MXPA99012017A - Absorption of minerals by intestinal cells - Google Patents

Abstract

A method for increasing or facilitating the absorption of minerals from the diet. A nutritional composition which contains lactobacilli is enterally administered to a mammal. The nutritional composition is suitable for the treatment or prophylaxis of subjects having mineral deficiencies, or to compensate for physiological deficiencies due to a diet low in minerals, or to satisfy major physiological requirements for minerals in young children, pregnant women, women who are breastfeeding and the elderly.

Description

ABSORPTION OF MINERALS THROUGH INTESTINAL CELLS DESCRIPTION OF THE INVENTION This invention relates to a method for facilitating or increasing the absorption, by mammals, of minerals from the general diet. In particular, this invention relates to a method that involves the administration of an enteric composition containing microorganisms La c t oba ci l l s s. Minerals are key elements in major physiological processes. Calcium is, for example, of vital importance for bones and teeth, muscle contraction and the synthesis of hormones. Calcium is also an essential secondary messenger in most cellular activation phenomena. The minerals, of which the diet is the main source, are assimilated by the body when crossing the intestinal mucosa to pass then into the bloodstream. The degree of assimilation (or absorption) of minerals by the body in fact depends on their solubility in the intestinal environment and the ability of intestinal cells to assimilate them and transfer them into the bloodstream (R. Wasserman et al., I Mineral Absorption in the Monogastric Gl Trac, Advances in Experimental Medicine and Biology, 249, 45-65, Plenum Press, New York, 1989). The location, efficiency and mechanisms of calcium absorption all along the intestine have been studied in rats and chickens for many years (Bronner F .. J. Nutr .. 122. 641-643, 1992; Schachter D ., a. J. Physiol., 196, 357-362, 1959). For obvious ethical and technical reasons, such studies have been limited in man (Hylander E. et al., Scand J. Gastroenterol, 25, 705, 1990) and only a few in vi tro studies have been undertaken (Elsherydah A et al., Gastroenterology, 109, 876, 1995; Feher JJ, Am. J. Physiol., 244, C303, 1983: Feher JJ, Cell Calcium, 10, 189, 1989). One of the most widely studied aspects of mineral absorption is the bioavailability of minerals depending on the composition of the daily diet (Bronner F., J. Nutr., 123, 797, 1993). However, many minerals that are highly bioavailable are also unstable and are not suitable for use in the diet. In addition, merely supplementing the diet with larger amounts of minerals often has a negative effect on the organoleptic properties of the diet. • A possible solution to the problem is to facilitate or improve the absorption of minerals from the diet. However, there have been few studies on the methods of facilitating or increasing the absorption of minerals from the diet and the results have not been consistent. Rasic and colleagues have reported that the minerals contained in dairy products are assimilated better than when these products are fermented. This effect is attributed to the presence of acids in fermented milk products (XP002052238: Jn Fermented Fresh Milk Product, volume 1, p.114-115, 1978). More recently, Yaeshima et al. Have shown an increase in calcium absorption in rats from a diet of wheat fortified with calcium when a combination of oligosaccharides and Biofi doba c t eri s is consumed (XP002052237: Bulletin of the International Dairy Fermentation, No. 313, 1996). However, Kot et al. Have reported that La c tuba ci l us a ci dophi ls naturally