FIELD OF THE INVENTION

The present invention pertains to a kit of parts for improving or stimulating the healthy gut microbiota and/or reducing the risk of developing health risks and/or prevention of disorders in infants delivered via caesarean section (C-section), wherein the kit of parts comprises composition(s) to be administered to the pregnant or lactating mother of the infant and/or to the infant directly. BACKGROUND OF THE INVENTION

At birth, human infants start accumulating intestinal microbiota until a relatively stable state is reached. Mode of feeding, the duration of gestation and mode of delivery all affect the rate and trajectory of acquisition of a gut microbiota, which in turn are believed to have a considerable impact on health later in life.

C-section delivered infants suffer from the consequences of a non-existent transmission of gut microbiota from the mother to the infant at birth, and where colonisation of the infant is impaired. EPl 940250 has clarified there is a complete lack of any detectable amounts of Bifidobacterium species in the gut of C-section infants compared to the presence of significant amounts of at least three different Bifidobacterium species of the group of B. longum, B. breve, B. infantis, B. catenulatum, B. pseudocatenulatum, B. adolescentis, B. animalis, B. gallicum, B. lactis and B. bifidum observed in infants born by natural birth. While vaginally delivered infants acquire bacterial communities resembling their own mother's vaginal and gut microbiota, C-section infants harbour bacterial communities similar to those found on the skin surface. All has been illustratively confirmed by Dominguez-Bello et al. "Delivery mode shapes the acquisition and structure of the initial microbiota across multiple body habitats in newborns" PNAS (2010) 107 no. 26 11971 - 11975.

The observations described above have been attributed to the direct transmission (or non- transmission) of gut and vaginal microbiota from the mother to the infant that occurs at the time of birth due to the mode of delivery. Vaginal delivery places the infant in direct contact with the mother's vaginal and gut microflora during the birth process. By comparison, C- section delivery effectively removes the infant from exposure to the maternal gut and vaginal microbiota, by bringing the infant into the world through incisions made in both the uterus and abdomen of the mother. Often, the C-section delivered infant's primary contact with bacteria is from skin as a result of being held after birth. However, it is only more recent that the field has become aware that mode of delivery also has indirect impacts on the infant's intestinal microbiota.

For instance, pregnant mothers with anticipated C-section deliveries of their infants are more and more often prescribed antibiotics before the day of delivery. If antibiotics are not already given before delivery, then certainly they are given on the day of delivery and at least for some period afterwards as they have been shown to be clearly advantageous in the prevention of maternal wound healing. However, the short and long-term consequences of these antibiotics on the infant are not so well understood. At least one study has shown that milk samples from women who received antibiotics during pregnancy or lactation contained significantly lower numbers of lactobacilli or bifidobacteria (Ana Soto et al. "Lactobacilli and Bifidobacteria in Human Breast Milk: Influence of Antibiotherapy and other Host and Clinical Factors" JPGN 2014;59: 78-88). It is also reported that there is the potential for antibiotics taken during breast-feeding to be transferred to the suckling infant (J L Matthew "Effect of maternal antibiotics on breast feeding infants" Postgrad Med J 2004;80: 196-200).

Furthermore, recent studies suggest that C-section birth also influences the microbial composition of human milk (see: Cabrera-Rubio R, et al, "The human milk microbiome changes over lactation and is shaped by maternal weight and mode of delivery. " The American Journal of Clinical Nutrition. 2012;96(3):544-51; and Khodayar-Pardo P, et al "Impact of lactation stage, gestational age and mode of delivery on breast milk microbiota. " Journal of Perinatology : official journal of the California Perinatal Association. 2014;34(8):599-605). Khodayar-Pardo and colleagues observed that Bifidobacterium was detected more frequently in the breast milk of mothers who delivered vaginally than the ones who gave birth by C-section. This observation suggests that breastfeeding does not compensate the lack of microbial inoculation at birth and provides a plausible explanation to substantiate the delayed colonisation of Bifidobacterium described in C-section infants who are breastfed (see: Makino H, et al. "Mother-to-infant transmission of intestinal bifidobacterial strains has an impact on the early development of vaginally delivered infant's microbiota. " PLoS ONE. 2013;8(1 l):e78331; and Hesla HM, et al. "Impact of lifestyle on the gut microbiota of healthy infants and their mothers-the ALADDIN birth cohort. " FEMS microbiology ecology. 2014;90(3):791-801. Yet, the worldwide rate of C-section deliveries has increased over the last decade, making it the most common surgical procedure performed in women of childbearing age today. While the WHO recommends that C-section deliveries be indicated in up to 15 % of all deliveries, in many countries the rate of C-section deliveries significantly exceeds this recommendation. Over the past years, the medical field has started to realise that a C-section delivery introduces health risks, and the obstetrician is thus advised to assess these long- and short-term health consequences for mother and baby, as well as weigh the risks associated with the procedure itself against not performing the procedure. Laubereau et al. "Caesarean section and gastrointestinal symptoms, atopic dermatitis and sensitisation during the first year of life" Arch. Dis. Child (2004) 89: 993-997 concluded that C-section delivery might be a risk factor for diarrhoea and allergic sensitisation in infants with family history of allergy. Similarly, Hakansson et al. "Caesarean section increases the risk of hospital care in childhood for asthma and gastroenteritis" Clin. Exp. Allergy (2003) 33 : 757-764 described that there was a significant increase of the risk for developing symptoms of asthma and/or gastroenteritis in C- section delivered children, and this increased risk was speculated to be linked to the disturbed intestinal colonisation pattern. These increased health risks associated with C-section delivery are reviewed in Cho et al. "Cesarean section and development of the immune system in the offspring" Am. J. Obstetrics & Gynecology (2012). Furthermore, C-section infants are believed to have an increased risk of developing obesity later in life.

