COMPOSITION COMPRISING LACTOBACILLUS AND A CARRIER

The invention relates to a food composition comprising Lactobacillus  which is capable of aggregating Helicobacter pylori under physiological conditions  and a carrier  wherein the carrier comprises milk serum and wherein the carrier is a product of a fermentation process. The invention also relates to methods for producing such food compositions  uses thereof and uses of carriers.

Background of the invention

Helicobacter pylori (H. pylori) is a bacterium of helical shape  which colonizes the stomach. In order to colonize the stomach  the bacteria enter the mucus  which covers the stomach""s epithelial cell layer. H. pylori are also found on the inner surface of the stomach epithelial cells and occasionally inside epithelial cells. The cells are attached to the stomach wall by specific molecules  such as adhesines. In contrast  H. pylori has a limited stability in the acidic lumen and is secreted easily therefrom. In most individuals  an infection with H. pylori is harmless. However  acute infections can develop which are associated with various diseases  such as gastritis or ulcers. Contact of the epithelial layer with the bacteria is considered to be a requirement of the development of such infectious diseases.

It is known in the art that Lactobacillus has advantageous effects against H. pylori. Lactobacillus is a genus of specific bacteria  which are capable of converting lactose and other sugars into lactic acid.

US 5 716 615 discloses pharmaceutical compositions comprising Lactobacillus and other active ingredients. The composition is useful for treating disorders in the gastrointestinal tract.

US 2005/0186190 A1 discloses a dietetic or pharmaceutical composition which comprises sphingomyelinase or Lactobacillus  which comprises sphingomyelinase. The composition is useful for the treatment of infections by H. pylori.

WO 2004/087891 A1 discloses specific strains of Lactobacillus  which are useful for preparing pharmaceutical or dietetic compositions for the treatment of infections of the gastrointestinal tract caused by H. pylori.

WO 2005/060937 A1 discloses tablet-like formulations comprising viable cells of Lactobacillus. The formulations are useful for oral administration and treatment of infections of the gastrointestinal tract caused by pathogens.

WO 2007/073709 discloses novel strains of Lactobacillus and their use against infections by H. pylori. In contrast to known Lactobacillus strains  the novel strains are capable of aggregating H. pylori under physiological conditions. The aggregates seem to be formed  because the Lactobacillus cells bind to the H. pylori and induce the formation of mixed aggregates. The relatively large aggregates are not capable of entering or penetrating the mucus any more. Consequently  the aggregated cells are not capable any more of contacting the epithelial cells of the stomach and infecting them. The aggregates are not attached to the inner walls of the gastrointestinal tract. They accumulate in the lumen  pass the gastrointestinal tract and are secreted naturally. The overall level of H. pylori is reduced and inflammatory reactions of the immune system are healed or prevented. Specifically  infections caused by H. pylori  such as gastritis and ulcers  are treated or prevented.

These novel strains described by WO 2007/073709 have a different mode of action and also a different efficiency compared to previously known Lactobacillus strains useful against H. pylori. Strains known in the art fail to form aggregates with H. pylori. Generally  known strains exert a beneficial effect against H. pylori by competing with H. pylori cells in the stomach and thereby slowly replacing the cells. The rapid formation and secretion of aggregates thus provides a novel approach in the treatment of H. pylori infections.

There is a basic need to improve such known Lactobacillus compositions further regarding efficiency and availability. Further  since H. pylori infections are widespread  it is desirable to provide efficient means for prevention and treatment  which are available easily and at low costs also for large groups of individuals.

Problem underlying the invention

The problem underlying the invention is to provide novel compositions  methods and uses which overcome the above mentioned problems and which are highly efficient in the treatment or prevention of diseases associated with H. pylori.

It is a specific problem underlying the invention to provide food compositions  which are highly effective against H. pylori. The level of H. pylori cells in the gastrointestinal tract of an animal  especially a human  shall be reduced rapidly and extensively. Further  the number of H. pylori cells in the stomach and intestine shall be maintained at a low level.

Further  the composition of the invention shall not have disadvantageous physiological side effects and shall be pharmacologically acceptable. The composition shall be manufactured and stored easily  efficiently and at relatively low costs. Thus  a convenient and efficient treatment of large numbers of individuals shall be possible.