internalizes Fe2 +, and oxidizes it to Fe3 +; which forms an insoluble form that is more difficult to assimilate (J. Agrie, Food Chem. 4_3, 1276-1282, 1995). Therefore, there is still a need for a means of facilitating or increasing the absorption of minerals present in the diet. Accordingly, this invention provides a method for increasing the absorption of minerals from the diet, the method comprising administering enterally to a mammal a nutritional composition containing a bacterium l a c t oba ci l l us. It has surprisingly been found, through the use of an i n vi tro model, that l a ct oba c y ls are able to directly facilitate or improve the absorption of minerals, especially calcium, by human intestinal cells. Without wishing to be bound by any theory, it is thought that this is linked to the induction of acidification in the microenvironment around the intestinal cells and the bacteria in contact with the intestinal cells. Bacteria and intestinal cells can participate in the induction of acidification. This localized acidification can thus play an active role in the solubilization of minerals, and therefore in the capacity of the body to assimilate them. In still another aspect, this invention provides the use of lactobacilli in the preparation of an enteral nutritional composition to facilitate or improve the absorption of minerals by the mammal. The enteral nutritional composition can be used for the treatment or prophylaxis of mineral deficiencies. The embodiments of the invention are now described, by way of example only, with Reference to the drawings in which: Figure 1 'represents the basal absorption of calcium by intestinal Caco-2 cells in the absence of lactobacilli; Figure 2 represents the influence of approximately 6.7 x 107 cfu / ml of various strains of lactobacilli on the absorption of calcium by intestinal Caco-2 cells; Figure 3 represents the influence of approximately 3.4 x 108 cfu / ml of various strains of lactobacilli on the absorption of calcium by intestinal Caco-2 cells. The invention relates to the enteral administration of a nutritional composition containing lactobacilli, to facilitate or improve the absorption of minerals present in a daily diet. Examples of minerals are calcium, magnesium, iron and / or zinc. The ingestion of lactobacilli increases the bioavailability of the minerals, that is, makes the minerals, which are frequently not very soluble in the intestine, more accessible to the intestinal cells. Any strain of food grade lactobacilli can be used. For example, the following lactobacilli may be used; Lactobacillus acidoph-lus r Lactobacillus crispatus r Lactobacillus amylovorous, Lactobacillus gallinarum, Lactobacillus gasseri and Lactobacillus j ohnsonii; Lactobacillus paracasei; Lactobacillus reuterii; Lactobacillus brevis; Lactobacillus fermentum; Lactobacillus plantarum; Lactobacillus casei especially L. casei subspecies casei and L. casei subsp. Rhamnosus. Lactobacillus delbruckii especially L. delbruckii subspecies lactis, L. delbruckii subspecies helveticus and L. delbruckii subspecies bulgaricus; and Leuconostoc mesenteroides especially L. mesenteroides subsp. cremoris, for example (Bergey's Manual of Systematic Bacteriology, vol.2, 1986; Fujisawa et al., Int.St. Bact. 42, 487-491, 1992). The cells may be able to adhere to the intestinal cells but do not necessarily need to. However, the molecules are preferably such that at least 50 bacteria, in particular at least 80 bacteria, are able to adhere to up to 100 intestinal cells. To select such an adherent type of bacteria, a culture of bacteria can be diffused on a confluent culture of an immortalized line of epithelial cells of the intestine.

