillus and prevent the growth and/or adhesion to the intestinal wall of other (pathogenic) bacteria. Hence, when an infant receives human milk, the infant's intestinal flora develops into a healthy flora rich in lactic acid producing bacteria. The presence of a healthy intestinal flora improves gut barrier maturation and/or gut barrier integrity, stimulates the formation of mucus and inhibits pathogen growth and stimulates the immune system.
WO2005039597 discloses a method for enhancing the immune system and the treatment and/or prevention of immune system related disorders in a mammal, particularly newborns, said method comprising the administration of acid oligosaccharide and neutral oligosaccharide.
Before birth the intestinal tract of the infant is normally sterile. During vaginal delivery the intestinal tract of the infant is inoculated with vaginal and/or fecal bacteria of the mother, resulting in a colonization of the infant's gastrointestinal tract by bacteria originating from the infant's environment.
Natren® produces the probiotic product Life Start® which is designed specifically for infants and suitable for infants delivered via caesarean section. Life Start® is made with Bifidobacterium infantis. Because the Life Start® product contains only one single Bifidobacteria species, the benefits for the infant will be very limited.

SUMMARY OF THE INVENTION

The present inventors have found that infants delivered via caesarean section have an intestinal flora which is different from the intestinal flora of infants born via the vaginal route. Particularly, infants born via caesarean section have a reduced rate of intestinal colonization by Bifidobacteria and have a less diverse Bifidobacterium intestinal flora regarding species than infants born via the vaginal route, particularly missing Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium infantis and Bifidobacterium bifidum (Example 1). It was further found that the intestinal flora of infants delivered via caesarean section have a lower content of Bifidobacteria compared to the intestinal flora of infants delivered vaginally. Additionally it was found that the intestinal flora of infants born via caesarean section has a high content of (undesirable) Escherichia coli 6 weeks after delivery.
Overcoming these deficiencies results in improved health and prevents and/or treats a variety of disorders.
A healthy and fully developed gastrointestinal flora has important physiological effects. One important aspect is that it reduces the occurrence of (gastrointestinal) infections. Because infants delivered via a caesarean section lack a healthy flora, preventing infection is particularly important for these infants. These infants are normally delivered in a hospital environment, which is a risk for pathogenic infection due to the occurrence of nosocomial bacteria. Additionally, the impaired development of a healthy intestinal flora results in faster colonization of pathogenic bacteria compared to a situation where the infants intestinal tract is inoculated by maternal bacteria.
The present invention particularly aims to decrease the number and severity of (gastrointestinal) infections in infants born via caesarean section, by (i) decreasing the adhesion of pathogenic bacteria to the intestinal epithelial cells and/or intestinal mucus; (ii) decreasing the growth of pathogenic bacteria; (iii) stimulating the growth of beneficial bacteria, preferably lactic acid producing bacteria and/or iv) modulate the immune system.
The present inventors have found that (gastrointestinal) infections can be prevented and treated by uronic acid oligosaccharides, particularly oligosaccharides derived from pectin. The present uronic acid oligosaccharides are suitable for enteral (particularly oral) administration, making it easy to include these in infant milk formula. The present uronic acid oligosaccharide reduces the occurrence of the infections, as well as reduces the severity of the infection due to its anti-adherence effects (see Example 2).
The occurrence and severity of infection in infants delivered via caesarean section can be even further reduced by providing, in addition to the present uronic acid oligosaccharide, one or more prebiotic oligosaccharides. These oligosaccharides stimulate the development of a healthy gastrointestinal flora. Galacto-oligosaccharides, and/or fructo-oligosaccharides are particularly suitable as these were found to stimulate the growth of B. iongum, B. breve and/or B. infantis. (Example 3).
In addition to reduced occurrence of infection, important other disorders can be prevented in the infants delivered via caesarean section by the present method. Disorders such as allergy and eczema are prevented by stimulating a healthy intestinal flora.
In a further aspect the present invention can be suitably brought to practice by incorporation of the present active ingredients in a nutritional composition. Such composition can be administered to the infant without posing a heavy burden on the infant delivered via caesarean section.

DETAILED DESCRIPTION OF THE PREFFERED EMBODIMENTS

The present method provides the use of a composition comprising uronic acid oligosaccharide for the manufacture of a composition for enteral administration to an infant delivered via caesarean section. In a further aspect the present method provides the use of a composition comprising uronic acid oligosaccharide for the manufacture of a composition for i) treatment and/or prevention of disorders in infants delivered via caesarean section and/or ii) the stimulation of health in infants delivered via caesarean section. In a further aspect the present invention provides a method for administering uronic acid oligosaccharides to an infant delivered via caesarean section, preferably a method for the prevention and/or treatment of infection and/or the prevention and/or treatment of allergy in infants delivered via caesarean section.
In a further aspect the present invention provides a method for stimulating the development of a healthy intestinal flora and/or decreasing the occurrence of intestinal pathogens in an infant born via caesarean section comprising the steps of:

- a) admixing I) a nutritionally or pharmaceutically acceptable liquid; and II) a dry composition, wherein the dry composition II comprises uronic acid oligosaccharides, preferably selected from the group consisting of guluronic acid oligosaccharide, iduronic acid oligosaccharide, riburonic acid oligosaccharide, mannuronic acid oligosaccharides, galacturonic acid oligosaccharides and glucuronic acid oligosaccharides; and
- b) administering the composition obtained in step a) to the infant.

In still a further aspect the present invention provides a method for stimulating the development of a healthy intestinal flora and/or decreasing intestinal pathogens in an infant delivered via caesarean section, comprising administering to the infant a composition comprising uronic acid oligosaccharides, preferably selected from the group consisting of guluronic acid oligosaccharides, iduronic oligosaccharides, riburonic oligosaccharides, mannuronic acid oligosaccharides, galacturonic acid oligosaccharides and glucuronic acid oligosaccharides.