Needless to say, in view of foregoing, the art is in need of a solution to support gut microbiota of C-section delivery infants in order to better reflect those corresponding with normal delivery infants. SUMMARY OF THE INVENTION

Infants delivered via C-section have an intestinal microbiota which is different from the intestinal microbiota of infants born via the vaginal route. This is once more confirmed further below in Figure 1 in the experimental part. Particularly, infants born via C-section have a reduced rate of intestinal colonisation by Bifidobacteria and have a less diverse Bifidobacterium intestinal microbiota with regards to species than infants born via the vaginal route, particularly lacking Bifidobacterium breve, Bifidobacterium longum subsp. longum, Bifidobacterium longum subsp. infantis and Bifidobacterium bifidum. Also, the intestinal microbiota of infants delivered via C-section have a high content of (undesirable) Enter ob acted aceae such as Klebsiella, and Escherichia coli, from birth.

As discussed earlier, the intestinal microbiota differences observed in C-section infants when compared to vaginally delivered infants was previously attributed to direct effects caused by different microbiota exposure caused by different mode of delivery. While this is still acknowledged to be a significant contributor to these differences, it is now also appreciated that there may be indirect effects influencing the intestinal microbiota of the infant from the situation of the pregnant and/or breast-feeding mother of the C-section infant. As evidenced in the experimental parts, particularly Figure 1 therein, the differences are most profound in first few weeks, , with a Bifidobacterium microbiota population that is 100 x lower for C-section delivery infants than for the reference normal delivery infants, before the Bifidobacterium population of C-section delivery infants eventually recovers, only after about 16 weeks. The inventors found that these C-section specific issues of lack of Bifidobacterium species in the first few weeks subsequent to birth can be addressed - and the Bifidobacterium microbiota population recovered, to achieve levels of microbiota in C-section infants that resemble those observed from the reference group of infants (being vaginally delivered).

In view thereof, the inventors propose a kit of parts for improving or stimulating the health of gut microbiota and/or reducing health risks and/or prevention of disorders in infants delivered via C-section, wherein the kit of parts comprises composition(s) to be administered to a pregnant woman carrying an infant to be born via C-section, to a lactating women providing human milk to an infant born via C-section and/or directly to the infant born via C-section, and wherein said kit of parts comprises one or more compositions(s) comprising a therapeutically effective amount of at least one probiotic Bifidobacterium species selected from the group consisting of Bifidobacterium breve, Bifidobacterium longum subsp. longum, Bifidobacterium longum subsp. infantis and Bifidobacterium bifidum, and optionally a therapeutically effective amount of at least one non-digestible prebiotic oligosaccharide. Suitably, the kit of parts comprises one or more, preferably at least two, more preferably at least three of the following:

(i) a first composition for administration to a pregant woman at a time prior to C- section delivery of the infant thus when the mother is pregnant with the infant,

(ii) a second composition for administration to the C-section infant,

(iii) a third composition for administration to the mother of the C-section infant at the time after delivery of the infant and when the mother is breast-feeding the infant;

(iv) a fourth composition for administration to the C-section infant different from said second composition,

wherein each composition comprises a therapeutically effective amount of at least one probiotic Bifidobacterium species selected from the group consisting of Bifidobacterium breve, Bifidobacterium longum subsp. longum, Bifidobacterium longum subsp. infantis and Bifidobacterium bifidum, and optionally a therapeutically effective amount of at least one non-digestible prebiotic oligosaccharide.

The invention also concerns a method for (a) improving or stimulating the health of gut microbiota and/or (b) reducing health risks and/or (c) prevention of disorders in infants delivered via C-section, said method involving providing a kit of parts according to the invention and administrating each of the one or more composition(s) to a pregnant woman carrying an infant to be born via C-section, to a lactating women providing human milk to an infant born via C-section and/or to an infant born via C-section.