(EP 0802257), the confluent culture is washed, and the number of bacteria adhering to the villi of the coating is measured. Probiotic bacteria are of particular interest. Some strains are in fact able to adhere to human intestinal cells, to exclude pathogenic bacteria that are on human intestinal cells, and / or to act on the human immune system by allowing it to react more strongly to external aggression (immunomodulation capacity) , for example by increasing the phagocytosis capacity of granulocytes derived from human blood (J. Of Dairy Science, 7_8, 491-197, 1995: the immunomodulation capacity of strain La-1 that was deposited by Nestec SA with the Budapest treaty in the National Collection of Cultures of Microorganisms (CNCM), 25 Route docteur Roux, 75724 Paris, June 30, 1992, where the deposit number CNCM 1-1225 was assigned). This strain is described in European Patent EP 0577904. By way of example, it is possible to use the probiotic strain Lactobacillus acidophilus CNCM I-1225. This strain was recently reclassified among the bacteria Lactobacillus johnsonii, subsequent to the new taxonomy, proposed by Fujisawa et al, which is now authoritative in the field of the taxonomy of lactobacillus acidophiles (Int. J. Syst. Bact., 42, 487-791, 1992). Other probiotic bacteria are also available, such as those described in European Patent EP0199535 (Gorbach et al.). US Patent US5296221 (Mitsuoka et al.), US Patent US556785 (Institut Pasteur) or US5591428 (Probi AB), for example. The nutritional compositions preferably comprise a sufficient amount of live lactobacilli for facilitated absorption of the intestinal minerals or cells, for example at least 10 6 cfu / ml, in particular 10 7 -10 11 cfu / ml, preferably 10 8 -10 11 cfu / ml, ( "cfu" means "colony forming unit." The nutritional composition may also contain other bacteria, as desired; for example other probiotic bacteria. The nutritional composition may also include a suitable protein source; for example a source of animal or vegetable proteins. Suitable protein sources are milk proteins, soy proteins, rice proteins, wheat proteins, sorghum proteins, and the like. The proteins can be in the intact or hydrolyzed form. The nutritional composition may also include a suitable carbohydrate source; for example sucrose, fructose, glucose, maltodextrin, and the like. The nutritional composition • may also include an adequate lipid source; for example a suitable animal or vegetable lipid source. Suitable lipid sources include milk fats, sunflower oil, rape seed oil, olive oil, safflower oil and the like. The nutritional composition can also be fortified with minerals and vitamins. It is especially preferred to fortify the nutritional composition with calcium. The nutritional compositions can be prepared in the form of food compositions intended for human or animal consumption. Suitable food compositions can be provided in the form of liquids, powders and solids. The nutritional composition can be fermented to obtain a sufficient amount of lactobacilli. Fermented milk-based compositions are thus particularly suitable. The term milk applies not only to animal milks, but also to what is commonly referred to as vegetable milk, ie an extract of treated or untreated plant materials such as legumes (soy, pea, lentil and the like) or oil seeds. (from rapeseed, soy, sesame, cotton and the like), whose extract contains proteins in solution or in colloidal suspension, which are coagulable by chemical action, by acid fermentation and / or by heat. It has been possible to attach these plant milks to thermal treatments similar to those for animal milks. It has also been possible to subject them to treatments that are specific to them, such as discoloration, deodorization, and treatments to suppress undesirable flavors. Finally, the word milk also designates mixtures of animal milks and vegetable milks. It is also possible to add, mix or coat the nutritional composition, during its preparation, with an appropriate amount of a lactobacillus culture in liquid, concentrated, dry or encapsulated form, according to the needs. It has also been found that the microencapsulation of lactobacilli has therapeutic advantages. First, microencapsulation significantly increases the survival of lactobacilli and therefore the number of live lactobacilli that reach the intestine. More importantly, lactobacilli are gradually released into the intestine, which allows the prolonged action of lactobacilli on the absorption of minerals by intestinal cells.

