WO2010117274A1 - Carbohydrates enhancing the production of a c5 and/or a c6 scfa - Google Patents

Abstract

The invention relates to a carbohydrate being fermented in at least 4 hours by a subject for use as a medicament for preventing, delaying and/or treating a disorder of the gastrointestinal tract. Said fermentation preferably induces a detectable increase of the production of a C5 and/or a C6 SCFA by a bacterium in the lumen of the GI tract of the treated subject. Said increase is more preferably detectable at least 10 hours after the administration of the carbohydrate.

Description

CARBOHYDRATES ENHANCING THE PRODUCTION OF A C5 AND/OR A C6 SCFA

Field of the invention

The invention relates to a carbohydrate which is able to induce a detectable increase of a C5 and/or a C6 Short Chain Fatty Acid (SCFA), said SCFA having a healthy effect on the gastrointestinal health of the subject treated.

Background of the invention The human large intestine harbors a complex diversity of micro-organisms, which exert both positive and negative effects on gut physiology. The gut microbiota affects the local and systemic immune system, has a role in the intestinal defense against pathogens and is important in the metabolism of nutrients and toxic compounds [I]. The two main types of bacterial metabolism that occur in the gut are saccharolytic and proteolytic fermentation. Saccharolytic fermentation is favorable for the host because it produces short chain fatty acids (SCFAs), such as butyrate, acetate and propionate. A variety of health promoting properties have been attributed to SCFAs. End products of proteolytic fermentation are less favorable, including nitrogenous metabolites, some of which are carcinogenic. The proximal colon is predominantly a site of saccharolytic fermentation, whereas in the distal colon, where fermentable carbohydrates are present only in minute amounts, mainly proteolytic fermentation takes place. This may partly explain why many gastrointestinal disorders may occur mainly in the distal colon. Dietary ingredients which can increase the amount of saccharolytic fermentation throughout the gut and reduce proteolysis as treatment for intestinal disorders are currently under investigation. There is growing interest in functional foods which affect the composition and activity of the gut microbiota. One approach is the ingestion of live microorganisms (probiotics) that enter the gut and persist long enough to have beneficial effects on the host. Another one is to use non-digestible carbohydrates (prebiotics) that are selectively fermented by indigenous beneficial bacteria [2]. The health-promoting effects that have been attributed to probiotics and prebiotics are modulation of the immune system, improved digestion and absorption, vitamin synthesis, protection against the development of colon cancer, cholesterol reduction and inhibition of the growth and colonization of potential pathogens [3]. Short chain fatty acids produced during bacterial fermentation of non-digestible carbohydrates were reported to have several beneficial effects [4].

Increased colonic butyrate formation has often been proposed as one of the protective mechanisms of high fiber diets [5-7]. Butyrate is the major energy source for colonocytes [2], and it may act as a signal metabolite affecting epithelial cell proliferation and differentiation [8]. Besides, there is some evidence that butyrate beneficially affects several inflammatory parameters including nuclear factor kappa B (NF-kB), secretory IgA (SIgA), cytokines and myeloperoxidase (MPO) activity [9]. Furthermore, butyrate stimulates intestinal mucus production supporting the mucosal barrier function [10, 11], possesses anti-oxidative capacity [12, 13], increases mucosal blood flow [14], and may decrease colonic epithelial permeability [15, 16]. Paradoxically, a rat study suggested that butyrate causes an increase in colonic sensitivity [17]. There is still a need for new ways of preventing, delaying or treating any gastrointestinal disorder in a subject. Surprisingly, the inventors discovered that a C5 and/or a C6 SCFA have an attractive healthy effect on the gastrointestinal tract and that the production of a C5 and/or a C6 SCFA effect could be advantageously induced by providing a specific carbohydrate to a subject as described herein.

Description of the invention A carbohydrate

In a first aspect, there is provided a carbohydrate being fermented in at least 4 hours by a subject for use as a medicament for preventing, delaying and/or treating a disorder of the gastrointestinal tract. In a preferred embodiment, said fermentation induces a detectable increase in the production of a C5 and/or a C6 SCFA by a bacterium in the lumen of said treated subject. More preferably, said increase is detectable at least 10 hours after the administration of the carbohydrate. Preferably, a carbohydrate is fermented in at least 4 hours by micro-organisms in the lumen of a subject. More preferably, a carbohydrate is fermented in at least 4 hours by micro-organisms in the colon of a subject.

A carbohydrate may be a non-digestible, non-digested or a digestible carbohydrate. A carbohydrate is preferably an oligosaccharide or a polysaccharide. An oligosaccharide is defined herein as a saccharide polymer with a small number (usually 3 to 10) of components sugars, also known as simple sugars. A polysaccharide is a relatively more complex carbohydrate than an oligosaccharide. Polysaccharides are polymers made up of many monosaccharides joined together by glycosidic bonds. A carbohydrate may be a non-digestible carbohydrate, or a non-digestible oligosaccharide (NDO) or a non- digestible polysaccharide. Within the context of the invention, "non-digestible" carbohydrate preferably means that the human being does not have the intestinal enzymes to digest this carbohydrate. However, this carbohydrate is expected to be fermented by (endogenous) intestinal bacteria. This fermentation mainly occurs in the colon.

Therefore in a preferred embodiment, a non-digestible carbohydrate is administered which will be converted at least partly into a C5 and/or a C6 SCFA in the colon of a subject by the endogenous intestinal bacteria. In another preferred embodiment, a non-digested or a digestible carbohydrate is used. When one refers herein to a digestible carbohydrate, it may be that under specific circumstances, this digestible carbohydrate is a non-digested carbohydrate. A non- digested carbohydrate is a carbohydrate which is potentially a digestible carbohydrate as defined herein but which is not digested due to certain circumstances such as transit through the upper intestines, inaccessibility by digestive enzymes (e.g. due to enclosure within a plant cell), or other reasons known to the skilled person in the art.

