Using lactobacillus plantarum for providing bacterial diversity

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to industrial microbiology, namely to creating of strain Lactobacillus plantarum specified in a group consisting of Lactobacillus plantarum 299, DSM 6595, Lactobacillus plantarum 299v, DSM 9843, Lactobacillus plantarum HEAL-9, DSM 15312, Lactobacillus plantarum HEAL-19, DSM 15313, and Lactobacillus plantarum HEAL-99, DSM 15316, for increasing the gastrointestinal bacterial diversity. The above strains are applicable to prevent developing low bacterial diversity (LBD) in a healthy individual, to prevent developing translocation, large intestinal bacterial overgrowth (LIBO) or small intestinal bacterial overgrowth (SIBO) in the individual with LBD, to prevent developing translocation in the individual with LIBO or SIBO.

EFFECT: using the above strains enables providing more effective prevention or treatment of gastrointestinal infections and a possibility to enlarge the bacterial diversity.

16 cl, 1 tbl, 1 dwg, 1 ex

 

The scope to which the invention relates

The invention relates to the use of one strain of Lactobacillus plantarum in order to obtain compositions that increase the diversity of bacteria in the gastrointestinal tract, and to the use of one strain of Lactobacillus plantarum in order to obtain a composition for prophylactic treatment.

Prior art

The General opinion of experts in the field of biology is that a large variety of bacteria (diversity of microorganisms of various types) plays a favorable role in the global ecosystem and ecosystem specific areas, and ecosystems of a particular individual. A large variety of microorganisms indicates that in this ecosystem supports a favorable bacterial balance. In contrast, the imbalance of the ecosystem or its breach or complete destruction lead to "excessive development of several microorganisms, to merge with this system, the occurrence of other disorders and to the development of new pathological conditions. This also applies to the ecosystem of the human gut.

The bacterial flora of the human gastrointestinal (LCD) tract is a complex ecosystem. Composition and activity of the bacterial flora plays an important role in maintaining the health of the person that is Bukovina the influence of such flora on the digestibility of food, development and continuous regulation of the immune system and resistance to colonization of bacteria. The gastrointestinal tract can be considered as a special tubular body, extending from the mouth to the anus. It is divided into several clearly defined anatomical areas, including the mouth, esophagus, stomach, small intestine (duodenum, skinny intestine and ileum) and large intestine (blind intestine, large intestine and rectum). The concentration of bacteria in the stomach and in the upper two thirds of the small intestine (duodenum and jejunum) are relatively low, due to the high acid environment of the stomach, a short period transfer its content, secretion of bile and pancreatic juice. These concentrations are usually in the range from 102up to 104colony forming units of bacteria (CoE) per ml of the contents of the stomach or intestines, and examples of typical bacteria living in these areas are Streptococcus and Lactobacillus. The distal part of the small intestine (the ileum) usually has a concentration of 107-108kOe per ml, and this part is usually dominated by the same types of bacteria that are present in the colon, there are different classes bacteria Firmicutes, Bacteriodetes, Fusobacteria, Verrucomicrobia, Proteobacteria and Bifidbacterium (WANG M., AHRNÉ s, JEPPSSON B. & MOLIN G. (2005). Comparison of bacterial diversity along the human intestinal tract by direct cloning and sequencing of 16S rRNA genes. FEMS Microbial Ecology 54:219-231). The highest concentration of bacteria found in the colon, due to a longer period transfer its contents (up to 60 hours). It was found that in the solid faeces bacterial biomass is 40-55%, and the concentration of live bacteria is usually about 1010-1011kOe per gram of intestinal content. In the contralateral, normal and healthy colon bacterial diversity reaches its peak. However, because of the extremely high concentrations of bacteria in the colon is the part of the gastrointestinal tract that is most prone to bacterial translocation, where live bacteria or toxic components of the bacteria pass through the mucous membrane in esentially lymph nodes and in other areas outside of the intestine, such as the spleen, liver, kidney, peritoneal cavity and the blood flow. The low diversity of bacteria (LBD) increases the risk of excessive bacteria in the colon (LIBO) and in the small intestine (SIBO), which can lead to their translocation.

Experts in the field to which the present invention was demonstrated that the gastrointestinal tract in patients with Crohn's disease n the observed low diversity of bacteria [Manichanh C, Rigottier-Gois L., Bonnaud e, Gloux K, Pelletier e, Frangeul L., Nalin R, Jarrin C, Chardon p, Marteau P., Roca J., and J. Doré (2006) Reduced diversity of faecal microbiota in Crohn''s disease revealed by a metagenomic approach. Gut 55:205-211], [Ott S.J. Musfeldt, M., Wenderoth D.F., Hampe J., O. Brant, U.R. Fölsch, Timmis K.N., and Schreiber S. (2004) Reduction in diversity of the colonic mucosa associated bacterial microflora in patients with active inflammatory disease. Gut 53:685-693].

It was also demonstrated that newborns with a small diversity of bacteria in the gastrointestinal tract increases the risk of developing allergies [Wang M., Karlsson C., Olsson C., Adlerberth I, Wold A., Strachan D.P., Martricardi P.M., Aberg N., Perkin M.R., Tripodi, S., Hesselmar b, Saalman R, Molin G. & Ahrné S. (2008). Reduced diversity in the early fecal microbiota of infants developing atopic eczema: Low diversity in early microbiota of infants developing atopy. Journal of Allergy and Clinical Immunology 121:129-134].

In addition, it was demonstrated that in female rats suffering from "excessive reproduction" (low diversity) of microorganisms in the gastrointestinal tract, babies are born with high levels of haptoglobin and immature intestine [FAK F., Ahrné s, Molin g, Jeppsson B. & Weström, B. (2008). Microbial manipulation of the rat dam changes bacterial colonization and alters properties of the gut in her offspring. American Journal of discrimination - Gastrointestinal and Liver discrimination 294:148-154].

Thus, it is obvious that there is a correlation between low diversity of bacteria in the gastrointestinal tract and the development of several physiological disorders in humans and other mammals.

It is known that antibiotic use red eye reduction is that the diversity of bacteria in the gastrointestinal tract. Because antibiotics are used almost all over the world, specialists in this field are faced with the necessity of finding new ways to solve the problems associated with low diversity of bacteria in the gastrointestinal tract, such as a low diversity of bacteria has a negative impact on the overall health of people.

In addition, generally known that the lifestyles of people in developed countries contributes to the development of many pathological disorders and conditions such as cardiovascular diseases, caused by, for example, stress and overweight. It is known that people with such disorders and conditions are often observed low diversity of bacteria in the gastrointestinal tract.