Preferably, in order to encapsulate lactobacilli, the lactobacilli are lyophilized or spray-dried (European Patent EP0818529), and these are incorporated into a gel consisting, for example, of solidified fatty acid, a sodium alginate, polymerized hydroxypropylmethylcellulose or polymerized polyvinylpyrrolidone. For this purpose, the teaching given in the French patent FR2,443,247 is incorporated by reference. The nutritional compositions do not need to contain carbohydrates necessary for active fermentation by lactobacilli in the intestinal environment. By contrast, the facilitated absorption of minerals is independent of the fermentative activity of lactobacilli, but rather seems to result from direct contact between lactobacilli and intestinal cells. It is thought that this induces the acidification of the microenvironment and therefore a better solubilization of the minerals. However, this may be desirable to provide for the specific renewal or multiplication of the lactobacilli in the intestinal environment, in order to prolong the facilitated absorption effect of the minerals. This can be achieved by the addition of fibers that facilitate the specific multiplication of the intestines in the intestinal environment to the nutritional composition. These fibers are soluble and fermentable. These fibers can be selected from, for example, vegetable pectins, quito-, fructo-, mobi-, galacto-, isomalto-, mano- or xylo-oligosaccharides or soy oligosaccharides, for example (Playne et al., Bulletin of the IDF 313. Group B42, Annual Session September 95, Vienna). Preferred pectins are polymers of α-1,4-D-galacturonic acids having a molecular weight of the order of 10 to 400 kDa, which can be purified from carrots or tomatoes, for example (Japanese Patent JP60164432). Preferred galacto-oligosaccharides comprise a saccharide moiety consisting of 2 to 5 repeating units of structure [-aD-Glu (1? 4) -β-D-Gal- (1-6) -] (Yakult Honsa Co., Japan). The preferred fructo-oligosaccharides are inulin-oligofructose extracted from chicory which may comprise, for example, 1 to 9 repetitive units of structure [-β-D-Fru- (1? 2) -β-D-Fru- (1? 2) -] (W094 / 12541; Raffinerie Tirlemontoise S.A. Belgium), or oligosaccharides synthesized from sucrose units which may comprise, for example, a saccharide moiety consisting of 2 to 9 repeating units of structure [-ad-Glu- (l- »2) -β-D- Fru (1-) 2- (Meiji Seika Kasiha Co., Japan) Preferred malto-oligosaccharides comprise a saccharide moiety consisting of 2 to 7 repeating units of structure [-DD-Gal- (1-4) -] (Nihon Shokuhin Kako Co., Japan) Preferred isomaltoses comprise a saccharide moiety consisting of 2 to 6 repeating units of structure [aD-Glu- (1- »6) -] (Showa Sangyo Co., Japan) . "The preferred gentio-oligosaccharides comprise a saccharide moiety consisting of 2 to 5 repeating units of structure [-β-D-Glu- (1-6) -] (Nihon Shokuhin Kako Co., Japan). Finally, the preferred xylo-oligosaccharides comprise a moiety that consists of 2 to 9 repeating units of structure [-β-xyl- (1-4) -] (Suntory Co., Japan), for example. The amount of fibers in the nutritional composition depends on their capacity to promote the development of the bacteria. As a general rule, the nutritional composition may contain from 1 to 50% of such fibers (by weight relative to the dry matter). The concentration of the compounds can be at least 10 3 CFU of 1 to 10 grams per gram of fiber, preferably 10 4 to 10 5 CFU / g of fiber. Yet another advantage, provided by the fibers, consists of the fact that the intestinal transit is retarded by the fibers. This is particularly the case if the amount of fibers is large, that is, in the order of 20 to 50% relative to the weight of the composition. The c obates, being truly eliminated by the action of the intestinal transit, it is possible, in this way, to prolong the beneficial action of the patients on the absorption of minerals by the intestine. The nutritional compositions may be in the form of any enterally administered, suitable food. For example, the nutritional composition may take the form of a fermented milk (European patent EP0577904), a baby food (European patent EP0827697), fresh cheese (PCT / EP97 / 06947), a matured cheese, an ice cream (WO 98/09535), a cream-filled sponge cake (European patent EP704164; European patent EP666031), a dry sauce and / or a pasta (European patent EP689769) . The nutritional compositions may also be in a form suitable for people who can not tolerate dairy products. These nutritional compositions will not contain allergenic dairy products. For example, for children who are allergic to milk proteins, the nutritional composition can be formulated to contain hypoallergenic dairy products. These dairy products can be in accordance with the European rule 96/4 / EC which states that in a hypoallergenic milk, the allergenic proteins must be immunologically at least 100 times less detectable than in a non-hydrolyzed milk (Off J. Europ. Comm.NoL49 / 12, Annexe 5.a. 196: fritsche et al., Arch. Aller and Appll, Imm. 93_, 289-293, 1990). The nutritional compositions are particularly suitable for the treatment or prophylaxis of people who have mineral deficiencies, or to compensate for physiological deficiencies due to a diet low in minerals, or to satisfy higher physiological requirements for minerals in children, pregnant women, women who are breastfeeding and in elderly people. This invention is now further described by means of specific examples. The percentages are given by weight, unless indicated otherwise. These examples are given by way of illustration only and in no way constitute a limitation of the invention. • Example 1 - Materials: 45CaCl2 is obtained from Amersham, yellow Lucifer from Sigma, collagen I from Centrix Pharmaceuticals. PBS, HEPES and the components of the Gibco cell culture medium, and the supports for Falcon cultivation.