As used herein a carbohydrate may be defined by at least one of the following parameters: has a DP of at least 10, is branched, comprises a charged molecule, is modified, comprises rhamnose, In a preferred embodiment, a carbohydrate has a degree of polymerisation (DP) of at least 10 . More preferably, the DP is of at least 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 50, 100, 200, 500, 700 or higher. Preferred examples of carbohydrate having a DP of at least 10 are: inulin, resistant starch. More preferred is FruTEX being an inulin having an average DP of 25 and which shows an attractive production level of a C5 SCFA. Average means that the DP may be ranged between 10 and 60. Within, the context of the invention, "branched" preferably means that a carbohydrate comprises a side chain attached to the linear backbone of a carbohydrate. Examples of sides chains include: Man (Mannose), Lignol, Gal (galactose), GIc (glucose), Ara (arabinose), Rham (rhamnose), Glue acid (gluconic acid) (see table 2 for examples of carbohydrate containing such branched molecule). Preferred examples of branched carbohydrate are: branched inulin from Agave, arabinogalactan. Within the context of the invention, "a charged molecule" preferably means a methyl-, an acetyl-, an ethyl, a carboxymethyl and other modifications known to the person skilled in the art. A preferred carbohydrate with a charged molecule includes a modified pectin: methylated or acetylated pectin, esterified pectin. Within the context of the invention, a carbohydrate is preferably modified. Preferred modifications include providing partial resistance to the endogenous microorganisms, or modifying the ratio mannose to galactose, or esterification. A preferred example of a resistant carbohydrate includes resistant starch, more preferably HiMaize as identified in the example. A preferred example of a carbohydrate having a modified ratio mannose to galactose includes a modified guar gum, more preferably Hindustan gum as identified in the example. A preferred examples of an esterified carbohydrate is esterified apple pectin. Esterification may be partly or total: ranged between 30 and 100%. Preferably esterification is at least 90%.

Preferred carbohydrates comprising rhamnose include Karaya gum and certain pectins as identified in the example.

Another preferred carbohydrate is arabinogalactan since it is known to be slowly fermented ( preferably 4, 6, 8, 10 hours or more) and can even be found in faecal excreta of individual with a fast intestinal transit time.

Without wishing to be bound by any theory, we expect that such a carbohydrate may lead to an induction of a higher detectable increase of a C5 and/or a C6 SCFA than corresponding induction for a carbohydrate with a lower DP (and/or being not branched, not comprising a charged molecule, being not modified, not comprising rhamnose).

Alternatively or in combination, we expect that such a carbohydrate might lead to an induction of a detectable increase of a C5 and/or C6 SCFA which may be detectable in a longer time period after the administration of said carbohydrate than corresponding induction for a carbohydrate with a lower DP (and/or being not branched, not comprising a charged molecule, being not modified, not comprising rhamnose). We expect this "slow fermentation" to be attractive for the reason mentioned below. The two major processes in terms of substrate fermentation in the colon are carbohydrate and protein fermentation. Most microorganisms prefer to ferment carbohydrate as they tend to incorporate the amino acids from protein into their own enzyme machinery, but when (fermentable) carbohydrate is depleted, they usually switch to protein fermentation. Therefore, in the proximal colon there is primarily saccharolytic fermentation (carbohydrate fermentation), whereas in the distal colon the major process going on is proteolytic fermentation (protein fermentation), with concommittent production of toxic metabolites such as ammonia, phenolic compounds and H₂S. Therefore delaying fermentation (what we call "slow fermentation") in itself would mean that one would extend carbohydrate fermentation to the distal colon, and one would reduce the amount of undesirable metabolites being produced there. In addition, this "slow fermentation" leads to the production of a C5 and/or C6 SCFA with healthy property. Besides, a so-called "slow fermentation" might be energetically beneficial for the microorganisms. If there is more than enough fast available substrate, they produced mostly lactate, formate and acetate. When substrate becomes limiting or is difficult to ferment, they need to switch to more energy conserving processes, and then produce the more longer-chain SCFA as C5 and C6. The production of lactate, formate and acetate is known to the skilled person as being detrimental for a microorganism since it lowers is capacity to produce ATP, i.e. its main energetic drive force. A carbohydrate as defined herein is preferably selected from the group consisting of: resistant starch such as RS HiMaize and other resistant starches, Locust bean gum, Guar gum, modified guar gum such as Hindustan S gum , arabinogalactan, pectin apple, pectin apple esterified. Each of these carbohydrates is commercially available as identified in the example.

A carbohydrate as provided herein is preferably being fermented in at least 4 hours by the microbiota or microorganism present in the lumen of a subject. This preferably means that it takes at least 4 hours or more for the endogenous bacteria of the intestine of the subject treated to ferment said carbohydrate. A carbohydrate is said to have been fermented when at least 50% of the ingested amount of carbohydrate has been fermented or at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%. In a preferred embodiment, a carbohydrate has been fermented when it is no longer detectable in the intestinal tract. The detection of a carbohydrate is carried out as later explained herein.

After at least 10 hours after ingestion of said carbohydrate, its presence in the intestinal tract is preferably no longer detectable. Two time periods are defined herein: the time needed to obtain a detectable increase of a C5 and/or a C6 SCFA starting from the ingestion of a carbohydrate by a subject. This first time period is of 10 hours or more. It may take 11, 12, 13 hours or more. The second time period is the time needed by the bacteria of the intestine to ferment a carbohydrate to produce a detectable increase of a C5 and/or a C6 SCFA. This second time period is of at least 4 hours. However, this second time period may be at least 5, 6, 7, 8, 9 or more than 9 hours. This second time period starts when an ingested carbohydrate has reached the colon wherein the microorganisms that will ferment it are present, which may be at least 6 hours after ingestion of a carbohydrate. The time needed by a subject to ferment a carbohydrate is preferably assessed as follows in an in vitro assay using a labeled substrate which can be fermented by the microbiota into a C5 and/or C6 SCFA. When a given substrate (for example starch) is no longer fermented, i.e. when the amount of labelled substrate is seen as stable or used up, that is, when the accumulation of SCFA reaches a plateau, it gives an indication of the time needed by said microbiota (for example a faecal inoculum) for fermenting said substrate. A C5/C6 is preferably quantified using GC- MS as referred to elsewhere. A substrate may be labelled using ¹³C isotope. Within the context of the invention, a C5 and/or a C6 source is preferably administered orally. However, other administration modes are not excluded. For other modes of administration, the first time period as defined herein may be the same as defined herein for oral administration or may be different. The first time period may be shorter than the first time period for oral administration. The second time period as defined herein is in principle not dependent on the administration mode since it reflects the time needed by the bacteria from the intestine to ferment a carbohydrate. Such a carbohydrate is attractive to be used as a medicament for preventing, delaying and/or treating a disorder of the gastrointestinal tract since such said carbohydrate is preferably able to induce a detectable increase of a C5 and/or a C6 SCFA. More preferably, said increase is detectable at least 10 hours after the oral administration of said carbohydrate. The induced detectable increase of the internal concentration of a C5 and/or a C6 SCFA in the lumen of a subject is preferably measured by comparison to the physiological concentration of a C5 and/or a C6 SCFA of a control subject as defined herein. The concentration of a C5 and a C6 SCFA in the lumen is preferably assessed using GC-MS (Gas Chromatograph- Mass Spectrometry) as described in 18.) and in the examples. More preferably, the increase is detectable when it is of at least 1%, 2%, 5%, 7%, 10%, 15%, 20%, 25%, or more. Usually, the physiological concentration of a C5 SCFA in the lumen of a subject is ranged between 0 (or undetectable concentration) and 10 mM and for a C6 SCFA between 0 (or undetectable concentration) and 10 mM. Therefore the administration of a carbohydrate as defined herein may lead to a concentration of a C5 SCFA in the lumen which is ranged between 1 and 50 mM or between 5 and 50 mM. Therefore the administration of a carbohydrate as defined herein may lead to a concentration of a C6 SCFA in the lumen which is ranged between 1 and 50 mM or between 2 and 50 mM or between 5 and 50 mM. An increase of a C5 and/or of a C6 concentration in the lumen of at least 1 mM is believed to be able to provide a desired healthy effect (protective, treating effect) in the gastrointestinal tract of a subject. The increase may be of at least 2 mM or more. A control subject may be a subject who has not yet been treated with a carbohydrate of the invention. In another preferred embodiment, the induction is detectable at least approximately 5, 6, 7 or 8 hours after the administration of the carbohydrate. It is also encompassed by the present invention, that the induction is detectable after a longer time period than 8 hours. The skilled person will understand that the type of carbohydrate administered may influence the time period at which an induction of a C5 and/or C6 SCFA is detectable.