Caesarean Section

The present invention relates to the enteral administration of a composition comprising uronic acid oligosaccharides to infants delivered via caesarean section. A caesarean section (c-section) is a surgical procedure where an infant is delivered through an incision made in the mother's abdominal wall, and then through the wall of the uterus. A caesarean section is usually performed when it is safer for the mother or the infant than a vaginal delivery. Alternatively, a woman may choose to have a caesarean section rather than deliver her infant vaginally.

Uronic Acid Oligosaccharides

The present composition comprises uronic acid oligosaccharide. The term uronic acid oligosaccharide as used in the present invention refers to an oligosaccharide wherein preferably at least 25%, preferably at least 50%, of the monosaccharide units present in the oligosaccharide is a uronic acid, preferably one selected from the group consisting of guluronic acid, iduronic acid, riburonic acid, mannuronic acid, galacturonic acid and glucuronic acid. In a preferred embodiment the uronic acid oligosaccharide comprises at least 50% galacturonic acid based on total uronic acid units in the uronic acid oligosaccharide. The uronic acid oligosaccharides used in the invention are preferably prepared from pectin, pectate, alginate, chondroitine, hyaluronic acids, heparine, heparane, bacterial carbohydrates, and/or sialoglycans, more preferably from pectin and/or alginate, even more preferably from pectin, most preferably polygalacturonic acid. The present uronic acid oligosaccharide is preferably a pectin degradation product and/or alginate degradation product. Preferably the pectin degradation product is a pectin hydrolysate (prepared by hydrolysis) and/or pectin lysate (prepared by beta-elimination). The pectin degradation product is preferably prepared from fruit or vegetable pectin, more preferably from apple pectin, citrus pectin and/or sugar beet pectin, more preferably from apple, citrus and/or sugar beet pectin. The pectin degradation product is preferably prepared with lyases and/or variations of the temperature and pressure, more preferably with pectin lysate, i.e. by beta-elimination. The pectin degradation product is preferably a pectin lysate
Preferably the present composition comprises uronic acid oligosaccharide with a degree of polymerization (DP) of 2 to 250, more preferably a DP of 2 to 100, even more preferably a DP of 2 to 50, most preferably a DP of 2 to 20. Preferably the present composition comprises between 25 and 100 wt. %, more preferably between 50 and 100 wt. % uronic acid oligosaccharide with a DP of 2 to 250 based on total weight of uronic acid in the composition, more preferably a DP of 2 to 100, even more preferably a DP of 2 to 50, most preferably a DP of 2 to 20.
The present uronic acid oligosaccharide is preferably obtainable by enzymatic digestion of pectin with pectin lyase, pectic lyase, endopolygalacturonase and/or pectinase. Preferable the pectin lysate and/or the galacturonic acid oligosaccharide is prepared from bacterial production.
In a preferred embodiment at least one of the terminal hexuronic acid units of the uronic acid oligosaccharide has a double bond, which is preferably situated between the C₄and C₅position of the terminal hexuronic acid unit. The double bond effectively protects against attachment of the pathogenic bacteria to the intestinal epithelial cells. Preferably one of the terminal hexuronic acid units comprises the double bond. The double bond at terminal hexuronic acid unit can for example be obtained by enzymatic hydrolysis of pectin with lyase.
In a further embodiment a mixture of uronic acid oligosaccharides is used, which have a different DP and/or comprise both unsaturated and saturated terminal hexuronic acid units. Preferably at least 5%, more preferably at least 10%, even more preferably at least 25% of the terminal hexuronic acid units of the uronic acid oligosaccharide is an unsaturated hexuronic acid unit. As each individual uronic acid oligosaccharide preferably comprises only one unsaturated terminal hexuronic acid unit, preferably less than 50% of the terminal hexuronic acid units is an unsaturated hexuronic acid unit (i.e. comprises a double bond). A mixture of uronic acid oligosaccharides preferably contains between 2 and 50% unsaturated terminal hexuronic acid units based on the total amount of terminal hexuronic acid units, preferably between 10 and 40%.
The uronic acid oligosaccharide can be derivatised. The uronic acid oligosaccharide may be methoxylated and/or amidated. In one preferred embodiment the uronic acid oligosaccharides are characterized by a degree of methoxylation above 20%, preferably above 30% even more preferably above 70%. As used herein, “degree of methoxylation” (also referred to as DE or “degree of esterification”) is intended to mean the extent to which free carboxylic acid groups contained in the uronic acid oligosaccharide have been esterified (e.g. by methylation). In another preferred embodiment the uronic acid oligosaccharides have a degree of methylation above 20%, preferably above 30% even more preferably above 70%.
The present composition preferably comprises between 0.01 and 10 grams uronic acid oligosaccharide with a DP of 2 to 250 per 100 gram dry weight of the present composition, more preferably between 0.05 and 6 gram, even more preferably 0.2 to 2 gram per 100 gram dry weight. The present composition preferably comprises between 0.01 and 10 grams galacturonic acid oligosaccharide with a DP of 2 to 250 (more preferably a DP of 2 to 100) per 100 gram dry weight of the present composition, more preferably between 0.05 and 6 gram, even more preferably 0.2 to 2 gram.
The uronic acid oligosaccharides are preferably non-digestible in the upper human intestinal tract and water-soluble (according to the method disclosed in L. Prosky et al, J. Assoc. Anal. Chem 71: 1017-1023, 1988). The uronic acid oligosaccharides are preferably fermentable by the intestinal flora. The uronic acid oligosaccharides of the invention advantageously reduce the adhesion of pathogenic micro-organisms to the intestinal epithelial cells, thereby reducing colonization of (nosocomial) pathogenic bacteria in the colon of the infant delivered by caesarean section. Furthermore, the uronic acid oligosaccharides of the present invention preferably stimulate the formation of a healthy intestinal flora and/or modulate the immune system and are fermented, resulting in a production of intestinal organic acids and a reduction of intestinal pH, which inhibit the growth of (nosocomial) pathogenic bacteria.
Non-Digestible Oligosaccharides other than Uronic Acid Oligosaccharides
