UA64159A - 1mb b-7019 bifidobacterium bifidum strain, the 1mb b-7020 bifidobacterium longum strain, the 1 mb b-7021 lactobacillus acidophilus strain and the 1mb b-7026 streptococcus thermophilus strain for production of bacterial concentrate, the bacterial concentrate "biocor-1" and the biologically active additive "biocorrectin" - Google Patents

Abstract

The 1MB B-7019 Bifidobacterium bifidum strain, the 1MB B-7020 Bifidobacterium longum strain, the 1 MB B-7021 Lactobacillus acidophilus strain and the 1MB B-7026 Streptococcus thermophilus strain for production of bacterial concentrates and food products correcting the microbial flora of human and animal digestive tract. A bacterial concentrate contains a mixture of frozen-dried cultures of Bifidobacteria and Lactobacteria strains. It contains the strains 1MB B-7019 of Bifidobacterium bifidum and 1MB B-7020 of Bifidobacterium longum as the Bifidobacteria. As the Lactobacteria, it contains the strains 1MB B-7021 of Lactobacillus acidophilus and 1MB B-7026 of Streptococcus thermophilus. A biologically active additive contains the bacterial concentrate and a carrier, which contains a fruit, citrus or vegetable powder or pineapple powder, or that of melons and gourds, or a mixture thereof, and also milk sugar and calcium stearate.

Description

Description of the invention

This group of inventions refers to medical and food biotechnology and can be used in 2 production of drugs and food products that correct the microflora of the digestive tract of people and animals, in medicine, food and dairy industry, agriculture.

The functions of normal microflora in the body are vital: synthesizing, regulatory, desensitizing, detoxifying, fermentative, protective, and immunostimulating. Recently, dysbacteriosis and secondary immunodeficiencies have been widely spread among the population. At 70, this manifests itself in severe sensitization of the body and activation of chronic bacterial and viral infections.

Numerous epidemiological studies prove that almost 9,095 people of the globe have violations of the qualitative and/or quantitative composition of the microflora in the human body. The causes of dysbacteriosis are antibiotic therapy, hormone therapy and chemotherapy, hypovitaminosis, poor and irrational nutrition, unfavorable socio-economic and environmental conditions. Therefore there was a need for active prevention and 12 effective correction of the disturbed microbiological balance. One of the most effective means of correction is probiotics. Among the probiotics, important are bifidobacteria Vydorasiegist Biact and Vydorasiegist Iopdit, as well as lactobacilli I asiorasiyz acidorpyyz and thermophilic streptococci Zigeriosossiv (Pegtorpyyy5, as well as bacterial concentrates and more complex compositions.

The well-known strain Vijdorasiegsht Biydit B-3300, used in the production of bacterial preparations, ferments milk for 18-24 hours, has antagonistic activity against 5. zoppei, Z. Lekhpegi, E. soii (220),

Z.atsgeiv, resistant to kanamycin, monomycin, sensitive to antibiotics of the penicillin series, kefzol, gentamicin, etc. (see patent BO 2023396, IPC A23C9/12, 1994).

The disadvantage of this strain is insufficiently high biological activity and manufacturability.

The known strain Vijdobrasiegisht Bidit M-4, deposited on April 5, 1993 in the Central Museum of Industrial 29 Microorganisms VNIYgenetica under number 5-1257 (patent of the Russian Federation 2108383, MPK9 сС12М1/20, "

A23С9/12, 1998). This strain was isolated from the intestinal contents of a healthy adult. Morphological and cultural signs. Cells of the M-4 strain grown on hydrolyzed milk media have the appearance of long thin granular rods located in clusters, very rarely - in the form of individual rods. The length of the cells is 3.5-5 μm, the thickness is 0.3-0.5 μm. Gram-positive, non-motile, do not form spores and capsules. In dense Ф 3о nutrient media (1.595 agar) they form paste-like colonies of light brown color in the form of "buckwheat grains" or discs up to 2.5 mm in diameter. Pigment is not formed. In liquid nutrient media, growth in the height of the nutrient medium is 90-9595. Physiological and biochemical signs. Non-pathogenic, spore-free obligate - anaerobic. Does not form catalase. The optimal temperature for growth is 36.5-38.0"C, the maximum temperature of the plant is 4120.57C, the minimum is 25-0.5"C; does not multiply at 46.57C. Optimal initial pH

From nutrient sterilized media 6.8-7.0 pH units. Gelatin does not dilute, does not reduce nitrates, does not form indole. It ferments lactose with the formation of acetic and lactic acids (does not produce gas). It forms more lactic acid than acetic acid on milk media; on liver media it forms more acetic acid than lactic acid. It curdles milk at the 59th dose of inoculation (the inoculum is prepared on two hydrolyzate-milk media) for 14-1 days, acidifying the milk to pH-(4.7-0.1) or to acidity -o (7555)7"T . The strain is resistant to 495 Mass, at 6b9o Mass in the medium, growth decreases by an order of magnitude, over 690 - by 2-4 times. Resistant to ristomycin, cormogrizin, kanamycin, neomycin, monomycin. Sensitive to tetracycline, :z" oxacycline, ampicillin, penicillin. Ferments glucose, galactose, lactose, sucrose; does not utilize arabinose, xylose, ribose, mannose, fructose, maltose, raffinose, sorbitol, inulin, mannitol, inositol, trehalose. Maximum 415 growth on nutrient media is (6-9)108CUO /ml, in milk - (4-6). 109CUOS/ml. The strain of B. riait M-4 is antagonistically active against pathogenic and conditionally pathogenic microorganisms - Shigella Zonne, Flexner, and enteropathogenic microorganisms. the desired technical result is insufficient resistance to - aggressive gastrointestinal environments about the tract

For the strain Viydobvasiegisht Biidit 1MV B-7019, which is applied for, the strain Voydobvasiegist o Biyidit 45 deposited in "NIIgenetica" under the number VKPM As-1621 (patent of the Russian Federation Mo2184146,