In a preferred embodiment, a carbohydrate as defined herein is fermented in at least 4 hours by the bacteria present in the lumen of a subject for use as a medicament for preventing, delaying and/or treating a disorder of the gastrointestinal tract, wherein the fermentation of said carbohydrate induces a detectable increase of the production of a C5 and/or a C6 SCFA by said bacteria in the lumen of the treated subject at least 10 hours after the administration of said carbohydrate.

More preferably, said carbohydrate is such that wherein at least 50% of the ingested carbohydrate is fermented in at least 4 hours by the bacteria present in the lumen of the treated subject.

In a preferred embodiment, preferably in combination with two previous preferred embodiments, a carbohydrate is such that : the carbohydrate has a DP of at least 100 and/or It comprises a mannose, lignol, galactose, glucose, arabinose, rhamnose and/or gluconic acid side chain and/or

It comprises a methyl-, acethyl- an ethyl, a carboxymethyl and/or It is an esterified carbohydrate. More preferably, a carbohydrate is: a branched inulin, preferably a branched inulin from agave, arabinogalactan, a methylated or acetylated pectin, an esterified pectin, preferably an esterified apple pectin, - Himaize, Hindustan gum, karaya gum, xanthan gum, gellan gum,.

- GOS (galactoligosaccharide) Alginate

Rice starch, waxy mais starch, tapioca-starch and/or carrageen C5 and C6 are both a SCFA with a carbon chain length of 5 and 6 respectively. The term C5 or C6 SCFA may encompass the linear unbranched C5 or C6 (n-valerate or n- caproate). In addition, the invention encompasses any form of branched chain and/or substituted valerate and/or caproate. C5 and C6 SCFA are molecules which have attractive properties for gastro intestinal health. C5 SCFA is known to have a protective effect on human colon cancer cells: an anti-proliferative, apoptotic effect and differentiating properties on human colon cancer cells has been demonstrated via histone hyperacetylation on human colon cancer cells (19). In addition, C5 seems to have an anti-inflammatory role. It has been shown that C5 is able to modulate the expression of ICAM-I (Intercellular Adhesion Molecule- 1) and E-selectin ( 20), said molecules being involved in gut inflammation.

In addition, the present invention shows that a C5 and/or a C6 SCFA, preferably a C5 SCFA is able to induce a detectable increase of the transepithelial resistance (TER) in a subject or in a cell. Therefore the invention also encompasses a carbohydrate as identified herein wherein the detectable C5 and/or C6 SCFA is able to induce a detectable increase of the transepithelial resistance (TER) in a subject or in a cell. A

TER may be assessed on a gastrointestinal cell from a subject or from a gastrointestinal control cell derived from a cell line. A preferred cell line is Caco2 as used in example 3. A detectable TER increase may be detected at least approximately 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,1 6, 17, 18, 19, 20, 21, 22, 23, or 24 hours after the detection of the presence of a C5 and/or C6 SCFA in a subject or in a cell. It is also encompassed by the present invention, that the induction is detectable after a longer time period than 24 hours. The skilled person will understand that the type of carbohydrate administered may influence the time period at which an induction of a C5 and/or C6 SCFA is detectable and therefore the time period at which a detectable increase of TER is detectable. Alternatively, a TER may be assessed as in example 3 by directly adding a C5 and/or a C6 SCFA to a cell.

Preferably, the TER increase is detectable when it is of at least 1%, 2%, 5%, 7%, 10%, 15%, 20%, 25%, or more. The increase is preferably measured by comparison to the TER of a cell or a subject before administration of a carbohydrate or before administration of a C5 and/or C6 SCFA. In view of the known healthy effects of a C5 SCFA, a C5 SCFA is a preferred SCFA in the context of the invention.

It is further encompassed by the present invention to administer one or more distinct carbohydrates as defined herein. The use of several distinct carbohydrates may be chosen to ensure a continuous induction of the production of a C5 and/or a C6 SCFA to optimise the healthy effect.

In a further preferred embodiment, a carbohydrate is administered orally, preferably in a slow-release and/or enteric formulation. To optimize the production of a C5 and/or a C6 SCFA, a carbohydrate is preferably formulated so that it will be as less as possible digested or degraded by the enzymes of the subject and will be as much as possible fermented by intestinal bacteria. Such formulations are known to the skilled person as enteric formulations, encapsulations (coatings), and/or slow-release formulations are more extensively described later herein.

As to the total quantity administered of carbohydrate, for example, quantities of 5 to 30 g may be orally administered on a daily basis. Preferably, 5 to 15 g a day taken 1-3 times a day. In a preferred embodiment, at least 5 g of carbohydrate is administered per day to induce a production of at least 1 mmol of a C5 SCFA, or at least 5 mmol, 10 mmol, 15 mmol, 20 mmol, 25 mmol, 30 mmol and/or at least 1 mmol of a C6 SCFA , or at least 5 mmol, 10 mmol, 15 mmol, 20 mmol, 25mmol or at least 30 mmol in the lumen of the treated subject. In a more preferred embodiment, at least 10 to 20 g of carbohydrate is administered per day to induce a production of at least 5 mmol, 10 mmol, 15 mmol, 20 mmol, 25 mmol, 30 mmol, 40 mmol, 50 mmol, 60 mmol, 70 mmol, 80 mmol, 90 mmol, 100 mmol, 110 mmol, 120 mmol of a C5 SCFA and/or at least approximately 5 mmol, 10 mmol, 15 mmol, 20 mmol, 25 mmol, 30 mmol, 40 mmol, 50 mmol, 60 mmol, 70 mmol, 80 mmol, 90 mmol, 95 mmol of a C6 of SCFA in the lumen o f the treated subj ect .