Disclosure of the invention

Surprisingly  the problem underlying the invention is solved by food compositions  methods and uses according to the claims. Further embodiments of the invention are outlined throughout the description.

Subject of the invention is a food composition comprising Lactobacillus  which is capable of aggregating Helicobacter pylori under physiological conditions  and a carrier  wherein the carrier comprises milk serum and wherein the carrier is a product of a fermentation process.

According to the invention  a "food composition" is any composition  which can be administered to an animal  specifically a human  as a food. Since the composition comprises an ingredient with activity against H. pylori  it is also a dietetic composition.

The Lactobacillus may be dead or viable. In a specific embodiment of the invention  the Lactobacillus or a major portion thereof are dead. It is assumed that the ability of the Lactobacillus to aggregate H. pylori is mediated by the membranes of the Lactobacillus  and that cell membranes or dead cells effectively aggregate H. pylori. In other embodiments  at least 10% of the total Lactobacillus cells  preferably more than 50% or more than 90% are living cells. In another embodiment  essentially all the cells are viable  i.e. more than 95% or more than 99% of the total Lactobacillus cells. The food composition is thus a probiotic.

In a preferred embodiment  the food composition is an end product  which is ready for consumption by a consumer. It thus can be bought  or obtained otherwise  by the consumer. However  the food composition may also be a basic component for the production of other foods.

In a preferred embodiment of the invention  the Lactobacillus is selected from Lactobacillus fermentum  Lactobacillus reuteri  Lactobacillus brevis and Lactobacillus pentosus.

Preferably  the Lactobacillus capable of aggregating H. pylori is a strain disclosed by WO 2007/073709  which is hereby expressively incorporated by reference. As already outlined above  this document discloses Lactobacillus strains which are capable of binding and aggregating Helicobacter pylori cells in the gastrointestinal tract under physiological conditions. The aggregates are not capable of entering or passing the mucus anymore and are secreted  thereby preventing an inflammatory reaction of the immune system. The Lactobacillus strains are thus highly efficient against diseases associated with H. pylori  such as gastritis or ulcers.

The strains are usable for preventing and/or treating an infection with H. pylori. Even when an infection has already occurred  the Lactobacillus prevents further infection with H. pylori bacterial cells and the existing infection can be treated more easily by elimination of the H. pylori  which have already passed the mucus. The elimination of cells  which have already caused the infection  is supported or mediated by the natural immune response of the individual. In addition  it is assumed that the Lactobacillus strains disclosed by WO 2007/073709 are also effective in inhibiting urease activity of H. pylori. Thereby  H. pylori cells within the formed aggregates lose their protection against gastric acid  which renders their elimination even more effective.

The term “under physiological conditions” refers to the conditions  under which the H. pylori to be aggregated dwell. If H. pylori in the gastrointestinal tract are to be treated  then physiological conditions are those in the gastrointestinal tract  i.e. in the presence of gastric acid.

The term “aggregate” refers to the formation of aggregates of bacteria  in which the bacterial cells bind or stick to each other. Such aggregates are visible under the microscope. Typically  the aggregates formed during aggregation have a diameter of approximately 1 to 10 µm  but they may be even larger in some instances. The aggregates comprise Lactobacillus and H. pylori cells. A useful aggregation assay for testing the formation of aggregates is outlined in the following working examples. Typically  culturing conditions are selected which mimic physiological conditions in the gastrointestinal tract of the respective animal  such as a human. A similar useful assay is disclosed in WO 2007/073709 in examples 2 and 3 and on pages 9 to 12.

In a preferred embodiment of the invention  the Lactobacillus is selected from Lactobacillus strains deposited as No. DSM 17648  DSM 17646  DSM 17647  DSM 17649  DSM 17650  DSM 17651  DSM 17652 or DSM 17653 at the DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH  Mascheroder Weg Ib  38124 Braunschweig  Germany). As outlined in WO 2007/073709  these strains are deposited for public use. The strains DSM 17646  17649  17652 and 17653 are Lactobacillus brevis. DSM 17650 is Lactobacillus pentosus. In a preferred embodiment of the invention  the Lactobacillus is DSM 17648.