Cell culture: the Caco-2 human cell line, isolated from colon adenocarcinoma, is obtained from the North American Culture Collection Species (American Type Culture Collection) (passage 41). The cells are placed in culture in a • amount of 4 x 104 cells / cm2 in DMEM containing 4.5 g / 1 of glucose, 20% fetal calf serum inactivated by heat, 1 mg / ml of fungizone. 100 U / ml penicillin / streptomycin, 20.0 μg / ml gentamicin and 1% non-essential amino acids. The cells are regularly trypsinized and placed in culture again 1/20. The cells used in the calcium transport experiments are placed in culture at 1 x 105 cells / cm2 in permeable inserts previously coated with a layer of collagen I at 50 μg / ml. In all cases, the cells are kept in an incubator with 10% CO2 / 90% air, and the medium is replaced every two days.

Feasibility of Caco-2 cells in order to exclude the possibility that the enhancement of calcium absorption by intestinal cells in the presence of lac tuba is due to cell damage, a portion of each sample that serves the Calcium assay was used for an assay of hexosaminidase activity (Landergren et al., J. Immunol Method, _6_2, 379-378, 1984). This colorimetric test makes it possible to quantify cell lysis and / or cell death by measuring the activity of hexosaminidase released into the supernatant from the cytosol of the damaged cells. The results show that in all experiments, the activity of hexosaminidase is equivalent in the presence of l a c t oba ci l os.

Permeability of the cell layer: the integrity of the layer formed by Caco-2 cells at the end of their development and their differentiation, is evaluated by measuring the transept telial electrical resistance (TEER) using the Millicell-ERS voltmeter / ohmmeter . The calcium absorption experiments are carried out when the resistance reaches at least 700 ohm x cm2. The permeability of the cell layer during the calcium absorption experiments is evaluated by measuring the diffusion level (in%) of Lucifer yellow, a molecule that does not cross the cell membrane.

Calcium transport: Caco-2 cells are grown on inserts for 3 to 5 weeks. On the day of the experiment, the cell layer is washed twice in PBS and then the compartment of the fundus of the insert that incorporates the serosa (basolateral pole of the cells) receives 2.5 ml of carrier buffer (140 M NaCl, 5.8 mM KCl, 0.34 mM NaH2P04, 0.44 M KH2P04, 0.8 mM MgSO4, 20 mM HEPES, 4 M glutamine, 25 mM glucose, pH 7.4) supplemented with 2.5 M CaCl2, while the upper compartment of the insert was incorporated into the intestinal lumen (apical pole of the cells) receives 1.5 ml of carrier buffer supplemented with 10 M CaCl2 and trace amounts of 15CaCl2 and Lucifer yellow. The inserts are then placed at 37 ° C and 50 μl of sample is removed in the upper and lower compartments at regular intervals. The radioactivity contained in these samples is evaluated by liquid scintillation counting and makes it possible to extrapolate on the amount of cold CaCl2 absorbed. Basal calcium transport is expressed as nmol of calcium transported to the lower compartment of the insert. The diffusion of Lucifer yellow detected by spectrofluorometry in the lower compartment is expressed in% of the amount introduced into the upper compartment.