The identity of the SCFA obtained (a C6 versus a C5 SCFA) can be controlled by the identity of the carbohydrate(s) used. For example, if one wishes to obtain more than 50%, or more than 60%, or more than 70%, or more than 80% or more than 90% of a C6 SCFA on the total amount of C5 and C6 SCFA produced, one would preferably use a carbohydrate comprising at least 50% of 90% esterifϊed pectin. Preferably, use is made of 90% esterifϊed pectin.

In another preferred embodiment, a C5 and/or a C6 SCFA is produced by the combined action of a bacterium and a carbohydrate, wherein both are preferably administered to a subject or wherein only a carbohydrate as defined herein is administered to a subject. Preferred bacteria include one or more of the following: a food-grade bacterium, a commensal bacterium. In one embodiment, a bacterium is a food-grade bacterium, particularly a gram-positive bacterium, and more preferably a lactic acid bacterium. A bacterium may also be a probiotic bacterium, which in itself has a beneficial effect when ingested by a subject.

A preferred bacterium belongs to a genus selected from the group consisting of Lactobacillus, Lactococcus, Leuconostoc, Carnobacterium, Streptococcus, Bifidobacterium, Bacteroides, Eubacterium, Clostridium, Fusobacterium,

Propionibacterium, Enterococcus, Staphylococcus, Peptostreptococcus, and Escherichia. A further preferred bacterium is a Lactobacillus or Bifidobacterium species selected from the group consisting of L. reuteri, L. fermentum, L. acidophilus, L. crispatus, L. gasseri, L. johnsonii, L. plantarum, L. paracasei, L. murinus, L. jensenii, L. salivarius, L. minutis, L. brevis, L. gallinarum, L. amylovorus, L. casei, B. bifidum, B. longum, B. infantis, B. breve, B. adolescente, B. animalis, B. gallinarum, B. magnum, and B. thermophilum. It is to be noted that each of these bacteria is probably not able to directly produce a C5 and/or a C6 SCFA. However, they indirectly induce their production in the colon by changing the colon environment, so that other bacteria would be able to produce a C5 and/or a C6 SCFA. It is for example known that lactic acid bacteria produce lactate, which will assist endogenous bacteria in a cross- feeding mechanism to produce SCFA. An amount of a bacterium ranged between 10⁷ and 10¹³ may be administered. Preferably, said amount is ranged between 10⁹ and 10¹¹.

Alternatively or in combination with earlier preferred embodiments, in another embodiment, a given quantity of a C5 and/or a C6 SCFA is directly introduced or administered into a subject. Depending on the desired effect, a quantity of a C5 and/or a C6 SCFA may be of at least 1 mmol per day administered to a human being or at least 5 mmo I/day, or at least 10 mmo I/day, or at least 15 mmo I/day, or at least 20 mmo I/day, or at least 25 mmo I/day, or at least 30 mmo I/day, or more.

Depending on the identity of the carbohydrate and optionally of bacterium used, the skilled person may have to adapt the quantity used to reach the desired effect (i.e. increased quantity of a C5 and/or a C6 SCFA produced in the lumen). Preferably, a carbohydrate and optionally a bacterium are suitable for convenient (oral) administration in one or more doses per day or per week.

In the context of the invention, an organism or an individual or a subject is preferably an animal, more preferably a mammal, even more preferably a human being. Even more preferably, a subject treated is suspected to have a high risk of developing a disorder or a condition affecting the health of its gastrointestinal tract due for example to potential genetic predisposition, and/or to the age of the subject and/or to the lifestyle of a subject (for example nutritional habit and/or to the absence of physical activity). Examples of disorder or conditions affecting a gastrointestinal tract include: inflammation due to infection with virus or pathogenic bacteria, inflammation due to genetic predisposition such as Irritable Bowel Diseases (IBD, inducing Crohn's disease and Ulcerative colitis), colon cancer, Pouchitis and Irritable Bowel Syndrome. Alternatively the invention may also be applied to any person having a high protein and/or low fibre diet. In this context high protein may mean more than 125g/day and low fibre may mean less than 15g/day.

The term "prevention" shall be understood to include complete prevention, prophylaxis, as well as statistically significantly decreasing the individual's risk of getting such a disorder or condition affecting his/her gastrointestinal tract. The term shall also be understood to include alleviation of any of the symptoms of such a disorder or condition affecting his/her gastrointestinal tract already developed.

The term "treatment" or "treating" is understood to be the management and care of a patient for the purpose of combating such disorders or conditions. The term "treating" is also herein synonymous of "delaying" the development of any of the symptoms associated with such conditions. Usually, one may expect that the delay in the development of a symptom is of approximately at least one day, one week or more.

In a preferred embodiment, a carbohydrate comprises at least one of: - at least 5 g of a carbohydrate as defined herein with or without non-digestible carbohydrate, and optionally between 10⁷ and 10¹³ bacteria.

Composition

In a second aspect, there is provided a composition, preferably a food composition or a pharmaceutical composition comprising a carbohydrate and optionally a bacterium as defined herein. More preferably, a pharmaceutical composition comprises a carbohydrate and optionally a bacterium and at least one inert excipient. In a preferred embodiment, a composition comprises at least one of: at least 5 g of a carbohydrate as defined herein, and optionally between 10⁷ and 10¹³ bacteria. Optionally, an additional active ingredient may be present in a food or pharmaceutical composition.

As additional active ingredient in a food or pharmaceutical composition, one may cite: an additional SCFA selected from the group comprising of acetate, propionate and butyrate.