The strains DSM 17647  17648 and 17651 are Lactobacillus fermentum. According to example 5 of WO 2007/073709  taxonomic determination of the strains was carried out by determining carbohydrate fermentation patterns according to the API50CH systems (BioMerieux  France). This test is based on the fermentation of 49 different carbohydrates and allows the identification of a strain as Lactobacillus fermentum with high certainty. In addition  the present inventors examined the strain DSM 17648 using a 16s DNA-based method. When using this approach  the strain was classified as Lactobacillus reuteri. Nowadays  the 16s DNA based method is considered more reliable. However  precise taxonomic determination of related strains of the same genus is generally difficult.

In a preferred embodiment of the invention  the amount of Lactobacillus in the composition is between 104 to 1015  preferably 106 to 1013  more preferably 108 to 1012 cells  most preferably 109 to 1011 cells. Specifically  it was found that an appropriate amount of cells in a single daily dose is 2x109. The total number refers to the total amount of living and dead cells. The number of cells can be determined by known methods  for instance with a cell counter. Reference is a single daily dose. However  reference can also be a packaging unit.

The inventive food composition comprises a carrier. The term "carrier" suggests that the Lactobacillus is distributed throughout the carrier. Thus the carrier is the main component of the food composition. Preferably  the total amount of Lactobacillus in the carrier is below 5%  more preferably below 2% or below 0.5% (w/w)  based on the combined amount of carrier and Lactobacillus. In a specific embodiment  the food composition consists of the Lactobacillus and the carrier. However  the composition may comprise additives. Preferably  such additives are also distributed throughout the carrier.

The carrier is the product of a fermentation process. According to the invention  this could mean that either the carrier itself was fermented  i. e.  that it is a direct product of a fermentation process. Alternatively  an intermediate product obtained during production of the carrier was fermented. After the fermentation of the intermediate product  it was then processed further  for example fractionated (such as yoghurt serum) and/or admixed with other components. In a specific embodiment  the carrier is a food by itself.

In a preferred embodiment of the invention  the carrier was not fermented by the Lactobacillus capable of aggregating Helicobacter pylori. In this embodiment  the carrier was fermented in a preceding fermentation process by a different microorganism. The Lactobacillus strain is then preferably grown separately and harvested  and subsequently mixed with the carrier to obtain the inventive food composition. In a preferred embodiment  in the inventive food composition the active Lactobacillus ingredient for aggregating H. pylori essentially does not ferment the carrier.

In another preferred embodiment  the carrier was fermented by the Lactobacillus capable of aggregating Helicobacter pylori. In this embodiment  the Lactobacillus could be grown in the food composition. Alternatively  a fermentation mixture could be used comprising the inventive Lactobacillus strain in combination with at least one additional strain  such as Lactobacillus bulgaricus or Streptococcus thermophilus. In a specific embodiment  the carrier is yoghurt or a yoghurt fraction and contains Lactobacillus delbrueckii ssp bulgaricus und Streptococcus thermophilus.

In a preferred embodiment of the invention  at least 50%  more preferably at least 90% or at least 98% of the H. pylori cells aggregate in an aggregation assay carried out in vitro  preferably under the conditions outlined in the working examples.

The carrier is a dairy product  i. e. a milk product. It is highly preferred that the milk is cow milk. The inventive carrier is milk serum or comprises milk serum. Milk serum is a milk fraction depleted of fat and depleted of the proteins  i.e. casein and whey protein. Milk serum can be obtained through ultrafiltration from skim milk. Milk serum is comprised in yoghurt serum. It is usually a colorless liquid. Milk serum contains essentially only the water soluble milk components  such as lactose  minerals and vitamins. Preferably  the carrier comprises less than 1%  preferably less than 0.5% or less than 0.2% (w/w) protein. Preferably  the inventive carrier comprises less than 5%  more preferably less than 3%  less than 1% or less than 0.5% (w/w) fat.

Milk contains about 3.3% total protein. There are two major categories of milk protein that are broadly defined by their chemical composition and physical properties. The casein proteins contain phosphorus and will coagulate or precipitate at pH 4.6. The casein family of protein consists of several types of caseins (a-s1  a-s2  ß  and 6) and each has its own amino acid composition  genetic variations  and functional properties. The casein proteins are suspended in milk in micelles.

Whey is a milk fraction obtained when casein micelles are removed when producing cheese. The whey proteins do not contain phosphorus and remain in solution in milk at pH 4.6. The whey protein family consists of approximately 50% ß-lactoglobulin  20% a-lactalbumin  blood serum albumin  immunoglobulins  lactoferrin  transferrin  and many minor proteins and enzymes. In cow""s milk  approximately 82% of milk protein is casein and the remaining 18% is whey protein. Preferably  the inventive carrier does not comprise whey proteins.

