Composition in powder form containing iron-milk protein complex and probiotic bacteria

Abstract

Provided is a composition comprising probiotic bacteria and an added iron source for use in the prevention, alleviation and / or treatment of iron deficiency, in which the viability of the probiotic bacteria is not impaired. The present invention provides a powdered composition comprising at least one probiotic bacterium and an iron source, wherein the iron source is an iron-milk protein complex, preferably casein, a casein derivative, a casein fraction, or a mixture thereof.

Description

[Technical Field]

[0001] The present invention relates to a composition in powder form comprising probiotic bacteria and at least one iron-milk protein complex. Advantageously, such iron-milk protein complex does not significantly reduce the viability of the bacteria, and therefore, such an iron source is advantageously used to fortify a composition in powder form comprising probiotic bacteria. [Background technology]

[0002] Beneficial bacteria, especially probiotic bacteria, are added to a wide range of products, namely, powder-form products that are reconstituted with liquid, such as nutritional compositions for infants, children, pregnant, lactating or pre-pregnant women, the elderly, or people who require specific nutrition due to adverse medical conditions.Such products usually contain a wide variety of macronutrients and micronutrients to meet the nutritional needs of consumers.For nutritional purposes, it is most important that consumers consume an appropriate amount of probiotic bacteria and other nutrients.

[0003] A particularly important micronutrient is iron. Worldwide, iron deficiency is one of the most widespread nutrient deficiencies. In humans, iron is essential for the functioning of numerous biological processes, such as oxygen binding and transport, gene regulation, neurological function, immune function, and the regulation of cell growth and differentiation. Iron deficiency can lead to anemia and various health problems, such as impaired thyroid, immune, and mental function, physical performance, cognitive development, increased sensitivity to insulin, and fatigue. Iron deficiency is particularly prevalent in pregnant and lactating women and children.

[0004] Food fortification with iron is one approach to address iron deficiency.Therefore, it is highly desirable to include an added iron source in dietary compositions or supplementary foods, especially dietary compositions or supplementary foods for infants, young children, pre-pregnant, pregnant and / or lactating women.Various iron compounds have been used as iron fortifiers in food products and dietary supplements.For example, ferrous sulfate is widely used due to its relatively low cost and high bioavailability.

[0005] However, the present inventors have found that some iron compounds, when used to fortify compositions containing probiotic bacteria, have a detrimental effect on the viability of the probiotic bacteria (see also co-pending International Application No. PCT / EP2016 / 063170 and European Patent Application Publication No. 16198292.1).

[0006] In most cases, the benefits of probiotic bacteria can only be obtained when the bacteria are alive at the time of consumption.Therefore, to compensate for the decrease in viable probiotic bacteria and ensure that consumers are provided with an adequate amount of live bacteria, probiotic bacteria are usually added in excess in products.However, this solution is not entirely satisfactory because the excess addition is very costly and causes waste.

[0007] It is therefore an object of the present invention to provide a composition comprising probiotic bacteria and an added iron source, wherein the viability of the probiotic bacteria is not impaired by the presence of the added iron source. Summary of the Invention

[0008] The present inventors have surprisingly found that iron-milk protein complexes, when used as an iron source in compositions containing probiotic bacteria, do not reduce the viability of the bacteria.

[0009] In a first aspect, the present invention provides a composition in powder form comprising at least one probiotic bacterium and an iron source, characterized in that the iron source is an iron-milk protein complex.

[0010] In a second aspect, the present invention relates to the use of an iron source for the fortification of a composition in powder form comprising at least one probiotic bacterium, characterized in that the iron source is an iron-milk protein complex.

[0011] In a third aspect, the present invention provides a method for producing a composition in powder form, the method comprising: a) providing a first composition in powder form; b) mixing a first composition in powder form with an iron source to form a second composition in powder form; c) mixing at least one probiotic bacterium into a first composition in powder form before or simultaneously with the iron source, or mixing at least one probiotic bacterium into a second composition in powder form after the addition of the iron source; and wherein the iron source is an iron-milk protein complex.

[0012] In a fourth aspect, the present invention provides a method for producing a product, the method comprising the step of reconstituting with a liquid a composition in powder form comprising at least one probiotic bacterium and an iron source, wherein the iron source is an iron-milk protein complex.

[0013] In a fifth aspect, the present invention provides a product obtainable or obtained by the method of the present invention.

[0014] In a sixth aspect, the present invention provides a method of providing nutrition to an individual comprising the step of feeding the individual a product of the invention that is an edible product.

[0015] In a seventh aspect, the present invention provides an edible composition in powder form comprising at least one probiotic bacterium and an iron source for use in the prevention, alleviation and / or treatment of iron deficiency in an individual, characterized in that the iron source is an iron-milk protein complex.

[0016] In an eighth aspect, the present invention provides a method for reducing and / or preventing loss of probiotic bacteria during reconstitution of a composition in powder form comprising at least one probiotic bacterium and an added iron source, characterized in that an iron-milk protein complex is used as the added iron source. [Brief explanation of the drawings]

[0017] [Figure 1] 1 is a graphical representation of the loss of viability of Lactobacillus rhamnosus (deposited under CGMCC 1.3724) in compositions containing 10 mg and 30 mg of iron per 100 g of composition, respectively. Two different iron sources are compared: spray-dried dissolved ferrous sulfate and an iron-casein complex. The loss of viable Lactobacillus rhamnosus is significantly lower for the iron-casein complex at both iron concentrations. [Figure 2] Effect of iron source on survival of B. longum. [Figure 3] Effect of iron source on B. longum in infant formula. [Figure 4] Effect of iron source on L. paracasei survival. [Figure 5] Effect of iron source on survival of L. paracasei in infant formula. [Figure 6] Effect of iron source on survival of L. rhamnosus in infant formula. DETAILED DESCRIPTION OF THE INVENTION

[0018] definition As used herein, the term "iron-milk protein complex" refers to a complex formed with iron cations chelated with milk protein. Milk protein is intended to include casein, whey protein, and mixtures thereof, as well as derivatives or fractions of casein or whey protein.

[0019] As used herein, the term "probiotic bacteria" refers to live bacterial cell preparations that have a beneficial effect on the health or well-being of the host [Salminen S, et al., "Probiotics: how they should be defined", Trends Food Sci. Zechnol, (1999), 10, 107-10].

