AU2015218488B2 - Method for stimulating the intestinal flora - Google Patents

Abstract

H:\SXD\Interwoven\NRPortbl\DCC\SXD\4977747_1.doc-7/03/2013 The present invention relates to methods for feeding and to compositions to be administered to infants delivered via caesarian section and in particular to the use of a) at least two different 5 microorganisms; or b) at least one microorganism and at least one indigestible oligosaccharide; or c) at least two different Bifodobacteria species, subspecies or strains for the manufacture of a composition for enteral administration to an infant delivered via caesarean section. Thereby it is possible to stimulate the healthy development of the intestinal flora of said infants.

Description

METHOD FOR STIMULATING THE INTESTINAL FLORA 2015218488 27 Aug 2015

This is a divisional of Australian patent application No. 2013201362, the entire contents of which are incorporated herein by reference. 5

FIELD OF THE INVENTION

The present invention relates to methods for feeding and to compositions to be administered to infants delivered via caesarian section.

10 BACKGROUND OF THE INVENTION

When formulating nutrition for infants, breast milk is used as the gold standard. Ingredients in particular of non-human origin are normally combined to mimic the compositional features and physiological effects of the human breast milk. 15 One important aspect of human milk is that it provides energy and fluids to the infant. Besides providing energy, human breast milk contains many additional components, which aim to maintain the health of the infant. Human breast milk for example contains prebiotic fiber, which stimulates the development of a healthy intestinal flora. A healthy intestinal flora has numerous positive effects on the infant, such as a reduced incidence of infections and a 20 strengthened immune system.

Several commercially available infant milk formulae contain ingredients that aim to stimulate the development of the intestinal flora. Infant formulae can for example contain prebiotic fibers or live probiotic organisms. The prebiotic fibers normally pass undigested 25 through the upper gastrointestinal tract and selectively stimulate the growth of beneficial bacteria in the colon. Live probiotic bacteria increase specific bacterial counts in the intestine. EP 1105002 describes a prebiotic carbohydrate mixture comprising one or more soluble 30 oligosaccharides and one or more soluble polysaccharides, with at least 80 wt. % of each being prebiotic. WO 2005039319 describes a preparation comprising Bifidobacterium breve and a mixture of non-digestible carbohydrates for non- or partially breast-fed infants as well as the 35 use thereof for the treatment or prevention of immune disorder in non- or partially breastfed infants.

SUMMARY OF THE INVENTION 2015218488 27 Aug 2015

Infant formulae are normally designed to mimic the development of an intestinal flora in an infant receiving human breast milk, with the implication that all infants react similar to human breast milk and infant formula. However, the present inventors have 5 found that a sub-population of infants, namely those infants delivered via caesarian section, will react differently because their intestinal flora at birth is completely different from the intestinal flora of infants bom via the vaginal route. In particularly, the profile and content of Bifidobacteria species of infants delivered via caesarean section is different from the intestinal profile and content of Bifidobacteria species of 10 infants delivered via the vaginal route.

The present inventors have analyzed the intestinal flora of newborns after caesarean delivery and newborns after vaginal delivery. It was surprisingly found that great differences exist between the two groups in composition of the intestinal flora. 15 Particularly infants bom via the vaginal route contain at least about three different Bifidobacteria species, whereas the infants bom via cesarean section lack the most important Bifidobacteria species.

It was also found that the infants bom via caesarian section particularly lack 20 Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis and Bifidobacterium longum. These species were present in the flora of most infants bom via the vaginal route. 25 Because the intestinal flora plays a crucial role in the development of the infant, in particularly the stimulation of the immune system and resistance against infections, it is of utmost importance to stimulate the healthy development of the intestinal flora of infants bom via cesarean section. 30 Furthermore the newborns delivered via caesarian section lacked a biodiversity in their intestinal flora. Only one or two different species of Bifidobacteria were detected in infants bom via c-section, while the intestinal flora of newborns delivered via the vaginal route normally contains several different species of Bifidobacteria. The present inventors believe that these observations are indicative for a general lack of a 2 species variety in the intestinal tract of infants delivered via caesarean section. The biodiversity is of great importance for achieving the desired physiological effects and optimally stimulate the health of the infant. 2015218488 27 Aug 2015

Natren® produces the probiotic product Life Start® which is designed specifically for infants and 5 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.

Particularly unexpected is also the observation that improvements of infant health can be achieved even when infants are fed with breast milk. In the case where infants are delivered via caesarean 10 section, breast milk is (in most cases) the best nutrition for the infant. However, the breast milk also does not instantaneously result in a similar flora as obtained in infant bom via vaginal delivery. The present method is therefore also advantageously used when infants receive human breast milk.

Hence, the present invention advantageously: a) increases the occurrence of particular species in the intestinal flora of infants bom via caesarean section; b) increases the biodiversity of microorganisms in 15 the intestinal flora; and/or c) stimulates the growth of beneficial microorganisms, particularly

Bifidobacteria.

In a first aspect, the present invention provides use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for enteral administration to an infant delivered via caesarean section, wherein the composition contains at least one species of 20 Bifidobacterium and wherein the composition is an infant milk formula.

In a second aspect, the present invention provides use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for enteral administration to an infant delivered via caesarean section, wherein the composition contains at least one species of Bifidobacterium and wherein the composition comprises an indigestible oligosaccharide selected from 25 the group consisting of transgalactooligosaccharides, indigestible dextrins, xylooligosaccharides, arabinooligosaccharides, glucooligosaccharides, mannooligo-saccharides, isomalto-oligosaccharide and ffuctopolysaccharides.