In a preferred embodiment of the invention  the fermentation was a bacterial fermentation. For example  the fermentation may be a fermentation carried out typically in the production of dairy products  such as yoghurt.

In fermentation processes with milk and milk products  is usually converted into lactate. Typically  fermented dairy products have a relatively high lactate content. In a preferred embodiment of the invention  the food composition comprises between 0.1 to 5%  preferably between 0.2 and 2%  more preferably between 0.4 and 2% (w/w) lactate. As used herein  the term "lactate" refers to the combined amount of lactic acid and lactate. It is preferred that the lactate concentration in the food composition is at least 0.2%  preferably above 0.5% or above 1%. In a preferred embodiment of the invention  the food composition comprises between 0.1 to 7%  preferably between 0.5 and 5% (w/w) lactose. In yoghurt or yoghurt serum  the lactose content may be between 2 and 7% (w/w) or between 3 and 5% (w/w). It is preferred that the lactate concentration in the food composition is below 5%  preferably below 3% or below 1%.

Without being bound to theory  lactose may inhibit attachment of the H. pylori cells to the Lactobacillus cells and thus decrease or inhibit aggregation. In a preferred embodiment  the carrier is a composition in which at least a portion of the lactose has been converted into lactate during the fermentation  preferably at least 10% or at least 20% of the lactose.

Typically  the fermented carrier of the present invention has an acidic pH value. The low pH is usually a result of the increased lactic acid concentration. In a preferred embodiment of the invention  the pH of the food composition is between 3 and 6  preferably between 3.5 and 5.5 or between 3.5 and 5.

In a preferred embodiment of the invention  the carrier is yoghurt serum; or comprises yoghurt serum. Yoghurt serum is the clear liquid trapped between the yoghurt solids. It is obtainable from yoghurt by methods such as filtration  centrifugation or straining  followed by separation of the non-soluble fraction. It was surprisingly found that yoghurt serum is highly efficient in enhancing the aggregation of H. pylori by specific Lactobacillus strains.

According to the invention  the carrier could be yoghurt. Yoghurt comprises yoghurt serum and thus milk serum. Yoghurt serum effectively enhances aggregation of H. pylori by the Lactobacillus strains on its own. The other yoghurt fraction may not improve aggregation further. It may even weaken the beneficial effect  for example simply by diluting the yoghurt serum. In a specific embodiment of the invention  the carrier is thus not yoghurt.

In a preferred embodiment of the invention  the carrier consists of
(a) 0.1 to 10 % (w/w) proteins 
(b) 0 to 5% (w/w) fat 
(c) 0 to 12%  preferably 0 to 2% carbohydrates 
(d) 0.1 to 5% (w/w) lactate 
(e) 0 to 2% minerals;
(f) 0 to 4% other dry matter and
(g) 75% to 96% (w/w) water.

In other preferred embodiments  the carrier in this above-mentioned composition comprises 0 to 2% (w/w) fat and/or 0.1 to 3% (w/w) proteins and/or 85% to 96% (w/w) water. Preferably  the lactate content is 0.2 to 2%.

The inventive food composition may consist of the Lactobacillus and the carrier. However  additives may be comprised  for example those providing a desired taste or functionality. For example  other microorganisms or compounds with activity against Helicobacter pylori could be added. Alternatively or in addition  other microorganisms or compounds  which are beneficial to the individual for other reasons  may be included  such as vitamins. The food composition may comprise typical ingredients for improving taste  flavour  stability and the like. Such ingredients may be fruits or fruit preparations flavour additives  preservatives  sweeteners and the like. In specific embodiments  the food composition (besides the carrier and Lactobacillus) might comprise less than 25%  less than 10% or less than 5% (w/w) additives  based on the total weight of the food composition. Preferably  the additives are distributed throughout the carrier.

It should be checked in an aggregation assay that the efficiency of the Lactobacillus is not decreased in such a mixture. Thus  additional ingredients should be selected not to interfere with the Lactobacillus or inhibit the Lactobacillus  affect the pH  etc.