Thus, specialists in this field it is necessary to solve the problem associated with physiological disturbances associated with low diversity of bacteria in the intestinal tract of individuals or caused such low diversity.

In WO 01/11077 A2 describes methods of diagnosis or treatment of irritable bowel syndrome and other disorders caused by excess production of bacteria in the small intestine (SIBO) by introducing an antimicrobial or probiotic agents, such as bifidobacteria (Bifidobacterium) or lactic acid bacteria (Lactobacillus), or by normalizing Cirrus is Tiki intestine using prokinetics tools.

Description of the invention

The above problems can be solved in accordance with the present invention.

In one of its aspects the present invention relates to the use of one strain of Lactobacillus plantarum in order to obtain a composition for enhancing bacterial diversity in the gastrointestinal tract by introducing a specified strain the individual to achieve his higher difference of the index of diversity of bacteria in comparison with the individual, which was introduced placebo.

In another aspect the present invention relates to the use of one strain of Lactobacillus plantarum in order to obtain a composition for carrying out preventive measures in healthy individuals to prevent the development of low bacterial diversity (LBD); individuals with LBD to prevent the development of his physiological disorders, excessively rapid growth of bacteria in the colon (LIBO) or overly intensive growth of bacteria in the small intestine (SIBO) and the individual with LIBO or SIBO to prevent the development of physiological disorders.

The present invention also relates to a method of increasing bacterial diversity in the gastrointestinal tract of the individual by introducing him one strain of Lactobacillus plantarum in order to achieve this the individual the mind, the higher the difference of indices of diversity of bacteria in comparison with the individual, which was introduced placebo.

The present invention also relates to a method of treating one or many physiological disorders due to low bacterial diversity (LBD) in the gastrointestinal (LCD) tract of the individual, induced, but not necessarily, the excessive growth of bacteria in the small intestine (SIBO) and/or in the colon (LIBO), where the method includes increasing and/or increase diversity of bacteria and preventing excessive growth of bacteria in the small intestine (SIBO) and in the colon (LIBO) of that individual.

In addition, the present invention also relates to the application of Lactobacillus plantarum in order to increase the difference of the indices of diversity and/or to the use of this strain for preventive treatment.

Brief description of the graphical material

The drawing shows the profile T-RFLP (restriction polymorphism of the lengths of terminal restriction fragments) Haelll-hydrolyzed genes 16S rRNA amplified from the mucosa of man.

Detailed description of the invention

The authors of the present invention, it was unexpectedly found that the introduction of a single strain of Lactobacillus plantarum leads to an increase in the diversity of bacteria in the gastrointestinal tract, i.e. the total number of bacteria of various types in the gastrointestinal tract ascending the melt after the introduction of only a single strain. Thus, in this case, there is not only increasing the number of input strain, but also the increase in the number of bacteria of other types. In addition, the introduction and colonization of the specified one strain of Lactobacillus plantarum gives rise to new groups of bacteria that were previously unable to grow in the gastrointestinal tract of the individual. This contradicts previously assume that only a mixture of different bacterial strains can cause the formation of a mixture of bacterial strains in the gastrointestinal tract, that is, to increase the diversity of bacteria. Thus, experts often doubt it considering in their assumptions that the introduction of only a single strain will lead to the reduction of biological diversity. In other words, the main argument in favor of obtaining a mixture of bacteria of various types is that such mixtures will satisfy the requirements of diversity.

Therefore, for the authors of the present invention was a complete surprise that the introduction of only a single strain of Lactobacillus plantarum leads to an increase in the diversity of bacteria.

As mentioned above, individuals suffering from one or more of the various physiological disorders, defined below, are often observed simultaneously LBD (low is e bacterial diversity), that is the cause of the subsequent occurrence of physiological disorders or which is specified physiological disorders. Itself LBD is not considered as a normal pathological condition, but over time it can lead to physiological disorders, including various diseases or LIBO (an overgrowth of bacteria in the colon), or SIBO (an overgrowth of bacteria in the small intestine). Although LIBO and SIBO do not necessarily result in LBD, however, in most cases, these effects are still observed. In the gastrointestinal tract of an individual with LIBO or SIBO occurs automatically LBD. Themselves LIBO and SIBO is also not considered to be pathological condition, but they all represent a significant risk for several disorders, including various diseases, defined below, such as the violation of translocation. Alternatively, LIBO and SIBO may be due to various physiological disorders. The composition according to the invention can be introduced healthy individual in order to prevent the development of LBD. In addition, the composition may also be administered to the individual with LBD, regardless of origin LBD, in order to prevent the development of one or more physiological disorders, SIBO or LIBO. Moreover, this composition may also be imposed is an individual with LIBO or SIBO, regardless of their origin, in order to prevent the development of one or more physiological disorders, such as impaired translocation of intestinal contents. In another embodiment of the invention the individuals with LDB and/or SIBO and/or LIBO and at the same time suffering from one or more of the above physiological disorders, may be entered in the specified composition in order to increase the diversity of bacteria in the gastrointestinal tract and thereby improve the overall health of the individual, as well as, but not necessarily, reduce the severity of these one or more physiological disorders.

The increased diversity of bacteria observed in accordance with the present invention, can be measured as an index of diversity, a certain well-known T-RFLP method, which uses restrictive enzyme Haelll, can be calculated indices of Shannon-Wiener and Simpson, described below. This increase is the difference between the indexes of diversity in that case, if you use the index of Shannon-Wiener, is at least about 0.15, preferably between 0.30, more preferably, equal to 0.45, and more preferably, 0,60.

Indicated an increased difference between the indexes of diversity in that case, if you are using the Simpson index is at least 0.02 and preferably of 0.04,more preferably, 0,06, and even more preferably 0.08 to.

The dierence of the indices of diversity (Didiff_total) carry out, in the first stage, by determining the difference between the indexes of diversity (Didiff) individuals, which was introduced a product containing one strain of Lactobacillus plantarum, i.e. determine the values of diversity index after injection (Diafter_product) strain and subtracting from it the value of the diversity index to product introduction (Dibefore_product), and then dividing the result by the value (nproduct), which represents the number of individuals who have been introduced this product. Then, from the values obtained in the first stage, subtract the difference of the indices of diversity for placebo (Diplacebo_total=Diafter_placebo-Dibefore_placebo)divided by the value (nplacebo), which represents the number of individuals who have been introduced to placebo. The equation for calculating the difference of the index of diversity is set out below:

Didiff_total=Σ(Diafter_product-Dibefore_product)/(nproduct)-

Σ(Diafter_placebo-Dibefore_placebo)/(nplacebo).

Of course, that the diversity index can be measured by other methods known in the art. In addition, restrictive enzyme Haelll may be replaced by any other known enzyme.