Ge MPK" С12М1/20, А23С9/12, АбИ1КЗ5/74, А23И1/03, 2002). The strain was isolated from the intestine of a healthy adult. Cultural and morphological characteristics. Polymorphic gram-positive rods are curved or with bifurcations at the ends, stationary, growing in liquid nutrient media throughout the entire volume, except for the aerobic zone. The culture of this strain does not form pigment. On Blauroch, hydrolyzate-milk, casein-hydrolyzate, and thioglycol media, the logarithmic phase of growth ends in 16-18 hours. Total acidity on the medium

R" Blaurok reaches 90-1007T in 24 hours. The optimal growing temperature is 37.5-40.57C. The best way to accumulate biomass is to grow a culture on a casein hydrolyzate medium. Sucrose-gelatin drying medium. In lyophilic form, it is stored from 1 year to 7 years. 60 Physiological and biochemical signs. Obligate anaerobe. It ferments cellobiose, lactose and glucose with the formation of acetic and | - lactic acid. Arabinose, xylose, ribose, mannitol, melicitose, salicin does not ferment. It ferments milk in 14-16 hours with the formation of a stable curd at an acidity of 90-1107T. Does not form catalase, does not dilute gelatin. Antagonistic properties. The strain of bifidobacteria B. biydit 45 VKPM As-1621 suppresses the growth of Shigella

Zonne, Flexner, enteropathogenic Escherichia coli, Staphylococcus aureus, Klebsiella, Proteus and other b5 conditionally pathogenic microorganisms. Antibiotic resistance of the strain. Strain V. Biydit 45 VKPM As-1621 is resistant to kanamycin, monomycin, gentamicin, chloramphenicol, kefzol, streptomycin, ampicillin. Adhesive properties of bifidobacteria. The adhesiveness of the strain was assessed by the average index of adhesion (SPA), which is equal to 9.8-10.

Bacteriocinogenic properties. The strain of bifidobacteria B.byact 45 VKPM As-1621 secretes bifidocins and forms a growth retardation zone of indicator cultures from 2 to 3.5 mm.

The reasons preventing the achievement of the desired technical result are that the Viiidobasiegiit rydtsit 45 VKPM As-1621 strain is not sufficiently resistant to the damaging factors of the cultivation media and the gastrointestinal tract, and therefore, its viability and colonizing properties are insufficient.

Bifidobacteria of the genus Vijdorasiegisht Iopduit are used in the composition of bacterial preparations for medical and preventive purposes and more complex compositions. For example, the well-known strain Vijdorasiegitst 70. Iopdit 274 (patent of the Russian Federation Mo2151606, IPK" AbIKZ5/74, С12М1/20, 2000). The strain was deposited in

The Central Museum of Industrial Microorganisms of the Institute "VNIJ Genetics" under the number As-1571 and is characterized by the following properties. Cultural and morphological properties. When cultivated in a column of 0.7595-th Blauroka agar, it forms colonies in the form of buckwheat grains or discs up to 0.5-1 mm in diameter within 2 days. When cultivated in a liquid medium, Blauroka under anaerobic conditions forms 75 uniform turbidity after 24 hours. On a dense Blauroka medium under anaerobic conditions, it grows in the form of 5-type colonies, round with even edges, 2 mm in diameter, opaque, shiny, they do not form pigment; the profile of the colonies is convex, easily removed with a loop from the surface of a dense medium. In smears, the cells are arranged in the form of "Chinese characters", have the appearance of straight or branched sticks with O-shaped bifurcations at one or both ends, sometimes forming thickenings at the ends. Immobile, they are arranged randomly in the smear.

Gram positive, sometimes granularity is found inside the cells. Spores do not form. Physiological properties.

Obligate anaerobe; grows in a tall column of agar in a test tube. In tubes with a liquid medium and on cups

Petri dish with dense medium grows under anaerobic conditions. During cultivation in a liquid environment under anaerobic conditions, 5.0.108-1.0.109 live microbial cells grow in 18-20 hours after sowing in one ml of the medium. Maximum growth at 37.57. Biochemical signs. Synthesizes the enzyme fructose-b6-phosphate phosphoketalase. It ferments glucose, lactose, sucrose, maltose, xylose, arabinose, melisitose, raffinose. Does not ferment glycerol, rhamnose, mannitol, salicin, cellobiose, mannose, sorbitol, trehalose. Does not form indole and catalase, does not reduce nitrates. Acid formation when grown in Blaurok's medium after 72 hours is 71.8"T. The Vijdorasiegsht Iopdit 24 strain has increased adhesive activity against vaginal epitheliocytes, significant antagonistic activity and acid-forming function, natural resistance to antibiotics. The strain is non-pathogenic for experimental animals (mice) ), as well as for humans (in experiments on volunteers). Sensitive to antibiotics: penicillin, oxacillin, ampicillin, cefazolin, erythromycin.

The reasons that prevent obtaining the desired technical result are the specific purpose of the strain for the treatment of vaginal microbiocenosis disorders. "

As a prototype for the strain Vijdobrasiegisht Iopdit 1MV B-7020, which is claimed, the strain was selected

From Vijdorasiegicht Iyupdit 17x (patent of the Russian Federation 2048517, mpke С12М1/20, А23С9/12, Аб1КЗ35/74, 1995 is), year. The strain has been deposited and stored in the All-Union Collection of Cultures of Industrial Microorganisms "VNIIGenetics" under number B-5813. The Viijdorasiegisht Iyuopdit 17x strain has the following morphological-cultural and physiological-biochemical characteristics. Liquid nutrient media (Blauroca, corn-lactose, hydrolyzate-milk). Two-day culture - straight and slightly branched rods with club-like thickenings, size 0.5x1.0, 3.6-5.2 microns, single, double, in the form of chains. On liquid media, o, with poured into test tubes, a high layer grows in the form of a "column" of turbidity with a zone of anaerobiosis. In milk, cells "" are rod-shaped, shortened, granular. During deep growth in an agar medium, the colonies have the appearance of "thickened needles" measuring 1x2, 2x4mm. The cells are gram-positive, weakly stained with methylene blue, rounded granules are visible at the edges of the cells, catalase-negative, do not produce gas, do not have proteolytic activity, do not reduce nitrates, nitrites. Relation to carbohydrates and alcohols. Absorbs fructose, galactose,

Contains glucose, ribose, mannose, lactose, maltose, melibiose, sucrose, trehalose, arabinose, xylose, inulin, their salicin, weakly sorbitol; does not absorb cellobiose, rhamnose, mannitol and dulcite. Relationship to organic acids.