[0020] Bacteria are considered "live" if they can grow under controlled culture conditions and form colonies or suspensions, or if the microbial metabolic activity and / or membrane integrity can be confirmed using methods known to those skilled in the art, such as flow cytometry. Dried probiotic bacteria are considered live if they can grow and / or if the membrane integrity can be confirmed as described above after reconstitution of the bacteria with a liquid, such as an aqueous liquid, preferably water.

[0021] In the present invention, "reconstitution" refers to the dissolution or suspension of a powder in a liquid, for example, an aqueous liquid, preferably water, a specific reconstitution medium such as those used in analytical microbiology, or a beverage such as milk or juice. The liquid used for reconstitution may be cold or hot. Preferably, it refers to reconstitution with an aqueous liquid, preferably water.

[0022] "Aqueous liquid" for the purposes of the present invention is intended as a liquid that comprises at least one aqueous phase. It can be, for example, an emulsion, an aqueous solution or dispersion, or water. Preferably, it is an oil-in-water emulsion, an aqueous solution or suspension, or water. More preferably, it is an aqueous solution or suspension, or water. Most preferably, it is water.

[0023] The term "iron" is used herein to mean Fe, unless otherwise specified, depending on the iron source used. 2+ or Fe 3+ It is intended to indicate either

[0024] "Additional iron source" is intended herein as a ferrous or ferric compound added to a composition solely for its iron supplementation effect. Depending on its nature, the composition may also contain iron from other ingredients, such as milk, fruit, vegetable, grain, or fiber components. Iron present in such ingredients is not intended herein as an "additional iron source" because it is naturally present in the ingredient to which it is added primarily for its overall nutritional value, rather than its iron content.

[0025] The iron source is intended for purposes of the present invention to be "substantially the only added source of iron" in the composition, provided that other added iron sources are used in sufficiently small amounts so as not to cause a statistically significant reduction in viable probiotic bacteria. Those skilled in the art can assess whether a statistically significant reduction in viable bacteria occurs by applying the methods described in the examples of this application and applying commonly known statistical methods for analyzing the results.

[0026] The term "exogenous," when referring to iron or phosphorus in the complex, refers to iron and / or phosphorus that was added during the manufacturing process of the complex and, therefore, refers to iron or phosphorus that was not naturally chelated by the milk protein.

[0027] The term "nutritional composition" refers to a product intended to provide complete or supplemental nutrition to an individual (i.e., to meet the essential nutritional needs of such an individual), the predominant purpose of which is to provide nutrition. Nutritional compositions are intended to provide specific nutrients to individuals with special nutritional needs, such as infants, children, pregnant or lactating women, the elderly, or people with adverse medical conditions, who require special foods (e.g., tube-fed or pediatric compositions). Products in which palatability is predominant and nutritional value is not of major importance are excluded from the term "nutritional product." Nutritional compositions preferably contain protein, fat, carbohydrates, and a variety of micronutrients.

[0028] In the present invention, the term "infant" refers to a child between birth and 12 months of age. The term "child" refers to a child between 12 months and 5 years of age, preferably between 12 months and 3 years of age.

[0029] As used herein, the expression "infant formula" refers to a food intended for specific nutritional use by infants and which, by itself, meets the nutritional requirements of this category of person (Article 2(c) of European Commission Directive 91 / 321 / EEC 2006 / 141 / EC of December 22, 2006 on infant and follow-on formulas). It also refers to nutritional compositions intended for infants and as defined in Codex Alimentarius (Codex STAN72-1981), and special infant foods (including foods for special medical purposes). Infant formulas can include starter infant formulas and follow-up or follow-on formulas. Generally, starter formulas are intended for infants from birth as breast milk substitutes. Follow-up or follow-on formulas are given from the sixth month onwards. They constitute the main liquid component in the increasingly diversified diet of infants falling into this category. It is understood that an infant can be fed infant formula alone or the infant formula can be used as a supplement or complement to human milk.

[0030] "Growing Up Milk" (or GUM) is offered from the first year onwards. It is generally a milk-based drink adapted to the specific nutritional needs of children.

[0031] The expression "infant food" means food intended for specific nutritional use by an infant or child, e.g., a toddler, during the first year of life.

[0032] The expression "infant cereal composition" means a cereal-based food product intended for specific nutritional use by an infant or child, e.g., a toddler, during the first year of life.

[0033] The term "fortifier" refers to a nutritional composition suitable for mixing with breast milk or infant formula. "Breast milk" is to be understood as mother's milk or mother's colostrum or donor's milk or donor's milk colostrum.

[0034] The term "food supplement" refers to a composition that can be used to supplement or add to the nutrition of an individual.

[0035] The term "prebiotic" refers to non-digestible carbohydrates that beneficially affect the host by selectively stimulating the growth and / or activity of healthy bacteria in the human colon (Gibson GR, Roberfroid MB. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics., J Nutr. 1995;125:1401-12).

[0036] composition The composition of the present invention is a composition in powder form comprising at least one probiotic bacterium and an iron source, the iron source being an iron-milk protein complex.

[0037] The inventors have surprisingly found that added iron in many components of complete nutritional compositions, such as infant formula, causes a significant reduction in probiotic bacteria in the reconstituted composition, and have determined that iron-milk protein complexes can be advantageously used as a nutritional source of iron in combination with probiotic bacteria without significantly compromising the viability of such probiotic bacteria.

[0038] The milk protein may be casein, a derivative or fraction of casein, whey, a derivative or fraction of whey, or a mixture thereof. Preferably, the protein is a phosphoprotein. In a preferred embodiment, the milk protein in the iron-milk protein complex comprises casein and / or a derivative or fraction of casein. Most preferably, the milk protein in the complex comprises casein. In a preferred embodiment, the milk protein present in the complex consists of casein and / or a derivative or fraction of casein. More preferably, the milk protein in the complex consists of casein.

[0039] Casein can be obtained from a variety of sources such as milk, sodium caseinate, potassium caseinate, ammonium caseinate, caseinate rennet, acid casein, e.g., lactated casein, and / or non-fat milk solids. Preferably, it is sodium caseinate, potassium caseinate, ammonium caseinate, and / or lactated casein.

[0040] In a preferred embodiment, the complex is a non-micellar complex.