Typically  such food compositions comprise sweeteners. Such sweeteners may be sugar or different from sugar. According to the invention  it was found that not only lactose  but also other sugars may inhibit or decrease aggregation by H. pylori. Thus  in case a sugar additive is added  it should be checked that the sugar does not inhibit or decrease aggregation of H. pylori. In a specific embodiment  the composition does not comprise a sugar additive and/or the composition comprises a sweetener  which is not a sugar. Common non-sugar sweeteners are stevia  aspartame  sucralose  neotame  acesulfame potassium and saccharin.

Another subject of the invention is the use of an inventive composition for reducing the level of H. pylori in the gastrointestinal tract of an animal. The inventive composition may be administered to animals suffering from H. pylori infections  or suspected of being suffering from H. pylori infections  or for preventing such infections. Preferably  the animal is a mammal  more preferably human. The animals may also be non-human  for example pets  such as cats or dogs  or companion animals  such as cattle  horses  pigs or sheep.

Another subject of the invention is the use of a carrier  wherein the carrier comprises milk serum and wherein the carrier is a product of a fermentation process  for enhancing the aggregation of Helicobacter pylori by Lactobacillus which is capable of aggregating Helicobacter pylori under physiological conditions. Preferably  the enhancement of aggregation by the carrier  compared to aggregation with the same amount of Lactobacillus without the carrier  is at least 5%  more preferably at least 10% or at least 50%. The enhancement can be determined by comparing the aggregation levels in aggregation tests.

Another subject of the invention is a method for producing a food composition capable of aggregating Helicobacter pylori  comprising the steps:
a) providing Lactobacillus  which is capable of aggregating Helicobacter pylori under physiological conditions 
b) providing a carrier  wherein the carrier comprises milk serum and wherein the carrier is a product of a fermentation process  and
c) mixing the Lactobacillus and the carrier to obtain the food composition.

In this production method  before  during or after step c)  further ingredients may be added. Preferably  Lactobacillus is initially cultivated in an appropriate fermentation medium. Conditions for growing Lactobacillus are known in the art and for example disclosed in WO 2007/073709. Preferably  the cells are separated from the cultivation medium by centrifugation  optionally washed and resuspended in a desired solution  for example water or a buffer. Washing and centrifugation steps may be repeated in order to avoid transfer of fermentation medium into the food composition. The capability of the cells and compositions for aggregating H. pylori may be routinely checked in aggregation assays during the production process.

Another subject of the invention is a method for reducing the level of Helicobacter pylori in the gastrointestinal tract of an animal  comprising administering to the animal an inventive composition. In other words  the method is for aggregating H. pylori in the gastrointestinal tract of an animal and/or secreting H. pylori from the gastrointestinal tract of an animal.

The inventive food composition solves the above-mentioned problems. Surprisingly  the aggregation of H. pylori by Lactobacillus strains capable thereof is boosted by a carrier  which is a fermentation product and which comprises milk serum. As shown in the working examples  the aggregation enhancement is significant and synergistic. The grade of enhancement of aggregation was unexpected in view of aggregation in the presence of Lactobacillus alone or the carrier alone.

The invention provides a simple and efficient method for treating and preventing infections associated with or caused by H. pylori colonization of the gastrointestinal tract. The inventive carrier is easily available in large amounts and at low costs. The inventive composition is thus useful for regularly large numbers of individuals at low costs. Specifically  it can be used for treatment or prevention of individuals  who do not have access  or have only limited access  to medical care. In this respect  it is advantageous that storage and consumption are not complicated and that the composition is pharmaceutically highly acceptable.

Explanation of the figures:

Figure 1 shows images of the microscopic examination of the aggregation test (negative controls with L. plantarum). Images a)  b)  c) and d) correspond to examples 3  8  4 and 9: a) L. plantarum (negative control); b) L. plantarum in yoghurt serum (negative control); c) L. plantarum (negative control  auto-aggregation test); d) L. plantarum in yoghurt serum (negative control  auto-aggregation test); e) H. pylori in AGJ with PBS (negative control).

Figure 2 shows images of the microscopic examination of the aggregation test (positive controls with DSM17648 L. reuteri). Images a)  b)  c) and d) correspond to examples 1  6  2 and 7: a) L. reuteri (positive control); b) L. reuteri in yoghurt serum (inventive composition); c) L. reuteri (positive control  auto-aggregation test); d) L. reuteri in yoghurt serum (positive control  auto-aggregation test).