In an embodiment of the first or second aspects, the composition comprises at least one species of 30 Bifidobacterium selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis, Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and 3

Bifidobacterium longum. In an embodiment, the composition comprises at least two species of Bifidobacterium selected from said group of species of Bifidobacterium. 2015218488 27 Aug 2015

In an embodiment of the first aspect, the indigestible oligosaccharide is selected from the group 5 consisting of galactooligosaccharides, indigestible dextrins, xylooligosaccharides, arabinooligosaccharides, glucooligosaccharides, mannooligo-saccharides, isomalto-oligosaccharide and fructopolysaccharides.

In an embodiment of the first or second aspects, the composition comprises between 5 and 25 en% 10 protein; between and 60 en% fat; between 30 and 70 en% carbohydrate. In an embodiment, the composition is administered to an infant receiving human breast milk. In an embodiment, the composition is administered to the infant within one week after birth.

In a third aspect, the present invention provides a method for the manufacture of an infant nutrition 15 suitable for infants bom via caesarean section comprising admixing: a) human breast milk; and b) a composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the composition contains at least one species of

Bifidobacterium. >0

In a fourth aspect, the present invention provides a use of a composition obtained by admixing: I) a nutritionally or pharmaceutically acceptable liquid; and II) a dry composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the composition contains at least one species of 25 Bifidobacterium for the manufacture of a composition to be administered to an infant bom via caesarean section for stimulating the development of a healthy intestinal flora.

In a fifth aspect, the present invention provides a use of a composition containing microorganisms and 30 indigestible oligosaccharides for the manufacture of a composition for stimulating the intestinal flora of an infant delivered via caesarean section, by administering said composition to the infant and within 100 hours after birth, wherein the composition contains at least one species of Bifidobacterium and wherein the composition is an infant milk formula. 35 In a sixth aspect, the present invention provides a use of a composition containing microorganisms and indigestible oligosaccharides for the manufacture of a composition for stimulating the intestinal flora 3a of an infant delivered via caesarean section by administering said composition to the infant within 100 hours after birth, wherein the composition contains at least one species of Bifidobacterium and the composition comprises an indigestible oligosaccharide selected from the group consisting of transgalactooligosaccharides, indigestible dextrins, xylooligosaccharides, arabinooligosaccharides, 5 glucooligosaccharides, mannooligo-saccharides, isomalto-oligosaccharide and fructopolysaccharides. 2015218488 27 Aug 2015

In a seventh aspect, the present invention provides a use of a composition comprising microorganisms and/or indigestible oligosaccharides for the manufacture of a composition for stimulating the intestinal flora of an infant delivered via caesarean section by rectally administering said composition to an .0 infant below 3 years of age, wherein the composition contains at least one species of Bifidobacterium.

In an eighth aspect, the present invention provides a method of treating an infant delivered by caesarean section comprising enterally administering to said infant a composition comprising at least one species of Bifidobacterium and at least one indigestible oligosaccharide, wherein the composition . 5 is an infant milk formula.

In a ninth aspect, the present invention provides a method of treating an infant delivered by caesarean section comprising administering to said infant a composition comprising at least one species of Bifidobacterium and at least one indigestible oligosaccharide, wherein the composition is admixed 10 with human breast milk.

In a tenth aspect, the present invention provides a method of stimulating development of a healthy intestinal flora of an infant delivered by caesarean section comprising administering to said infant a dry composition comprising at least one species of Bifidobacterium and at least one indigestible 25 oligosaccharide, wherein the composition is admixed with a nutritionally or pharmaceutically acceptable liquid.

In an embodiment of the eighth, ninth of tenth aspect, the Bifidobacterium species is selected from the group of species of Bifidobacterium consisting of Bifidobacterium breve, Bifidobacterium infantis, 30 Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis, Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum. 3b H:\fmt\Interwoven\NRPortbl\DCC\FMT\12223314_l.docx-14/12/2016 2015218488 14 Dec 2016

In an embodiment, the indigestible oligosaccharide is selected from the group consisting of transgalactooligosaccharides, indigestible dextrins, xylooligosaccharides, arabinooligosaccharides, glucooligosaccharides, mannooligo-saccharides, isomalto-oligosaccharide and fructopolysaccharides.

In an embodiment, the composition comprises at least two Bifidobacterium species selected from said group of species of Bifidobacterium species.

In an embodiment, the composition is administered to said infant within 100 hours of birth.

In another aspect the present invention provides a method for stimulating the healthy development of the intestinal flora of infants bom via caesarean section by administering a composition containing: - at least two different microorganisms; - at least one microorganism and at least one indigestible oligosaccharide; or - at least two different Bifidobacteria species, subspecies or strains.

In another aspect the present invention provides use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the development of a healthy intestinal flora in an infant delivered via caesarean section, wherein the composition is for enteral administration and the microorganism is Bifidobacterium breve and the indigestible oligosaccharide is galactooligosaccharide.

In another aspect the present invention provides a method for the manufacture of an infant nutrition suitable for infants born via caesarean section comprising admixing: a) human breast milk; and b) a composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the composition contains at least one species of Bifidobacteria selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum. 3c H:\fmt\Interwoven\NRPortbl\DCC\FMT\12223314_l.docx-14/12/2016 2015218488 14 Dec 2016

In another aspect the present invention provides use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the development of a healthy intestinal flora in an infant born via caesarean section, wherein the composition is obtained by admixing: I) a nutritionally or pharmaceutically acceptable liquid; and II) a dry composition comprising the at least one microorganism and the at least one indigestible oligosaccharide, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligo s accharide.

In another aspect the present invention provides use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the intestinal flora of an infant delivered via caesarean section, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligosaccharide; and wherein the composition is administered to the infant within 100 hours after birth.