In addition to the uronic acid oligosaccharide, the present composition preferably comprises non-digestible oligosaccharide other than uronic acid oligosaccharides (hereinafter referred to as “non-digestible neutral oligosaccharide”), which preferably stimulates the growth of the intestinal lactic acid producing bacteria, particularly Bifidobacteria and/or the Lactobacilli.
The term “non-digestible oligosaccharide” as used in the present invention refers to oligosaccharides/carbohydrates which are not or only partially digested in the intestine by the action of acids or digestive enzymes present in the human upper digestive tract (small intestine and stomach) but which are fermented by the human intestinal flora. Preferably the present non-digestible oligosaccharide comprises an oligosaccharide with a degree of polymerization (DP) of 2 to 250, more preferably 2 to 100, even more preferably a DP 2 to 60, even more preferably 2 to 10. For example, sucrose, lactose, maltose and the maltodextrins are considered digestible. Preferably the non-digestible oligosaccharide other than uronic acid oligosaccharide is a non-digestible neutral oligosaccharide. The term “neutral oligosaccharide” as used in the present invention refers to oligosaccharides wherein more than 75% of the saccharide units are selected from the group consisting of glucose, fructose, fucose, galactose, mannose, ribose, arabinose, rhamnose, and xylose, preferably more than 85%, more preferably more than 95%, even more preferably more than 99%. Preferred non-digestible neutral oligosaccharides are transgalacto-oligosaccharides and fructo-oligosaccharides.
Preferably the present non-digestible oligosaccharide is a prebiotic oligosaccharide. The term “prebiotic oligosaccharide” refers to non-digestible oligosaccharide that beneficially affects the host by selectively stimulating the growth and/or activity of one or a limited number of bacterial species in the colon.
Preferably the present non-digestible oligosaccharide is soluble. The term “soluble” as used herein, when having reference to a polysaccharide, fibre or oligosaccharide, means that the substance is soluble according to the method described by L. Prosky et al., J. Assoc. Off. Anal. Chem. 71, 1017-1023 (1988).
Preferably the present composition comprises at least one oligosaccharide selected from the group consisting of galacto-oligosaccharides, non-digestible dextrins, xylo-oligosaccharides, arabino-oligosaccharides, gluco-oligosaccharides (including gentio-oligosaccharides and cyclodextrins), chito-oligosaccharides, fuco-oligosaccarides, manno-oligosaccharides, isomalto-oligosaccharides, glucomanno-oligosaccharides, galactomanno-oligosaccharides, arabinogalacto-olgosaccharides, and fructo-oligosaccharides, more preferably galacto-oligosaccharides and fructo-oligosaccharides, most preferably galacto-oligosaccharides.
The term “fructo-oligosaccharide” as used herein refers to a carbohydrate comprising a chain of at least 3 β-linked fructose units, with a DP between of 2 to 250, preferably between 7 to 100, more preferably 20 to 60. Preferably inulin is used. Inulin is available under the tradename “Raftilin HP®”, (Orafti). The average DP of the present fructo-oligosaccharide is preferably at least 7, more preferably at least 10, preferably below 100. The inulin used preferably has the fructose units linked with a β(2→1) linkage. Fructo-oligosaccharides in the present invention include compounds to which names such as inulin, fructopolysaccharide, polyfructose, fructans, or oligofructose are given. The present composition preferably comprises fructo-oligosaccharides with a DP of 2 to 100.
Non-digestible polydextrins refer to digestion-resistant (malto)dextrins or digestion-resistant polydextrose which have a DP of 10 to 50, preferably between 10 and 20. The non-digestible polydextrins comprise α(1→4), α(1→6) glucosidic bonds and 1→2 and 1→3 linkages Non-digestible polydextrins are for example available under the tradename “Fibersol 2®” from Matsutami Industries or Litesse® from Danisco.
The present inventors found that galacto-oligosaccharides can be advantageously used in the present composition, because these oligosaccharides where particularly effective in stimulating the growth of Bifidobacteria. Hence, in a preferred embodiment the present composition comprises galacto-oligosaccharides. The term “galacto-oligosaccharide” as used herein refers to a non-digestible oligosaccharide, wherein at least 30% of the saccharide units are galactose units, preferably at least 50%, more preferably at least 60%. The present composition preferably comprises galacto-oligosaccharides with a DP of 2 to 100, more preferably a DP of 2 to 10. Preferably the saccharides of the galacto-oligosaccharide are β-linked, as is the case in human milk oligosaccharide-core structures.
Preferably the present composition comprises a galacto-oligosaccharide selected from the group consisting of transgalacto-oligosaccharides, lacto-N-tetraose (LNT) and lacto-N-neotetraose (neo-LNT). In a particularly preferred embodiment the present method comprises the administration of transgalacto-oligosaccharide ([galactose]_n-glucose; wherein n is an integer between 1 and 60, i.e. 2, 3, 4, 5, 6, . . . , 59, 60; preferably n is 2, 3, 4, 5, 6, 7, 8, 9 and/or 10). Transgalacto-oligosaccharides (TOS) are for example sold under the trademark Vivinal™ (Borculo Domo Ingredients, Netherlands). Preferably the saccharides of the galacto-oligosaccharides are β-linked.
The present composition preferably comprises 0.5 to 75 grams of the non-digestible oligosaccharides per 100 gram dry weight, preferably between 0.5 and 50 grams. The present composition preferably comprises 0.1 to 95 grams of the galacto-oligosaccharides per 100 gram dry weight, preferably between 0.1 and 50 grams.
The present method preferably comprises the administration of a serving comprising between 0.05 and 25 grams non-digestible oligosaccharide, preferably between 0.1 and 5 grams. The present method preferably comprises the administration of a serving comprising between 0.05 and 25 grams galacto-oligosaccharides, preferably between 0.1 and 5 gram galacto-oligosaccharides.
The present inventors have also found that a mixture of a long chain non-digestible oligosaccharides and short chain non-digestible oligosaccharides synergistically stimulates the growth of a healthy intestinal flora, particularly Bifidobacteria and reduces the occurrence of E. coli in infants delivered via caesarean section.
The present composition thus preferably comprises at least two non-digestible neutral oligosaccharides with different average degrees of polymerization (DP). Preferably the weight ratios:

- a. (non-digestible neutral oligosaccharides with DP 2 to 5): (non-digestible neutral oligosaccharides with DP 6, 7, 8, and/or 9)>1; and/or
- b. (non-digestible neutral oligosaccharides with DP 10 to 60): (non-digestible neutral oligosaccharides with DP 6, 7, 8, and/or 9)>1.
  Preferably both weight ratios are above 2, even more preferably above 5.