In other words, the invention also concerns the use of one or more composition(s) for the manufacture of a kit of parts for (a) improving or stimulating the health of gut microbiota and/or (b) reducing health risks and/or (c) prevention of disorders in infants delivered via C- section, wherein each of the one or more composition(s) comprises a therapeutically effective amount of at least one probiotic Bifidobacterium species selected from the group consisting of Bifidobacterium breve, Bifidobacterium longum subsp. longum, Bifidobacterium longum subsp. infantis and Bifidobacterium bifidum, and wherein each of the one or more composition(s) is administered to a pregnant woman carrying an infant to be born via C- section, to a lactating women providing human milk to an infant born via C-section and/or to an infant born via C-section. In other words, the invention also concerns a kit of parts according to the invention for use in(a) improving or stimulating the health of gut microbiota and/or (b) reducing health risks and/or (c) prevention of disorders in infants delivered via C-section, wherein each of the one or more composition(s) is administered to a pregnant woman carrying an infant to be born via C-section, to a lactating women providing human milk to an infant born via C-section and/or to an infant born via C-section.

Suitably, the first composition is administered to the pregnant mother when the infant is anticipated to be delivered by C-section. Suitably, the first composition is administered during the third trimester, more suitably during the second trimester and the third trimester, most suitably during the first trimester, second trimester and third trimester. Suitably, the first composition is administered at least one day prior to delivery, more suitably at least two days prior to delivery, even more suitably at least three days prior to delivery, most suitably at least one week prior to delivery. In some embodiments, the first composition may be administered before, during or after the pregnant woman is prescribed or given antibiotic therapy in anticipation of C-section delivery. Where the pregnant woman receives antibiotic therapy, the first composition may suitably be administered at a time of the day different from the time of day when antibiotic therapy is administered. In a separate aspect, the invention also pertains to the use of a composition for improving or stimulating the health of gut microbiota and/or reducing health risks and/or prevention of disorders in infants delivered via C-section, said composition being characterized as the first composition (i.e. composition (i)) herein. Herein, the first composition is administered to the pregnant women expecting to deliver the infant by C-section.

Suitably, the second composition is administered to the infant delivered via C-section starting at least in the first sixteen weeks after birth, preferably at least within twelve weeks after birth, even more preferably at least within eight weeks after birth, most preferably at least within four weeks after birth. Suitably, the second composition is administered to the infant delivered via C-section starting at least in the first two weeks after birth, preferably at least in the first week after birth, more preferably at least within 5 days after birth, even more preferably at least within 3 days after birth, most preferably at least within 2 days after birth. The same holds for the optional fourth composition intended for C-section infant, and which may be in a different administration form and/or as a subsequent form, i.e. a form for use after the second composition.

Suitably, the third composition is administered to the breast-feeding mother starting at least in the first sixteen weeks after birth, preferably at least within twelve weeks after birth, even more preferably at least within eight weeks after birth, most preferably at least within four weeks after birth. Suitably, the composition is administered to the breast-feeding mother starting at least in the first two weeks after birth, at least in the first week after birth, preferably at least within 5 days after birth, even more preferably at least within 3 days after birth, most preferably at least within 2 days after birth. It is of utmost importance to improve and/or accelerate developing the appropriate bifidobacteria population and Bifidobacterium species diversity in the gastrointestinal tract of C-section delivered infants.

The consequences of improving the gastrointestinal Bifidobacterium population in terms of numbers and diversity of species according to the invention are a reduced risk of occurrence or development of allergy, preferably food allergy, eczema (e.g. atopic dermatitis), asthma, allergic rhinitis and/or allergic conjunctivitis in infants delivered by C-section. Improvement of the gastrointestinal Bifidobacterium population in terms of numbers and diversity of species may also result in a reduced risk of infections, including gastrointestinal infections, and reducing the occurrence and/or severity of infections including gastrointestinal infections. C-section has also been associated with increased risks of developing allergies and obesity later in life, and the improvements in terms of swift microbiota restoration achieved by the invention thus also relate to reducing the risks of developing allergies and obesity later in life. BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows a comparison of the reference group and the control group in terms of total Bifidobacteria levels over the time period from day 3-5 until week 22, demonstrating that C- section infants (without any supplemented intervention directly or to the pregnant / breastfeeding mother) experience delayed colonisation of Bifidobacteria compared to vaginally delivered and breast-fed infants.

Figure 2 shows the same graph from Figure 1 with the addition of the results from the prebiotic group and the synbiotic group. In these results, it can be observed that levels of bifidobacteria in the synbiotic group were achieved that resemble those observed from the reference group of infants.

Figure 3 shows the percentage of bifidobacteria in each group relative to total bacteria at day 3-5, demonstrating that the effect of the synbiotic group generally described in Figure 2 could be observed from very early days of life.

DETAILED DESCRIPTION OF THE INVENTION

The invention pertains to infants born 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. The infant delivered via C-section may be either term or preterm human infant, where a "preterm infant" is a human infant born before 37 weeks of gestation and a "term infant" is a human born at 37 weeks or later of gestation.

Kit of parts

The kit of parts according to the invention comprises one or more composition(s) to be administered to a pregnant woman carrying an infant to be born via C-section, to a lactating women providing human milk to an infant born via C-section and/or directly to the infant born via C-section, and wherein said kit of parts comprises one or more compositions(s) comprising a therapeutically effective amount of at least one probiotic Bifidobacterium species selected from the group consisting of Bifidobacterium breve, Bifidobacterium longum subsp. longum, Bifidobacterium longum subsp. infantis and Bifidobacterium bifidum, and optionally a therapeutically effective amount of at least one non-digestible prebiotic oligosaccharide.