In another aspect the present invention provides use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the intestinal flora of an infant delivered via caesarean section, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligosaccharide; and wherein the composition is administered rectally to an infant below 3 years of age.

Associated methods of stimulating the development of a healthy intestinal flora in an infant delivered by caesarean section are also provided.

The microorganism will increase the biodiversity of the infants flora, while the indigestible oligosaccharides stimulate the development and growth of inherent and administered microorganisms.

The composition to be administered in the present method preferably contains multiple different bacterial species, preferably multiple Bifidobacteria species. This 3d H:\TRNMnterwoven\NRPortbl\DCC\TRNY73 5 5 281_l.docx.-2/03/2015 2015218488 27 Aug 2015 results in the optimal stimulation of the intestinal flora of the caesarean delivered infants.

The present invention provides the use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the 5 development of a healthy intestinal flora, stimulating the immune system and/or providing, resistance against infections in an infant delivered via caesarean section, wherein the composition is for enteral administration and contains at least one species of Bifidobacterium selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis 10 Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum and wherein the composition is an infant milk formula. 15 In a further aspect the present invention provides the use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the development of a healthy intestinal flora, stimulating the immune system and/or providing resistance against infections in an infant delivered via caesarean section, wherein the composition is for enteral administration and contains at least one species of Bifidobacterium 20 selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum and wherein the composition comprises an 25 indigestible oligosaccharide selected from the group consisting of transgalactooligosaccharides, indigestible dextrins, xylooligosaccharides, arabinooligosaccharides, glucooligosaccharides, mannooligo-saccharides, isomalto-oligosaccharide and fructopolysaccharides. 30 In still a further aspect the present invention provides a method for the manufacture of an infant nutrition suitable for infants bom via caesarean section comprising admixing: a) human breast milk; and 4 H:\TRNMnterwoven\NRPortbl\DCC\TRNY73 5 5 281_l.docx.-2/03/2015 2015218488 27 Aug 2015 b) a composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the composition contains at least one species of

Bifidobacterium selected from the group consisting of Bifidobacterium breve,

Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, 5 Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum,

Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum.

In still a further aspect, the present invention provides a use of a composition obtained by 10 admixing: I) a nutritionally or pharmaceutically acceptable liquid; and II) a dry composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the composition contains at least one species of

Bifidobacterium selected from the group consisting of Bifidobacterium breve, 15 Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum,

Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum for the manufacture of a composition to be administered to an infant bom via caesarean 20 section for stimulating the development of a healthy intestinal flora.

The present invention also provides the use of a composition containing microorganisms and indigestible oligosaccharides for the manufacture of a composition for stimulating the intestinal flora of an infant delivered via caesarean section, by administering said composition 25 to the infant and within 100 hours after birth, wherein the composition contains at least one species of Bifidobacterium selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum,

Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, 30 Bifidobacterium thermacidophilum and Bifidobacterium longum and wherein the composition is an infant milk formula. 4a H:\TRNMnterwoven\NRPortbl\DCC\TRNY73 5 5 281_l.docx.-2/03/2015 2015218488 27 Aug 2015

The present invention also provides the use of a composition containing microorganisms and indigestible oligosaccharides for the manufacture of a composition for stimulating the intestinal flora of an infant delivered via caesarean section by administering said composition to the infant within 100 hours after birth, wherein the composition contains at least one species 5 of Bifidobacterium selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum and the composition 10 comprises an indigestible oligosaccharide selected from the group consisting of transgalactooligosaccharides, indigestible dextrins, xylooligosaccharides, arabinooligosaccharides, glucooligosaccharides, mannooligo-saccharides, isomalto oligosaccharide and fructopolysaccharides. 15 In a further aspect the present invention provides the use of a composition comprising microorganisms and indigestible oligosaccharides for the manufacture of a composition for stimulating the intestinal flora of an infant delivered via caesarean section by rectally administering said composition to an infant below 3 years of age, wherein the composition contains at least one species of Bifidobacterium selected from the group consisting of 20 Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum. 25 In still a further aspect the present invention provides a method method of treating an infant delivered by caesarean section comprising enterally administering to said infant a composition comprising at least one species of Bifidobacterium selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium 30 gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum and at least one indigestible oligosaccharide, wherein the composition is an infant milk formula. 4b H:\TRN\Interwoven\NRPortbl\DCC\TRN\7355281_l .docx-2/03/2015 2015218488 27 Aug 2015

In a further aspect the present invention provides a method method of treating an infant delivered by caesarean section comprising administering to said infant a composition comprising at least one species of Bifidobacterium selected from the group consisting of 5 Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum and at least one indigestible oligosaccharide, wherein the composition is admixed with human breast 10 milk.

In still a further aspect the present invention provides a method method of stimulating development of a healthy intestinal flora of an infant delivered by caesarean section comprising administering to said infant a dry composition comprising at least one species of 15 Bifidobacterium selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum and at least one indigestible oligosaccharide, 20 wherein the composition is admixed with a nutritionally or pharmaceutically acceptable liquid.

DETAILED DESCRIPTION OF THE PREFFERED EMBODIMENTS

The present invention provides a method for feeding infants, said method comprising enterally administering a composition to an infant delivered via caesarean section, said composition comprising: a) at least two different microorganisms; or b) at least one microorganism and at least one indigestible oligosaccharide; or c) at least two different Bifidobacteria species, subspecies or strains. 4c H:\TRN\Interwoven\NRPortbl\DCC\TRN\735528 l_l.docx-2/03/2015 2015218488 27 Aug 2015

In a further aspect the present invention provides a method for stimulating the health of an infant delivered via caesarean section, comprising administering to the infant and within 100 hours after birth a composition containing microorganisms and/or indigestible oligosaccharides.