For further improvement, the present non-digestible oligosaccharide preferably has a relatively high content of short chain non-digestible oligosaccharides, as these strongly stimulate the growth of Bifodobacteria. Hence, preferably at least 10 wt. % of the non-digestible oligosaccharides in the present composition has a DP of 2 to 5 (i.e. 2, 3, 4, and/or 5) and at least 5 wt. % has a DP of 10 to 60. Preferably at least 50 wt. %, more preferably at least 75 wt. % of the non-digestible oligosaccharides have a DP of 2 to 9 (i.e. 2, 3, 4, 5, 6, 7, 8, and/or 9).
To improve the biodiversity and stimulate the growth of multiple intestinal organisms, the present composition preferably comprises two non-digestible oligosaccharides with a different structure. The present composition comprises at least two different non-digestible oligosaccharides, wherein the non-digestible oligosaccharides have a homology in saccharide units below about 90%, preferably below 50%, even more preferably below 25%, even more preferably below 5%. The term “homology” as used in the present invention is the cumulative of the percentage of same saccharide unit in the different non-digestible oligosaccharides. For example, oligosaccharide 1 (OL1) has the structure fruc-fruc-glu-gal, and thus comprises 50% fruc, 25% gal and 25% glu. Oligosaccharide 2 (OL2) has the structure fruc-fruc-glu, and thus comprises 66% fruc, 33% glu. The different non-digestible oligosaccharides thus have a homology of 75% (50% fruc+25% glu).
The present composition preferably comprises galacto-oligosaccharides and fructo-oligosaccharides, more preferably transgalacto-oligosacharides with a DP of 2-7 and fructo-oligosaccharides with a DP of 10-100.

Lactic Acid Producing Bacteria

The present composition preferably comprises lactic acid producing bacteria, either living or dead. Dead bacteria can for example be prepared by inactivating living bacteria by heat treatment and/or sonication. Living lactic acid bacteria are advantageously incorporated to stimulate fast colonization of the infant's intestinal tract. Lactic acid producing bacteria are preferably provided as a mono- or mixed culture of live microorganisms. Lactic acid producing bacteria are preferably provided as a mono- or mixed culture of live microorganisms. The present composition preferably comprises 10²to 10¹³colony forming units (cfu) of lactic acid producing bacteria per gram dry weight of the present composition, preferably 10²to 10¹², more preferably 10⁵to 10¹⁰, most preferably from 10⁴to 5×10⁹cfu.
Preferably the present composition comprises bacteria of the genus Lactobacillus or Bifidobacterium. Preferably the composition comprises at least one Bifidobacterium selected from the group consisting of B. longum, B. breve, B. infantis, B. catenulatum, B. pseudocatenulatum, B. adolescentis, B. animalis, B. gallicum, B. lactis and B. bifidum, more preferably B. breve, B. infantis, B. bifidum, B. catenulatum, B. Iongum, more preferably B. iongum and B. breve, more preferably B. breve. Preferably the composition comprises at least two different Bifidobacterium species, subspecies or strains. The present composition preferably comprises at least one, more preferably at least two, even more preferably at least three, most preferably at least four different Bifidobacterium species. The present composition preferably comprises at least one, more preferably at least two, even more preferably at least three, most preferably at least four different Bifidobacterium strains. Preferably the present composition comprises at least B. longum and B. breve. The above-mentioned combinations commonly aim to increase the diversity and/or the quantity of microorganisms in the intestine of the caesarean section delivered infant. This beneficially affects the infant, proving numerous health benefits.
Preferably the present composition comprises a Lactobacillus selected from the group consisting of L. casei, L. reuteri, L paracasei, L. rhamnosus, L. acidophilus, L. johnsonii, L. lactis, L. salivarius, L. crispatus, L. gasseri, L. zeae, L. fermentum and L. plantarum, more preferably L. casei, L. paracasei, L. rhamnosus, L. johnsonii, L. acidophilus, L. fermentum and most preferably L. paracasei. Even more preferably the present composition comprises Bifidobacterium breve and/or Lactobacillus paracasei, because the growth of these bacteria in impaired in the intestine of formula fed infants compared to breast fed infants. The further increased biodiversity will have a stimulatory effect on health of the newborn delivered by caesarean section.