[0041] In one embodiment, the milk protein is derived from a dairy raw material having a protein-to-calcium ratio of at least 45:1. Preferably, the protein / calcium w / w ratio in the dairy raw material is at least 58:1, more preferably between 58:1 and 640:1, and most preferably between 70:1 and 95:1. This represents a significant reduction in the amount of calcium in the dairy raw material, since cow's milk typically has a protein / calcium w / w ratio of 26:1. In another preferred embodiment, the milk protein is derived from a dairy raw material from which at least 70% w / v of calcium has been removed. Such dairy raw materials are preferably selected from whole milk, skim milk, low-lactose milk, ultrafiltration retentate, concentrated milk, and combinations thereof. In one embodiment, the milk is bovine milk. Removal of calcium from the dairy raw material is advantageous in that it aids in the binding of a large amount of iron to the milk protein, i.e., by removing chelated calcium and allowing it to be replaced with iron.

[0042] In another embodiment, the iron-milk protein complex comprises exogenous phosphorus, more preferably exogenous orthophosphate. Preferably, the w / w ratio of protein to phosphorus (preferably orthophosphate) is between 64:1 and 6.25:1, preferably between less than 32:1 and 6.25:1, preferably between less than 32:1 and 8:1, more preferably between 28:1 and 8:1, even more preferably between 25:1 and 8:1, and most preferably between 20:1 and 8:1. The presence of phosphorus is advantageous in that it aids in the efficient binding of iron to the milk protein and therefore contributes to the binding of a large amount of iron in the complex.

[0043] In another preferred embodiment, the complex is soluble in solutions at physiological pH, preferably pH 6.6 to 6.9, which is beneficial in avoiding the formation of insoluble precipitates in liquid solutions at such pHs and also contributes to good bioavailability of the complex.

[0044] Preferably, the iron in the complex is exogenous iron. Because only small amounts of iron are naturally bound to milk proteins, such as casein, it is advantageous for the complex to contain exogenously added iron. Preferably, the complex contains greater than 1% w / w bound iron, more preferably 1-20% w / w bound iron, even more preferably 1-8% w / w%, and most preferably 4-8% w / w bound iron. More preferably, the w / w ratio of protein to iron (preferably exogenous iron) is 92:1-19.5:1, preferably 90:1-19.5:1, more preferably 80:1-19.5:1, and most preferably 50:1-19.5:1. Being able to have such high amounts of iron bound to milk proteins in the complex is particularly advantageous, because the higher the iron loading in the complex, the less complex is required to fortify the product.

[0045] In a preferred embodiment, the iron cation in the complex is Fe 3+ is.

[0046] In a preferred embodiment, the complex is provided in the form of a powder.

[0047] Particularly preferred conjugates are those described in U.S. Patent Application Publication Nos. 2015 / 0164123 and WO 00 / 51446, both of which are incorporated herein by reference. Such documents provide detailed descriptions of conjugates, such as those described above, and methods that can be used to prepare conjugates of certain embodiments that can be advantageously used for purposes of the present invention.

[0048] The most preferred are the complexes described in US Patent Application Publication No. 2015 / 0164123. These are particularly advantageous in that they are soluble in aqueous liquid food products or beverages, even when a large amount of iron is bound to milk proteins in the complex. Therefore, the complexes advantageously do not form insoluble precipitates when added to clear beverages and solutions, which can cause haze and give food products and beverages an undesirable gritty texture. Further advantages of the above complexes can be found in the cited US Patent Application Publication No. 2015 / 0164123.

[0049] Here, the inventors have found that most iron sources, such as the widely used ferrous sulfate heptahydrate and spray-dried dissolved ferrous sulfate, both cause a significant decrease in the viability of probiotic bacteria, while no significant decrease in the viability of probiotic bacteria is observed with the iron-milk protein complex described herein. Without being bound by theory, the inventors have determined that probiotic bacteria become sensitive to free iron when the powder composition is reconstituted with a liquid. It is believed that the slow release of free iron when the powder composition is reconstituted with a liquid reduces the decrease in viability of probiotic bacteria, as it provides the probiotic bacteria with sufficient time to rehydrate before encountering the free iron.

[0050] This positive effect is observed when the iron-milk protein complex is used as such, and when such complex is dispersed in an amorphous matrix, provided that the complex structure is maintained. Such a component having the complex dispersed in the matrix can be obtained by mixing the complex with a carrier solution and spray-drying the carrier solution. Suitable carriers include, for example, maltodextrin. Those skilled in the art can determine the iron content by combining protein identification evaluations such as SDS-PAGE with mineral analysis using ICP atomic emission spectroscopy, as well as by determining the iron content of the milk protein complex. 2+ and Fe 3+Whether a product or ingredient contains iron-milk protein complexes can be routinely assessed by performing a "free iron" test using a compound such as potassium hexacyanoferrate, which involves a color change in the presence of iron.

[0051] The use of the above-mentioned iron-milk protein complex is particularly advantageous in that it is characterized by both good bioavailability and low adverse effects on the viability of probiotic bacteria. Such iron-milk protein has been shown to have a bioavailability similar to that of ferrous sulfate, the gold standard for bioavailability in humans. See, for example, U.S. Patent Application Publication No. 2015 / 0164123 and WO 00 / 51446.

[0052] In preferred embodiments, at least 50% by weight of the added iron is in the form of an iron-milk protein complex as defined above. Even more preferably, the iron-milk protein complex as defined above is substantially the only added source of iron in the composition. Most preferably, the iron-milk protein complex as defined above is the only added source of iron in the composition.

[0053] The added iron source is preferably present in an amount to provide from 1 to 300 mg, preferably from 1 to 250 mg, more preferably from 1 to 200 mg, more preferably from 1 to 100 mg, even more preferably from 1 to 75 mg, and most preferably from 1 to 50 mg of iron per 100 g of composition, based on the total dry weight of the composition.

[0054] Any live probiotic bacteria can be used in the compositions of the present invention. The examples provided in this application show that the above-mentioned complex does not impair the viability of probiotic bacteria. The effect is not strain-specific and can be applied to a wide range of bacterial strains.

[0055] Examples of probiotic bacteria that may be present in the compositions of the present invention include bifidobacteria, lactobacilli, lactococci, enterococci, streptococci, leuconostoc, Escherichia, propionibacteria, or combinations thereof, preferably bacteria of the genus Lactobacillus or Bifidobacterium.