Other additional active ingredients in a pharmaceutical composition may be an ingredient which is normally classically used for treating a disorder or a condition of the gastrointestinal tract such as 5-ASA (5 -aminosalicylic acid), an anti-acid, a steroid or combination thereof. In a preferred embodiment, a food or a pharmaceutical composition is suited for oral administration. Pharmaceutical compositions will usually comprise a pharmaceutical carrier in addition to a carbohydrate and optionally a bacterium. The formulation of a composition depends on the intended mode of administration and (therapeutic) application, on the carbohydrate chosen and on the presence of a bacterium. A pharmaceutical carrier can be any compatible, non toxic substance suitable to deliver a C5 and/or C6 source to the GI-tract of a subject. E.g. sterile water, or inert solids or excipient may be used as the carrier usually complemented with pharmaceutically acceptable adjuvants, buffering agents, dispersing agents, and the like. Compositions will either be in liquid, e.g. a stabilized suspension of a carbohydrate, or in solid and/or dry forms, e.g. a powder of lyophilized bacteria. E.g. for oral and rectal administration, the bacteria can be administered in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions. In this case, a carbohydrate is preferably in the form of a solution or emulsion. The bacteria as used in the invention may be dried and later on mixed with inactive ingredients and/or excipients and optionally encapsulated with for example gelatin to form gelatine capsules. Alternatively, the bacteria may be tabletted. Inactive ingredients or inert excipients are such as flavouring agents, stabilizers, sugars or other energy sources, buffering agents, thickeners, diluents, dispersing aids, emulsifiers, and/or binders. Examples of inactive agents are: glucose, lactose, sucrose, mannitol, starch, cellulose or cellulose derivatives, magnesium stearate, stearic acid, sodium saccharin, talcum, magnesium carbonate and the like. Some of these components are carbohydrate and may also be fermented into a SCFA, possibly into a C5 and/or a C6 SCFA. In one embodiment, a bacterium is not formulated with a carbohydrate. In another embodiment, it is advantageously that a bacterium is formulated with a carbohydrate, since it may increase the formation of a C5 and/or a C6 SCFA any further. The bacteria may also be first added in a liquid form, after which the combination is dried. The use of a carbohydrate enriched media such as a dairy product, preferably milk may be used to this end.

A preferred composition according to the invention is suitable for consumption by a subject, preferably a human. Such compositions may be in the form of a food supplement or a food or food composition, which besides a carbohydrate and optionally a bacterium also contains a suitable food base. A food or food composition is herein understood to include liquids for human consumption, i.e. a drink or beverage. The food or food composition may be a solid, semi-solid and/or liquid food or food composition, and in particular may be a dairy product, such as a fermented dairy product, including but not limited to a yoghurt, a yoghurt-based drink or buttermilk. A wide range of food products is suitable as a carrier for a carbohydrate of the present invention to be consumed to obtain the beneficial health effects.

Examples of such food products are cereal bars, chocolate bars, cookies and biscuits, confectionery products, condiments, confectionary, beverages, desserts, snacks, spreads like margarine or low fat margarines or dairy spreads, ice cream, dressings, mayonnaise, sauces, bakery products like bread, shortenings, cheese (soft cheese, hard cheese), soups, dairy drinks, fruit drinks or juices, vegetable drinks or juices, combinations of dairy, and/or fruit, and/or vegetable drinks, cocoa drinks, and especially dairy mini-drinks. Other examples of food products are chocolate, and ice cream. It is also envisaged that a food product according to the invention comprises a chocolate composition, that may be employed as a filling, ingredient and/or coating for a confectionary product. For example, the chocolate may be used to coat ice confections (such as ice cream , sorbets, water ices and the like) and/or the chocolate may be dispersed within an ice confection.

In case the food product is a beverage, more specifically a fruit drink, or combination of fruit and dairy drink, it preferably comprises at least 10% by weight of the composition of a fruit component, wherein the fruit component is selected from fruit juice, fruit concentrate, fruit juice concentrate, fruit puree, fruit pulp, comminuted fruit, fruit puree concentrate, and combinations thereof. Examples of such fruit components are orange juice, apple juice, grape juice, peach pulp, banana pulp, apricot pulp, concentrated orange juice, mango pulp, concentrated peach juice, raspberry puree, strawberry puree, apple pulp, raspberry pulp, concentrated grape juice, concentrated aronia juice, concentrated elderberry juice. Preferably such a beverage comprises at least 30% by weight of the beverage of said fruit component, more preferred at least 40% by weight of the beverage of said fruit component. These amounts are calculated as if undiluted, non-concentrated fruit juices and purees and the like are used. Thus, if 0.5% by weight of a 6-fold fruit concentrate is used, the actual amount of fruit component incorporated is 3% by weight of the beverage. Any commonly available fruit component might be used in the beverages according to the invention, and may be selected from one or more of the following fruit sources: citrus fruit (e.g. orange, tangerine, lemon or grapefruit); tropical fruit (e.g. banana, peach, mango, apricot or passion fruit); red fruit (e.g. strawberry, cherry, raspberry or blackberry), or any combination thereof.

In a further preferred embodiment the food product is a spread such as water-in-oil emulsions, for example a margarine or low fat margarine type food product. A spread may also be an oil- in- water emulsion, like dairy spreads. Suitably the total triglyceride level of such a spread may range from about 10% by weight to 85% by weight of the composition, more preferred from 20% to 70% by weight, most preferred from 30% to 60% by weight of the composition.

A unit amount of a food product is a quantity of a food product which is usually consumed as a single serving. The unit amount or serving size of such food products depends on the specific product. A few non- limiting examples of typical serving sizes are: milk, yoghurt: 200 mL natural cheese: 43 gram processed cheese: 57 gram fruit juice: 177 mL soft drink: 200 mL bread: 1 slice, 35 gram coffee: 125 mL tea: 15O mL cereal bar, candy bar: 50 gram chocolate: 30 gram ice cream: 100 mL spread: 15 gram soup: 250 mL cocoa beverage: 200 mL

A unit amount of a food product in the context of the present invention may be packed and sold as a single portion. For example, ice cream may be packed as individual units, therewith making such an individual portion a unit amount in the context of the present invention. The actual weight or volume of such an individually packed product may be higher or lower than indicated above for a standard serving size. For example probiotic dairy drinks are consumed from small bottles, individually packed, having a volume of about 10O mL.

The food product may be dried and contain less than 40% water by weight of the composition, preferably less than 25%, more preferably from 1 to 15%. Alternatively, the food may be substantially aqueous and contain at least 40% water by weight of the composition, preferably at least 50%, more preferably from 65 to 99.9%.

The food or food product preferably comprises nutrients including carbohydrate (including sugars and/or starches), protein, fat, vitamins, minerals, phytonutrients (including terpenes, phenolic compounds, organosulfides or a mixture thereof) or mixtures thereof. The food may be low calorie (e.g. have an energy content of less than 100 kCal per 100 g of the composition) or may have a high calorie content (e.g. have an energy content of more than 100 kCal per 100 g of the composition, preferably between 150 and 1000 kCal). The food may also contain salt, flavours, colours, preservatives, antioxidants, non-nutritive sweetener or a mixture thereof.