Suitably, the kit of parts comprises one or more, preferably at least two, more preferably at least three of the following:

(i) a first composition for administration to a pregant woman at a time prior to C- section delivery of the infant thus when the mother is pregnant with the infant,

(ii) a second composition for administration to the C-section infant,

(iii) a third composition for administration to the mother of the C-section infant at the time after delivery of the infant and when the mother is breast-feeding the infant; (iv) a fourth composition for administration to the C-section infant different from said second composition,

wherein each composition comprises a therapeutically effective amount of at least one probiotic Bifidobacterium species selected from the group consisting of Bifidobacterium breve, Bifidobacterium longum subsp. longum, Bifidobacterium longum subsp. infantis and Bifidobacterium bifidum, and optionally a therapeutically effective amount of at least one non-digestible prebiotic oligosaccharide.

In one embodiment, the kit of parts comprises one or more, preferably at least two, more preferably at least three of the following:

(i) a first composition for administration to an unborn infant to be born via C-section, wherein the first composition is to be administered to the woman pregnant with the infant,

(ii) a second composition for administration to the infant born via C-section, wherein the infant is 0 - 7 days of age, preferably 1 - 5 days of age, via direct administration to the infant,

(iii) a third composition for administration to the infant born via C-section, wherein the infant is 0 - 6 months of age, preferably 0 - 1 months of age, via administration to the lactating mother of the infant;

(iv) a fourth composition for administration to the C-section infant, wherein the infant is 3 days - 6 months of age, preferably 5 days - 1 month of age, via direct administration to the infant,

wherein each composition comprises a therapeutically effective amount of at least one probiotic Bifidobacterium species selected from the group consisting of Bifidobacterium breve, Bifidobacterium longum subsp. longum, Bifidobacterium longum subsp. infantis and Bifidobacterium bifidum, and optionally a therapeutically effective amount of at least one non-digestible prebiotic oligosaccharide.

The compositions comprised in the kit of parts are conveniently individually packaged, where appropriate comprising unit dose forms. As will be appreciated by the skilled person, the optimal packaging form depends on the form taken by a specific composition. The individually packaged compositions are preferably physically interconnected, such as by a single holder-packaging or single wrapper, enabling the kit to be recognised as a single unit. Preferably the holder-packaging is a carton box. Alternatively, the nutritional compositions of the kit of parts are unified by that each of the individually packaged nutritional compositions contains labelling or indicia, said labelling or indicia specifying the other nutritional compositions of the kit, or alternatively, the said labelling on the individually packed nutritional compositions making reference to the present method described hereinbelow.

In one embodiment, the kit comprises (i) and at least one of (ii) and (iii). In one embodiment, the kit comprises (i), (ii) and optionally (iv). In one embodiment, the kit comprises (i) and (iii). In one embodiment, the kit comprises (i), (ii), (iii) and optionally (iv). Composition (iv) is conveniently only present in case composition (ii) is present, and is different in composition or administration form from composition (ii). In one embodiment, the kit comprises at least (ii) and (iv).

Some examples of envisaged combinations of compositions that are comprised in the kit of parts according to the invention are:

(1) a first composition being a pregnancy product (supplement for pregnant women), a second composition being a booster (e.g. small volume format) and a fourth composition being an infant formula or stick sachet for addition to human milk, infant formula or combination thereof;

(2) a first composition being a pregnancy product and a second composition being stick sachet for addition to human milk, infant formula or combination thereof;

(3) a first composition being a pregnancy product and a third composition being a lactation product (supplement for lactating women). One or more compositions in the present kit of parts may be the same or all compositions in the kit of parts may be different, in terms of form, dosage, volume, additional ingredients, and any other variation possible. Suitably, each composition is formulated in a form that is optimal for the subject to which the composition is to be administered. It is thus preferred that the first composition is a composition, such as a nutritional supplement, that is especially beneficial for pregnant women, and which preferably comprises further components that are known in the art to be beneficial in this respect. It is thus preferred that the second composition is a composition, such as a complete nutrition or a nutritional supplement, that is especially beneficial for newly born infants, and which preferably comprises further components that are known in the art to be beneficial in this respect. It is thus preferred that the third composition is a composition, such as a nutritional supplement, that is especially beneficial for lactating women, and which preferably comprises further components that are known in the art to be beneficial in this respect. It is thus preferred that the fourth composition is a composition, such as a complete nutrition or a nutritional supplement, that is especially beneficial for newly born infants, and which preferably comprises further components that are known in the art to be beneficial in this respect. Preferred forms of the compositions will be described further below. It is preferred that the fourth composition is provided in another form as the second composition.