In accordance with the present invention General is the state of people's health can be improved by increasing the index of diversity of bacteria in their gastrointestinal tract. As discussed above, many physiological disorders associated with low diversity index. Because one strain of Lactobacillus plantarum is easily absorbed in the form of a solid or liquid preparation, a food product, such as product, discussed below, can help improve the health of individuals.

In addition, described herein is an increased diversity index can be used in order to eliminate the negative effects that occur in modern society and in wealthy countries. So, for example, as discussed above, a huge number of antibiotics that take people around the world, upset the balance of substances in the gastrointestinal tract of these people. In accordance with the present invention such balance may be normal, and the person's state of health will improve as a result of introduction of one strain of Lactobacillus plantarum.

In addition, it is clear that many physiological disorders, which affects people in modern society, can be prevented as a result of continuous infusion of one strain of Lactobacillus plantarum in accordance with the present invention.

In one embodiment of the invention the specified composition is a liquid composition or a solid composition, where the specified solid composition selected from the group consisting of tablets, pills on the I-sucking sweets, chewing tablets, chewing gums, capsules, sachets, powders, granules, particles coated and coated tablets, pills and capsules with intersolubility coating and consumable strips and films, as specified liquid composition selected from the group consisting of oral fluids, suspensions, emulsions and syrups. The composition according to the invention may be introduced by any suitable methods. However, this strain is preferably administered orally.

In one embodiment of the invention, this composition contains a carrier material, where the specified carrier material independently selected from the group consisting of liquid porridge made from oat flour, food products obtained by the fermentation of lactic acid, resistant starch, dietary fiber, carbohydrates, proteins and glycated proteins.

In one embodiment of the invention, this composition is a dietary food product, functional food, dietary Supplement, nutritional product or food preparation. For example, Lactobacillus plantarum can be administered to the individual in many different forms. Specified food product may be selected from the group consisting of beverages, yogurts, juices, ice cream, bread, biscuits, products from cereals, environmentally friendly product is and pasty products. Thus, it is clear that this composition can easily be introduced in the form of food for daily consumption. For example, the General health status may improve with the use of the composition according to the invention.

Lactobacillus plantarum is present in the composition in amounts of about from 1×106up to 1×1014CoE (colony forming units), preferably from about 1×108up to 1×1012CoE, and more preferably, from about 1×109up to 1×1011CoE

The expression "increased the difference between the indices of diversity among individuals compared with individuals who were introduced placebo"used in the description of the present application, means that the change in bacterial diversity in individuals, which were introduced strains of Lactobacillus plantarum was comparable with changes in other individuals, in which the research process has introduced a similar product that does not contain Lactobacillus plantarum. Tests of this type are called test "blind" method, i.e. during the study, neither the volunteers nor the personnel performing the test and analyzing the results, I don't know about what the individual received placebo and which was given medicinal product. This eliminates many sources of error and false results.

The expression "esbatech the initial growth of bacteria in the colon (LIBO)", used in the description of the present application, means that the bacteria of one or more types largely dominate the bacterial flora of the large intestine, i.e. they are present in much larger quantities than most other bacteria types.

The expression "an overgrowth of bacteria in the small intestine (SIBO)"used in the description of the present application, means that the bacteria of one or more types largely dominate the bacterial flora of the small intestine, i.e. they are present in much larger quantities than most other bacteria types.

The expression "low bacterial diversity (LBD)"used in the description of this application, means a bacterial imbalance in the digestive tract of the individual, which can be determined from the difference of the indices of diversity, part at least of 0.15, in accordance with the diversity index of Shannon-Wiener, or at least 0,02, in accordance with the diversity index Simpson. It should be noted that in the description of the present application, the term "diversity" sometimes means "bacterial diversity".

The expression "physiological disorders", as used in the description of this application, means any poor health or condition, associiruemyi is consistent with the presence of LBD and/or LIBO or SIBO or caused by their presence, such as gastrointestinal disorders, such as impaired translocation, Crohn's disease, ulcerative colitis, irritable bowel syndrome and pathological conditions caused by use of antibiotics. In addition, LBD, SIBO and LIBO can be negative factors contributing to the development of coronary heart disease, diseases of "fatty liver", not caused by alcohol, type 2 diabetes, allergies, atopic eczema, and autoimmune diseases.

The expression "to increase the diversity of bacteria in the gastrointestinal tract"as used in the description of the present application, means that the increase of the flora of microorganisms in the gastrointestinal tract is achieved in the presence of bacteria of various types. This means that the risk of excessive growth of some harmful bacteria is reduced, resulting in improved overall health of individuals. For example, in accordance with the present invention, the excessive growth of some bacteria in the gastrointestinal tract is reduced, and therefore achieve the desired balance.

The diversity index measure T-RFLP method using restricteduser enzyme Haelll, where compute diversity indices of Shannon-Wiener and Simpson.

As mentioned above, the present invention also relates to the treatment of one or many physiological disorders asso is eireannach with low bacterial diversity (LBD) in the gastrointestinal (LCD) tract of the individual, induced, but not necessarily, the excessive growth of bacteria in the small intestine (SIBO) and/or in the colon (LIBO), where said treatment includes increasing and/or increase diversity of bacteria and the removal of excessive growth of bacteria in the small intestine (SIBO) and in the colon (LIBO). Treatment LBD is carried out by introduction of one strain of Lactobacillus plantarum selected from the group consisting of Lactobacillus plantarum 299, DSM 6595, Lactobacillus plantarum 299v, DSM 9843, Lactobacillus plantarum HEAL-9, DSM 15312, Lactobacillus plantarum HEAL-19, DSM in 15,313, and Lactobacillus plantarum HEAL-99, DSM 15316.

The bacterium L. plantarum belongs to a huge and relatively diverse genus Lactobacillus, which includes about 90 officially named species or subspecies. Bacteria Lactobacillus spp. traditionally divided into three functional groups based on their fermentability, namely obligate bacteria, sbrasivaya one substance (group I), facultative bacteria, sbrasivaya various substances (group II), and obligate bacteria, sbrasivaya various substances (group III). Bacteria group I fermented exclusively hexose up of lactic acid and can not brivati gluconate or pentoses, and the bacteria group II is fermented not only hexose to lactic acid, but also have the added ability to brivati pentoses and/or gluconate. Bacteria group III of fermented hexose to dairy is acid, acetic acid and/or ethanol and carbon dioxide. The bacterium L. plantarum is a facultative microorganism, spaziali different substances. The strain L. plantarum of this type was designated ATCC 14917.