Such foods or food compositions may be prepared in a manner known per se, e.g. by adding a carbohydrate and optionally a bacterium to a suitable food or food base, in a suitable amount. In such carbohydrate and optionally bacterium comprising composition intended for oral administration, at least part or all of a carbohydrate may be formulated in a capsule, an enteric formulation and/or a slow-release formulation Such capsule and/or formulation are well known to the skilled person (21, 22, 23, 24). This is a preferred embodiment, since it will protect at least part or all of a carbohydrate from degradation in the intestinal tract before reaching the intestine wherein it will be converted into a C5 and/or a C6 SCFA. If a digestible carbohydrate is present as carbohydrate, it is preferred that it is formulated in such a capsule, enteric formulation and/or slow release formulation.

In a further preferred embodiment, the bacteria are micro-organisms that are used in or for the preparation of a food or food composition, e.g. by fermentation. Examples of such bacteria include lactic acid bacteria, such as probiotic lactic acid bacteria strains as earlier exemplified herein. In doing so, the bacteria as used in the invention may be used in a manner known per se for the preparation of such fermented foods or food compositions, e.g. in a manner known per se for the preparation of fermented dairy products using lactic acid bacteria. In such methods, the bacteria as used in the invention may be used in addition to the micro-organism usually used, and/or may replace one or more or part of the micro-organism usually used. For example, in the preparation of fermented dairy products such as yoghurt or yoghurt- based drinks, a food grade lactic acid bacterium as used in the invention may be added to or used as part of a starter culture or may be suitably added during such a fermentation or may be added at the end of a fermentation. Preferably, a lactic acid bacterium is added at the end of the fermentation.

Use In a further aspect, there is provided the use of a carbohydrate or of a composition as defined herein, for the manufacture of a medicament for preventing, delaying and/or treating a disorder of the gastrointestinal tract of the subject. Said carbohydrate or composition is preferably fermented by a bacterium in the lumen of said subject and thereby induces a detectable increase of a C5 and/or a C6 SCFA in the lumen of a subject. More preferably, said increase is detectable at least 10 hours after the administration of the carbohydrate. All features of this use have already be defined herein. Method

Accordingly, in a further aspect, there is provided a method for preventing, treating and/or delaying a disorder of the gastrointestinal tract comprising administration of a carbohydrate or a composition as defined herein. Said carbohydrate or composition is preferably fermented by a bacterium in the lumen of said subject and thereby induces a detectable increase of a C5 and/or a C6 SCFA in the lumen of a subject. More preferably, this increase is at least 10 hours after oral ingestion. All features of this method have already be defined herein.

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 the verb "to consist" may be replaced by "to consist essentially of meaning that a carbohydrate or a product or a composition as defined herein may comprise additional component(s) than the ones specifically identified, said additional component(s) not altering the unique characteristic of the invention. 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". All patent and literature references cited in the present specification are hereby incorporated by reference in their entirety. Each embodiment as identified herein may be combined together unless otherwise indicated.

The following examples are offered for illustrative purposes only, and are not intended to limit the scope of the present invention in any way.

Description of the figures

Figure 1. Cumulative production of valerate and caproate after 72 h in TIM-2 with addition of 5 gram substrate per day, except for potato starch, which was dosed at 10, 15 and 20 gram. Figures 2, 3, 4. The increase in TER seen on administration of C4 (butyrate, Figure 2), C5 (valerate, Figure 3) and C6 (caproate, Figure 4) at 2mM within the first 24 hours of addition to confluent Caco2 cell monolayer.

Examples Example 1 Materials and methods

TNO' s in vitro model of the proximal large intestine (TIM-2) simulates the average conditions in the lumen of the human proximal colon (25). The model consisted of a number of linked glass units with flexible walls inside. Body temperature and peristaltic movements were achieved by pumping water at 37 degrees C into the space between the glass jacket and the flexible wall at regular intervals. The computer controlled the sequential squeezing of the walls, causing the chyme to be mixed and transported through the system. The model was equipped with ho How- fibre semipermeable membranes inside the lumen of the model to remove water and fermentation products, such as SCFA. In addition, they maintained physiological concentrations of small molecules, such as electrolytes, and prevented product inhibition of enzymes due to accumulation of microbial metabolites. A constant volume of the luminal content was maintained by water absorption controlled by a level sensor. The environment in the model was kept strictly anaerobic by flushing with gaseous nitrogen, to allow for the growth of a dense, complex microbiota, comparable to that found in humans in the first part of the colon (caecum/proximal colon). The pH of 5.8 in the proximal colon was controlled via the computer by using a pH sensor in combination with 2M NaOH secretion. At the start of each experiment the model was inoculated with approximately 30 ml of a standardized, cultivated faecal microbiota (29). The microbiota was allowed to adapt to the model conditions for 16 h, after which the experiments started. Addition of the various substrates started after the adaptation period. The substrates were added to the standard ileal delivery medium without the standard carbohydrate mixture (see below) and fed to the system at a rate of 5 g of substrate per day for a period of 3 days, except for potato starch which was tested in a dose range of 10, 15 and 20 gram/day for 3 days. Every 24 h a 25-ml sample was removed from the model to simulate passage of material from the proximal part to the distal colon. In these luminal samples and samples of the dialysate taken at the same moment in time the concentrations of the SCFA were determined.

Gibson et al. (28) described a medium which simulates the material passing the ileo- caecal valve in humans (ileal delivery). This medium was modified for the experiments in TIM-2 concerning the following components (g/1): 4.7 pectin, 4.7 xylan, 4.7 arabinogalactan, 4.7 amylopectin, 23.5 casein (all from Sigma- Aldrich, Zwijndrecht, The Netherlands), 39.2 starch (BDH, Amsterdam, The Netherlands), 17 Tween-80 (Merck, Amsterdam, the Netherlands), 23.5 bactopeptone (Oxoid, Haarlem, the Netherlands), 0.4 bile (Oxoid). The test susbstrates were added to this medium with the omission of the standard carbohydrates listed above.

SCFAs (acetic, propionic, butyric, valeric, and caproic acid) were analysed in samples collected from the lumen and dialysis of TIM-2 as described by Van Nuenen (26). Briefly, analysis was performed on a gas chromatograph (GC; Chrompack CP9001, Varian, Bergen op Zoom, The Netherlands) according to the method described by Jouany (27). After centrifugation of the TIM-2 samples, the supernatant was diluted (7% by volume) with a mixture of methanol, internal standard (2 mg/ml 2-ethyl butyric acid) and formic acid (20%). Of this mixture 0.5 ml was loaded onto a 'wide-bore' GC column (Stabilwax-DA; length 15 m; ID 0.53 mm; film thickness 0.1 mm; Restek, Bad Homburg, Germany) using a Chrompack CP9050 automatic sampler (Varian).