Long-Chain Polyunsaturated Fatty Acids

The present composition preferably comprises long chain polyunsaturated fatty acids (LC-PUFA). LC-PUFA are long chain fatty acids or acyl chains with a length of 22 to 24, preferably 20 and 22, carbon atoms comprising two or more double bonds. The present composition more preferably comprises eicosapentaenoic acid (EPA, n-3), docosahexaenoic acid (DHA, n-3) and/or arachidonic acid (ARA, n-6), since these LC-PUFA effectively reduce intestinal tight junction permeability. Reduced tight junction permeability reduced the occurrence of infection and/or reduces passage of allergens. Hence incorporation of EPA, DHA and/or ARA in the present composition improves intestinal barrier integrity, which is of utmost important for babies delivered via a caesarean section since these babies have a less developed intestinal flora and hence a slower maturing gut barrier. The incorporation of these LC-PUFA's will further contribute (synergistically) to the reduced occurrence and severity of infections and allergy.
Since low concentration of ARA, DHA and/or EPA are already effective in reducing tight junction permeability, the content of LC-PUFA with 20 and 22 carbon atoms in the present composition preferably does not exceed 15 wt. % of the total fat content, preferably does not exceed 10 wt. %, even more preferably does not exceed 5 wt. % of the total fat content. Preferably the present composition comprises at least 0.1 wt. %, preferably at least 0.25 wt. %, more preferably at least 0.6 wt. %, even more preferably at least 0.75 wt. % LC-PUFA with 20 and 22 carbon atoms of the total fat content. For the same reason, the EPA content preferably does not exceed 5 wt. % of the total fat, more preferably does not exceed 1 wt. %, but is preferably at least 0.03 wt. %, more preferably at least 0.05 wt. % of the total fat. The DHA content preferably does not exceed 5 wt. %, more preferably does not exceed 1 wt. %, but is at least 0.1 wt % of the total fat. As ARA was found to be particularly effective in reducing tight junction permeability, the present composition comprises relatively high amounts, preferably at least 0.1 wt. %, even more preferably at least 0.25 wt. %, most preferably at least 0.35 wt. % of the total fat. The ARA content preferably does not exceed 5 wt. %, more preferably does not exceed 1 wt. % of the total fat. When the present enteral composition comprises ARA, EPA and DHA are advantageously added to balance the action of ARA, e.g. reduce the potential pro-inflammatory action of ARA metabolites. Excess metabolites from ARA may cause inflammation. Hence, the present composition preferably comprises ARA, EPA and DHA, wherein the weight ratio ARA/DHA preferably is above 0.25, preferably above 0.5, even more preferably above 1. The ratio is preferably below 25, more preferably 15. The weight ratio ARA/EPA is preferably between 1 and 100, more preferably between 5 and 20.
The present composition preferably comprises between 5 and 75 wt. % polyunsaturated fatty acids based on total fat, preferably between 10 and 50 wt. %.
The content of LC-PUFA, particularly the LC-PUFA with 20 and 22 carbon atoms, preferably does not exceed 3 wt. % of the total fat content as it is desirable to mimic human milk as closely as possible. The LC-PUFA may be provided as free fatty acids, in triglyceride form, in diglyceride form, in monoglyceride form, in phospholipid form, or as a mixture of one of more of the above. The present composition preferably comprises at least one of ARA and DHA in phospholipid form.

Nucleotides

Preferably the present composition comprises nucleotides and/or nucleosides, more preferably nucleotides. Preferably, the composition comprises cytidine 5′-monophospate, uridine 5′-monophospate, adenosine 5′-monophospate, guanosine 5′-monophospate, and/or inosine 5′-monophospate, more preferably cytidine 5′-monophospate, uridine 5′-monophospate, adenosine 5′-monophospate, guanosine 5′-monophospate, and inosine 5′-monophospate.
Preferably the composition comprises 5 to 100, more preferably 5 to 50 mg, most preferably 10 to 50 mg nucleotides and/or nucleosides per 100 gram dry weight of the composition. The presence of nucleotides and/or nucleotides advantageously enhance gut growth and maturation in the infant, which is of crucial importance in infants delivered by caesarean section. The nucleotides and/or nucleosides further stimulate the immune system thereby enhancing protection against a high load of intestinal pathogens such as E. coli. The nucleotides and/or nucleosides are deemed to act synergistically with the other ingredients of the present composition.

Formulae

The present composition is preferably enterally administered, more preferably orally.
The present composition is preferably a nutritional formula, preferably an infant formula. The present composition can be advantageously applied as a complete nutrition for infants. The present composition preferably comprises a lipid component, protein component and carbohydrate component and is preferably administered in liquid form. The present invention includes dry food (e.g. powders) which are accompanied with instructions as to admix said dry food mixture with a suitable liquid (e.g. water).
The present invention advantageously provides to a composition wherein the lipid component provides 5 to 50% of the total calories, the protein component provides 5 to 50% of the total calories, the carbohydrate component provides 15 to 90% of the total calories. Preferably, in the present composition the lipid component provides 35 to 50% of the total calories, the protein component provides 7.5 to 12.5% of the total calories, and the carbohydrate component provides 40 to 55% of the total calories. For calculation of the % of total calories for the protein component, the total of energy provided by the proteins, peptides and amino acids needs to be taken into account.
The present composition preferably comprises at least one lipid selected from the group consisting of animal lipid (excluding human lipids) and vegetable lipids. Preferably the present composition comprises a combination of vegetable lipids and at least one oil selected from the group consisting of fish oil, animal oil, algae oil, fungal oil, and bacterial oil. The present composition comprising non-digestible oligosaccharides excludes human milk.
The protein component used in the nutritional preparation are preferably selected from the group consisting of non-human animal proteins (preferably milk proteins), vegetable proteins (preferably soy protein and/or rice protein), hydrolysates thereof, free amino acids and mixtures thereof. Cow's milk derived nitrogen source, particularly cow's milk protein proteins such as casein and whey proteins are particularly preferred. Preferably the protein comprises intact proteins, more preferably intact bovine whey proteins and/or intact bovine casein proteins. As the present composition is suitably used to reduce the allergic reaction in an infant, the protein of is preferably selected from the group consisting of hydrolyzed milk protein. Preferably the present composition comprises hydrolyzed casein and/or hydrolyzed whey protein, vegetable protein and/or amino acids. The use of these proteins further reduced the allergic reactions of the infant. The use of these hydrolysed proteins advantageously improves the absorption of the dietary protein component by the immature intestine of the infant delivered by caesarean section.
The digestible carbohydrates used in the present composition are preferably selected from the group consisting of sucrose, lactose, glucose, fructose, corn syrup solids, starch and maltodextrins, and mixtures thereof, more preferably lactose.
Stool irregularities (e.g. hard stools, insufficient stool volume, diarrhoea) are an important problem in babies delivered via caesarean section. This may be caused by the high content of E. coli in the feces. It was found that stool problems may be reduced by administering the present uronic acid oligosaccharides in liquid food with an osmolality between 50 and 500 mOsm/kg, more preferably between 100 and 400 mOsm/kg. The reduced stool irregularities enhance the colonization and development of a healthy intestinal flora.
In view of the above, it is also important that the liquid food does not have an excessive caloric density, however still provides sufficient calories to feed the subject. Hence, the liquid food preferably has a caloric density between 0.1 and 2.5 kcal/ml, even more preferably a caloric density of between 0.5 and 1.5 kcal/ml, most preferably between 0.6 and 0.8 kcal/ml.