Influence of lactobacilli strains Lactobacillus johnsonii Lal (CNCM 1-1225), Lal7, La22, La31; Lactobacillus acidophilus LalO, Lal8, La31; Lactobacillus bulgaricus Lfi5, YL8; Lactobacillus paracasei ST11; Lactobacillus gasseri LGA7; Lactobacillus reuteri LR7 and Streptococcus thermophilus Sfi20, YS4 (Nestec Collection, Lausanne, Switzerland) are placed in culture under anaerobic conditions in MRS broth for Lactobacillus or Mil for Streptococcus for two 24 hour periods, washed in PBS and resuspended in carrier buffer before being inserted inside the upper compartment of the inserts. The ratio of Caco-2: bacteria is then approximately 1: 100 according to the tests (6.7 x 107 or 3.4 x IO8 cfu / ml in the upper compartment of the inserts for the tests presented in Figures 2 and 3) j The absorption of calcium is evaluated according to the aforementioned protocol.

- Results of basal calcium transport: a calcium gradient was established in the inserts when CaCl2 2.5, mM was introduced into the lower compartment, which corresponds to the normal concentration of human plasma, and 10 mM CaCl2 is arbitrarily placed inside the upper compartment , which could correspond to the calcium content of a diet. As shown by the results of a representative experiment illustrated in Figure 1, the basal absorption of calcium by Caco-2 cells increases with time to reach 600 nmol / insert, comprised approximately 3 x 10 6 cells after 4 hours. As a check for the integrity of the cell layer during the experiment, the diffusion of Lucifer yellow was measured and proved to be less than 2%.

Measurement of the influence of lactobacilli: in Figures 2 and 3 the absorption of calcium by Caco-2 cells is significantly increased in the presence of strains of Lactobacillus j ohnsonii adherent Lal and La22, in the presence of non-adherent strains of Lactobacil us acidophilus LalO and Lal8, and in the presence of the strains L. paracasei (ST11), L. gasseri (LGA7) and L. reuteri (LR7). The ability of bacteria to adhere to intestinal cells does not therefore seem to correlate directly with their ability to increase calcium absorption by these same cells. In all these experiments, the diffusion of Lucifer yellow is modulated in a similar way, but it remains negligible. A decrease in pH in the upper compartment of the inserts is also observed when the Caco-2 cells are in the presence of lactobacilli, no matter the strain, except with the strain Sfi20 (Table 1). There is therefore no correlation between the increase in calcium absorption and this decrease in pH. However, certain bacterial strains capable of increasing the absorption of calcium are not able to acidify the experimental medium in the absence of Caco-2. This means that acidification in the presence of Caco-2 and bacteria requires a collaboration between the two types of bacteria. organisms and could be due to Caco-2 cells.

Table 1 Influence of lactobacilli on the pH of the experimental medium in the absence or presence of Caco-2 cells.

Example 2 Tests similar to those carried out in Example 1 were carried out to determine the influence of lactobacilli on the absorption of calcium by intestinal cells in the presence of marked inulin (3H-inulin, Amersham, prebiotic fiber tracker). The results confirm that lactobacilli increase in vitro the absorption of minerals by intestinal cells.

Example 3 Tests similar to those carried out in Example 1 were carried out in order to determine the influence of lactobacilli on the magnesium, iron and zinc solution by the intestinal cells. The results confirm that lactobacilli increase in vitro the absorption of minerals by intestinal cells.

Example 4 Encapsulation of lactic acid bacteria In a 100 liter tank, 80 liters of culture medium having the following composition are prepared, in% The inoculation was carried out with 1 liter of a 20-hour culture of La ct obacil l us j ohnsonii Lal (CNCM 1.1225). The medium is incubated for 12 hours at 30 ° C. The culture broth is centrifuged and 240 grams of cells are recovered. This is diluted in 250 ml of skim milk supplemented with 7% lactose. The mixture is frozen using liquid nitrogen. The lyophilization is carried out at 40 ° C all night. A 5% dispersion of the obtained powder is prepared in hydrogenated vegetable fat having a melting point of 42 ° C and liquefying at 45 ° C. The dispersion is injected at 45 ° C under a pressure of 4 bar, at the same time as liquid nitrogen, in an amount of one part of dispersion per 5 parts of nitrogen, in the upper part of a vertical cylinder of 1.5 m in diameter and 10 m high. A container is placed in the bottom of the cylinder, which contains liquid nitrogen in which the microspheres containing the bacteria are collected, whose diameter varies between 0.1 and 0.5 mm. The microspheres are then placed in a fluidized bed and an alcoholic solution containing 8% zein is sprayed onto the bed, in an amount such that the zein layer formed around the microspheres represents 5% of its weight. The microspheres are then incorporated into a food composition aimed at facilitating the absorption of minerals by the intestinal cells.