[0056] Preferably, the probiotic bacteria are Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus salivarius ... salivarius, Lactobacillus rhamnosus, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus reuteri, Lactococcus lactis, Streptococcus thermophilus, Lactococcus diacetylactis, Lactococcus cremoris, Lactobacillus delbrueckii subsp. bulgaricus), Lactobacillus delbrueckii subsp.lactis, Lactobacillus helveticus, Escherichia coli, Enterococcus faecium, Leuconostoc pseudomesenteroides, Bifidobacterium bifidum, Lactobacillus gasseri, Lactobacillus sakei, Streptococcus salivarius species, and / or mixtures thereof.

[0057] More preferably, the probiotic bacteria are selected from the species Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus helveticus, Bifidobacterium bifidum, Lactobacillus gasseri, Lactobacillus sakei, and mixtures thereof.

[0058] Examples of bacterial strains that may be advantageously present in the composition include Bifidobacterium longum (deposited under ATCC BAA-999), Bifidobacterium longum (deposited under CNCM I-2618), Bifidobacterium breve (deposited under CNCM I-3865), Bifidobacterium lactis (deposited under CNCM I-3446), Lactobacillus johnsonii (deposited under CNCM I-1225), Lactobacillus paracasei (deposited under CNCM I-2116), Lactobacillus rhamnosus (deposited under CGMCC 1.3724), Streptococcus thermophilus (deposited under CNCM I-1422), Streptococcus thermophilus (deposited under CNCM I-4153), Streptococcus thermophilus (deposited under CNCM I-4153), Streptococcus thermophilus (deposited under CNCM I-4153), Streptococcus rhamnosus (deposited under CGMCC I-4153), Streptococcus spp. (deposited under CNCM ... I-1985), Streptococcus thermophilus (deposited as CNCM I-3915), Lactobacillus casei (deposited as CNCM I-1518), Lactobacillus casei (deposited as ACA-DC 6002), Escherichia coli Nissle (deposited as DSM 6601), Lactobacillus bulgaricus (deposited as CNCM I-1198), Lactococcus lactis (deposited as CNCM I-4154), or a combination thereof.

[0059] More preferred bacterial strains include Bifidobacterium longum (deposited as ATCC BAA-999), Bifidobacterium longum (deposited as CNCM I-2618), Bifidobacterium breve (deposited as CNCM I-3865), Bifidobacterium lactis (deposited as CNCM I-3446), Lactobacillus johnsonii (deposited as CNCM I-1225), Lactobacillus paracasei (deposited as CNCM I-2116), Lactobacillus rhamnosus (deposited as CGMCC 1.3724), Lactobacillus casei (deposited as CNCM I-1518), Lactobacillus casei (deposited as ACA-DC 6002), Streptococcus thermophilus (deposited as CNCM I-2116), Lactobacillus casei (deposited as CGMCC I-2116), Lactobacillus casei (deposited as ACA-DC 6002 ... I-3915), and Lactobacillus bulgaricus (CNCM I-1198), or a combination thereof.

[0060] In a further preferred embodiment, the probiotic bacteria are selected from Bifidobacterium longum (deposited under ATCC BAA-999), Lactobacillus rhamnosus (deposited under CGMCC 1.3724) and Lactobacillus paracasei (deposited under CNCM I-2116) and mixtures thereof.

[0061] The probiotic bacteria are preferably present in the composition in an amount of at least 5E+06 CFU per gram of composition, preferably 5E+06 to 1E+12 CFU per gram of composition, more preferably 5E+06 to 5E+1 CFU per gram of composition, and most preferably 5E+06 to 5E+10 CFU per gram of composition, on a dry weight basis.

[0062] The selected probiotic bacteria can be cultured according to any suitable method and prepared for inclusion in the composition by known techniques, such as freeze-drying or spray-drying. Alternatively, bacterial preparations can be purchased from specialist suppliers such as DSM, DuPont Danisco, Morinaga, Institut Rosell, Christian Hansen and Valio, already prepared in a form suitable for inclusion in the composition in powder form.

[0063] The composition in powder form may be in the form of a free-flowing powder or in the form of a compressed powder, such as in the form of a tablet. Preferably, the composition in powder form is not intended to be used in powder form, but is intended to be reconstituted with a liquid, preferably an aqueous liquid, most preferably water, before use.

[0064] Preferred compositions in powder form of the present invention include food or beverage products, animal feed products, human or animal nutritional supplements, pharmaceutical compositions or cosmetic compositions.

[0065] In another preferred embodiment, the composition in powder form is an edible composition.

[0066] Food and beverage products include all products intended for oral consumption by humans for the purpose of providing nutrition and / or pleasure. In a preferred embodiment, the product is a nutritional composition. More preferably, it is a nutritional composition selected from infant formula, growing-up milk, infant food, infant cereal composition, fortifier, food supplement, and nutritional composition for pregnant or lactating women. More preferably, it is selected from infant formula, growing-up milk, infant cereal composition, and nutritional composition for pregnant or lactating women. Even more preferably, it is selected from infant formula, growing-up milk, and infant cereal composition. Most preferably, it is infant formula or growing-up milk.

[0067] The product may also be in the form of an animal feed product or an animal nutritional supplement. Preferably, the animal is a mammal. Examples of animals include primates, cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, etc.

[0068] Powdered dietary supplements include free-flowing or compressed powder supplements, and typically include supplements that are dissolved in water or sprinkled on wet food or beverages. Such supplements are intended to provide additional nutrients and / or health benefits to the subjects who consume them, as well as other beneficial ingredients, including probiotic bacteria and iron. The dietary supplements according to the present invention can be used to provide nutrients and / or health benefits to humans and animals, as defined above. Dietary supplements include, for example, powdered supplements that are added to breast milk, for example, for premature or low-birth-weight infants. This also includes supplements for women before pregnancy, during pregnancy, and / or during lactation.

[0069] A pharmaceutical composition is a composition intended to treat or prevent an adverse medical condition in a subject in need thereof.

[0070] Cosmetic compositions are typically intended for aesthetic effects on the body and can be administered preferably by the oral route.