In still a further aspect the present invention provides a method for the manufacture of infant nutrition suitable for infant bom via cesarean section comprising admixing: A) human breast milk; and B) a composition comprising: i) at least two different microorganisms; or ii) at least one microorganism and at least one indigestible oligosaccharide; or iii) at least two different Bifidobacteria species, subspecies or strains.

In still a further aspect, the present invention provides a composition comprising at least four Bifidobacteria species, subspecies or and/or strains selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis, Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum.

The present invention also provides a nutritional composition comprising between 5 and 25 en% protein; between 25 and 60 en% fat; between 30 and 70 en% 4d 2015218488 27 Aug 2015 carbohydrate; at least two different species of Bifidobacteria and at least one species of Lactobacillus.

The present invention also provides a container with a liquid volume between 0.5 and 5 50 ml containing a composition to be administered to an infant comprising at least two different microorganisms; or at least one microorganism and at least one indigestible oligosaccharide; or at least two different Bifidobacteria species, subspecies or strains. 10 In a further aspect the present invention provides a method for stimulating the development of a healthy intestinal flora in an infant comprising the step of: A) admixing I) a nutritionally or pharmaceutically acceptable liquid; and II) a dry composition, wherein the dry composition II comprises at least two different microorganisms; or at least one microorganism and at least one indigestible 15 oligosaccharide; or at least two different Bifidobacteria species, subspecies or strains; and B) administering the composition obtained in step a) to an infant bom via caesarean section.

The composition used in the present invention are preferably nutritional and/or 20 pharmaceutical compositions and suitable for administration to infants.

Caesarian section

The present invention relates to the enteral administration of a composition containing a microorganism to infants delivered via caesarean section. A caesarean section (c-25 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. Other times, a woman may choose to have a caesarean section rather than deliver her infant vaginally. 30

Diversity

The present composition contains at least two different microorganisms; or at least one microorganism organism and at least one indigestible oligosaccharide; or at least two different Bifidobacteria species, subspecies or strains. The microorganisms are 5 preferably bacteria and/or yeasts. Preferably the microorganisms used in the present invention are probiotic, i.e. when applied to man or animal, it beneficially affects the host by improving the properties of the indigenous microflora. The above-mentioned combinations commonly aim to increase the diversity and/or the quantity of 5 microorganisms in the intestine of the cesarean section delivered infant. This beneficially affects the infant, proving numerous health benefits. 2015218488 27 Aug 2015

The term “different microorganisms” as used for the present invention refers to microorganism belonging to a different genus and/or species. Preferably the different 10 microorganisms are different bacteria. Preferably, the different microorganisms are different species. For example and preferably, the present composition comprises or consists of Bifidobacterium breve (B. breve) and Bifidobacterium catenulatum (B. catenulatum). B. breve and B. catenulatum are herein considered as two different bacterial species. Bacterial species are not considered different if these are different 15 subspecies. For example, Lactobacillus delbrueckii subspecies delbrueckii and Lactobacillus delbrueckii subspecies bulgaricus are not considered two different species. The present composition contains at least two different microorganisms, preferably at least two different bacterial species. Preferably the present composition contains at least three different species, more preferably at least four different species. 20 Preferably the microorganism or bacterial species used are probiotic.

In one embodiment the present composition contains at least two different Bifidobacteria species, subspecies or strains. The present composition preferably comprises at least one, more preferably at least two, even more preferably at least 25 three, most preferably at least four different Bifidobacterium 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 Bifidobacteria subspecies. For example Bifidobacterium animalis subspecies animalis and Bifidobactrium animalis subspecies lactis are different subspecies. B. breve M16V en 30 B. breve R0070 are different strains.

Microorganisms

Preferably the present composition contains at least one microorganisms selected from the group consisting of lactic acid bacteria, bacilli and yeasts, preferably at least 6 one lactic acid bacteria selected from the group consisting of Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Oenococcus, Pediococcus, Streptococcus, Tetragenococcus, Vagococcus and Weissella and Bifidobacteria. The present composition preferably contains at least two, more preferably at least three, 5 most preferably at least four different microorganisms. 2015218488 27 Aug 2015

Preferably the present composition contains at least one species selected from the genus Bifidobacteria, more preferably at least two, even more preferably at least three, more preferably at least four, most preferably at least five species from the 10 genus of Bifidobacteria. Bifidobacteria are Gram-positive, anaerobic, rod-shaped bacteria. The present Bifidobacterium species preferably have at least 95 % identity of the 16 S rRNA sequence when compared to the type strain of the respective Bifidobacterium species, more preferably at least 97% identity as defined in handbooks on this subject for instance Sambrook, J., Fritsch, E.F., and Maniatis, T. 15 (1989), Molecular Cloning, A Laboratory Manual, 2nd ed., Cold Spring Harbor (N.Y.) Laboratory Press. The Bifodobacteria preferably used are also described by Scardovi, V. Genus Bifidobacterium, p.1418 - p.1434. In: Bergey’s manual of systematic Bacteriology. Vol. 2. Sneath, P.H.A., N.S. Mair, M.E. Sharpe and J.G. Holt (ed.). Baltimore: Williams & Wilkins. 1986. 635 p. 20

The present composition preferably comprises at least one, more preferably at least two, even more preferably at least three, most preferably at least four Bifidobacterium species preferably selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, 25 Bifidobacterium adolescentis, Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum. More preferably the present composition preferably comprises at least one, more preferably at least two, even more preferably at least three, most preferably at least 30 four Bifidobacterium species preferably selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis and Bifidobacterium longum. Most preferably the present composition contains Bifidobacterium breve and/or Bifidobacterium catenulatum. 7 2015218488 27 Aug 2015

In a further preferred embodiment, the present composition comprises at least one, more preferably at least two, even more preferably at least three, most preferably at least four Bifidobacterium subspecies. In still a further preferred embodiment, the 5 present composition comprises at least one, more preferably at least two, even more preferably at least three, most preferably at least four Bifidobacterium strains.