Results

Figure 1 shows the cumulative production of valerate and caproate on the different substrates, compared to the standard carbohydrate mix. The results show that there is a correlation between speed of fermentation of the substrates and the production of valerate and caproate. For instance, lactose and inulin with a low degree of polymerization (DP; <DP>=3 and <DP>=9) show no valerate and caproate production. Increasing the DP to 25, and thus decreasing fermentation speed, results in the production of valerate. Also branching of inulin (from Agave) leads to some valerate production. Since not all micro-organisms can produce valerate or caproate, both SCFA are not necesarily expected on all substrates. HiMaize, a resistant fraction of starch, leads to much higher production of valerate and caproate than potato starch, which also contains regular digestible fractions and thus is expected to be fermented more easily than HiMaize. Guar gum leads to higher valerate and caproate production than hydro lyzed guar gum (Sunfϊbre AG and R), confirming the hypothesis of increased DP leading to more C5 and C6, although the production of valerate and caproate is already low on guar gum. The modified guar gums indicated by Hindustan gums have a higher ratio of mannose to galactose compared to regular guar gum, and lead to higher production of valerate and caproate indicating that besides degree of polymerization, monosaccharide composition is also important. Arabinogalactan, a slow fermentable substrate leads to relatively large quantities of valerate and caproate. Pectin from apple also shows large quantities of both SCFA, while pectin from citrus shows no detectable production. Therefore, the source of pectin is very important. Modification of pectin by esterification leads to variable results, with 30% esterification leading to an increase in the two SCFA, whereas further esterification to 60% reduces production, likely because the substrate cannot be completely fermented. In contrast, 90% esterification leads to more valerate and caproate than the 60% esterified pectin. Importantly, it leads to the highest ratio of caproate:valerate of all the substrates tested. Karaya gum, a substrate containing rhamnose, like pectin also leads to production of valerate and caproate. In conclusion, degree of polymerization (DP), as well as branching and monosaccharide composition dictate the amount and ratio of valerate and caproate being produced by the intestinal microbiota. This seems to be related to speed of fermentation, as well as the specific micro-organisms involved in fermentation of the substrates. The identity of the SCFA obtained can be controlled by the identity of the substrate(s) used. For example, if one wishes a high C6 SCFA, one would use 90% esterified pectin.

Example 2

Breads were baked using the constituents as given in table 3. In the recipe for the brown bread, use was made of the fibre materials rice bran, wheat germ and soya grit. The advantage of using these raw materials is that a brown bread is made containing the same or larger amount of the minerals (Fe and Zn) and vitamins (Vitamin Bl, B3 and B6) as a wholemeal bread. In a normal diet wholemeal bread contributes in relevant amounts to the daily intake of these vitamins and minerals. The white bread may optionally be fortified or restored with mineral and vitamin preparations. On an industrial scale (100 kg flour batches) the doughs could be processed without problems during dividing, proofing and moulding (no stickiness; normal dough properties). During baking the doughs showed no wild oven spring. Inulin was taken as a representative example of an attractive carbohydrate for the production of a C5 and/or C6 SCFA.

Example 3

The effects of SCFA's (butyrate, propionate and acetate) on increasing transepithelial resistance (TER) in Caco2 cell line, an established model of coloncytes, have been shown by others (30 and 31). An increase in TER is regarded as beneficial as it shows that the epithelia have an enhanced barrier function and a concomitant decrease in permeability. This increase in TER has been shown to be caused by remodelling of tight junction proteins. The effects of longer chain SCFA valerate and caproate were studied in the Caco2 system as explained below. Cells were treated with 2mM of the SCFA and the TER over the next 24 hours constantly monitored. Protocol SFCA C4, C5 and C6 - Caco-2 - TER -Caco-2 cells were cultured in 24 wells inserts for 2 weeks. -The 70% alcohol was removed the cellzcope unit and PBS was added to each well to remove traces of alcohol.

-The Cellzscope unit (nanoAnalytics GmbH, Mϋnster,Germany http://www.nanoanalytics.com/en/hardwareproducts/cellzscope/index.php) was placed in the CO₂ incubator for at least 1 hour before adding inserts. This allowed the unit to equilibrate up to 37⁰C to avoid temperature effects on the cells.

-The Cellzscope unit was placed in the safety cabinet and PBS was removed from each well and the filters inserts were transferred from the 24 wells tissue culture plate to the Cellzscope unit with sterile tweezers. 350 μl prewarmed DMEM was added to the apical compartment and 800 μl was added to the basolateral compartment. -Cellzscope unit was placed in the CO₂ incubator and connected to the amplifier and computer. Software was set to continuous scanning and started. Cells on filter inserts were left for at least 3 hours to calibrate in the Cellzscope in the CO₂ incubator.

-After the calibration period, the cellzscope was disconnected from the amplifier and placed in the safety cabinet. 50 μl 16 mM C4, C5 or C5 was added to apical compartment to reach a final 2 mM C4, C5 or C6 concentration. -The Cellzscope unit was placed in CO₂ incubator and reconnected. The TER was recorded for at least 20 hours. The data was transferred to excel and normalized to the TER values when the SCFA's were added.

CULTURING CACO-2 CELLS

Medium: IxDMEM (InVitrogen, cat. 61965-026) containing 10% foetal bovine serum (FBS, cat. Al 5- 151 , PAA, Pasching, Austria), heat-inactivated at 56 ⁰C, 30 min. 100 U/ml penicillin/ 100 μg/ml streptomycin, Sigma, cat. P0781)

Remove old medium by aspiration

Wash monolayer in flasks with 10 ml PBS for 2 min Remove PBS by aspiration

Add ImI of 0.25 % trypsin-EDTA

Incubate the monolayer with trypsin at 37 ⁰C for 10-15 minutes. Stop trypsinization if the monolayer starts displaying "cracks".

Add 8 ml of DMEM medium containing 10% FBS and resuspend cells; the FBS will block trypsin activity.

SEEDING CELLS ON FILTERS IN TRANS-WELL PLATES.