L. plantarum is different from many other strains of Lactobacillus spp. the following characteristics:

1) L. plantarum has a relatively large genome, which indicates its ability to accept many different conditions.

2) L. plantarum has an amazing ability to brivati many different carbohydrates.

3) Bacteria L. plantarum, for its rapid growth requires manganese, and this bacteria can accumulate manganese in high intracellular levels. Manganese protects the bacterium L. plantarum from the toxic effects of oxygen through the restoration of oxygen radicals to H2About2. Then produced H2About2can turn into About2and water under the action of the enzyme-cofactor manganese-pseudocatalase.

4) L. plantarum has a high resistance to low pH values. The fact that the bacterium L. plantarum often spontaneously prevails in food, prepared by fermentation of lactic acid, where the pH is usually below 4.0, and is preserved when passing through the acidic environment of the human stomach, indicates a high level of resistance of this bacterium to acid is the context.

5) the Bacterium L. plantarum may have tennesee activity, as well as the ability to metabolize phenolic acids.

L. plantarum often spontaneously present in large quantities in most foods, prepared by fermentation of lactic acid, in particular food products, prepared from materials of vegetable origin, for example in the salty olives, capers (caper berries), sauerkraut, salt the West Indies pickles, sourdough, Nigerian cakes (made from maize or sorghum), in the Ethiopian coho (prepared from starch Ensete ventricosum), in the Ethiopian sourdough made from TEF (Eragrostis tef), and cassava. Thus, it is obvious that individuals who use herbal products made by fermentation of lactic acid, also consume a large number of L. plantarum. In addition, L. plantarum is present in grape juice and wine. L. plantarum is often present in the mucosa of the gastrointestinal tract of man, extending from the mouth to the rectum [Molin g, Jeppsson B., Ahrné, S., Johansson, M.-L., Nobaek, S., Stahl, M., and Bengmark, S. (1993). Numerical taxonomy of Lactobacillus spp. associated with healthy and diseased mucosa of the human intestines, J. Appl. Bacteriol. 74:314-323; Ahrné s, Nobaek s, Jeppsson b, Adlerberth I, Wold A., and Molin, G. (1998). The normal Lactobacillus flora of healthy human rectal and oral mucosa, J. Appl. Environ. 85:88-94)]. It follows that the species L. plantarum,present in the food of man and in his intestines, occupy a unique place, and for human consumption of food prepared by fermentation of lactic acid, the process of digestion from the very beginning is spontaneous, provided that these herbal products are stored under pressure in an enclosed space, for example in the cellar, and this means that, in the future, people will consume microorganisms that normally prevail in this environment and play a key role in the human intestine.

Examples

Defining diversity of bacteria

The analysis by the method of the polymorphism of the lengths of terminal restriction fragments (T-RFLP)is a common method of fingerprinting"based on the hydrolysis of fluorescently labeled at the ends of PCR products under the action of restrictive endonuclease, and this method allows to obtain data on the presence of known and unknown populations of bacteria. T-RFLP-patterns generated in a series of stages. Briefly, the DNA of this community of bacteria extracted directly from the sample. Interest genes subjected to PCR amplification using primers, one of which is fluorescently labeled. After purification of PCR products hydrolyzing restricteduse the endonuclease, usually an enzyme, comprising 4 bases. Hydrolyzed product is mixed with fluores into labeled DNA standard size, and then the fragments separated by electrophoresis using gel or capillary system, equipped with a laser detector, so were visualized fluorescently labeled terminal fragments. The result of this analysis present in two forms: 1) electropherogram illustrating the profile of the bacterial community as a series of peaks of different height, 2) table created using the automated program based on the analysis of the fragments, including the most important parameters, such as size, mating grounds, and the height (or area) of each peak. T-RFLP profiles for the compared samples can be numbered using statistical methods. For comparison of microbial communities used several statistical methods.

In recent years, T-RFLP analysis of microbial communities in environmental studies is on the rise. It is reproducible and gives high resolution.

Results

Individuals and sampling

Individuals involved in the research, were men with good physical condition, but with a certain well-controlled cardiovascular disease. These individuals were subjected to screening using flexible sigmoidoscopy before and after consumption of the test solutions in four weeks. iptat took the standard method of the mucosa of the lower sigmoid colon for the subsequent analyses.

In studies on the diversity of bacteria were included 16 volunteers, recruited from a large group of individuals included in the randomized placebo-controlled trials conducted double-blind. Nine individuals were using 100 ml of therapeutic product daily for four weeks, which corresponded to a daily consumption of 1011colony forming units of L. plantarum in the day. Seven individuals were consuming 100 ml of the same product in a day, not containing bacteria within four weeks.

Extraction of DNA

Sampling mucosa was treated in an ultrasonic bath for 5 minutes, and then intensively stirred for 2 minutes. Then the sample was transferred into a UV-treated for 1.5-ml tubes and centrifuged at 9000 rpm for 7 minutes. To the precipitate was added 380 ál buffer G2 and 30 μl of proteinase K (Qiagen, Hilden, Germany). These samples were processed in a water bath at 56°C until until they are fully dissolved. Then the suspension was subjected to disintegration by shaking together with 12-15 glass beads (diameter 2 mm) for 45 minutes at 4°C in Eppendorf mixer (model 5432, Eppendorf, Hamburg, Germany). After centrifugation at 5000 rpm for one minute and the supernatant was transferred into two different 2-ml test tubes for samples (200 ál in each tube). The stage is niteline purification was performed on the device BioRobot® EZ1 using DNA sample tissue EZ1 DNA and DNA tissue EZ1 (Qiagen, Hilden, Germany) according to manufacturer's instructions. DNA was suirable 200 ál.

PCR amplification, purification and concentration measurement

Gene 16S rRNA amplified using universal direct primer Cy5-ENV1 (5'-AGA GTT TGA TII TGG CTC AG-3'), fluorescently labeled with Cy5 at the 5'-end, and the reverse primer ENV2 (5'-CGG ITA CCT TGT TAC GAC TT-3'), which hybridized with 8-27 softwares 1511-1492 BP, respectively. PCR reaction mixture contained 0.2 μm of each primer, 0.2 mm of each deoxynucleotide (Roche Diagnostics, Indianapolis, IN), 5 μl 10 × PCR reaction buffer (100 mm Tris-HCl, 500 mm KCl, pH 8,3), and 2.5 units/μl Taq polymerase (Roche Diagnostics, Mannheim, Germany) and 0.2-10 ál of the matrix, in the final volume of 50 µl. Amplification was carried out in thermoacetica Eppendorf Mastercycler (Hamburg, Germany) using the following program: one cycle was carried out at 94°C for 3 min, then 32 cycles were performed at 94°C for 1 minute, at 50°C for 45 seconds, and 72°C for 2 minutes and an additional cycle of elongation was performed at 72°C for 7 minutes.