Examples:

Comparative examples 1 to 5: aggregation tests and controls

The aggregation test aims at testing the ability of Lactobacillus reuteri to aggregate Helicobacter pylori under similar conditions as found in the stomach. A working cell culture (WCC) of H. pylori DSM 21031 was prepared by growing the strain on Brucella medium supplemented with 5% defibrinated horse blood (Oxoid) at 37°C under microaerophilic conditions. The horse blood was pre-treated by freezing at -20°C and then thawing to ensure the lysis of the blood cells  which release their growth factors into the medium. At the end of the growth phase the culture was mixed with sterile glycerol to achieve an end concentration of 50%. This working cell culture was left for 2 – 3 hours at room temperature before storing at -80°C. This WCC (500µl) was used to inoculate Brucella medium supplemented with 10% foetal calf serum which was incubated at 37°C for 72h. The active test culture was characterized by motile  curved to spiral rods. If the culture was characterized by the coccoid  non-motile form then this culture was discarded as it is not suitable for the aggregation test. After growth a solution of H. pylori with an optical density of 2.0 (? = 600nm) was prepared by centrifuging the culture at 4500g for 5 min. The supernatant was discarded and the cells washed in phosphate buffered saline  pH 7.0 (PBS). The pellet was carefully re-suspended in this buffer and then centrifuged again at 4500g for 5 min. This supernatant was discarded and the pellet was carefully re-suspended in artificial gastric juice (AGJ; fresh solution of 5g/L NaCl solution  pH4.0 plus 3g/L pepsin). It is important that solution of artificial gastric juice and H. pylori is prepared just prior to the aggregation test.

In addition to the H. pylori test culture fresh cultures of the L. reuteri DSM17648 (classified as L. fermentum in WO2007/073709 by carbohydrate patterns)  which is capable of aggregating with H. pylori (positive control) and Lactobacillus plantarum DSM20205  which does not aggregate with H. pylori (negative culture)  were required. Both these cultures were grown on MRS medium in closed tubes overnight at 37°C without shaking. Analogue to the harvesting of H. pylori (described above) the cells from these two cultures were centrifuged  the pellets washed in PBS  re-centrifuged and then the pellets were re-suspended in PBS instead of artificial gastric juice to achieve solutions with an optical density of 4.0 (? = 600 nm).

Experimental conditions and results of various control tests are summarized in Table 1. All experiments were carried out in the absence of the inventive carrier. Example 1 was a positive control with L. reuteri. Example 2 was a control to exclude that L. reuteri auto-aggregates. Example 3 was a negative control with L. plantarum. Example 4 was a control to exclude that L. plantarum auto-aggregates. Example 5 was a control to exclude that H. pylori auto-aggregates. The samples were mixed carefully and the results read after about 10 min at room temperature. The co-aggregation is judged both macroscopically and microscopically (400 times magnification). The respective images are shown in figures 1 and 2. In tables 1 and 2  “+” refers to normal aggregation and “++” refers to strong aggregation. The results show that aggregation is only observed when H. pylori is combined with the L. fermentum strain.

Table 1: Comparative examples 1 to 5
Ex. H.pylori. in AGJ L.reuteri. in PBS L. plant. in PBS PBS AGJ Aggreg. Fig.
1 750µl 750µl - - - + 2a
2 - 750µl - - 750µl - 2c
3 750µl - 750µl - - - 1a
4 - - 750µl - 750µl - 1c
5 750µl - - 750µl - - 1e