Probiotic Bifidobacterium species

The kit of parts according to the invention comprises compositions comprising a probiotic Bifidobacterium species. In the context of the present invention, the term "probiotic" refers to a strain of bacteria which is perceived as probiotic bringing a health benefit to the targeted infants delivered via C-section. Probiotic Bifidobacteria are known in the art. It should be noted that it is preferably neither the object nor the effect of the treatment of the invention to promote colonisation by the species of probiotic that is administered but rather to promote colonisation with other bifidobacteria species so as to achieve an early bifidogenic intestinal microbiota comparable with that found in healthy, breast-fed, vaginally-delivered infants. Thus, in one embodiment, the improvement or stimulation of a healthy gut microbiota concerns the promotion of the bifidobacterial colonization of gut of C-section infants, preferably the colonization of at least one Bifidobacterium species other than the one(s) present in the composition according to the invention is promoted. Thus, in an especially preferred embodiment, the compositions comprised in the kit of part according to the invention comprise B. breve and the improvement or stimulation of a healthy gut microbiota concerns the promotion of the intestinal colonization of at least one Bifidobacterium species other than B. breve.

The compositions of the present kit of parts preferably comprise at least one Bifidobacterium selected from the group consisting of B. breve, B. longum subsp. longum, B. longum subsp. infantis, B. bifidum and B. catenulatum, even more preferably at least one Bifidobacterium selected from the group consisting of B. breve and B. longum subsp. longum, most preferably at least B. breve. In one embodiment, the composition comprises at least two different Bifidobacterium species, preferably including at least one, more preferably two of the above list, more preferably B. breve and B. longum. In one embodiment, the compositions comprise at least three, most preferably at least four different Bifidobacterium species. In a preferred embodiment, the composition comprises a maximum of one or two Bifidobacterium species.

In one embodiment, the probiotic species in the compositions of the kit consists of one or two, preferably one of the above Bifidobacterium species, and the probiotics in the compositions preferably consists of B. breve and B. longum subsp. longum, most preferably consists of B. breve.

In one embodiment, the compositions of the present kit of parts further comprise 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.

The probiotic(s) is(are) present in the compositions of the kit of parts in a therapeutically effective amount or "amount effective for treating" in the context of the invention. The optimal content of probiotics aill vary depending on the form of the composition and the target group, but in general terms it is preferred that the probiotic is included in an amount of

2 13 5 12 1

10 - 10 cfu per g dry weight of the composition, suitably 10 - 10 cfu/g, most suitably 10 - 10 ¹⁰ cfu/g in case the composition is a complete nutrition, in particular an infant formula. In case the composition according to the invention is in the form of a supplement to be added to a nutritional product or to be ingested as such, the content of the probiotics therein may be somewhat higher, such as 10 ⁴ to 10 ¹⁶ cfu/g, suitably 10 ⁷ to 10 ¹³ cfu/g, more suitably 10 ¹⁰ to 10 ¹² cfu/g, based on dry weight of the supplement. Non-digestible prebiotic oligosaccharides

Preferably the compositions of the present kit of parts comprises at least one non-digestible prebiotic oligosaccharide. The term "oligosaccharide" as used in the present invention refers to saccharides with an average degree of polymerization (DP) of 2 to 250, preferably an average DP 2 to 100, more preferably 2 to 60.

In the context of the present invention, the term "prebiotic" refers to one or more non- digestible oligosaccharides. Advantageously, the non-digestible oligosaccharide is water- soluble (according to the method disclosed in L. Prosky et al, J. Assoc. Anal. Chem. 71 : 1017- 1023, 1988). Non-digestible oligosaccharides are not digested in the intestine by the action of digestive enzymes present in the upper digestive tract (small intestine and stomach) of the C- section infant but instead are fermented by the intestinal microbiota of said infant, thus conferring benefits upon the host wellbeing and health. Preferably the growth of at least bifidobacteria and preferably also Lactobacilli is stimulated. An increased content of bifidobacteria and/or lactobacilli stimulate the formation of a healthy intestinal microbiota..

Suitable non-digestible oligosaccharides are selected from the group consisting of fructo- oligosaccharide, non-digestible dextrin, galacto-oligosaccharide, xylo-oligosaccharide, arabino-oligosaccharide, arabinogalactooligosaccharide, gluco-oligosaccharide, glucomannooligosaccharide, galactomanno-oligosaccharide, mannanoligosaccharide, chito- oligosaccharide, uronic acid oligosaccharide, sialyl-oligosaccharide and fuco-oligosaccharide. In some embodiments, the non-digestible oligosaccharides comprise those obtainable from human milk.

The present non-digestible oligosaccharide is preferably selected from the group consisting of galacto-oligosaccharides and fructo-oligosaccharides (including fructo-polysaccharides such as inulin). Preferably at least 50 wt.% of the present non-digestible oligosaccharides have an average degree of polymerisation of 2 to 60. In one embodiment, the non-digestible oligosaccharide comprises galacto-oligosaccharides, in particular β-galacto-oligosaccharides. The galacto-oligosaccharides are preferably [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, a good example being tram'-galacto-oligosaccharides. The galacto-oligosaccharides preferably comprise saccharides with an average DP of 2 to 10 (scGOS). (Trans)galactooligosaccharide is for example available under the trade name Vivinal®GOS (Borculo Domo Ingredients, Zwolle, Netherlands), Bimuno (Clasado), Cup-oligo (Nissin Sugar) and 01igomate55 (Yakult). Fructo-oligosaccharides may be inulin hydrolysate products having an average DP within the aforementioned (sub-)ranges. Such fructo-oligosaccharide products are for instance short-chain fructooligosacharides (scFOS) commercially available such as the product sold under the trade mark Beneo® P95 or Raftilose P95 (Orafti). A particular type of long-chain fructo-oligosaccharide (lcFOS) is inulin, such as Raftilin FIP.