[0071] The composition, preferably the nutritional composition, preferably comprises proteins, carbohydrates, fats, vitamins and / or other minerals. Preferably, it comprises all of these types of nutrients.

[0072] The proteins may be intact or hydrolyzed (extensively or partially hydrolyzed).

[0073] The nutritional composition according to the present invention generally contains a lipid source. This is particularly appropriate when the nutritional composition of the present invention is an infant formula. In this case, the lipid source may be any lipid or fat suitable for use in infant formula. Some suitable fat sources include palm oil, high-oleic sunflower oil, and high-oleic safflower oil. The essential fatty acids linoleic acid and alpha-linolenic acid may be added, as well as small amounts of long-chain polyunsaturated fatty acids (LC-PUFAs), such as arachidonic acid (ARA), docosahexaenoic acid (DHA), and / or eicosapentaenoic acid (EPA). Such LC-PUFAs may be provided in the form of fish oil or microbial oil. The iron-milk protein complexes described herein advantageously cause little or no oxidation of LC-PUFAs.

[0074] The composition according to the present invention may contain a carbohydrate source such as lactose, maltodextrin, starch, and mixtures thereof. The composition according to the present invention may also contain a specific type of carbohydrate: prebiotics. Prebiotics that can be used according to the present invention include, but are not limited to, all food substances that promote the growth of probiotics or health-beneficial microorganisms in the intestine. Preferably, they can be selected from the group consisting of: oligosaccharides optionally containing fructose, galactose, and mannose; dietary fiber, especially soluble fiber, soy fiber; inulin; or mixtures thereof. Some examples of prebiotics are fructooligosaccharides (FOS), galactooligosaccharides (GOS), isomaltooligosaccharides (IMO), xylooligosaccharides (XOS), arabinoxylooligosaccharides (AXOS), mannanoligosaccharides (MOS), inulin, polydextrose, glycosylsucrose (GS), lactosucrose (LS), lactulose (LA), palatinose oligosaccharides (PAO), maltooligosaccharides, gums and / or their hydrolysates, pectin and / or their hydrolysates. In certain embodiments, the prebiotic may be fructooligosaccharides and / or inulin. Suitable commercial products that can be used include a combination of FOS and inulin, such as the product sold by BENEO under the trademark Orafti, or polydextrose sold by Tate & Lyle under the trademark STA-LITE®.

[0075] The prebiotic may also be a BMO (bovine milk oligosaccharide) and / or a HMO (human milk oligosaccharide), such as an N-acetylated oligosaccharide, a sialylated oligosaccharide, a fucosylated oligosaccharide, and any mixture thereof.

[0076] A particular example of a prebiotic is a mixture of galactooligosaccharide(s), N-acetylated oligosaccharide(s), and sialylated oligosaccharide(s), wherein the N-acetylated oligosaccharide(s) represent 0.5 to 4.0% by weight of the oligosaccharide mixture, the galactooligosaccharide(s) represent 92.0 to 98.5% by weight of the oligosaccharide mixture, and the sialylated oligosaccharide(s) represent 1.0 to 4.0% by weight of the oligosaccharide mixture. For example, a composition for use according to the invention may contain 2.5 to 15.0% by weight of CMOS-GOS on a dry matter basis, provided that the composition comprises at least 0.02% by weight of N-acetylated oligosaccharides, at least 2.0% by weight of galactooligosaccharides, and at least 0.04% by weight of sialylated oligosaccharides. WO2006087391 and WO2012160080 provide some examples of the production of such oligosaccharide mixtures.

[0077] The composition of the present invention may also contain all vitamins, minerals, and other micronutrients that are considered essential and nutritionally significant in daily diet.Minimum requirements for certain vitamins and minerals have been established.Examples of minerals, vitamins, and other nutrients that may optionally be present in the composition of the present invention include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorus, iodine, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine, and L-carnitine.Minerals are usually added in salt form.The presence and amount of specific minerals and other vitamins vary depending on the intended target group.

[0078] Iron-milk protein complexes as described herein advantageously cause little or no oxidation of vitamins, such as vitamin A and vitamin C. It is also preferred that the compositions of the present invention include polyphenols, as iron-milk protein complexes as described herein cause little or no oxidation of polyphenols.

[0079] Method for producing a composition in powder form In one embodiment, the present invention relates to a method for producing a composition in powder form, the method comprising: a) providing a first composition in powder form; b) mixing an iron source with a first composition in powder form to obtain a second composition in powder form; c) mixing at least one probiotic bacterium into a first composition in powder form before or simultaneously with the iron source, or mixing at least one probiotic bacterium into a second composition in powder form after the addition of the iron source; and wherein the iron source is an iron-milk protein complex.

[0080] The composition in powder form, the iron source and the probiotic bacteria are as defined in any of the embodiments above in the "Composition" section.

[0081] The powder form of the first composition can be prepared by any method known in the art, such as spray drying. In this method, the components of the composition are mixed into a wet mixture and sprayed in a drying tower to form a powder. Reactive ingredients such as minerals or vitamins can also be added in dry form, for example, before, simultaneously with, or after the addition of an iron source and / or probiotic bacteria. For example, an iron source is typically added to the powder form of the first composition as a trace element premix containing various minerals and vitamins.

[0082] The probiotic bacteria are preferably added in a premixed form comprising a carrier and, optionally, a protectant. Carriers for probiotic bacteria are well known to those skilled in the art of incorporating probiotic bacteria into products. The carrier is preferably a carbohydrate, such as maltodextrin. A wide variety of protectants are known to those skilled in the art. Particularly suitable protectants are those described in the co-pending patent application of the same applicant, International Application No. PCT / EP16 / 065359.

[0083] The steps of mixing the probiotic bacteria and the iron source can be carried out using any method known in the art for mixing such types of ingredients with a powder composition. For example, mixing can be carried out by dry mixing, or by direct addition in a spray drying tower, or by direct addition in a fluidized bed, coater, or agglomerator, or by direct addition in the final stage of extrusion or roller drying. Preferably, the probiotic bacteria and / or the iron source are mixed with the powdered composition by dry mixing or by adding to a fluidized bed. Most preferably, the probiotic bacteria and / or the iron source are mixed with the powdered composition by dry mixing.

[0084] In preferred embodiments, at least 50% by weight of the added iron in the composition is in the form of an iron source as defined above. Even more preferably, the iron source as defined above is substantially the only added iron source in the composition. Most preferably, the iron source as defined above is the only added iron source in the composition.