They produce lactic and acetic acids. Relation to products of intestinal metabolism. Resistant to the content in the medium of 4090 bile, 0.595 phenol, 2.596 Mass. The optimal growth temperature is 37-39"С, does not grow at 45750. s 20 Resistance of the strain to antibiotics. Minimum inhibitory concentration, μg/ml: erythromycin - 0.05; oleandomycin - 0.1; benzylpenicillin - 0.4; ampicillin - 2 .0, oxacillin, tetracycline - 3.0, gentamicin, methicillin - 10.0;

Me; streptomycin - 24.0; chloramphenicol - 65.0; levorin, nystatin - 2060.0. Antagonistic activity. Relative to pathogenic, conditionally pathogenic microorganisms, when co-cultivated 1:11 Viijdorasiegisht Iyupdit 17x, in Blaurok's liquid medium, inhibited growth after 48 hours, E. soii - 100; bZaItopeiIa (Urpitogisht - 100; h Rg. mcyYidagiv - 99.75; Re. aegidipova - 76; K. rpeytopiae - 94-95. The strain is non-pathogenic. When introduced into Vijdorasiegicht Iopdut 17x in milk in the amount of 5-6b95, fermentation occurs after 14- 16 hours, acidity 85-1102T, titer of bifidobacteria reaches 5.109-1.1019, The strain is recommended for the preparation of bifidobacterial preparations and medicinal fermented milk products.

The disadvantage of this strain is insufficiently high biological activity and manufacturability. 60 Lactobacteria of the genus Iasioraciiyshz acidorpyyz are widely used in various branches of the national economy. So, for example, the strain Iasiorasiy5 asidorpyyiz SMSM 1-1225, registered in

National collection of cultures of microorganisms (SMSM) of the Pasteur Institute (patent of the Russian Federation

Mo2126446, MPKU S12M1/20, AbIKZ5/74, 1999; patent of Ukraine Mo26430, MPKU C12M1/20, AbI1KZ5/74, 1999), proposed for the treatment of diarrhea. The proposed culture and composition based on it have a higher ability to competitively displace pathogenic bacteria from intestinal cells, especially bacteria responsible for diarrhea.

The reasons that prevent obtaining the desired technical result are the narrowly specific purpose of the strain.

The patent of the Russian Federation Мо2103354, МПКУ сС12М1/20, А23С9/123, 1998 describes the strain of lactic acid bacteria - I asiorasiyi5 asidorpyshe-13 (VKPM B) for the production of medicinal and dietary fermented milk products. According to the authors, the strain is capable of actively colonizing the mucous membrane of the digestive tract and colon, resistant to the action of magnetic fields, and retains active biochemical properties.

The reasons that prevent obtaining the desired technical result are the specific purpose of the 70 strain.

Lactobacteria of the genus Bigeriosossis (Pegtorpiz) are widely used in various sectors of the national economy. So, for example, in the patent U/O 82/03971, IPC? sazei in the production of cheese with a low fat content. In the patent EP 0148299, IPC C12M01/20, 17.07.1985, 75 information is given on the use in the production of yogurt Zigeriosossisis (pegtorpyiz as part of a bacterial culture.

The use of the Zigeriosossuz strain (teptorpiyiz SSM in the production of fermented milk products with a fat content from 0.05 to 30956 and a maximum acidity of 207T is given in the patent MUO 93/16600, IPC

A23С09/123, 02.09.1993. A mixture of Zigeriosossiv (pepporpiyiz 0OM-001 147, 5igeriosossiv (pegtorpiyiz 0M-001 339, and

And asiorasiiiz BiYidagisiz UOM-100 182 proposed for fermentation of dairy food products in the patent

ER 0794707, MPKU A23С09/123, 17.09.1997,

For the Zygeriosossis strain (Ppegtorpiyiz 1MV B-7026, which is claimed, the Zygeriosossiv strain was selected as the prototype

The specimen is deposited in the National Collection of Cultures of Microorganisms of France under the number 1-1477, (ER 0783566, IPC S12M01/20, 16.07.1997). The Sigeriosossiv strain (pegtorpytse 1-1477) is proposed for milk fermentation. 29 The reasons that prevent obtaining the desired technical result are insufficient resistance to "aggressive environments of the gastrointestinal tract.

Most modern bacterial preparations (bacpreparations) contain several strains. Lactobacterin, acylact, bifidumbacterin, colibacterin, bificol, biosporin, sporobacterin, etc., are well-known bacterial preparations that are able to normalize the microflora of the body (Vorobyev et al. Dysbacteriosis - actual medical problems/ Vestnik RAMN - 1997, Mo3Z, p. 3-5). The well-known bacterial preparation "Biolactin", described in the patent of

of Ukraine Mo20506, МПК9 сС12М1/20; A23C9/12, 27.02.98, Bull. My. "Biolactin" contains bacteria from genera

Vijdorasiegisht, I asiorasii5 and Iasiosossiv. From the genus Vijdorasiegitst, it contains strains of the species: V. riyadit, -

V. opdit, V. iptapiivg, from the genus Iasiorasiyy5 - strains of species: 1bB. Peimeyisiv, IB. sazei, 15. riapiagit. During the preparation of seed material, the previously obtained consortium of lactic acid bacteria is combined with the consortium of bifidobacteria in a ratio of 1:3. oh