PCR products (5 µl) were identified on a 1.5% (wt./about.) agarose gel in 1 × TBE buffer (89 mm Tris, 89 mm boric acid, 2.5 mm EDTA) after staining ethidiumbromid.

PCR products of the three reactions were combined in order to reduce the standard error by PCR in order to obtain sufficient DNA for T-RFLP analysis. Amplic the us was purified and concentrated using a set of PCR-purification MinElute (Qiagen, Hilden, Germany) according to the manufacturers Protocol. Elution was performed 30 ál of sterile distilled water.

The concentration of purified DNA was measured by spectrophotometer FlouroMax-2 using software DataMax for WindowsTM(ISA Jobin Yvon - Spex Instruments S.A., Inc., New Jersey) using a reagent Quant-iTTMPicoGreen® (Invitrogen, Eugen, Oregon, USA), which is built of double-stranded DNA. Reagent Quant-iTTMPicoGreen® used in accordance with the manufacturer's instructions. Excitation was performed at a wavelength of 480 nm.

T-RFLP-analysis

Aliquots of 200 ng of purified PCR products separately hydrolyzed for 16 hours at 37°C with 15% restrictive endonuclease Haelll (Sigma-Aldrich, St Louis, USA) in a total volume of 10 µl. After hydrolysis enzymes iactiveaware by heating at 65°C for 15 minutes. The hydrolysates were mixed with 1 µl of the internal standard sizes and 4 ál formamide boot dye (3,3 μl of deionized formamide, and 0.7 μl of 25 mm EDTA and 5% wt./about. dextranomer blue) and the mixture was denaturiruet at 94°C for 3 minutes, and then before loading on the polyacrylamide gel was immediately placed on ice.

Internal size standards contained primer Cy5-ENV1 (20 BP, as described above) and 697 BP PCR product, amplificatory from ATCC 11775 E. coli using primer 685r (5'-TCT ACG CAT TTC ACC GCT AC-3'; E. coli, the provisions 705-685) and Cy5ENV1. External standards of size, consisting of a classifier sizes ALFexpress 50-500 (GE Healthcare, Uppsala, Sweden) and Cy5-labeled 697 BP PCR product were loaded on polyacrylamide gels, containing the sample, to estimate the lengths of the T-RF (terminal restriction fragments). Fluorescently labeled fragments were separated and detected using a DNA sequencing machine ALFexpress Il 7% gel ReproGel Long Read (GE Healthcare, Uppsala, Sweden) for 700 minutes under the following conditions: 1500 V, 60 mA and 55°C.

Statistical analysis

The peak areas of fluorescently labeled T-RF was estimated using the program for analysis of fragments ALFwinTMFragment Analyser 1.03 (Amersham Biosciences, Uppsala, Sweden). The relative amount of each T-RF in this T-RFLP-pattern was calculated as the peak area of the corresponding T-RF, divided by the total peak area of all T-RF, rectified by the length of the fragment between 20 and 697 BP Indexes Simpson (D) and Shannon-Wiener (H') was calculated according to the formula: D=TPMi2and H'=-Σ pi, ln piwhere pirepresents the relative amount of the i-th peak in this bacterial community (Magurran, A., 1996, Ecological diversity and its measurement, Chapman and Hall, London). The use of the diversity index Simpson (1-D) instead of the original formula of Simpson index ensures that the value of this index will increase with increasing diversity of bacteria. For each of the individual indices calc is for samples before and after treatment. The difference of the index of diversity was calculated by subtracting the value of the index before processing of the index values after processing. The difference between the indices of bacterial diversity in individuals probiotic group and placebo group was calculated using the criterion of the sum of ranks Mann-Whitney (SigmaStat, Systat Software, Point Richmond, USA). The value of p<0,05 was considered statistically significant.

When analyzing data for T-RFLP profiles was found that a statistically significant difference of the index of diversity of the microbiota in the gut of individuals treated with strain L. plantarum 299v in the past four weeks, was comparable to the difference between the indices of the individuals who received placebo. Restricteduse endonuclease Haelll used as restricteduser enzyme, and the average difference between the diversity indices measured for the Shannon index, was 0,2305803 for the probiotic group and -0,3929243 for the placebo group (p=0,026).

The calculation of the diversity index Simpson confirmed a higher diversity of bacteria after administration of L. plantarum 299v. The enzyme Haelll used as restricteduser enzyme, and the result was statistically significant, i.e. the average difference for index of diversity Simpson was 0,0367907 for the probiotic group and -0,04792 for the placebo group (p=0,026).

The average difference of the index the s diversity

Index ShannonIndex Simpson
Dibefore 299v-Diafter 299v0,23058030,0367907
Dibefore placebo-Diafter placebo-0,3929243-0,04792
Ditotal0,62350460,0847107
The importance of DitotalP=0.026P=0.026

The above results showed that the bacterial diversity in the human intestine increased with the introduction of one strain of Lactobacillus plantarum, i.e. individuals, which was introduced strain Lactobacillus plantarum, bacterial diversity increased as compared with individuals who were introduced placebo.

1. The use of one strain of Lactobacillus plantarum selected from the group consisting of Lactobacillus plantarum 299, DSM 6595, Lactobacillus plantarum 299v, DSM 9843, Lactobacillus plantarum HEAL-9, DSM 15312, Lactobacillus plantarum HEAL-19, DSM in 15,313, and Lactobacillus plantarum HEAL-99, DSM 15316, to increase diversity of bacteria in the gastrointestinal tract by introducing a specified strain the individual to achieve his higher once the barb indexes diversity of bacteria in comparison with the individual, which was introduced placebo, where indicated increased difference between the diversity indices measure the T-RFLP method using Haelll as enzyme and diversity index of Shannon-Wiener index and Simpson.

2. The use according to claim 1, where this increased difference between the indexes of diversity in that case, if you use the index of Shannon-Wiener, is at least about 0.15, preferably between 0.30, more preferably, equal to 0.45, and most preferably, 0,60.

3. The use according to claim 1, where this increased difference between the indexes of diversity in that case, if you are using the Simpson index is at least 0.02 and preferably of 0.04, more preferably 0.06 to, and most preferably 0.08 to.

4. The use according to any one of claims 1 to 3, where the specified strain is administered in the form of liquid compositions or solid composition.

5. The use according to claim 4, where this solid composition is selected from the group consisting of tablets, tablets for sucking, sweets, chewing tablets, chewing gums, capsules, sachets, powders, granules, particles and coated tablets, pills and capsules with intersolubility coating and raspravlyalsya strips and films.