[0085] Use of an iron-milk protein complex for the enrichment of compositions in powder form Iron-milk protein complexes can be advantageously used to enrich compositions in powder form containing at least one probiotic bacterium. Such iron sources advantageously provide bioavailable iron while not causing a significant decrease in the viability of the probiotic bacteria during reconstitution of the powder with a liquid such as water.

[0086] As noted above, iron-milk protein complexes with bioavailability similar to that of ferrous sulfate are particularly useful for fortifying food products.

[0087] In another embodiment, the present invention relates to a method for enriching a composition in powder form comprising at least one probiotic bacterium, the method comprising: a) providing a first composition in powder form; b) mixing a first composition in powder form with an iron source to produce a second composition in powder form; c) mixing at least one probiotic bacterium into a first composition in powder form before or simultaneously with the iron source, or mixing at least one probiotic bacterium into a second composition in powder form after the addition of the iron source; wherein the iron source is an iron-milk protein complex.

[0088] The first composition in powder form, the second composition in powder form, and the method steps are as defined above in the section relating to the method for producing the composition of the present invention, and the composition in powder form, the iron source, and the probiotic bacteria are as described above in any embodiment in the "Composition" section.

[0089] In preferred embodiments, at least 50% by weight of the added iron in the composition is in the form of the above-defined iron source, more preferably at least 60% by weight, more preferably at least 70% by weight, more preferably at least 80% by weight, and even more preferably at least 90% by weight. Even more preferably, the above-defined iron source is substantially the only added iron source in the composition. Most preferably, the above-defined iron source is the only added iron source in the composition.

[0090] Compositions for use in methods of preventing, alleviating, and / or treating iron deficiency As noted above, since the compositions of the invention are fortified with a highly bioavailable iron source, the invention also relates to compositions for use in methods of preventing, alleviating, and / or treating iron deficiency in an individual.

[0091] Product manufacturing method The present invention relates to a method for producing a product, preferably an edible product, comprising the step of reconstituting with a liquid a composition in powder form comprising at least one probiotic bacterium and an iron source, characterized in that the iron source is an iron-milk protein complex.

[0092] The composition in powder form, the probiotic bacteria and the iron source are as defined in any of the above embodiments.

[0093] The use of iron-milk protein complexes is particularly advantageous when the powdered composition is reconstituted with a liquid. The inventors have found that probiotic bacteria are particularly reactive upon rehydration, and that the loss of viability is particularly severe when the powdered composition is reconstituted with a liquid such as water. The inventors have also found that iron-milk protein complexes, in contrast to common iron sources such as ferrous sulfate heptahydrate or dissolved ferrous sulfate in spray-dried form, do not significantly reduce the viability of probiotic bacteria at this critical stage.

[0094] Reconstitution is preferably carried out with an aqueous liquid, most preferably with water. The amount of liquid, preferably water, added for reconstitution varies depending on the type of product being prepared. For the purposes of the present invention, what is important is that sufficient liquid is added to reconstitute the probiotic bacteria to their natural water content.

[0095] product The present invention also relates to a product, preferably an edible product, obtainable or obtainable by the above-described method. The product comprises a composition, as defined in any embodiment of the present invention, in powder form to be reconstituted with a liquid. Preferably, it is reconstituted with an aqueous liquid, preferably water.

[0096] Preferably, the product is a nutritional product obtained by reconstitution of the nutritional composition described above.

[0097] How to provide nutrition Also contemplated is a method of providing nutrition to an individual, comprising the step of feeding the individual an edible product of the present invention, preferably a nutritional product of the present invention. The product used in this method is a food composition or a beverage composition. Preferably, such a composition is a nutritional composition as defined above. Such a product is particularly advantageous because it contains a bioavailable iron source and a stable concentration of probiotic bacteria, since the added iron source does not cause a significant reduction in the probiotic bacteria.

[0098] In one embodiment, the method comprises: a) reconstituting an edible composition in powder form according to any of the embodiments of the present invention; b) administering the reconstituted composition to the individual; Includes.

[0099] In one embodiment, the individual has iron deficiency or is at risk of developing iron deficiency.

[0100] In another preferred embodiment, the edible composition is a nutritional composition.

[0101] Methods for preventing or reducing the loss of probiotic bacteria The present invention relates to a method for reducing or preventing the loss of probiotic bacteria during the reconstitution of a composition in powder form comprising at least one probiotic bacterium and an added iron source, characterized in that an iron-milk protein complex is used as the added iron source.

[0102] The probiotic bacteria, added iron source, and composition in powder form are as described in any embodiment in the "Composition" section.

[0103] In preferred embodiments, the iron-milk protein complex represents at least 50% by weight of the added iron in the composition, more preferably at least 60% by weight, more preferably at least 70% by weight, more preferably at least 80% by weight, and even more preferably at least 90% by weight. More preferably, the iron-milk protein complex is substantially the only added iron source used in the composition. Most preferably, the iron-milk protein complex is the only added iron source in the composition. In other words, the composition does not contain other ferrous or ferric compounds added as iron sources in the composition.

[0104] The inventors have shown that by using an iron-milk protein complex in place of other commonly added iron sources such as ferrous sulfate heptahydrate or dissolved ferrous sulfate in spray-dried form, the loss of probiotic bacteria upon reconstitution of the composition can be prevented or at least significantly reduced.

[0105] Although added iron sources have a significant effect on the survival of probiotic bacteria, iron sources present as part of ingredients that are not primarily intended for iron supplementation have a much smaller effect due to the complexity of such ingredients, among other reasons: the iron they contain is typically not found as free iron when the composition is reconstituted, thus reducing any potential harmful effects on the probiotic bacteria.

[0106] The present invention will now be described in further detail by way of the following examples.

[0107] Example 1: Effect of iron source on the survival of Lactobacillus rhamnosus Four powder compositions (Samples A to D) were prepared, each containing dried Lactobacillus rhamnosus (deposited under CGMCC 1.3724), an iron source, and 100 g of maltodextrin. The composition of the samples is summarized in Table 1.