In a particular preferred embodiment the compositions of the present kit of parts comprise at least galacto-oligosaccharides and fructo-oligosaccharides oligosaccharides and/or fructopolysaccharides, preferably scGOS and lcFOS (preferably having an average DP > 15), suitably in a weight ratio 5: 1 - 20: 1, even more suitably 7: 1 - 15: 1, even more suitably 8: 1 - 10 : 1 , most suitably about 9: 1.

The prebiotics are present in therapeutically effective of prebiotic amounts. The compositions of the present kit of parts suitably comprise 0.05 to 20 wt% of said non-digestible oligosaccharides, more suitably 0.5 to 15 wt%, even more suitably 1 to 10 wt%, most suitably 2 to 10 wt%, based on dry weight of the compositions of the kit.

LCPUFAs

The compositions of the present kit of parts may further comprise long chain polyunsaturated fatty acids (LC-PUFA). LC-PUFA are fatty acids wherein the acyl chain has a length of 20 to 24 carbon atoms (preferably 20 or 22 carbon atoms) and wherein the acyl chain comprises at least two unsaturated bonds between said carbon atoms in the acyl chain. More preferably the present compositions comprises at least one LC-PUFA selected from the group consisting of eicosapentaenoic acid (EPA, 20:5 n3), docosahexaenoic acid (DHA, 22:6 n3), arachidonic acid (ARA, 20:4 n6) and docosapentaenoic acid (DP A, 22:5 n3), comprising at least DHA. LC-PUFA further have anti-inflammatory effects and promote the adhesion of lactic acid producing bacteria to mucosal surfaces, thereby stimulating the development of a healthy microbiota, which are further advantages for use in C-section delivered infants. 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 compositions preferably comprises at least one of ARA and DHA in phospholipid form. Administration

The compositions in the present kit of parts are enterally administered, and the term "enteral" is intended to refer to the delivery directly into the gastrointestinal tract of the infant (e.g., orally or via a tube, catheter or stoma), but it also encompasses rectal or anal administration. Suitably, the compositions are intended for oral administration.

The compositions in the present kit of parts may be in any form known in the art, such as in solid form, in semi-solid form or in liquid form. Preferably, the compositions are a nutritional composition or a nutritional supplement, preferably in the form of a powder, capsule or tablet. Preferably, the compositions comprise the probiotic in freeze-dried form, which is especially suitable when the compositions are in powder, capsule or tablet form. Preferably, the compositions, when in powder, capsule or tablet form, especially when in powder form, are contained within a container, preferably a stick or stickpack or a sachet. The second or fourth composition to be administered to the C-section infant may be in the form of complete nutrition. For instance, the second or fourth composition may comprise an infant formula, preferably in powder form suited to be reconstituted with water. In one preferred embodiment, the second or fourth composition is an infant formula. An infant formula for use according to the present invention may contain a protein source in an amount of not more than 2.0 g/lOOkcal, preferably 1.8 to 2.0 g/lOOkcal. The protein preferably provides 5 to 15% of the total calories. Preferably the composition comprises protein that provides 6 to 12% of the total calories. More preferably protein is present in the infant formula below 9% based on calories. The source of the protein should be selected in such a way that the minimum requirements for essential amino acid content are met and satisfactory growth is ensured. Hence protein sources based on cow's milk proteins such as whey, casein and mixtures thereof and proteins based on soy, potato or pea are preferred. Preferably, the infant formula comprises casein and whey protein. In case whey proteins are used, the protein source is preferably based on acid whey or sweet whey, whey protein isolate or mixtures thereof and may include alpha-lactalbumin and alpha-lactoglobulin. More preferably, the protein source is based on acid whey or sweet whey from which caseino- glyco-macropeptide (CGMP) has been removed. Preferably the infant formula comprises casein, preferably it comprises at least 3 wt.% casein based on dry weight. Preferably the casein is intact and/or non-hydrolyzed. For the present invention protein includes peptides and free amino acids. As far as whey proteins are concerned, the protein source may be based on acid whey or sweet whey or mixtures thereof and may include alpha-lactalbumin and beta- lactoglobulin in whatever proportions are desired. The proteins may be intact or hydrolysed or a mixture of intact and hydrolysed proteins. The infant formula may contain a carbohydrate source. Any carbohydrate source conventionally found in infant formulae such as lactose, saccharose, maltodextrin, starch and mixtures thereof may be used although the preferred source of carbohydrates is lactose. Preferably the carbohydrate sources contribute between 35 and 65% of the total energy of the formula. The infant formula may also contain a source of lipids. The lipid source may be any lipid or fat which is suitable for use in infant formulas. Suitably the infant formula contains at least one, suitably at least two lipid sources selected from the group consisting of rape seed oil (such as colza oil, low erucic acid rape seed oil and canola oil), high oleic sunflower oil, high oleic safflower oil, olive oil, marine oils, microbial oils, coconut oil, palm kernel oil and milk fat. The lipid component of the infant formula suitably provides 2.9 to 6.0 g, more suitably 4 to 6 g per 100 kcal of the composition. When in liquid form, the infant formula suitably comprises 2.1 to 6.5 g lipid per 100 ml, more suitably 3.0 to 4.0 g per 100 ml. Based on dry weight the infant formula suitably comprises 12.5 to 40 wt% lipid, more suitably 19 to 30 wt%. The fat source preferably has a ratio of n-6 to n-3 fatty acids of about 5: 1 to about 15: 1; for example about 8: 1 to about 10: 1. The infant formula may also contain all vitamins and minerals understood to be essential in the daily diet and in nutritionally significant amounts. Minimum requirements have been established for certain vitamins and minerals. Examples of minerals, vitamins and other nutrients optionally present in the infant formula include vitamin A, vitamin Bl, vitamin B2, vitamin B6, vitamin B 12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorous, iodine, iron, magnesium, copper, zinc, manganese, chloride, potassium, sodium, selenium, chromium, molybdenum, taurine, and L-carnitine. Minerals are usually added in salt form. The infant formula may optionally contain other substances which may have a beneficial effect such as lactoferrin, nucleotides, nucleosides, and the like.