6. The use according to claim 4 where a specified liquid composition selected from the group consisting of oral fluids, suspensions, emulsions and syrups.

7. The application of l is the Boma one of claims 1 to 3, where the specified strain is administered in the form of medicinal food, functional food, dietary Supplement, nutritional product or food preparation.

8. The use of one strain of Lactobacillus plantarum selected from the group consisting of Lactobacillus plantarum 299, DSM 6595, Lactobacillus plantarum 299v, DSM 9843, Lactobacillus plantarum HEAL-9, DSM 15312, Lactobacillus plantarum HEAL-19, DSM in 15,313, and Lactobacillus plantarum HEAL-99, DSM 15316, for prophylaxis in healthy individual development low bacterial diversity (LBD).

9. The use of one strain of Lactobacillus plantarum selected from the group consisting of Lactobacillus plantarum 299, DSM 6595, Lactobacillus plantarum 299v, DSM 9843, Lactobacillus plantarum HEAL-9, DSM 15312, Lactobacillus plantarum HEAL-19, DSM in 15,313, and Lactobacillus plantarum HEAL-99, DSM 15316, for prevention in individuals with LBD development translocation, excessive growth of bacteria in the colon (LIBO) or in the small intestine (SIBO).

10. The use of one strain of Lactobacillus plantarum selected from the group consisting of Lactobacillus plantarum 299, DSM 6595, Lactobacillus plantarum 299v, DSM 9843, Lactobacillus plantarum HEAL-9, DSM 15312, Lactobacillus plantarum HEAL-19, DSM in 15,313, and Lactobacillus plantarum HEAL-99, DSM 15316, to prevent the individual from LIBO or SIBO development translocation.

11. Use p-10 to eliminate low bacterial diversity (LBD) in the gastrointestinal (LCD) tract of the individual, not necessarily induced by excessive growth of bacteria in the small intestine (SIBO) and/or in the colon (LIBO), DG is the specified application includes increasing and/or an increase in bacterial diversity and the elimination of excessive growth of bacteria in the small intestine (SIBO) and in the colon (LIBO).

12. Use PP-10, where the specified strain is administered in the form of liquid compositions or solid composition.

13. The application indicated in paragraph 12, where this solid composition is selected from the group consisting of tablets, tablets for sucking, sweets, chewing tablets, chewing gums, capsules, sachets, powders, granules, particles and coated tablets, pills and capsules with intersolubility coating and raspravlyalsya strips and films.

14. The application indicated in paragraph 12, where this liquid composition is selected from the group consisting of oral fluids, suspensions, emulsions and syrups.

15. The use according to any one of p-10, 13, 14, where the specified strain is administered in the form of medicinal food, functional food, dietary Supplement, nutritional product or food preparation.

16. The application indicated in paragraph 15, where the specified food preparation selected from the group consisting of beverages, yogurts, juices, ice cream, bread, biscuits, products from cereals, organic products and pastry products.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to a preparation for babies for reduction or prevention of inflammation in a baby. The preparation for babies includes a source of protein, providing from 1 to 5 g of protein per 100 kkal of the preparation, a source of fat or lipids, providing from 3 to 7 g of fat or lipids per 100 kkal of the preparation, a source of carbohydrates, providing from 8 to 12 g of carbohydrates per 100 kkal of the preparation, a source of long-chain polyunsaturated fatty acids, including docosahexaenoic acid. The preparation also includes from 1×104 CFU to 1×1010 CFU of Bifidobacterium longum AH1206 NCIMB 41382 per a gram of the preparation. Also claimed are probiotic baby food and a method of reduction or prevention of inflammation in the baby or child with application of the said food.

EFFECT: invention ensures induction of anti-inflammatory response, makes it possible to reduce secretion of anti-inflammatory cytokines.

14 cl, 19 dwg, 2 tbl, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to biotechnology and represents a method of producing a bacterial ghost preparation, a pharmaceutical composition, which is a vaccine or an adjuvant, a method of inactivating live bacterial cells in the bacterial ghost preparation and application of beta-propiolactone. The method includes obtaining the bacterial ghost preparation and processing the bacterial ghost preparation with beta-propiolactone in a final concentration from 0.01% to 1% (vol/vol), at which the quantity of live bacterial cells in the said ghost preparation reduces by at least 103-104. The pharmaceutical composition includes efficient quantity of the bacterial ghost preparation, processed with beta-propiolactone in a final concentration from 0.01% to 1% (vol/vol), and a pharmaceutically acceptable carrier, a diluent and/or an adjuvant.

EFFECT: claimed invention makes it possible to obtain the bacterial ghost preparation, which in fact does not contain live bacterial cells.

18 cl, 13 dwg, 7 ex

FIELD: medicine.

SUBSTANCE: 1 glass of an aqueous infusion of swallowwort herb or garden sage herb is orally administered daily on an empty stomach in the morning 1-2 hours before meals for 10 days. That is followed by administering an aqueous infusion of camomile blossom daily on an empty stomach for 10 days also. For the further 10 days, an aqueous infusion of plantain leaves is administered daily on an empty stomach; the infusions of chamomile blossom and plantain leaves are taken in a dose of 1/2 glass 3-5 times a day 30 minutes before meals. Administering the aqueous infusions is combined with orally daily administering an infusion of Saint-John's-wort for 30 days orally in a dose of 50 drops per 1/2 glass of water 3 times a day 30 minutes before meals. After that, the liquid biocomplex Normoflorin is orally administered 3 times a day daily for 4 weeks 30 minutes before meals in age doses. Normoflorin L is used for the first and second administration, and Normoflorin B - for the third one.

EFFECT: effective treatment ensured by Helicobacter pylori growth inhibition, antiseptic, anti-inflammatory herb action improving gastric mucosa trophism in a combination with the positive effect on the nervous system.

1 ex

FIELD: chemistry.

SUBSTANCE: group of inventions relates to field of biotechnology. Strain Bifidobacterium breve MCC 1274 FERM BP-11175 demonstrates low coefficient of conversion of linoleic acid into conjugated linoleic acid. Strain demonstrates coefficient of conversion of linoleic acid into conjugated linoleic acid not higher than 10%. To reduce or prevent obesity or to improve tolerance to glucose said strain is introduced in efficient quantity to subject requiring it. Also claimed are food product and drink, which contain preparation based on said strain.

EFFECT: strain is used to reduce or prevent obesity or improve tolerance to glucose as component of pharmaceutical composition or preparation.

22 cl, 4 dwg, 2 tbl, 7 ex

FIELD: biotechnologies.