24 well filters: add 1 ml of cell suspension medium from above to 16 ml DMEM medium. Add 0.5 ml cell suspension to the apical side of the transwell filter in a 24-wells plate; add 1 ml to a 12 wells filter; add 2 ml to a 6 wells filter. Add 1, 2, 3 ml medium to the basal compartment of a 24, 12, 6 wells filter respectively. Incubate plates at 37 ⁰C, 5% CO₂, change medium the next day. Change then the medium each two days. The transepithelial resistance should reach 800 Ω.cm² after 2 weeks of culture. Density is approximately 2.6*10⁵ cells

The results of figures 2, 3 and 4 show that C5 (valerate) and C6 (caproate) have an increased ability over butyrate to increase TER

Table 1: valerate and caproate produced from a given substrate (mmol)

Valerate Caproate

#la potato starch 10 g/d 0,873128698 0,07452472

#la potato starch 15 g/d 0,637776012 0,016893828

#la potato starch 20 g/d 0,584631506 -0,052015594

#lb RS HiMaize260 , 10 g/d 5,256658656 0,639323687

#2, lactose 0,021264 -0,110428729

#3, FOS dp3 (P92) -0,038307 -0,063618616

#4, FruIQ dp9 -0,026781 -0,105384069

#5, FruTEX dp25 2,539477388 -0,093014257

#6, Agave (branched inulin) 0,13287756 -0,036385372

#7, Locust Bean Gum 1,649443436 0,44205079

#8, Guar gum 0,130634528 0,161740563

#9, Sunfibre AG 0,036642 0,0666773

#10, Sunfibre R 0,083705 0,093414273

#11, Hindustan S Gum

CC 1666 2,17838822 0,452713414

#12, Hindustan S Gum

CC1504 2,60370812 0,229567406

#16, Arabinogalactan 4,256515236 0,266396866

#17, Pectine apple 1,855706072 0,233356314

#18, Pectine citrus 0,087927344 0,071151449

#19, Pectin 30% esterified 4,027625944 1,229590434

#20, Pectin 60% esterified 0,417113296 0,090432558

#21, Pectin 90% esterified 0,788773182 1,187125689

#22, Karaya Gum 0,661813952 0,085118766 standard ileal effluent medium -0,005894 0,012574822 Table 2 : Structural information of some of the carbohydrates used

Table 3: constituents of bread

Reference lists

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5. Kanauchi, O., et al, Dietary fiber fraction of germinated barley foodstuff attenuated mucosal damage and diarrhea, and accelerated the repair of the colonic mucosa in an experimental colitis. J Gastroenterol Hepatol, 2001. 16(2): p. 160-8.

6. Jacobasch, G., et al., Dietary resistant starch and chronic inflammatory bowel diseases. Int J Colorectal Dis, 1999. 14(4-5): p. 201-11.

7. Tsukahara, T., et al., Stimulation of butyrate production in the large intestine of weaning piglets by dietary fructooligosaccharides and its influence on the histological variables of the large intestinal mucosa. J Nutr Sci Vitaminol

(Tokyo), 2003. 49(6): p. 414-21.

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15. Bordin, M., et al., Histone deacetylase inhibitors up-regulate the expression of tight junction proteins. MoI Cancer Res, 2004. 2(12): p. 692-701. 16. Venkatraman, A., et al., Amelioration of dextran sulfate colitis by butyrate: role of heat shock protein 70 and NF-kappaB . Am J Physiol Gastrointest Liver Physiol, 2003. 285(1): p. G177-84.

17. Tarrerias, A.L., et al., Short-chain fatty acid enemas fail to decrease colonic hypersensitivity and inflammation in TNBS-induced colonic inflammation in rats. Pain, 2002. 100(1-2): p. 91-7.

18 . Venema K, et al (2005), Microbial Ecology in Health and Disease, 17:47-5719. Hinnebusch B. F., Meng S., Wu J.T., Archer S.Y. and Hodin R.A., J. Nutrition, 2002, 132: 1012-1017.

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Pharm. 2002 Jan l;231(l):83-95.

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26. Van Nuenen HMC, Meyer PD, Venema K. The effect of various inulins and Clostridium difficile on the metabolic activity of the human colonic microbiota in vitro. Microb Ecol Health Dis 2003; 15: 137-44. 27. Jouany JP. Volatile fatty acids and alcohols determination in digestive contents, silage juice, bacterial culture and anaerobic fermentor contents. Sci Aliments 1982; 2: 131-44

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Claims

Claims

1. A carbohydrate being fermented in at least 4 hours by the bacteria present in the lumen of a subject for use as a medicament for preventing, delaying and/or treating a disorder of the gastrointestinal tract, wherein the fermentation of said carbohydrate induces a detectable increase of the production of a C5 and/or a

C6 SCFA by said bacteria in the lumen of the treated subject at least 10 hours after the administration of said carbohydrate.

2. A carbohydrate according to claim 1, wherein at least 50% of the ingested carbohydrate is fermented in at least 4 hours by the bacteria present in the lumen of the treated subject.

3. A carbohydrate preferably according to claim 1, wherein: the carbohydrate has a DP of at least 100 and/or - is a carbohydrate comprising a mannose, lignol, galactose, glucose, arabinose, rhamnose and/or gluconic acid side chain and/or comprises a methyl-, acethyl- an ethyl, a carboxymethyl and/or is an esterified carbohydrate.

4. A carbohydrate according to any one of the preceding claims, wherein the carbohydrate is: a branched inulin, preferably a branched inulin from agave, arabinogalactan, a methylated or acetylated pectin, an esterified pectin, preferably an esterified apple pectin,

Himaize, Hindustan gum, karaya gum, xanthan gum, gellan gum,. - GOS (galactoligo saccharide)

Alginate

Rice starch, waxymais starch, tapioca-starch and/or - Carrageen.

5. A carbohydrate according to any one of claims 1 to 4, wherein the induction is detectable at least 11 hours after the administration of the carbohydrate.

6. A carbohydrate according to any one of claims 1 to 5, wherein the detectable C5 and/or C6 SCFA is able to induce a detectable increase of the transepithelial resistance (TER) in a subject.

7. A carbohydrate according to any one of claims 1 to 6, wherein the carbohydrate is administered orally, preferably in a slow-release and/or enteric formulation.

8. A composition comprising a carbohydrate as defined in any one of claims 1 to 7.

9. A composition according to claim 8, wherein the composition is a food composition.

10. A composition according to claim 8, wherein the composition is a pharmaceutical composition.

11. A composition according to claim 9 or 10, wherein the composition comprises at least 5 g of a carbohydrate as defined in any one of claims 1 to 7.

12. Use of a carbohydrate or of a composition as defined in any one of claims 1 to 11, for the manufacture of a medicament for preventing, delaying and/or treating a disorder of the gastrointestinal tract of the subject.

13. A use according to claim 12, wherein said carbohydrate or composition is fermented by a bacterium in the lumen of said subject and thereby induces a detectable increase of a C5 and/or a C6 SCFA., preferably wherein said increase is detectable at least 10 hours after the administration of the carbohydrate

14. A method for preventing, delaying and/or treating a disorder of the gastrointestinal tract of a subject .comprising administration of a carbohydrate or composition as defined in any one of claims 1 to 11.

15. A method according to claim 14, wherein the carbohydrate has a DP of at least 10 and/or is branched and/or comprises a charged molecule and/or is modified and/or comprises rhamnose.

16. A method according to claim 14 or 15, wherein the carbohydrate is administered orally.