Concentrated Non-Digestible Oligosaccharides

According to a further preferred embodiment, the present inventions provides a method for providing nutrition to an infant delivered via caesarean section, wherein a composition containing uronic acid oligosaccharide is admixed to the nutrition to be administered to the infant born via caesarean section.
The present invention thus provides a method for providing nutrition to an infant delivered via caesarean section, said method comprising the steps of:

- a) admixing a composition comprising uronic acid oligosaccharide with a nutrition to be administered to the infant delivered via caesarean section; and
- b) administering the mixture obtained in step a) to the infant delivered via caesarean section.
  Preferably the nutrition as referred to in step a) of the method is infant milk formula or human breast milk. The composition comprising non-digestible oligosaccharides as used in step a) is a concentrated composition of non-digestible oligosaccharides or in other words is a composition comprising non-digestible oligosaccharides in high amounts.

The composition as used in step a) in the above-described method, preferably comprises at least 5 wt. %, preferably at least 10 wt. %, more preferably at least 25 wt. % uronic acid oligosaccharide based on dry weight of the composition.
The present invention also provides a composition which can be suitably used in the present method for admixing to a nutrition, i.e. a composition which comprises a high concentrations of non-digestible neutral oligosaccharide. The composition comprises preferably at least 5 wt. % (preferably at least 10 wt. %) non-digestible oligosaccharide based on dry weight of the composition, wherein the non-digestible oligosaccharide is preferably selected from the group consisting of galacto-oligosaccharides, non-digestible dextrins, xylo-oligosaccharides, arabino-oligosaccharides, gluco-oligosaccharides (including gentio-oligosaccharides and cyclodextrins), chito-oligosaccharides, fuco-oligosaccharides, manno-oligosaccharides, isomalto-oligosaccharide and fructo-oligosaccharide (including inulins); and at least 1 wt. % (preferably at least 10 wt. %) of the present uronic acid oligosaccharide based on dry weight of the composition. The composition preferably contains galactooligosaccharides and/or fructooligosaccharides.
The composition is preferably designed to be added to a single serving of infant nutrition. The cumulative weight of the uronic acid oligosaccharide in the present composition is preferably between 0.01 and 10 gram, preferably between 0.2 and 5 gram per serving. The composition is preferably packed per serving, i.e. in a unit dose of one serving, preferably in the form of a sachet. On serving of the composition preferably has a dry weight of 0.5 to 25 grams, preferably between 1 and 10 grams.

Application

The present invention provides i) the treatment and/or prevention of disorders in infants delivered via caesarean section and/or ii) the stimulation of health in infants delivered via caesarean section. The disorders are preferably selected from the group consisting of disorders caused by low bifidogenic flora. Preferably the disorder is selected from the group consisting of infection and allergy. Preferably the present invention is used to modulate the immune system in infants born via caesarean section. Administration of a composition comprising the present uronic acid oligosaccharides results in a decreased adherence and/or growth of (nosocomial) pathogenic micro-organisms in the underdeveloped intestinal flora of the infant born via caesarean section. Hence the present invention provides a method for the prevention and/or treatment of infections and/or infection disorders, particularly gastrointestinal infections, more preferably the treatment and/or prevention of infections selected cause by one or more micro-organisms selected from the group consisting of Escherichia coli, Klebsiella, Enterobacter, Aeromonas, Acinetobacter, Proteus, Pseudomonas (especially P. aeruginosa), Enterococcus, Streptococcus, (especially Streptococcus group B), Staphylococcus (especially S. epidermidis, S. aureus, S. haemolyticus), Bacillus (especially B. subtilis) and Clostridium (especially Clostridium difficile), preferably Escherichia coli (E. coli). Preferably, the present composition is used in a method for treatment or prevention of intestinal infection, urinary tract infection, intestinal inflammation and/or diarrhoea in infants delivered by caesarean section. Preferably the present composition is used in a method modulate the immune system to protect from allergies.
The present composition is preferably administered to the infant delivered via caesarean section in the first year of life, preferably within 3 months after birth, even more preferably within two weeks after birth, even more preferably within 100 hours, more preferably within 72 hours, most preferably within 48 hours after birth.
The present invention thus also provides a method for stimulating the development of a healthy intestinal flora in an infant comprising step A: admixing I) an in particular nutritionally or pharmaceutically acceptable liquid; and II) a dry composition, wherein the dry composition II comprises uronic acid oligosaccharides; and step B) administering the composition obtained in step A to an infant born via caesarean section.
Administration of the present composition results in an improved intestinal flora and/or in the formation of organic acids as metabolic end products of microbial fermentation. An increased amount of organic acids results in an increased mucus production, improves gut maturation and/or and increases gut barrier function. Hence, in a further aspect, the present invention therefore provides a method for treatment and/or prevention of allergy (preferably food allergy), atopic eczema (e.g. atopic dermatitis), asthma, allergic rhinitis, allergic conjunctivitis in infants delivered by caesarean section, said method comprising administering to the infant a composition comprising the present uronic acid oligosaccharides.
Furthermore, administration of the present composition strengthens the immune system. In a further aspect, the present invention therefore provides a method for treatment and/or prevention of systemic infections, otitis and/or respiratory infections in infants delivered by caesarean section.