In a further preferred embodiment, the present composition contains a lactic acid bacteria, preferably at least a bacteria selected from the group consisting of 10 Lactobacilli, Lactococci and Streptococci. More preferably the present composition contains at least one bacteria selected from the group consisting of Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus johnsonii, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus fermentum, Lactobacillus lactis, Streptococcus thermophilus and Lactobacillus paracasei. The 15 further increased biodiversity will have a stimulatory effect on health of the newborn.

The present composition preferably contains between 101 and 1013 colony forming units (cfu) microorganisms per gram dry weight of the present composition, preferably between 102 and 1012, more preferably between 103 and 1010. Preferably, 20 the present composition contains between 101 and 1013 colony forming units (cfu) Bifidobacteria per g dry weight of the present composition, more preferably between 102 and 1012, most preferably between 103 and 1012.

The present method preferably comprises the administration of a serving containing 25 between 101 and 1013 cfu microorganisms more preferably between 102 and 1011, most preferably between 103 and 1010. The present method preferably comprises the administration of a serving containing between 101 and 1013 cfu Bifidobacteria, more preferably between 102 and 1012, most preferably between 103 and 1011. 30 Oligosaccharides

The term “indigestible oligosaccharides” 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 8 digestive tract (small intestine and stomach) but which are fermented by the human intestinal flora. Preferably the present indigestible oligosaccharide has a degree of polymerisation (DP) of 2 to 100, preferably a DP 2 to 50. 2015218488 27 Aug 2015

Preferably the present indigestible oligosaccharide is a prebiotic fiber. The term 5 "prebiotic fiber" refers to non-digestible fibers 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 indigestible oligosaccharide is soluble. The term "soluble" as used herein, when having reference to a polysaccharide, fibre or oligosaccharide, 10 means that the substance is at least 50% soluble according to the method described by L. Prosky et al., J. Assoc. Off. Anal. Chem. 71,1017-1023 (1988).

Preferably the present composition contains at least one oligosaccharide selected from the group consisting of galactooligosaccharides, indigestible dextrins, 15 xylooligosaccharides, arabinooligosaccharides, glucooligosaccharides, mannooligo-saccharides, isomalto-oligosaccharide and fructopolysaccharide.

The term “fructopolysaccharide” as used herein refers to a polysaccharide carbohydrate comprising a chain of at least 3 β-linked fructose units, with a DP 20 between 3 and 300, preferably between 20 and 150. Preferably inulin is used. Inulin is available under the tradename “Raftilin HP®”, (Orafti). The average DP of the fructopolysaccharide is preferably at least 15, more preferably at least 20 or more, up to 300. In inulin the fructose units are linked with a β(2—>1) linkage. 25 Indigestible 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 indigestible polydextrins comprise a(l->4), a(l-»6) glucosidic bonds and l->2 and 1—>3 linkages Indigestible polydextrins are for example available under the tradename “Fibersol 2®” from Matsutami Inductries or Litesse® from Danisco. 30

The present inventors found that galactooligosaccharides can be advantageously used in the present composition, because these oligosaccharides where particularly effective in stimulating the growth of Bifidobacteria. Hence, in a preferred 9 2015218488 27 Aug 2015 embodiment the present composition contains galactooligosaccharides. The term “galactooligosaccharide” as used herein refers to an indigestible saccharide, wherein at least 30 % of the saccharide units are galactose units, preferably at least 50%, more preferably at least 60%. Preferably the saccharides of the galactooligosaccharide are 5 β-linked, as is the case in human milk.

Preferably the present composition contains a galactooligosaccharide selected from the group consisting of transgalactooligosaccharides, lacto-N-tetraose (LNT) and lacto-N-neotetraose (neo-LNT). In a particularly preferred embodiment the present 10 method comprises the administration of transgalactooligosaccharide ([galactose]n-glucose; wherein n is an integer between 1 and 60, i.e. 2, 3, 4, 5, 6, ...., 59, 60; preferably n is selected from 2, 3,4, 5, 6, 7, 8, 9, or 10). Transgalactooligosaccharides (TOS) are for example sold under the trademark Vivinal™ (Borculo Domo Ingredients, Netherlands). Preferably the saccharides of the 15 transgalactooligosaccharides are β-linked.

The present composition preferably contains 0.5 to 75 grams of the indigestible soluble 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 20 galactooligosaccharides per 100 gram dry weight, preferably between 0.1 and 50 grams.

The present method preferably comprises the administration of a serving containing between 0.05 and 25 grams indigestible oligosaccharide, preferably between 0.1 and 5 25 grams. The present method preferably comprises the administration of a serving containing between 0.05 and 25 grams galactooligosaccharides, preferably between 0.1 and 5 gram galactooligosaccharides.

The present inventors have also found that a mixture of a long chain indigestible 30 oligosaccharides and short chain indigestible oligosaccharides synergistically stimulate the growth of a healthy intestinal flora, particularly Bifidobacteria. Hence the present composition preferably contains at least two oligosaccharides with different average degrees of polymerization (DP). Preferably the weight ratios: 10 2015218488 27 Aug 2015 a. (indigestible oligosaccharides with DP 2 to 5) : (indigestible oligosaccharides with DP 6, 7, 8, and/or 9) > 1; and b. (indigestible oligosaccharides with DP 10 to 60) : (indigestible oligosaccharides with DP 6,7,8, and/or 9) > l 5 Preferably both weight ratios are above 2, even more preferably above 5.