In other embodiments, at least one of the second or fourth compositions of the kit of parts of the invention are not in the form of complete nutrition. For instance, the second compositions may be intended to be administered to the infant directly and in addition to breast-milk or infant formula ingested by the infant. Such compositions are typically of small size, e.g. 0.5 - 5 ml or 0.5 - 5 g, and may be referred to as supplement or fortifier. In one aspect, the invention thus pertains to a method for the manufacture of a nutritional product for infants born via caesarean section comprising admixing: (a) human breast milk, infant formula or a mixture thereof; and (b) the second composition according to the invention. In a further embodiment, the invention concerns a method for providing nutrition to an infant, said method comprising the admixing of (a) and (b) to manufacture a nutritional product as defined above and administering the nutritional product thus obtained to an infant born via caesarean section, preferably in the aforementioned short time frames after birth.

The 'small volume' compositions may also be administered to the infant with a syringe, pipette or tube, thus especially useful for C-section infants in hospital settings. In case of oral administration, the composition is preferably administered in liquid form having a volume between 0.5 to 5 ml. The compositions may be provided in the form of unit dose form, which refers to individual single-use packages. The composition may be present in a container containing one single or more unit dose(s).

In some other embodiments, the second composition is intended to be added to another product before ingestion by the infant. Preferably, the present composition is accompanied with instructions to add the composition to a nutritional product before ingestion. The nutritional product to which the present composition is to be added may be any suitable nutritional product fro C-section infants, such as breast-milk or infant formula.

In a further preferred embodiment, the composition of the present kit of parts is rectally or anally administered to the mother and/or infant, preferably in the form of a suppository, pill or tablet. Any base components commonly used for suppositories can be used as a base component of the suppository of the present invention, including oils and fats, waxes, and the like of animal, vegetable or mineral origins. They may be partially or totally synthesised materials. Applications

The kit of parts of the present invention is for improving or stimulating the healthy gut microbiota and/or reducing the risk of developing health risks and/or prevention of disorders in infants delivered via C-section.

In one aspect, the invention pertains to a method for

(a) improving or stimulating the health of gut microbiota and/or

(b) reducing health risks and/or

(c) prevention of disorders

in infants delivered via C-section, comprising providing a kit of parts according to the invention, said method involving at least two, preferably at least three of the following:

administering said first composition to a pregnant woman carrying an infant to be born via C-section delivery;

administering said second composition to the C-section delivery infant;

- administering said third composition to the mother of the C-section infant at a time after delivery of the infant and when the mother is breast-feeding the infant mother; administering said optional fourth composition to the C-section delivery infant.

In the context of the invention, the embodiments (b) and (c) involve risks and disorders associated with the impaired gut microbiota balance, particularly the lack of bifidobacteria microbiota, of C-section delivery infants at the onset of life.

It is of utmost importance to improve and/or accelerate developing the appropriate Bifidobacteria population and Bifidobacterium species diversity in the gastrointestinal tract of C-section delivered infants before and at the onset of life outside the womb, and that will contribute to create a favourable gut ecosystem milieu through their metabolic capability. It is thus preferred that the treatment is targeted at the infant delivered via C-section at least in the first two weeks after birth, preferably within the first week after birth, more preferably at least within 5 days after birth, even more preferably at least within 3 days after birth, most preferably at least within 2 days after birth. It shows from Figures 2 and 3 that the impact is most dramatic in the early onset.