EXAMPLES

Example 1

Molecular Characterization of Intestinal Microbiota in Infants Born by Vaginal Delivery vs. Caesarean Delivery

In the present study the influence of mode of delivery (caesarean delivery versus vaginal delivery) on the intestinal microbial composition at the third day of life by was studied using by PCR amplification with species-specific primers for ten Bifidobacterium species, three Ruminococcus species and one Bacteroides species.
The microbial DNA was extracted and analysed according to Favier et al, Environ Microbiol 2002; 68:219-226 and Satokari et al, Appl Environ Microbiol 2001; 67:504-513; Satorkari et al System Appl Microbiol 2003; 26:572-584.
The results of the Bifidobacterium and other species detected in faecal samples of 21 newborns after caesarean delivery obtained at the 3rd day of life are given in Table 1. Table 2 gives the Bifidobacterium and other species detected in faecal samples of 21 newborns after vaginal delivery obtained at the 3rd day of life. No signal specific for the species B. dentium, B. angulatum, B. lactis, Ruminococcus bromii, Ruminococcus callidus and Ruminococcus obeum was observed in the faeces of infants delivered by caesarean section as well as of vaginally delivered infants.
It can be concluded that the microbial flora of an infant born via caesarean section differs from that of an infant born via the vaginal route. Not only is the amount of bifidobacteria and other species quantitatively much lower, also on a species level the flora of caesarean section delivered infants is less diverse. Since Bifidobacterium is the dominant genus in infant's flora these results can also be generalised to less intestinal bacteria and a less diverse intestinal flora in ceasarean section delivered infants, leaving the intestine more susceptible to colonisation by (nocosomial) pathogens.
These results are indicative for the advantageous use of the composition and method according to the present invention, e.g. a method for feeding babies born via caesarean section, decreasing intestinal pathogens, stimulating a healthy intestinal flora and consequently preventing infection, stimulating a healthy immune system, and stimulating gut maturation.

TABLE 1

Caesarean section delivered infants

	B.	B.	B.	B. catenu-	B.	B.	B.	Bacteroides
NEWBORN	breve	infantis	bifidum	latum group	adolescentis	longum	gallicum	fragilis

1	−	−	−	−	−	−	−	−
2	−	−	−	−	−	++	−	−
3	−	−	−	−	−	−	−	−
4	−	−	−	−	−	−	−	−
5	−	−	−	−	−	−	−	−
6	−	−	−	−	−	−	−	−
7	−	−	−	−	−	−	−	−
8	−	−	−	−	−	−	−	−
9	−	−	−	−	−	−	−	−
10	−	−	−	−	−	−	++	−
11	−	−	−	−	−	−	−	−
12	−	−	−	−	−	−	−	−
13	−	−	−	−	−	−	−	−
16	−	−	−	−	−	−	−	−
17	−	−	−	−	−	−	−	−
18	−	−	−	−	−	−	−	−
19	−	−	−	−	−	−	−	−
20	−	−	−	−	−	−	−	−
21	−	−	−	−	−	−	−	−
22	−	−	−	−	−	−	−	−
23	−	−	−	−	−	−	−	−

(−) = no amplification;
(+/−) = weak amplification;
(+) = positive amplification;
(++) = strong amplification

TABLE 2

Vaginally delivered infants

	B.	B.	B.	B. catenu-	B.	B.	B.	Bacteroides
NEWBORN	breve	infantis	bifidum	latum group	adolescentis	longum	gallicum	fragilis

1a	−	+	−	−	−	++	−	−
2a	+/−	−	++	++	−	++	−	−
3a	−	−	−	+	−	−	−	−
4a	+/−	−	−	++	+	+	−	−
5a	+/−	−	++	++	−	++	++	−
6a	−	−	+/−	++	−	++	−	−
7a	−	−	+/−	++	++	−	−	−
8a	++	++	−	+	++	−	−	−
9a	−	−	−	+	++	+	−	−
10a	++	−	−	+	+	−	−	−
11a	++	−	++	++	−	++	−	+
12a	+	+	+	+	−	++	−	+
13a	+/−	−	−	+	−	+	−	−
16a	−	−	−	++	−	+	−	−
17a	+/−	−	+	+	−	+	−	−
18a	+/−	−	+	+	−	+	−	−
19a	+	−	−	+	−	+	−	−
20a	−	−	−	+	−	+	−	−
21a	−	−	−	+	++	+	−	−
22a	−	+	−	++	−	+	−	−
23a	+	−	++	++	−	+	−	−

(−) = no amplification;
(+/−) = weak amplification;
(+) = positive amplification;
(++) = strong amplification

Example 2

Anti-Adherence effect of Uronic Acid Oligosaccharides

The effects of pectin oligosaccharides (39 mol unsaturated galacturonic acid per 100 mol reducing oligosaccharides in the galacturonic acid molecules; and a degree of methylation of 41%) on enteroinvasive E. coli (EIEC 4608-58) was tested. In an invasion assay with cultured human enterocytes (Caco-2) an anti-infective effect of the pectin derived oligosaccharides was demonstrated on the enteroinvasive E. coli. The results indicate a reduced adherence and invasion: 1 hours post exposure bacterial invasion of E. coli into enterocytes was 49% in the presence of pectin oligosaccharide (compared to 100% in the control). This was most likely due to a decreased adherence of the E. coli to enterocyes (46% compared to control). Results also indicated a reduced extracellular growth of the E. coli in the presence of the pectin oligosaccharides compared to the control.

Example 3

Bifidogenic effect of Non-Digestible Oligosaccharides on the Flora in Caesarean Section Delivered Infants

The bifidobacterial content in the feces was determined. The percentage of the genus Bifidobacterium as a total of total bacteria in the first week was 4.3% in caesarean section delivered infants (n=44) versus 19.8% in vaginally delivered infants (n=28). The percentage E. coli after 6 weeks was 11.8% in the caesarean delivered infants.
These results indicate the desirability of the administration of the present uronic acid oligosaccharides to infants born via caesarean section.