The reasons preventing the obtaining of the desired technical result are the excessive complexity of the bacterial concentrate, and therefore the high probability of the lack of a technological process for the production of the final product, which uses a significant number of strains. « du For the "Biocor-1" bacterial concentrate, the bacterial concentrate "Start" (patent of Ukraine 22655, MPKO С12М1/20; А23С23/00, 30.06.98. Byul. Mo3) was chosen as the prototype based on the largest number of common essential features. with "Start" contains bifidobacteria and lactic acid streptococci, from bifidobacteria it contains the strain Viidobasiegiit:z» Iopdit MKRM 5-1514, from lactic acid streptococci Zigeriosossis (pegporpyiz - strains VKPM 8-4463 and VKPM

B-3161, or VKPM 8-4464 and VKPM 8-7331 strains, or VKPM 8-7329 and VKPM 8B-7330 strains at the ratio

Culture (5-8):111. At the same time, the strains of microorganisms used according to their taxonomic affiliation would correspond to the species composition of the human intestinal microflora. Features in common with the claimed invention are the presence of a mixture of lyophilized cultures of bifidobacteria and lactobacilli strains of bifidobacteria and lactobacilli Viidobasiegit Iopdit and Zigeriossis (Pepporpiyiv. - Reasons that prevent obtaining the desired technical result are the moderate properties of the "Start" bac concentrate. o In addition to bac concentrates containing lyophilized biomass of bifidobacteria and lactobacteria, for treatment

Ge) of dysbacteriosis use more complex compositions, which include carriers (various medicinal, flavoring, food additives and fillers). So, for example, in the patent of Ukraine Mo 20509, IPC

Ab1KZ35/74; S12M1/20, 27.02.98, Bull. I have given data on the tablet form of the bacterial preparation, which contains lactose as a carrier (filler), in addition, the tablets contain polyvinylpyrrolidone, stearic acid or carboxymethylcellulose. Common features with the claimed invention are the presence

P» of baking concentrate and lactose. The reasons that prevent obtaining the desired technical result are the other purpose of analogue tablets.

According to the largest number of common essential features, the composition described in examples 3 and 4 of the patent of Ukraine Mo24636, IPC cS12M1/20 was chosen as the prototype for the biologically active additive "Biocorrectin";

A23С9/12, 30.10.98, Bull. Mob5, consisting of a bac concentrate (a lyophilized mixture of Vigdobasiegiit strains

Iopdit MKRM 5-1514, 5igeriosossiz (pegpporpyiz: VKPM B-4463 and

B-2707) and carrier (dry adapted mixture "Detolact" or mixture of sucrose, glucose). This composition also contains barley malt extract, ascorbic acid and calcium stearate. Common features with the invention that 65 is claimed are the presence of the following components in the composition of the prototype: baking concentrate, carrier and calcium stearate.

Reasons that prevent obtaining the desired technical result are the moderate properties of the compositions according to the prototype.

The group of claimed inventions is based on the task of finding such strains of bifidobacteria and lactobacilli, which, while maintaining the effectiveness of the prototype, would be in a more stable equilibrium, and would be more adapted to the conditions of Ukraine, to create an effective dry bac concentrate "Biokor-1"7 from the strains , as well as the biologically active additive "Biocorrectin".

The specified task is solved by the Vijdorasiegisht Byydit 1MV B-7019 strain for the production of bacterial concentrates and food products that correct the microflora of the digestive tract of humans and animals.

The specified task is solved by the Vijdorasiegitst Iopdit 1MV B8-7020 strain for the production of bacterial 7/0 concentrates and food products that correct the microflora of the digestive tract of humans and animals.

This task is solved by the Iasiorasiy5 asidorpishz 1MV B8-7021 strain for the production of bacterial concentrates and food products that correct the microflora of the digestive tract of humans and animals.

This problem is solved by the strain of bacteria Zigeriosossuiz (peptorpyiz 1MV 8-7026 for the production of bacterial concentrates and food products that correct the microflora of the digestive tract of humans and animals.

The specified problem is solved by the fact that the bac concentrate, which contains a mixture of lyophilized cultures of bifidobacteria and lactobacilli strains, according to the invention, it contains from bifidobacteria strains Viiidobvasiegiit

Biyacst 1MV B8-7019 and Vitidobasiegisht Yuyupdit 1MV B-7020, and as lactobacilli contains strains of I aciorasiSh5 acidorpiiv 1MV B-7021 and Bigeriosossiuv (Ppepporpyiz 1MV B-7026 at this ratio, 9o wt.: bifidobacteria 40-80 lactobacilli 20-60

The specified problem is solved by the fact that the biologically active supplement, which contains a bac concentrate and a carrier, according to the invention, as a bac concentrate, it contains the bac concentrate "Biokor-1", which consists of re bifidobacteria strains Vijdorasiegisht Biibut 1MV B-7019 and Vijdobasiegicht Iopdit 1MV B- 7020 in the amount of 107-10 12 CFU/g and lactobacilli strains I asiobasic acidorpytse 1MV B-7021 and Zigeriosossiv (teptorpyv 1MV B-7026 " in the amount of 107-10'9 CFU/g, as a carrier it contains fruit, citrus or vegetable powder, or powder of pineapples or melon crops, or their mixture, as well as milk sugar and calcium stearate at this ratio of components, 90 wt.: Ge»!

From baking concentrate 10-25 IU) milk sugar 20-35 "- calcium stearate 1-2 carrier the rest "! ! ! ! with, (Se)

According to the invention, the biologically active supplement additionally contains vitamins per kg of product: vitamin C 1.0-20g vitamin B1 /15-8Omg « 20 vitamin Bb 15-8Omg - c vitamin B12 1.0-4.Og vitamin RR 10 -20 mg. and vitamin A /2.0-B,O0mg vitamin O 10-BOmMg vitamin E 5.0-10g (22)

According to the invention, the biologically active additive as a carrier contains a powder selected from the following list: apple powder, pear powder, grape powder, citrus powder, pineapple powder, carrot powder, beetroot powder, garlic powder, pumpkin powder, watermelon powder , melon powder, cabbage powder, parsley powder, dill powder, spinach powder, Jerusalem artichoke powder. o According to the invention, the biologically active additive as a carrier contains a mixture of fruit, vegetable powders,

He melon and citrus crops.