[0108] [Table 1]

[0109] Samples were prepared as follows: Maltodextrin, dried probiotic powder, and iron source were added together in a 500 mL plastic beaker and mixed for 5 minutes at 34 rpm using a Turbula® T2F dry mixer (WAB, Switzerland). A 25 g quantity of the freshly mixed sample was diluted 1 / 10 with aqueous tryptone salt solution supplemented with antifoam at 37°C. After 5 minutes of homogenization using a Masticator® (IUL, Spain), decimal dilutions were performed. Appropriate dilutions yielding 30–300 colonies after incubation were then transferred to Petri dishes and mixed with MRS agar according to the pour plate method. Once solidified, the plates were incubated at 37°C under aerobic conditions. After 48 hours of incubation, the colonies present on the plates were counted.

[0110] The results are summarized in Figure 1. From this graph, it is clear that a significant reduction in live L. rhamnosus was observed in the reconstituted composition when the spray-dried form of dissolved ferrous sulfate was used. In contrast, no significant reduction in L. rhamnosus was observed when the iron-casein complex was used under the same conditions and with the same amount of L. rhamnosus and iron.

[0111] Example 2: Effect of iron source on the survival of Bifidobacterium longum Five powder compositions (Samples E to I) were prepared, each containing dried Bifidobacterium longum (deposited under ATCC BAA-999), an iron source, and 100 g of maltodextrin. The compositions of the five samples are summarized in Table 2 below.

[0112] [Table 2]

[0113] Samples were prepared as follows: Maltodextrin, dried probiotic powder, and iron source were added together in a 500 mL plastic beaker and mixed for 5 minutes at 34 rpm using a Turbula® T2F dry mixer (WAB, Switzerland). A 25 g quantity of the freshly mixed sample was diluted 1 / 10 with aqueous tryptone salt solution supplemented with antifoam at 37°C. After 5 minutes of homogenization using a Masticator® (IUL, Spain), decimal dilutions were performed. Appropriate dilutions yielding 30–300 colonies after incubation were then transferred to Petri dishes and mixed with RCA agar according to the pour plate method. Once solidified, the plates were incubated at 37°C under anaerobic conditions. After 48 hours of incubation, the colonies present on the plates were counted.

[0114] The results are summarized in the graph of Figure 2. From this graph, it is clear that a significant reduction in live B. longum was observed in the reconstituted composition when the spray-dried form of dissolved ferrous sulfate was used. In contrast, no significant reduction in B. longum was observed when the iron-casein complex was used under the same conditions and with the same amount of B. longum and iron.

[0115] Example 3: Effect of iron source on the survival of Bifidobacterium longum Five powder compositions (Samples I–M) were prepared, each containing dried Bifidobacterium longum (deposited under ATCC BAA-999), an iron source, and 100 g of infant formula. The compositions of the five samples are summarized in Table 3 below.

[0116] [Table 3]

[0117] Samples were prepared as follows: Infant formula, dried probiotic powder, and iron source were added together in a 500 mL plastic beaker and mixed for 5 minutes at 34 rpm using a Turbula® T2F dry mixer (WAB, Switzerland). A 25 g quantity of the freshly mixed sample was diluted 1 / 10 with aqueous tryptone salt solution supplemented with antifoam at 37°C. After 5 minutes of homogenization using a Masticator® (IUL, Spain), decimal dilutions were performed. Appropriate dilutions yielding 30–300 colonies after incubation were then transferred to Petri dishes and mixed with RCA agar according to the pour plate method. Once solidified, the plates were incubated at 37°C under anaerobic conditions. After 48 hours of incubation, the colonies present on the plates were counted.

[0118] The results are summarized in the graph of Figure 3. From this graph it is clear that when the iron source used was iron-casein, there was significantly less loss of live B. longum in the reconstituted composition than when a spray-dried form of dissolved ferrous sulfate was used under the same conditions and with the same amount of B. longum and the same amount of iron.

[0119] Example 4: Effect of iron source on the survival of Lactobacillus paracasei Five powder compositions (Samples N to R) were prepared, each containing dried Lactobacillus paracasei (deposited as CNCM I-2116), an iron source, and 100 g of maltodextrin. The compositions of the five samples are summarized in Table 4 below.

[0120] [Table 4]

[0121] Samples were prepared as follows: Maltodextrin, dried probiotic powder, and iron source were added together in a 500 mL plastic beaker and mixed for 5 minutes at 34 rpm using a Turbula® T2F dry mixer (WAB, Switzerland). A 25 g quantity of the freshly mixed sample was diluted 1 / 10 with aqueous tryptone salt solution supplemented with antifoam at 37°C. After 5 minutes of homogenization using a Masticator® (IUL, Spain), decimal dilutions were performed. Appropriate dilutions yielding 30–300 colonies after incubation were then transferred to Petri dishes and mixed with MRS agar according to the pour plate method. Once solidified, the plates were incubated at 37°C under aerobic conditions. After 48 hours of incubation, the colonies present on the plates were counted.

[0122] The results are summarized in the graph of Figure 4. From this graph, it is clear that a significant reduction in live L. paracasei was observed in the reconstituted composition when the spray-dried form of dissolved ferrous sulfate was used. In contrast, when the iron-casein complex was used under the same conditions with the same amount of L. paracasei and the same amount of iron, no significant reduction in L. paracasei was observed.

[0123] Example 5: Effect of iron source on the survival of Lactobacillus paracasei Five powder compositions (Samples S–W) were prepared, each containing dried Lactobacillus paracasei (deposited as CNCM I-2116), an iron source, and 100 g of infant formula. The compositions of the five samples are summarized in Table 5 below.

[0124] [Table 5]

[0125] Samples were prepared as follows: Infant formula, dried probiotic powder, and iron source were added together in a 500 mL plastic beaker and mixed for 5 minutes at 34 rpm using a Turbula® T2F dry mixer (WAB, Switzerland). A 25 g quantity of the freshly mixed sample was diluted 1 / 10 with aqueous tryptone salt solution supplemented with antifoam at 37°C. After 5 minutes of homogenization using a Masticator® (IUL, Spain), decimal dilutions were performed. Appropriate dilutions yielding 30–300 colonies after incubation were then transferred to Petri dishes and mixed with MRS agar according to the pour plate method. Once solidified, the plates were incubated at 37°C under aerobic conditions. After 48 hours of incubation, the colonies present on the plates were counted.