SUBSTANCE: method provides for cultivation at 37±1°C of strains of lactobacilli and bifidobacteria in the medium and packing of liquid product with account of daily dose necessary for patients. The medium contains components in the following quantities: caseine hydrolysate dissolved by distilled water, 0.33-0.4 g/l, sodium chloride 5 g/l, fructose 10 g/l, peptone 2 g/l, agar-agar 0.75 g/l for lactobacilli, for bifidobacteria - 1.0 g/l, ascorbic acid 0.25 g/l, distilled water 0.67-0.6 g/l. Strains-producers are Lactobacillus plantarum 8 RA-3, Lactobacillus fermentum 39, Lactobacillus fermentum 90 TC-4, Bifidobacterium bifidum 791, Bifidobacterium longum 379, Bifidobacterium bifidum 1. For lactobacilli and bifidobacteria they prepare accordingly media with different content of agar-agar, amine nitrogen in caseine hydrolysate 160-170 and 180-200 mg% and medium pH 7.8-8.0 and 8.5-8.6. Starting from the first generation the bifidobacteria and lactobacilli are cultivated for 24±1 hours, two strains together, one separately. The produced biomass is mixed with the fresh nutrient medium at the ratio of 1:1000, and strains of lactobacilli are cultivated 24±1 hours, strains of bifidobacteria - 48±1 hours. Upon completion of cultivation of biomass of strains they are mixed at the ratio of 2:1:2:1.

EFFECT: production of a product with high content of live microbial cells.

3 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to application of probiotic bacterial strain for production of a probiotic composition for reducing sleep disorders and/or improvement of sleep quality in people and animals. As a bacterial strain used is Lactobacillus reuteri DSM 17938 or Bifidobacterium longum NCC 3001 (ATCC BAA-999).

EFFECT: invention ensures normalisation of sleep pattern in an individual by reduction of active sleep time, increase of slow sleep time and reduction of a number of awakening incidents.

14 cl, 2 dwg, 3 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to versions of medication for reduction of halitosis, versions of compositions based on said medications and versions of application of said medications. Device for reduction of halitosis represents strains of microorganisms Lactobacillus acidophilus, selected from group Lactobacillus acidophilus DSM 19825, Lactobacillus acidophilus DSM 19826 and Lactobacillus acidophilus DSM 19827. Version of said medication represents culture supernatant of said strains. Also claimed are compositions, which contain said microorganisms, and their application for reduction of halitosis.

EFFECT: invention makes it possible to considerably reduce concentration of peptides in saliva, depleting in this way substrate, used by anaerobic microorganisms of oral cavity microflora, which are halitosis-causing agents.

10 cl, 7 dwg, 19 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to paediatrics and neonatology, and can be used for treating small premature infants at the hospital stage of developmental care. A therapeutic complex comprises administering a probiotic preparation into the newborns. The preparation is presented with a liquid probiotic containing E.faecium L3 109 CFU in 1 ml. If the enteral nutrition volume of the newborn is 5 ml or more, this preparation is orally administered in a dose of 0.5 ml 3 times a day for 14 days.

EFFECT: method is effective in children with a very low body weight, promotes normalising the intestinal microflora and reducing a rate of manifestations of infectious complications.

2 ex, 3 dwg, 3 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine and biotechnology, and represents a method for preparing a combined antibacterial preparation for treating acute intestinal infections. The invention is to prepare a biological ingredient biomass representing a complex of immunoglobulins or bifidus bacteria biomass, to mix it with an antibiotic substance in specified proportions, and differs from the known analogues by the fact that mixing the two ingredients of the preparation is preceded by grinding the biological ingredient only to be ground to the greatest maximum bulk density of the ground material.

EFFECT: invention improves the antibacterial activity of the combined preparation, which leads to reducing the length of treating an acute intestinal infection.

2 cl, 1 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: group of inventions relates to biotechnology and medicine. Disclosed is a polysaccharide which is isolated from the Bifidobacterium infantis NCIMB 41003 strain and has the structure [-β(1,3)-D-GalpNAc-β(1,4)-D-Glcp-]n, where said disaccharide unit repeats n times, which yields a polysaccharide with molecular weight greater than 100000 Da. The polysaccharide exhibits immunomodulating activity and is used in preparing medicinal agents for treating or preventing undesirable inflammatory activity, undesirable gastrointestinal inflammatory activity, rheumatoid arthritis and autoimmune disorders.

EFFECT: pharmaceutical composition for treating and preventing inflammatory disorders and a food product containing the isolated polysaccharide are disclosed.

9 cl, 6 dwg, 3 ex

FIELD: biotechnology, microbiology, medicine.

SUBSTANCE: invention relates to the strain Lactobacillus paracasei CNCM I-2116 used for diarrhea prophylaxis causing by pathogenic microorganisms. Supernatant of this strain culture elicits ability to prevent colonization of intestine with pathogenic microorganisms causing diarrhea also and this strain is designated for preparing agent used for prophylaxis and/or treatment of disorders associated with diarrhea. Agent for oral administration represents therapeutically effective dose of the strain L. paracasei CNCM I-2116 or supernatant of its culture and acceptable foodstuff. Invention provides the enhanced viability of the strain in its applying and effectiveness in prophylaxis of adhesion to intestine cells and invasion to intestine cells of pathogenic microorganisms causing diarrhea.

EFFECT: valuable medicinal properties of strain.

5 cl, 8 dwg, 10 ex

FIELD: biotechnology, microbiology, veterinary science.

SUBSTANCE: for preparing a preparation cells of microorganism Escherichia coli VKM CR-322D is cultured in nutrient medium containing Hottinger's broth, glucose, yeast extract, manganese sulfate, potassium hydrophosphate, sodium chloride and tap water in the content of amine nitrogen 125-155 mg%. Glucose is added by batch portions in the process of culturing cells that is carried out at temperature 30-31oC at stirring and aeration for 10-12 h. Prepared cultural fluid containing 3 x 109 bacterial cells/ml is mixed with protective sucrose-gelatin medium and subjected for lyophilic drying. Dried mass is stored under nitrogen that enhances safety of viable cells in the preparation. Applying the preparation for prophylaxis and treatment of agricultural and domestic animals and poultries with gastroenteric diseases provides its effectiveness.

EFFECT: improved preparing method, valuable veterinary properties of preparation.

17 cl, 8 ex

FIELD: biotechnology, food and medicinal industry, microbiology.