Example 5 Prepare a concentrated base for ice cream, by mixing, at 60-65 ° C, for 20 minutes of approximately 11% lactic fat, 8.8% milk solids (non-solid fat), 25% sucrose, 5% of glucose syrup and 0.6% Emulstab® SE30. The base is homogenized at 72-75 ° C and 210 bar (2 stages at 210/50 bar), pasteurized at 85 ° C for 22 seconds (pasteurizer APV, France, Evreux, 400 l / h), cooled to 4 ° C and 40% of milk acidified by Lactobacillus j ohnsonii La-1 (5 x 108 cfu / ml) and Bi idobacterium longum Bil6 (3 x 108 cfu / ml) are added to it. The composition of this concentrated base is presented in the following table.

After maturing the cream for 12 hours at 5 ° C, it freezes to an overflow of 95% "by volume (Crepaco Freezer, France, Evreux, 160 lt of product / hour) A wafer-shaped dough It is prepared containing 10% of Raftilose® L30 fructo-oligosaccharide (Raffineries Tirlemontoise SA BE), according to the recipe reproduced in the table below.After baking, the wafer is conventionally formed into a cone.After cooling, the inner part The cones are coated with a greasy film, and then the cones are filled with the ice-cream milkshake described above For a wafer cone weighing 11.5 grams, 130 ml of whipped cream (approx. 5 g of chocolate (sprinkling on the cream).

Ingredient Weight (g) Supplier Ordinary wheat flour 55 52 Starch 0.2 Fructo-oligoscaride 10 Raffinerie Tirlemontoise Raftilose® L30 S.A. BE Sugar 27.8 Emulsifying grease, 1.5 Salt 0.5 Total: recipe for wafer 100 1. 1 g of fibers and approximately 108 cfu / g of lactobacilli are thus provided by ice cream cone. The fibers, by promoting the specific development of lactobacilli in the intestinal tract, promote in this way the assimilation of minerals.

Claims (10)

RE IVINDICATIONS

1. Use of lactobacilli in the preparation of an enteral nutritional composition to facilitate or improve the absorption of minerals by a mammal.

2. Use according to claim 1, wherein the lactobacillus is a Lactobacillus bacterium that is capable of adhering to intestinal cells.

3. Use of compliance with claim 2, wherein the lactobacillus is Lactobacillus j ohnsonii strain CNCM 1-1255.

4. Use according to claim 1, wherein the enteral nutritional composition contains 10 7 to 10 11 cfu of lactobacillus.

5. Use in accordance with the claim 1, in which the enteral nutritional composition facilitates the absorption of calcium, magnesium, iron and / or zinc.

6. Use according to claim 1, wherein the enteral nutritional composition contains milk proteins.

7. Use in accordance with claim 6, wherein the. enteral nutrition composition is a infant formula that includes hydrolysates of hypoallergenic milk protein.

8. Use in accordance with the claim 1, in which the enteral nutritional composition further comprises prebiotic fibers.

9, Use of proteins in preparation of an enteral nutritional composition for the treatment or prophylaxis of mineral deficiencies.

10. A method to increase the absorption of minerals from the diet, the method comprises enterally administering to a mammal a nutritional composition containing l a c t oba cles.