[0126] The results are summarized in the graph of Figure 5. When each of the iron sources was added in an amount providing 10 mg of iron, no significant reduction in probiotics was observed, but when the spray-dried form of dissolved ferrous sulfate was used in an amount providing 30 mg of iron, a clear effect on the viability of L. paracasei was observed in the reconstituted composition. In contrast, when the iron-casein complex was used under the same conditions with the same amount of L. paracasei and the same amount of iron, no significant reduction in L. paracasei was observed.

[0127] Example 6: Effect of iron source on the survival of Lactobacillus rhamnosus Five powder compositions (Samples X through BB) were prepared, each containing dried Lactobacillus rhamnosus (deposited under CGMCC 1.3724), an iron source, and 100 g of infant formula. The compositions of the five samples are summarized in Table 6 below.

[0128] [Table 6]

[0129] Samples were prepared as follows: Infant formula, dried probiotic powder, and iron source were added together in a 500 mL plastic beaker and mixed for 5 minutes at 34 rpm using a Turbula® T2F dry mixer (WAB, Switzerland). A 25 g quantity of the freshly mixed sample was diluted 1 / 10 with aqueous tryptone salt solution supplemented with antifoam at 37°C. After 5 minutes of homogenization using a Masticator® (IUL, Spain), decimal dilutions were performed. Appropriate dilutions yielding 30–300 colonies after incubation were then transferred to Petri dishes and mixed with MRS agar according to the pour plate method. Once solidified, the plates were incubated at 37°C under aerobic conditions. After 48 hours of incubation, the colonies present on the plates were counted.

[0130] The results are summarized in the graph of Figure 6. From this graph it is clear that a significant reduction in live L. rhamnosus was observed in the reconstituted composition when the spray-dried form of dissolved ferrous sulfate was used. In contrast, when an iron-casein complex was used under the same conditions with the same amount of L. rhamnosus and the same amount of iron, the reduction in L. rhamnosus was significantly reduced.

Claims

1. A composition in powder form comprising at least one live probiotic bacterium and an iron source, characterized in that the iron source is an iron-milk protein complex, said milk protein being casein, said live probiotic bacteria being present in an amount of 5E+06 to 5E+10 CFU per gram of said composition on a dry weight basis, A composition wherein said live probiotic bacteria are selected from the group consisting of Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei and mixtures thereof.

2. 2. The composition of claim 1, wherein the iron-milk protein complex comprises exogenous iron.

3. 3. The composition according to claim 1 or 2, characterized in that the iron-milk protein complex is a non-micellar complex.

4. The composition according to any one of claims 1 to 3, characterized in that the iron-milk protein complex contains exogenous phosphorus.

5. 5. The composition of claim 4, wherein the iron-milk protein complex comprises exogenous orthophosphorus.

6. The iron in the iron-milk protein complex is Fe ３＋ 6. The composition according to claim 1, wherein the composition is in the form of

7. Composition according to any one of claims 1 to 6, characterized in that the composition in powder form is a nutritional composition.

8. A composition according to any one of claims 1 to 7, characterized in that the iron source is present in an amount to provide from 1 to 300 mg of iron per 100 g of composition.

9. 1. Use of an iron source in a powder form composition for iron fortification comprising at least one live probiotic bacterium, characterized in that the iron source is an iron-milk protein complex, said milk protein being casein, said live probiotic bacteria being present in an amount of 5E+06 to 5E+10 CFU per gram of said composition on a dry weight basis, said live probiotic bacteria being selected from the group consisting of Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei and mixtures thereof.

10. A dietary composition in powder form comprising at least one live probiotic bacterium and an added iron source for use in the prevention, alleviation and / or treatment of iron deficiency in an individual, comprising: the added iron source is an iron-milk protein complex and the milk protein is casein; the live probiotic bacteria are present in an amount of 5E+06 to 5E+10 CFU per gram of the edible composition on a dry weight basis; 10. An edible composition, characterized in that the live probiotic bacteria are selected from the group consisting of Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei and mixtures thereof.

11. 1. A method for producing a composition in powder form, comprising: a. providing a first composition in powder form; b. mixing a source of iron with said first composition in powder form to obtain a second composition in powder form; c. mixing at least one live probiotic bacterium into said first composition in powder form prior to or simultaneously with said iron source, or mixing at least one live probiotic bacterium into said second composition in powder form after the addition of said iron source; wherein the iron source is an iron-milk protein complex, the milk protein being casein, and the live probiotic bacteria are present in an amount of 5E+06 to 5E+10 CFU per gram of the powdered composition on a dry weight basis; 13. A method according to claim 12, wherein the live probiotic bacteria are selected from the group consisting of Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei and mixtures thereof.

12. 1. A method for manufacturing a product, comprising: reconstituting a composition in powder form comprising at least one live probiotic bacterium and an iron source with a liquid; 13. A method according to claim 12, wherein the iron source is an iron-milk protein complex, the milk protein being casein, the live probiotic bacteria being present in an amount of from 5E+06 to 5E+10 CFU per gram of the powdered composition on a dry weight basis, and the live probiotic bacteria being selected from the group consisting of Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei and mixtures thereof.

13. 1. A liquid product comprising at least one live probiotic bacterium and an iron source, said iron source being an iron-milk protein complex, said milk protein being casein, said live probiotic bacteria being present in an amount of from 5E+06 to 5E+10 CFU per gram of said product on a dry weight basis, said live probiotic bacteria being selected from the group consisting of Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei and mixtures thereof.

14. 14. The liquid product of claim 13 which is a dietary supplement.

15. 1. A method for reducing or preventing loss of live probiotic bacteria during reconstitution of a composition in powder form comprising at least one live probiotic bacteria and an added iron source into a liquid product, comprising: an iron-milk protein complex is used as the added iron source, the milk protein being casein, and the live probiotic bacteria are present in an amount of 5E+06 to 5E+10 CFU per gram of the powdered composition on a dry weight basis; 13. A method according to claim 12, wherein the live probiotic bacteria are selected from the group consisting of Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei and mixtures thereof.

16. A composition according to any one of claims 1 to 8, wherein the viability of said live probiotic bacteria is not compromised by the presence of said iron source.

17. A composition according to any one of claims 1 to 8, wherein the iron-milk protein complex is a complex formed of iron cations chelated with milk protein.

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