For further improvement, the oligosaccharide preferably has a relatively high content of short chain oligosaccharides, as these strongly stimulate the growth of Bifidobacteria. Hence, preferably at least 10 wt.% of the oligosaccharides in the 10 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 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, 15 the present composition preferably comprises two oligosaccharides with a different structure. The present composition comprises at least two different oligosaccharides, wherein the 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 20 percentage of same saccharide unit in the different oligosaccharides. For example, oligosaccharide 1 (OL1) has the structure fruc-fruct-glu-gal, and thus comprises 50% fine, 25% gal and 25% glu. Oligosaccharide 2 (OL2) has the structure fruc-fruc-glu, and thus comprises 66% fruc, 33% glu. The different oligosaccharides thus have a homology of 75% (50% fruc + 25% glu). 25

Preferably the present composition contains galactooligosaccharides and fructopolysaccharides. AOvlication 30 The present composition is preferably enterally administered, more preferably orally. The present composition is therefore preferably a liquid. The term "enteral" here used also encompasses a rectal or anal administration. 11 2015218488 27 Aug 2015

When infants receive infant milk formula, the present composition with microorganisms and optionally including at least one indigestible oligosaccharide, is preferably included in the nutritional formula. The probiotic and/or prebiotic may be separately added to the infant formula. When newborns receive nutrition via a tube, 5 the microorganisms can be suitably included in the nutrition administered via tube. Stimulating diversity is of great importance, thus the present invention also provides a nutritional composition comprising between 5 and 25 en% protein; between 25 and 60 en% fat; between 30 and 70 en% carbohydrate; at least two different species of Bifidobacteria and at least one species of Lactobacillus. 10

When infants receive human breast milk, the present composition can be suitably admixed with human breast milk. The present invention consequently also provides a method for the manufacture of infant nutrition suitable for infant bom via cesarean section comprising admixing; A) human breast milk; and B) a composition 15 comprising; (i) at least two different microorganisms; or (ii) at least one microorganism and at least one indigestible oligosaccharide; or (iii) at least two different Bifidobacteria species, subspecies or strains. The preferred characteristics of composition B) are as provided hereinabove. 20 In a further preferred embodiment the present composition is administered to the infant in a very small volume, e.g. by “inoculating” the infant. Preferably, the present composition is administered to the infant with a syringe, pipette or tube, preferably directly after birth. In a further preferred embodiment, the present composition is rectally or anally administered to the infant delivered via caesarean section, preferably 25 in the form of a suppository, pill or tablet. Hence, the present invention also provides a suppository, pill or table suitable for rectal administration to a infant with the age below one year, wherein said suppository, pill or tablet contains microorganisms and/or indigestible oligosaccharides, preferably the present composition as described hereinabove. 30

In a further preferred embodiment the present invention provides a method stimulating the intestinal flora of an infant comprising rectally administering to an infant with the age below 3 year a composition comprising microorganism and/or indigestible oligosaccharide. Preferably the infant has an age below 1 year, more 12 2015218488 27 Aug 2015 preferably below 2 weeks. Preferably the infant is delivered via caesarean section. Preferably this composition for rectal administration contains the above-described composition. 5 This procedure has the advantage that it does not interfere with the normal breastfeeding practice and has high resemblance with vaginal inoculation, which occurs during birth. The present invention also provides a method for stimulating the health of an infant delivered via caesarean section, comprising administering to the infant and within 100 hours after birth, preferably within 72 hours after birth, most 10 preferably within 48 hours after birth, a composition containing microorganisms and/or indigestible oligosaccharides.

The composition is preferably suitable for administration directly after birth. Hence, in a further preferred embodiment, the present invention provides a container 15 comprising a liquid composition with a volume between 0.5 and 50 ml, which contains the present composition. The liquid with the present microorganism, optionally combined with indigestible oligosaccharides, can be suitably used in the present method. Preferably the liquid has a volume between 0.5 and 25 ml. This volume is preferably small, because it otherwise could interfere with the appetite and 20 drinking behaviors of the infant.

Similarly and encompassed by this invention is a container with a reconstitutable dry composition containing the present composition, wherein the container has a volume of between 0.5 and 50 ml. This container is preferably accompanied with instruction 25 to reconstitute the powder in a small volume of liquid, e.g. water.

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 Π) a dry composition, wherein 30 the dry composition Π comprises at least two different microorganisms; or at least one microorganism and at least one indigestible oligosaccharide; or at least two different Bifidobacteria species, subspecies or strains; and step B) administering the composition obtained in step a) to an infant bom via caesarean section. 13 2015218488 27 Aug 2015

The present composition is preferably administered to the infant in the first year of life, preferably within two weeks after birth, even more preferably within one week after birth, most preferably within 48 hours after birth.

5 EXAMPLES

Example 1: Molecular characterization of intestinal microbiota in infants bom by vaginal delivery vs. caesarean delivery 10 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 using PCR-Denaturing Gradient Gel Electrophoresis (DGGE) and PCR-Temperature Gradient Gel Electrophoresis (TGGE). Both DGGE and TGGE analyses have been utilized, together with the specific amplifications for ten Bifidobacterium species. 15

After written informed consent had been obtained from the parents twenty-three newborns after caesarean delivery and vaginal delivery have been enrolled in the study. The faecal samples were obtained on the third day of life.