The technical result of the claimed group of inventions is the adaptability of the properties of biocorrectin to the conditions

of Ukraine, as well as ensuring concentration and reproduction in the lumen of the intestines of bacteria included in the composition of the bac concentrate.

Below is information on the strains being claimed. Strain Vijdorasiegisht Bijast MV B-7019

R" was identified in the scientific and methodical center "Biokorektsia" in Kyiv, Uzviz Protasiv Yar. The Vijdorasiegiit riydit strain is deposited in the collection of the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine under the number 1МВ В-7019 in 200Ор. 60 Pedigree of the strain. The strain was isolated from intestinal contents. The method of obtaining the strain. The strain was cultivated in Blaurok's semi-liquid nutrient medium by reseeding from individual colonies 1-2 times a week with exposure in a thermostat at 38:20.57"С. Cloning was carried out by twenty passages from isolated colonies from a dense growth dilution, which corresponds to 1:100 million. Strain showed antagonism to common opportunistic and pathogenic microbes, has a high level of adhesion to the intestinal mucosa, a biomass accumulation rate sufficient for the production of 65, high acidity indicators.

Cultural-morphological and physiological-biochemical features of the strain Morphology. Strain cells

Gram-positive polymorphic rods, 4-5 μm long, about 0.5 μm thick, may have bifurcations at one or both ends; are located in clusters of 7 or more cells, or singly, in pairs.

Cultural properties. Requires anaerobic conditions, use of regenerated media. In laboratories, it is well cultivated in columns of semi-liquid medium of Blaurok and others, provided that the height of the column is not lower than 5 cm. It is sown with pipettes with a capacity of 1-2 ml, with the introduction of the material to the bottom of the test tube, followed by mixing without bubbling. After a day, with a dense seeding, a strip of transparent non-encrusted liquid directly under the meniscus becomes visible. The culture requires 1-2 days of cultivation to reach maximum concentration. During the full cultivation period, the concentration can reach 1O0Omln. per ml and more. 7/0 The strain quickly forms striped elongated colonies, which on the second day become coarser and look like "rocket launches" with characteristic branching in the lower part. On the surface of the agar media, the colonies are translucent, rounded, slightly blurred with a diameter of 2 mm.

Physiological and biochemical properties. The test results are shown in Table 1.

And small 10000000 baht bayan 10011101 cheap

Celluveya 10000000 Dlyuya 00000100 z nn IN no

Mel 31711020 Dam 11110113 yuyu 01000001 te 01011111 lLayuovyazdyud

Dulcite 77711111 about zhelamn | does not dilute/ | "

Milk sours, does not form a clot.

The strain is intended for use as part of starter cultures, bacterial concentrates for the production of mixtures and preparations for the purpose of correcting microbiocenosis disorders and treating intestinal diseases of humans and domestic animals.

Products synthesized by the strain. The strain synthesizes acids, in particular acetic and lactic, factors of microbial MO antagonism. "--

Activity and industrial indicators of the strain. Antagonistic activity of the strain. To determine the antagonistic activity of the strain, the method of growing it on agar plates with layering of the culture was used.

Zegaia tagsezsepe, as an oxygen absorber. The strain was sown with a line on the surface of the agar, and after a day, the tested strains were sown perpendicular to it at a distance of no more than 1-2 mm with tapes. The composition of the agar for testing the antagonistic effect corresponded to the composition of the casein-yeast-hydrolyzate medium with the addition of agar to 295. The results are shown in Table 2.

F

2 sh si s

Specific activity in the intestine (level of adhesion of the strain to the surface of the mucous membrane (level of histaadhesion) of experimental animals). » The level of adhesion (histaadhesion) of cells of intestinal microbes, in particular bifidobacteria, is a direct determination of their colonization activity. Tested on freshly excised intestines according to the method of V. P. Zhalko-Tytarenko,

V.M. Bondarenko, A.V. Grigorieva, L.G. Kupchinsky. The method consists of 9 separate procedures: cooking in a microbial suspension, preoperative preparation of the animal, euthanasia, preparation of the intestine, its cannulation and introduction of the microbial suspension, keeping at a low temperature, washing, tryptic disintegration and seeding. The results were calculated according to the formula of V. Zhalko-Gitarenko:

Stakh - tu, ve de: Stdh "7 the level of histaadhesion in the number of colony-forming units per square mm of the mucous surface, i.e. the concentration of microbes in the disintegrate,

M is the volume of the disintegrate, 1 is the length of the exposed segment of intestine, and is the width of the segment of intestine.

Histadhesion activity of the strain depends on the concentration of bifidobacteria in the intestinal cavity - K, according to the ratio:

Styak MK where: "U is the coefficient of histaadhesion, which is calculated based on the results of the experiment according to the formula: ц- Stakh

Kk

The specific, colonization (histadhesion) activity of the strain is shown in Table 3. iv

To determine the competitive ability of the strain for the adhesion sites in the intestine, the competition index was calculated compared to the virulent Shigella of Fleck-sner, having O0-0.03. Competition index: (Mshtamu) KYshigel)70.36:0.03-12.

This means that the strain exceeds the hystaadhesive power of the causative agent of dysentery by almost 12 times.

Reproductive activity. The strain reaches its maximum concentration during reproduction in vials with a casein-yeast-hydrolyzate medium on the third day of cultivation. The initial rate of reproduction at concentrations of 10,000-1,000,000 CFU per ml is t-0.2 per hour. The levels of maximum concentrations and, accordingly, acidity according to Turner are given in Table 4. (22) zov - pnnin Po NI poto z zv F

Long-term storage of the strain. For long-term storage of the strain, it is necessary to lyophilize it according to the "technology used in the production of preparations of the type "Bifidumbacterin dry". Temporary extended storage for a week, up to a month inclusive, is allowed in test tubes with Blaurok's medium and other under a layer of vaseline oil in a refrigerator with a temperature not lower than 57C. For lyophilization, it is necessary to have a culture of the "" strain grown in a semi-liquid environment at 397C for 2 days. Freshly prepared drying medium in the amount of 33965 is added to the taken culture volume. The drying medium consists of three equal parts of 5 ml each: skimmed milk - "reverse"; a solution of 31.5 g of sucrose in distilled water; solution

Ge") 4.5 g of gelatin in distilled water (gelatin is kept in warm water for 1-2 hours to swell). The components are sterilized separately in an autoclave. at 1107C, mixed in a warm state and added to the culture at 37-39".