Improvement of the gastrointestinal Bifidobacterium population in terms of numbers and diversity of species results in a reduced risk of occurrence of allergy, preferably food allergy, eczema (e.g. atopic dermatitis), asthma, allergic rhinitis and/or allergic conjunctivitis in infants delivered by C-section.

Improvement of the gastrointestinal Bifidobacterium population in terms of numbers and diversity of species may also result in a reduced risk of infections, including gastrointestinal infections, and including reducing the occurrence and/or severity of infections, such as infections due to reduction of intestinal concentrations of pathogens particularly Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus haemolyticus, Streptococcus, Clostridium difficile, Bacilus subtilis, Pseudomonas aeruginosa, Enterobacter, Klebsiella, Acinetobacter, Proteus, Aeromonas and Escherichia coli. This is achieved by (i) stimulating the growth of the lactic acid producing bacteria, (ii) decreasing the growth of pathogenic bacteria; and/or (iii) decreasing the adhesion of pathogenic bacteria to the intestinal epithelial cells and/or intestinal mucus. C-section has also been associated with increased risks of developing allergies and obesity later in life, and the improvements in terms of swift microbiota restoration achieved by the invention thus also relate to reducing the risks of developing allergies and obesity later in life, meaning at an age exceeding the age at which the infant receives the composition, suitably exceeding the age by at least 12 months, more suitably by 24 months, by 36 months, by 5 years, most suitably by 8 years. Suitably, "later in life" means at toddler age, childhood age, adolescence age, or adult age.

In this document and in its claims, the verb "to comprise" and its conjugations is used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. In addition, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that more than one of the element is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article "a" or "an" thus usually means "at least one".

EXAMPLES Example 1 Method & Materials: In a multi- country, double-blind, randomised controlled study, infants born by C-section were randomly assigned to receive (1) a standard infant formula (control group, n = 45), (2) the standard formula supplemented with scGOS/lcFOS (about 9: 1 weight ratio) (0.8g/100ml) (prebiotic group, n = 39), and (3) the standard formula supplemented with scGOS/lcFOS (about 9: 1 weight ratio) (0.8g/100ml) and B. breve M-16V (7.5 lO ⁸ CFU/lOOml) (i.e. synbiotic group, n = 45), from birth until age 16 weeks. 'scGOS' are short- chain galactooligosaccharides (Vivinal-GOS™; Borculo Domo Ingredients, Netherlands; Degree of Polymerisation [DP] 2 - 8), 'lcFOS' are long-chain fructooligosaccharides (Raftiline HP™, Orafti, Tienen, Belgium; average DP 22 - 25).

One further group of infants delivered vaginally and breastfed as long as possible were included as the reference group (n = 28).

Stool samples were collected at day 3-5, week 2, week 4, week 8, week 12, week 16, and week 22 (6 weeks post-intervention). Bifidobacteria levels were analysed by q-PCR. Faecal pH and short chain fatty acids (SCFA) were assessed.

Results: The results are shown in the figures and described below.

Figure 1 shows a comparison of the reference group and the control group in terms of total Bifidobacteria levels over the time period from day 3-5 until week 22, demonstrating that C- section infants (without any supplemented intervention directly or to the pregnant / breastfeeding mother) experience delayed colonisation of bifidobacteria compared to vaginally delivered and breast-fed infants.

Figure 2 shows the same graph from Figure 1 with the addition of the results from the prebiotic group and the synbiotic group. In these results, it can be observed that levels of bifidobacteria in the synbiotic group were achieved that resemble those observed from the reference group of infants.

Figure 3 shows the percentage of bifidobacteria in each group relative to total bacteria at Day 3-5, demonstrating that the effect of the synbiotic group generally described in Figure 2 could be observed from very early days of life.

In terms of the faecal pH and SCFA analysis, it was noted that the synbiotic group resulted in a favourable pH intestinal milieu in the C-section delivered infants (data not shown), and that the lower pH in the synbiotic group could be attributed to a high production of acetate (data not shown).

Example 2: Kits of parts according to the invention

Each of the kits are accompanied from instructions directed to the treatment of C-section delivery infants.

Kit 1 comprises:

(i) a first composition being a pregnancy product (supplement for pregnant women);

(ii) a second composition being a booster (volume of 5 ml) for C-section delivery infant, and (iv) a fourth composition being an infant formula.

Kit 2 comprises:

(i) a first composition being a pregnancy product (supplement for pregnant women);

(ii) a second composition being a booster (volume of 5 ml) for C-section delivery infant, and (iv) a fourth composition being a stick sachet for addition to human milk, infant formula or combination thereof.

Kit 3 comprises:

(i) a first composition being a pregnancy product (supplement for pregnant women); and

(ii) a second composition being an infant formula.

Kit 4 comprises:

(i) a first composition being a pregnancy product (supplement for pregnant women); and (ii) a second composition being stick sachet for addition to human milk, infant formula or combination thereof.

Kit 5 comprises:

(i) a first composition being a pregnancy product (supplement for pregnant women); and (iii) a third composition being a lactation product (supplement for lactating women).

All compositions in these exemplary kits of part comprise B. breve and a mixture of galaco- oligosaccharides and fructo-oligosaccharides.