Example 4

Composition for Babies Born via Caesarean Section

An infant formula composition comprising comprises per 100 ml ready to feed formula: 1.6 g protein, 3.6 g fat, 6.4 g digestible carbohydrates (mainly lactose), 0.8 g non-digestible carbohydrates of which 0.54 g transgalacto-oligosaccharides, 0.06 g inulin, and 0.2 g pectin oligosaccharide (prepared by lyase hydrolysis of citrus pectin). The package and/or supporting material accompanying the product indicates that the product can be suitable used to a) prevent and/or treat infection in infants delivered via caesarean section; b) prevent and/or treat infection with E. coli in infants delivered via caesarean section; c) prevent and/or treat allergy and/or atopic dermatitis in infants delivered via caesarean section; and/or d) modulate the immune system

Example 5

Method for Feeding Babies Born via Caesarean Section

Sachet which contains:

- 0.2 g uronic acid oligosaccharide (about 2 wt. % based on dry weight);
- 2 g galacto-oligosaccharide and 0.3 grams fructo-oligosaccharides (about 20 wt. % based on dry weight);
- Bifiodobacterium breve and Bifidobacterium longum, about 5* 10⁹cfu of each strain;
- Bulking agent

Claims

1-16. (canceled)

17. A composition comprising:

(a) at least 5 weight (wt.) % uronic acid oligosaccharide; and

(b) at least 5 wt. % non-digestible oligosaccharide selected from the group consisting of a galacto-oligosaccharide and fructo-oligosaccharide,

both of said wt. % of (a) and (b) based on dry weight of the composition;

wherein the cumulative weight of the uronic oligosaccharide and the non-digestible oligosaccharide is between 0.1 and 10 gram per serving of the composition.

18. A composition according to claim 17 wherein

(a) the uronic acid oligosaccharide is present at least at 10 wt. %; and

(b) the non-digestible oligosaccharide is present at least at 10 wt. %.

19. A composition comprising

(a) at least 5 wt. % uronic acid oligosaccharide; and

both of said wt. % of (a) and (b) based on dry weight of the composition; and

(c) 5-100 mg nucleotides per 100 g dry weight of the composition.

20. A composition according to claim 19, wherein:

(a) the uronic acid oligosaccharide is present at least at 10 wt. %, and

(b) the non-digestible oligosaccharide is present at least at 10 wt. %.

21. A composition comprising

(a) at least 5 wt. % uronic acid oligosaccharide;

(b) at least 5 wt. % non-digestible oligosaccharide selected from the group consisting of a galacto-oligosaccharide and fructo-oligosaccharide

both of said wt. % of (a) and (b) based on dry weight of the composition; and

(c) 10²to 10¹³colony forming units of lactic acid-producing bacteria per gram dry weight of the composition.

22. A composition according to claim 21, wherein:

(a) the uronic acid oligosaccharide is present at least at 10 wt. %, and

(b) the non-digestible oligosaccharide is present at least at 10 wt. %.

23. A composition according to claim 17 further comprising a long chain polyunsaturated fatty acids and/or a nucleotide.

24. A composition according to claim 17 that comprises:

(i) a lipid component that provides 5 to 50% of total calories of the composition;

(ii) a protein component that provides 5 to 50% of total calories of the composition; and

(iii) a carbohydrate component that provides 15 to 90% of the total calories of the composition.

25. A method for treating an infant delivered via caesarean section, comprising administering feeding the infant by enteral route feeding the infant an effective amount of a composition comprising a uronic acid oligosaccharide.

26. A method according to claim 25, wherein the composition further comprises non-digestible oligosaccharides selected from the group consisting of galacto-oligosaccharides and/or fructo-oligosaccharides.

27. A method according to claim 25, wherein the composition further comprises long chain polyunsaturated fatty acids and/or nucleotides.

28. A method according to claim 25, wherein the composition comprises a lipid component, a protein component and a carbohydrate component, wherein said lipid component provides 5 to 50% of the total calories, said protein component provides 5 to 50% of the total calories and said carbohydrate component provides 15 to 90% of the total calories.

29. A method for treating and/or preventing a disease or disorder in an infant delivered by caesarean section, comprising feeding an infant in need thereof an effective amount of a composition comprising a uronic acid oligosaccharide, wherein the disease or disorder is one or more of:

(a) an infection or an infectious disease,

(b) asthma,

(c) a food allergy,

(d) atopic eczema or dermatitis,

(e) allergic rhinitis, and

(f) allergic conjunctivitis.

30. A method according to claim 29 wherein the infant has an infection or infectious disease.

31. A method according to claim 29 wherein the uronic oligosaccharide is selected from the group consisting of a pectin degradation product with a degree of polymerization (DP) of 2 to 250 and an alginate degradation product with a DP of 2 to 250.

32. A method according to claim 29, wherein the

(i) the composition comprises between 0.01 and 10 grams uronic acid oligosaccharide per 100 gram dry weight of the composition,

(ii) the uronic acid oligosaccharide has a DP of 2 to 250.

33. A method according to claim 29, wherein the uronic acid oligosaccharide comprises 2 to 50% unsaturated terminal hexuronic acid units based on the total terminal hexuronic acid units present.

34. A method according to claim 29 wherein the composition is administered to the infant within one week after birth.

35. A method according to claim 31 wherein the composition is administered to the infant within 100 hours after birth.

36. A method according to claim 25, wherein the composition is prepared by admixing

(I) a nutritionally or pharmaceutically acceptable liquid; and

(II) a dry composition comprising said uronic acid oligosaccharide which is selected from the group consisting of guluronic acid oligosaccharide, iduronic acid oligosaccharide, riburonic acid oligosaccharide, mannuronic acid oligosaccharides, galacturonic acid oligosaccharide and glucuronic acid oligosaccharide.

37. A method according to claim 36 wherein administration of said composition stimulates development of healthy intestinal flora and/or decreases the occurrence of intestinal pathogens in said infant.

38. A method for stimulating development of healthy intestinal flora in an infant delivered by caesarean section, the method comprising feeding an infant in need thereof an effective amount of a composition comprising a uronic acid oligosaccharide selected from the group consisting of guluronic acid oligosaccharide, iduronic acid oligosaccharide, riburonic acid oligosaccharide, mannuronic acid oligosaccharide, galacturonic acid oligosaccharide and glucuronic acid oligosaccharide.