The method, conditions and composition of the medium for strain cultivation. The strain is cultivated in semi-liquid media according to known standard formulations. The basis of cultivation is a number of test tubes with the appropriate medium

Ge) Oml, which is desirable to regenerate beforehand. The colony of the strain is introduced into the first test tube of the row, thoroughly mixed by suction without bubbling. The material from the first test tube is further transferred to the following 1 ml (serial tenfold dilutions). Cultivation temperature 38:20.57"C. To obtain the strain from the vials provided by the Depository, the culture is revived by rehydration by adding 5 ml of sterile distilled water, physiological sodium chloride solution or nutrient medium. After that, the material

P is serially diluted by iml in rows with 9 test tubes of Uml medium in each and thermostated with daily check for 4 days. Genetic features of the strain. The strain is genetically homogeneous. The strain is sensitive to some antibiotics: gentamicin, tetracycline, ampicillin, carbenicillin, rifampicin, cefazolin, because Monomycin, kanamycin. The strain is resistant to nitroxolin, nalidixic acid.

Strain Vojdobasiegisht Iyupdit 1MV B-7020 deposited in the Depository of the Institute of Microbiology and Virology

NAS of Ukraine on October 25, 2000. The specific name of the culture is Vijdorasiegisht Iopdyt. Lineage of the strain - The strain was isolated from intestinal contents. Cultivated in Blaurok's semi-liquid nutrient medium by transplanting from individual colonies 1-2 times a week with exposure in a thermostat at 38:2-0.57С. Cloning was carried out by 65 twenty passages from isolated colonies from the breeding of dense growth, which corresponds to 1:100 million. The strain showed antagonism to common opportunistic and pathogenic microbes, has a high level of adhesion to the intestinal mucosa, a biomass accumulation rate sufficient for production, and high acidity in Turner degrees. The strain was identified in the scientific and methodical center "Biocorrection" in Kyiv, Uzviz

Protasiv Yar, 4.

Cultural-morphological and physiological-biochemical features of the strain. Morphology. Strain cells

Gram-positive polymorphic rods, 2-3.5 µm long, about 0.5 µm thick, may have bifurcations at one or both ends; are located in clusters of 7 or more cells, or singly, in pairs.

Cultural properties. Requires anaerobic conditions, use of regenerated media. On enriched media, it forms barely noticeable growth under aerobic conditions. It is well cultivated in columns of 7/o semi-liquid medium of Blaurok and others, provided that the height of the column is not lower than 5 cm. It is sown with pipettes with a capacity of 1-2 ml, with the introduction of the material to the bottom of the test tube, followed by mixing without bubbling. After a day, with a dense seeding, a strip of transparent non-encrusted liquid directly under the meniscus becomes visible. The culture requires 1-2 days of cultivation to reach maximum concentration. During the full cultivation period, the concentration can reach 100 million/ml or more. The strain quickly forms striped elongated colonies, which become coarser on the second day: they look like "rocket launches" with characteristic branching in the lower part. On the surface of the agar media, the colonies are translucent, rounded, slightly blurred with a diameter of 2 mm. Physiological and biochemical properties. The test results are shown in table 5. drabnyuya 00020000 bek 0001003 san | 0 howl yatlyu, oh

Melcityuu| 5 (ema 171 tet 0000000 Layuozanazdyu!

Dulcite 7771-11 0 gelatin | dilutes | F

Milk sours, forming a characteristic clot. According to the list of enzymatic features, strain M is identified as a variant of Vijdorasiegiit Iopdit. "--

The field of use of the strain. The strain is intended for use as part of starter cultures, bacterial concentrates for the production of mixtures and preparations for the purpose of correcting microbiocenosis disorders and treatment "

Intestinal diseases of humans and domestic animals. co

The product synthesized by the strain. The strain synthesizes acids, in particular acetic and lactic, factors of microbial antagonism.

Antagonistic activity of the strain. To determine the antagonistic activity of the strain, the method of growing it on agar plates with sub-layering of the culture as an oxygen absorber was used. The strain was sown with a line "0" on the surface of the agar, and after a day perpendicular to it at a distance of no more than 1-2 mm, ribbons were sown - with the tested strains. The composition of the agar for testing the antagonistic effect corresponded to the composition of the casein-yeast-hydrolyzate medium with the addition of agar to 295. The results are shown in Table 6, in which the names of the tested microbial species and their strains are also given. and n and

F z sh y s s

CorpeotolveAtostya: 00

Bure freshly harvested 6 ov »

Specific activity in the intestine, the level of adhesion of the strain to the surface of the mucous membrane (level of hystaadhesion) of experimental animals (white mice).

The level of adhesion (histaadhesion) of cells of intestinal microbes, in particular bifidobacteria, is a direct determination of their colonization activity. It is tested on freshly excised intestines according to the method of V.P.

Zhalko-Tytarenko, V.M. Bondarenko, A.V. Grigorieva, L.G. Kupchinsky. The method consists of 9 separate procedures: preparation of the microbial suspension, preoperative preparation of the animal, euthanasia, preparation of the intestine, its cannulation and introduction of the microbial suspension, kept at a low temperature, washing, tryptic disintegration and seeding. The results are calculated according to the formula of V.P. Zhalko-Titarenko:

t.

Sttvk - and: where: Stdh "7 the level of histaadhesion in the number of colony-forming units per square mm of the mucosal surface, t - the concentration of microbes in the disintegrate,

M is the volume of disintegrate, 1 is the length of the exposed segment of intestine, and is the width of the segment of intestine.