The microbial DNA was extracted and analysed according to Favier et al, Environ 20 Microbiol 2002; 68:219-226 and Satokari et al, Appl Environ Microbiol 2001; 67:504-513; Satorkari etal System Appl Microbiol 2003; 26:572-584.

The results of the Bifidobacteria detected in faecal samples of newborns after caesarean delivery obtained at the 3rd day of life are given in Table 1. Table 2 gives 25 the Bifidobacteria detected in faecal samples of newborn after vaginal delivery obtained at the 3rd day of life.

It can be seen that the microbial flora of a infant bom via caesarean section shows strong differences with a infant bom via the vaginal route. Recognition of these 30 differences on species level enabled the present inventors to design the present compositions and methods. 14

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 bom via caesarean section, stimulating a healthy intestinal flora and consequently preventing infection and stimulating a healthy immune system. 2015218488 27 Aug 2015 5

Table 1: Caesarean NEWBORN B. breve B. infantis B. dentium B. angulatum B. bifidum B. lactis B. catenulatum group B. adolescentis B. longum B. gallic, 1 - - - . . . _ 2 - - - - - _ • ++ 3 - . _ . _ 4 - . _ . . . . 5 - . _ • • . . . 6 - . _ . . • - . 7 - • _ « . . . • 8 - . - • . • • 9 - . . _ _ • 10 - . _ - _ _ . · _ - ++ 11 - _ _ . • • • - 12 - - _ . . _ - 13 - _ _ _ _ . _ 16 - _ . . - . - - 17 - _ _ . _ 18 - . - _ _ _ . - 19 - . - _ _ _ - - 20 - _ _ _ _ - 21 - • - _ _ - - 22 - . _ _ _ _ _ . 23 - - - - - . - - - - (-) = no amplification 10 (+/-) = weak amplification ( + )= positive amplification (++) = strong amplification 15 15 2015218488 27 Aug 2015

Ta ble 2: Norma B. B. B. B. B. B. B. catenulatum B. B. B. NEWBORN breve infantis dentium angulatum bifldum lactis group adolescentis longum gallicum la - + - - - - - - ++ - 2a +/- - - - ++ - ++ - ++ - 3a - - - - - - + - - - 4a +/- - - - - - ++ + + - 5a +/- - - - ++ - ++ - ++ ++ 6a - - - - +/- - ++ - ++ - 7a - - - - +/- - ++ ++ - - 8a ++ ++ - - - - + ++ - - 9a - - - - - - + ++ + - 10a ++ - - - - - + + - - 11a ++ - - - ++ - ++ - ++ - 12a + + - - + - + - ++ - 13a +/- - - - - - + - + - 16a - - - . . - ++ - + - 17a +/- - - - + - + - + - 18a +/- - - - + - + - + - 19a + . . - - - + - + - 20a - . _ . - - + - + 21a - . - - - - + ++ + - 22a - + - . - - ++ - + - 23a + - - - ++ - ++ - + - 5 (-) = no amplification (+/-) = weak amplification ( + )= positive amplification (++) = strong amplification 10

Example 2: In vitro fermentation of Galacto-oligosaccharide by infant’s faeces

Aim: The capacity of galactooligosaccharides (GOS) and a combination of GOS and 15 inulin to favor the activity of lactic acid bacteria was evaluated using an in vitro semi dynamic batch fermentation system using infants faeces. The amount of lactate was determined, since this is the fermentation product of lactic acid bacteria including Bifidobacteria. 16 2015218488 27 Aug 2015

Method: Fresh faeces was obtained from healthy bottle-fed babies. Fresh faecal material from babies ranging 1 to 4 months of age was pooled and put into a preservative medium (buffered peptone 20.0 g/1, L-Cysteine-HCl 0.5 g/1, Sodium 5 thioglycollate 0.5 g/1, resazurine tablet 1/1, pH 6.7) within 1 h and stored at 4 °C for at most 2 h before the fermentation experiment was started.

The preserved solution of faeces was centrifuged at 13,000 rpm for 15 min. The supernatant was removed and the faces was mixed with McBain & MacFarlane 10 medium (Buffered peptone water 3.0 g/1, yeast extract 2.5 g/1, mucin (brush borders) 0.8 g/1, Tryptone 3.0 g/1, L-Cysteine-HCl 0.4 g/1, bile salts 0.05 g/1, K2HPO4.3H2O 2.6 g/1, NaHC03 0.2 g/1, NaCl 4.5 g/1, MgS04.7H20 0.5 g/1, CaCl2 0.228 g/1, FeS04 0.005 g/1), which is representative for the intestinal environment, in a weight ratio of 1:5. 15 At t = 0 15 ml of the faecal suspension was combined with 500 mg prebiotic in a bottle and mixed thoroughly. As a control no prebiotic was added. The prebiotics added were as followed:

Media: 1 2 3 GOS: 0 500 450 Inulin 0 0 50

As a source of GOS, Vivinal GOS (Borculo Domo) was used. As a source of inulin RaftilinHP (Orafti) was used. 25 15 ml was transferred into a dialysis tube in a 250 ml bottle filled with 250 ml of buffered medium (K2HP04.3H20 2.6 g/1, NaHCOa 0.2 g/1, NaCl 4.5 g/1, MgS04.7H20 0.5 g/1, CaCfe 0.228 g/1, FeS04.7H20 0.005 g/1, pH 6.3(7)). The bottle was closed and incubated at 37 °C. Samples of 1 ml were taken from the dialysis tube and from the 30 dialysis buffer with a hypodermic syringe after 3 h and stored at -18 °C. Experiments were performed in duplo.