SUBSTANCE: the strain Bifidobacterium longum 379-IN is obtained by selection without using methods of genetic modification of the strain Bifidobacterium longum B379M and distinct by ability to utilize insulin. The strain is deposited in GKNM GU "MNIIEM named for G. N. Gabrichevskiy Russia Ministry of Public Health" at № 172. The strain shows high technological effectiveness, accumulates biomass with substrates of vegetable origin and artificial nutrient media for short periods with concentration of bifidobacteria, it elicits acid-forming and antagonistic properties with respect to pathogenic and putrid microflora. This allows its using in manufacturing bacterial preparations, biologically active supplements for food, fermented-dairy and nonfermented-dairy foodstuffs, ferments, hygienic and cosmetic agents providing probiotic effect and normalization of microbiocenosis in human body, among them in gastroenteric and urogenital tracts, cutaneous and mucosa integuments. Invention can be used in manufacturing bacterial preparations, biologically active supplements for food, fermented-dairy and nonfermented-dairy foodstuffs, hygienic and cosmetic agents.

EFFECT: valuable properties of strain, expanded assortment of similar agents.

6 ex

FIELD: biotechnology, medicine, infectious diseases, medicinal microbiology.

SUBSTANCE: invention relates to a composition designated for treatment and prophylaxis of infections caused by Neisseria microorganism that comprises the following components: (a) protein with amino acid sequence similar by 65% and above with the natural Neisseria protein of a single species (the first group of amino acid sequences is given in the text) and/or its fragment consisting of 10 and more amino acids and eliciting antigen properties; (b) the second protein with amino acid sequence similar by 65% and above with the natural Neisseria protein of another species (the second group of amino acid sequences with even numbers is given in the text), and/or its fragment consisting of 10 or more amino acids and eliciting antigen properties; in particular, the second protein represents NspA. The composition comprises additionally adjuvant. The composition is used both a medicinal agent and for manufacturing the medicinal agent. Applying the invention provides enhancing the effectiveness of prophylaxis or treatment due to the universal effect of the composition (vaccine). Invention can be used in medicine for treatment of infections.

EFFECT: valuable medicinal properties of composition.

8 cl, 137 dwg, 5 tbl, 12 ex

FIELD: biotechnology, medicinal microbiology.

SUBSTANCE: invention relates, in particular, to a method for preparing nutrient medium used for production of bacteriophages. Nutrient medium for production of bacteriophages contains acidic casein hydrolyzate as a base with hydrolysis degree 0.6-0.7 and vitamins: nicotinic acid, folic acid, calcium pantothenate, riboflavin, thiamine bromide and biotin, and distilled water. Also, invention relates to a method for preparing nutrient medium used for production of bacteriophages involving hydrolysis of natural casein mixed with water using hydrochloric acid under pressure 0.2 ± 0.05 MPa up to attainment of hydrolysis degree 0.6-0.7 being isoprecipitation and treatment with carbon are carried out simultaneously. Invention provide enhancing growth properties of medium and quality of bacteriophages produced and reducing the cost.

EFFECT: improved preparing method.

3 cl, 1 ex

FIELD: medicine, surgery, urology.

SUBSTANCE: one should apply bactisporin probiotic preparation introduced per os at 1-10 x 109 microbial cells twice or thrice for 10-20 d in combination with staphylo-proteic-pyocyanic adsorbed (SPPA) vaccine; moreover, the mentioned combination of preparation should be applied, also, for instillation of wounds, serous cavities, cavities of urinary and biliary ducts at the quantity of 1-5 x 109 microbial cells in isotonic sodium chloride solution. The present innovation provides better humoral and cellular immunity to already developed infectious process, moreover, combined application of SPPA vaccine and bactisporin leads to increased body immune response to antigens of associated vaccine at the background of secondary immunodeficient state induced by infectious process.

EFFECT: higher efficiency of therapy.

1 cl, 5 ex, 3 tbl

FIELD: biotechnology, veterinary science, microbiology.

SUBSTANCE: the strain of bacterium Bacillus subtilis VKM B-2287 is isolated from soil. Bacterial cells are gram-positive with aerobic type of respiratory, don't form capsule but form round spores. The strain hydrolyzes glucose, mannitol and lactose but it doesn't ferment sucrose, inositol, sorbitol and maltose, it doesn't form gas in fermentation process and inhibits the growth of staphylococcus, Escherichia coli, enterobacteria, citrobacteria and aeromonas. The strain is used as industrial one for preparing a preparation named by authors as "Subtilis+". The preparation normalizes function of digestive tract in agricultural animals, poultries and fishes and can be used in treatment and prophylaxis of bacterial infections. Invention can be used for preparing the probiotic preparation.

EFFECT: valuable properties of strain and preparation.

1 tbl, 3 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: the strain Lactobacillus paracasei CNCM I-2116 (Ncc 2461) is able to attach with mammal intestine mucosa and to grow in the presence of up to 0.4% of bile acid salts and to prevent the infection of intestine epithelial cells with rotaviruses. Agent shows the content from 1 x 105 to 1 x 1012 CFU of the strain L. paracasei I-2116 (NCC 2461) and can be used for preparing an agent and foodstuff taken among milk, yogurt, cheese, dry milk, children's nutrition products or a pharmaceutical preparation taken among a liquid bacterial suspension, dry oral supplement, liquid oral supplement, dry product for feeding through a tube or liquid product for feeding through a tube. Invention provides the enhanced viability of the strain in its using and effectiveness in prevention of adhesion to intestine cells and invasion of pathogenic microorganism cells in intestine causing diarrhea. Invention can be used in food industry and medicine in prophylaxis and/or treatment of diarrhea-associated diseases.

EFFECT: valuable medicinal properties of strain and agent.

3 cl, 5 dwg, 7 ex

FIELD: medicine, gastroenterology.

SUBSTANCE: invention relates to methods for treatment of chronic helicobacter pylori-associated gastritis. Method is carried out by monotherapy with the probiotic "Laminolakt" in the dose 3 dragees per 24 h for 1 month. Method provides elimination of Helicobacter pylori cells on the background of activation of the immune response in stomach mucosa by effect on microflora and the colon intestine immune system.

EFFECT: enhanced effectiveness of treatment.

2 tbl, 1 ex

FIELD: medicine, veterinary science, food industry.

SUBSTANCE: invention relates to the preparation-synbiotic eliciting the biological activity. The biologically active preparation comprises the preparation "Laktobakterin" or "Bifidum-bakterin", or "Koli-bakterin", and water-soluble chitosan succinate, and water-soluble antioxidant as special supplements taken in the definite ratio of components of the preparation. Invention provides enhancing effectiveness of preparations based on microorganism cultures and their storage time. Invention can be used as a curative preparation correcting the damaged normal microflora of digestive tract, in therapy of inflammatory processes of urogenital tract and mouth cavity, for enhancing the general resistance of body.

EFFECT: enhanced and valuable properties of preparation.

8 tbl

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