Histadhesion activity of the strain depends on the concentration of bifidobacteria in the intestinal cavity - K, according to the 70 ratio:

Stakh MK where: ЧУ - coefficient of histaadhesion, which is calculated based on the results of the experiment according to the formula: ц- Stakh

Kk y y no: no y y

The specific, colonization (histaadhesion) activity of the strain is shown in Table 7.

To determine the competitive ability of the strain for adhesion sites in the intestines, the competition index was calculated compared to virulent Flexner shigellae, which have O-0.03: the competition index: (strain )HMshigel)70.307:0.03-10.05

This means that the strain exceeds the hystaadhesive power of the causative agent of dysentery by almost 10 times.

Reproductive activity. The strain reaches its maximum concentration during reproduction in vials with a casein-yeast-hydrolyzate medium on the third day of cultivation. The initial rate of reproduction at concentrations of 10,000-1,000,000 CFU per ml is t-0.706 per hour. The levels of maximum concentrations and, accordingly, acidity according to Turner are given in table 8. - « z5 F

Ов яко0116 « вд тмн000010000в6 о З с хз»

Long-term preservation of the strain. For long-term preservation of the strain, it is necessary to lyophilize it according to the technology used in the production of "Dry Bifidumbacterin" type preparations. Temporary extended

It can be stored for a week, up to a month, in test tubes with Blaurok's medium and others under a layer of vaseline oil in a refrigerator with a temperature not lower than 57C. For lyophilization, it is necessary to have a strain culture grown in a semi-liquid environment at 397 for 2 days. Freshly prepared drying medium in the amount of 33965 is added to the volume of the culture. a solution of 31.5 g of sucrose in distilled water; solution of 4.5 g of gelatin in distilled water (gelatin is kept in warm water for 1-2 hours to swell). Components c are sterilized separately in an autoclave ЗОхв. at 1107C, mixed in a warm state and added to the culture at 37-39".

Drying mode: freezing under vacuum to -457C, gradual increase in temperature by 57C per hour to 22 - 4247C, drying - 12 hours. 99 The method, conditions and composition of the medium for strain cultivation. The strain is cultivated in semi-liquid media according to known standard recipes. The basis of cultivation is a number of test tubes with the appropriate medium

Oml, that it is desirable to regenerate beforehand. The colony of the strain is introduced into the first test tube of the row, thoroughly mixed by suction without bubbling. The material from the first test tube is then transferred to the following ml (serial tenfold dilutions). Cultivation temperature 38:20.5"C. To obtain a strain from the provided 60 vials of the Depository, the culture is revived by rehydration by adding 5 ml of sterile distilled water, physiological sodium chloride solution or nutrient medium. After that, the material is serially diluted by ml in rows with 9 tubes per OUml of medium in each and thermostated with daily check for 4 days.

Genetic features of the strain. The strain is genetically homogeneous. The strain is sensitive to some antibiotics: gentamicin. tetracycline, ampicillin, carbenicillin, rifampicin. The strain is resistant to nitroxolin, nevigramon, polymyxin, chloramphenicol, streptomycin, monomycin.

Strain I asiorasiyiz acidorpyyiz 1MV B-7021 deposited in the Depository of the Institute of Microbiology and Virology

NAS of Ukraine on October 25, 2000. The specific name of culture I asiobasiyiz5 asidorpyshv5. Pedigree of the strain - isolated from a healthy 7-year-old child in 1999 by employees of the "Biocorrection" scientific and medical center, Kyiv, Uzviz

Protasiv Yar, 4.

Cultural-morphological and physiological-biochemical features of the strain. Morphology. Strain cells

Gram-positive rods of regular shape, measuring (1.4-1.5)X(3.0-6.0)μm (sometimes filamentous), are arranged in 1-2 or short chains. Spores do not contain, are not mobile. Mesodiaminopimelic acid was not detected in the cell walls. Colonies - whitish with uneven edges, opaque, rough, 1-2 mm in size. 70 Cultural properties. Requires rich complex nutrient media for growth. The optimal pH for growth is 5.5-6.2, the optimal temperature is 40:27, there is no growth at 157C. Facultative anaerobe.

Physiological and biochemical properties. Does not synthesize catalase, indole, NoZ, cytochromes (benzidine test is negative), does not dilute gelatins, does not reduce nitrates to nitrites. Grows at 4095th concentration of bile in the medium, grows in milk with the formation of a clot and restores milk with litmus or methylene blue.

Ferments carbohydrates to lactic acid, does not form CO from glucose. Ferments: fructose, galactose, glucose, lactose, maltose, sucrose, mannose, salicin. Does not ferment: rhamnose, ribose, trehalose, xylose, arabinose, sorbitol, mannitol, gluconate, glycerol. The field of use of the strain. The strain is intended for use as part of starter cultures and bacterial concentrates, for the preparation of drugs and food products that correct the microflora of the digestive tract of humans and animals. The product synthesized by the strain. The strain synthesizes lactic acid, which is a factor in microbial antagonism. Antagonistic activity of the strain. To determine the antagonistic activity of the strain, the method of growing it on milk agar and Lactoagar plates in the absence of oxygen in a desiccator was used. The results of the research are shown in Table 9.

Evorepolassoioets 00008

F zo yu -

Zaitopea urni 562 freshly isolated 6

Revuetiavtyaya 00000105 from zv F

The method, conditions and composition of the medium for cultivation and storage of the strain. For long-term storage of the strain, it is necessary to lyophilize it according to the technology used in the production of preparations such as "Bifidumbacterin dry." Temporary extended storage for 1-3 weeks, up to a month inclusive, let's assume pt) s in test tubes with milk agar and under a layer of petroleum jelly in a refrigerator with a temperature not lower than 570.