Lactate was determined enzymatically, using a L-lactate acid detection kit with D-and L-lactate-dehydrogenase (Boehringer Mannheim, Mannheim, Germany). 17 2015218488 27 Aug 2015

Results and Conclusions: The results are expressed as amounts of lactate formed per g of prebiotic added. The results show that the amount of lactate is increased when an infant’s microflora ferments GOS or a mixture of GOS and inulin. When no prebiotic was added, lactate production is not observed. With GOS alone 0.1801 mmol/g 5 prebiotic lactate is produced. When a combination of GOS and inulin is added the lactate produced is 0.2119 mmol per g prebiotic.

The results indicate that the oligosaccharides stimulate the growth and/or activity of lactic acid bacteria in infants, including Bifidobacteria. These results are indicative for 10 the advantageous use of the indigestible oligosaccharides, particularly galactooligosaccharides, in the present composition and method according to the present invention, i.e. in a method for feeding babies bom via caesarean section.

Example 3: Method for feeding babies bom via caesarean section 15 Within two days after the infant is bom via caesarean section, a nutritional composition is administered which contains per 100 ml ready to feed formula: 1.6 gram protein, 3.6 gram fat, 6.4 gram digestible carbohydrates (mainly lactose), 0.8 gram non-digestible carbohydrates of which 0.60 gram transgalactooligosaccharides, 0.07 gram inulin, 1 x 109 cfu Bifidobacterium breve, 1 x 109 cfu Bifidobacterium 20 catenulatum, 1 x 109 cfu Bifidobacterium longum and 1 x 109 cfu Lactobacillus paracasei.

Example 4: Packaged composition

Packaged composition wherein the package indicates that the product is particular 25 suitable for oral feeding infants bom via Caesarean section, said composition comprising Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis, Bifidobacterium longum, galactooligosaccharides and inulin. 30 Example 5: Packaged composition

Composition as in Example 4, wherein the composition has a volume of 10 ml and is packed in a syringe. 18

Example 6: Rectal administration 2015218488 27 Aug 2015

Pll suitable for rectal administration to an infant within two weeks after birth, comprising galactooligosaccharides and Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum. 5 Although a preferred embodiment of the present invention has been described in the foregoing detailed description, it will be understood that the invention is not limited to the embodiment disclosed, but is capable of numerous rearrangements, modifications and substitutions without departing from the scope of the invention. 0 Throughout this specification the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps. 5 All publications mentioned in this specification are herein incorporated by reference. Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed in Australia or elsewhere 0 before the priority date of each claim of this application. 19

Claims (14)

1. THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:

   1. Use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the development of a healthy intestinal flora in an infant delivered via caesarean section, wherein the composition is for enteral administration and the microorganism is Bifidobacterium breve and the indigestible oligosaccharide is galactooligosaccharide.
2. 2. Use according to claim 1, wherein the composition is administered orally and is in the form of an infant milk formula or a reconstitutable dry composition; or wherein the composition is administered rectally and is in the form of a suppository, pill or tablet.
3. 3. Use according to claim 1 or 2, wherein the composition further comprises a Bifidobacterium selected from the group consisting of Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis, Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum.
4. 4. Use according to any one of claims 1-3, wherein the composition comprises Bifidobacterium breve and Bifidobacterium catenulatum.
5. 5. Use according to any one of claims 1-4, wherein the composition is administered to an infant receiving human breast milk.
6. 6. Use according to any one of claims 1-5, wherein the composition is administered to the infant within one week after birth.
7. 7. Method for the manufacture of an infant nutrition suitable for infants born via caesarean section comprising admixing: a) human breast milk; and b) a composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the composition contains at least one species of Bifidobacteria selected from the group consisting of Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium catenulatum, Bifidobacterium adolescentis Bifidobacterium thermophilum, Bifidobacterium gallicum, Bifidobacterium animalis, Bifidobacterium angulatum, Bifidobacterium pseudocatenulatum, Bifidobacterium thermacidophilum and Bifidobacterium longum.
8. 8. Use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the development of a healthy intestinal flora in an infant bom via caesarean section, wherein the composition is obtained by admixing: I) a nutritionally or pharmaceutically acceptable liquid; and II) a dry composition comprising the at least one microorganism and the at least one indigestible oligosaccharide, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligosaccharide.
9. 9. Use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the intestinal flora of an infant delivered via caesarean section, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligosaccharide; and wherein the composition is administered to the infant within 100 hours after birth.
10. 10. Use of at least one microorganism and at least one indigestible oligosaccharide for the manufacture of a composition for stimulating the intestinal flora of an infant delivered via caesarean section, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligosaccharide; and wherein the composition is administered rectally to an infant below 3 years of age.
11. 11. A method of stimulating the development of a healthy intestinal flora in an infant delivered by caesarean section comprising enterally administering to said infant a composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligosaccharide.
12. 12. A method of stimulating the development of a healthy intestinal flora in an infant delivered by caesarean section comprising administering to said infant a composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the composition is obtained by admixing: I) a nutritionally or pharmaceutically acceptable liquid; and II) a dry composition comprising the at least one microorganism and the at least one indigestible oligosaccharide, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligosaccharide.
13. 13. A method of stimulating the development of a healthy intestinal flora of an infant delivered by caesarean section comprising administering to said infant a composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligosaccharide; and wherein the composition is administered to the infant within 100 hours after birth.
14. 14. A method of stimulating the development of a healthy intestinal flora of an infant delivered by caesarean section comprising administering to said infant a composition comprising at least one microorganism and at least one indigestible oligosaccharide, wherein the at least one microorganism is Bifidobacterium breve and the at least one indigestible oligosaccharide is galactooligosaccharide; and wherein the composition is administered rectally to an infant below 3 years of age.