Examples 6 to 9: Effects of yoghurt serum addition

In addition to the controls described in examples 1 to 5  the effect of the yoghurt serum (pH 4.3) on aggregation of H. pylori by L. reuteri was tested in the following manner. L. plantarum strain (negative control) and the L. reuteri strain (positive control) were suspended in the clear yoghurt serum instead of PBS to give an optical density of 4.0 (? = 600nm). The solutions of these strains were then combined with the H. pylori solution and the co-aggregation judged microscopically (400 times magnification) after about 10 min. The conditions and results are summarized in table 2 below. When the negative control  L. plantarum  was suspended in PBS or yoghurt serum and then combined with the H. pylori solution  no aggregation was visible (Ex. 3 and 8; Fig. 1a  1b). There was also no auto-aggregation observed when the L. plantarum was suspended in PBS or yoghurt serum (Ex. 4 and 9; Fig. 1c  1d). In the case of the positive control L. reuteri  the strain suspended in PBS aggregated  as expected  when combined with the H. pylori solution (Ex. 1; Fig. 2a) and no auto-aggregation was observed (Ex. 9; Fig. 2c). When the L. reuteri was suspended in yoghurt serum instead of PBS  some auto-aggregation was observed (Ex. 7; Fig. 2d). However  when the same solution was combined with the H. pylori solution in inventive example 6  the aggregation was significantly stronger (Fig. 2b)  indicating a synergetic effect when the aggregating L. reuteri strain is combined with yoghurt serum. The synergistic effect is also evident from comparing inventive example 6 with comparative example 1 (Fig. 2a  2b). The aggregation of H. pylori in the presence of L. reuteri is significantly more pronounced in yoghurt serum than in PBS.

Table 2: Inventive example 6 and comparative examples 7 to 9
Ex. H. pylori in AGJ L. reuteri in yoghurt serum L. plantarum in yoghurt serum Aggreg. Shown inFigure
6 750µl 750µl - ++ Fig. 2b
7 - 750µl - + Fig. 2d
8 750µl - 750µl - Fig. 1b
9 - - 750µl - Fig. 1d

WE CLAIM :

1. A food composition comprising Lactobacillus  which is capable of aggregating Helicobacter pylori under physiological conditions  and a carrier  wherein the carrier comprises milk serum and wherein the carrier is a product of a fermentation process.
2. The food composition of claim 1  wherein the Lactobacillus is selected from Lactobacillus fermentum  Lactobacillus reuteri  Lactobacillus brevis and Lactobacillus pentosus.
3. The food composition of at least one of the preceding claims  wherein the Lactobacillus is selected from Lactobacillus strains deposited as No. DSM 17648  DSM 17646  DSM 17647  DSM 17649  DSM 17650  DSM 17651  DSM 17652 or DSM 17653.
4. The food composition of at least one of the preceding claims  wherein the amount of Lactobacillus in the composition is between 104 to 1015  preferably 108 to 1012  more preferably 109 to 1011 cells.
5. The food composition of at least one of the preceding claims  wherein the fermentation was a bacterial fermentation.
6. The food composition of at least one of the preceding claims  wherein the pH of the composition is between 3.5 and 5.5.
7. The food composition of at least one of the preceding claims comprising 0.1 to 5% (w/w) lactate.
8. The food composition of at least one of the preceding claims  wherein the carrier comprises yoghurt serum.
9. The food composition of at least one of the preceding claims  wherein the carrier consists of
(a) 0.1 to 10 % (w/w) proteins 
(b) 0 to 5% (w/w) fat 
(c) 0 to 12% carbohydrates 
(d) 0.1 to 3% (w/w) lactate 
(e) 0 to 2% minerals;
(f) 0 to 4% other dry matter and
(g) 75% to 96% (w/w) water.
10. A method for producing a food composition of at least one of the preceding claims  comprising the steps:
a) providing Lactobacillus  which is capable of aggregating Helicobacter pylori under physiological conditions 
b) providing a carrier  wherein the carrier comprises milk serum and wherein the carrier is a product of a fermentation process  and
c) mixing the Lactobacillus and the carrier.
11. Use of a composition of any of the preceding claims for reducing the level of Helicobacter pylori in the gastrointestinal tract of an animal.
12. Use of a carrier  wherein the carrier comprises milk serum and wherein the carrier is a product of a fermentation process  for enhancing the aggregation of Helicobacter pylori by Lactobacillus  wherein the Lactobacillus is capable of aggregating Helicobacter pylori under physiological conditions
13. A method for reducing the level of Helicobacter pylori in the gastrointestinal tract of an animal  comprising administering to the animal a composition of any of claims 1 to 9.

COMPOSITION COMPRISING LACTOBACILLUS AND A CARRIER

ABSTRACT OF THE INVENTION

Subject of the invention is a food composition comprising Lactobacillus  which is capable of aggregating Helicobacter pylori under physiological conditions  and a carrier  wherein the carrier comprises milk serum and wherein the carrier is a product of a fermentation process. Subject of the invention are also methods for producing such food compositions and uses thereof.