For lyophilization, the culture is grown in a semi-liquid environment at 39-407"C for two days. 3395 of a freshly prepared protective medium for drying of the following composition is added to the culture: skimmed milk (reverse) BOml b 15 31.5956th sucrose solution in distilled water BOml 4.595 -th solution of gelatin in distilled water (preliminarily let the gelatin swell in warm water for 1-2 hours) BHOml

ЧК» -z Components are sterilized separately in an autoclave ЗОхв. at 1107C and mixed before use. Drying mode: freezing to -45"C, drying under vacuum from -457C for 10-12 hours with a temperature increase of 50 degrees at 57C per hour to 22-24"C and further drying at this temperature for 6-12 hours. The strain is cultivated on the commercial medium "Laktoagar" (Russia), agar with milk, hydrolyzate-milk or

Fo corn-lactose medium.

Milk agar. 295 agar and 20965 skimmed milk (reconstituted commercial dry milk with an acidity of 16-187T in the amount of 100g per 9U0Oml of distilled water). Agar and milk are sterilized separately at (121-41)7С 1 Ov. 59 When using, add 2095 of skim milk to melted agar, mix thoroughly, and pour into Petri dishes.

Hydrolyzate-milk environment. Hydrolyzed milk is diluted with drinking water 1:1. In a small amount of diluted hydrolyzate, agar is melted at the rate of (2,540.5) g per Yakub.dm of the prepared medium. 20g of peptone, 3.5g of sodium chloride are added to the remaining hydrolyzate, the mixture is heated to 805-27C, and then mixed with molten agar. After setting the pH to 7.50.1, boil for 15 minutes, settle, filter, add hot distilled water to the given volume and add 10.0g of lactose, 0.15g of cysteine hydrochloride. The medium is poured into tubes of 10 cubic cm and sterilized with steam at 1122417C for 30 minutes. in a heated autoclave, the pH of the finished medium is 7.102.

Corn-lactose medium. Melt agar in a small amount of distilled water at the rate of (2.5:20.5) g per cubic meter of the prepared medium. (10:1) g of peptone is added to the remaining distilled water,

(40:41) g or cubic cm of an aqueous solution of corn extract diluted 1:6, (6-0.5) g sodium citric acid trisubstituted, (0.12-0.02) g magnesium sulfate, (250.1) g of disubstituted potassium phosphate. The mixture is heated to (80-2)"С, after which it is combined with molten agar, (10:0.5) g of lactose, (0.15--0.05) g of bolyanoKacidic cysteine or (0.52- 0.1)g of ascorbic acid. The pH of the finished medium is 7.0-0.2. To obtain the strain from the vials provided by the Depository, the culture is revived by rehydrating the dry material by adding bml of sterile distilled water, physiological sodium chloride solution, or liquid nutrient medium. After that the suspension is serially diluted by iml in rows with 9 tubes with uml of medium in each and thermostated with a daily check for 4 days. Serial dilutions of dry biomass can be inoculated on milk agar or Lactoagar in Petri dishes and thermostated.

Features of the strain. The strain is genetically homogeneous. The strain is sensitive to some antibiotics: ampicillin, cephalothin, monomycin, penicillin, clindamycin, kanamycin, gentamicin, resistant to nitroxolin, 5-NOK, nevigramon.

The Zigeriosossis strain (Pegtorpyiz 1MV 8-7026) was deposited in the Depository of the Institute of Microbiology and 7/5 of Virology of the National Academy of Sciences of Ukraine on June 18, 2001.

The specific name of the microorganism: Zigeriosossisis (Tegtorpiy5. The lineage of the strain is isolated from fermented milk products. The strain was identified in the "Biocorrection" laboratory, Kyiv.

Cultural-morphological and physiological-biochemical features. Facultative anaerobe. Requires rich media with carbohydrates and organic nitrogen sources (peptone, casein hydrolyzate) for growth. When growing on the surface of nutrient agar media (AMR), whitish round shiny colonies with a diameter of 0.5-1.0 mm are formed. Deep colonies in the form of white discs with a diameter of up to 1 mm. A homogeneous turbidity and an amorphous sediment form in the broth. Cocci-shaped gram-positive cells 0.8-1 µm, sometimes up to 1.2 µm in diameter, are located in pairs, groups and chains (the latter prevail when growing in a liquid and semi-liquid environment). Immobile, do not form spores. Gelatin is not diluted, ammonia is formed from arginine. Do not contain catalase. Milk is fermented, litmus in milk is restored, milk with 0.196 methylene blue is not restored. They do not grow at 4095 mol, at 6.595 Mass, at рHO,b, at i-0, 4-15 "С. They grow at 2.595 Mass, at 457"С and weakly at 5070.

Good growth in the zone of 25-457C, optimum 40"C. The optimal value for growth is рННб, 0-6.5. They survive in a liquid medium at рНУ, after warming up at 60"С during ЗОхв. Carbohydrates are fermented to lactate without the production of carbon dioxide. They ferment: glucose, lactose (the ability to ferment lactose can often be lost during cultivation), fructose, sucrose; do not ferment mannitol, glycerin.

The field of use of the strain. The strain is most likely to be used in the food industry. at

The product synthesized by the strain. A complex of biologically active substances is produced, in particular lactic acid, antibiotic-like substances. Strain activity. Sterile skimmed milk (reverse) ferments after 4-6 hours of cultivation at a temperature of 40°C. Method, conditions and composition of the medium for strain cultivation.

The strain is cultivated at 37-40"C for 24-48 hours on the surface of agar or in liquid media.

MRS, MPA or MPB media with 195 glucose, sterile skimmed milk, reconstituted 1095 skimmed milk are used. Culture is transplanted once a month. Composition of MRS medium (g/l): peptone 10 " meat water 100 - yeast autolysate 100 s liver extract 100 s glucose 20 sodium acetate B potassium hydrogen phosphate 2

He») magnesium sulfate 02 manganese sulfate 0.05 o tween 80 1 sha distilled water to 10001 sl 50 rn-v,v

Ge Method, conditions and composition of the medium for long-term storage of the strain. The culture is grown under optimal conditions in 1095 reconstituted milk, sterile skimmed milk, semi-liquid media (see above) with 0.7g/l agar. Store at 4"C for a month. Cells can be freeze-dried afterwards

Freeze to -357C and store for 3-5 years. For lyophilization, the culture is grown for 24 hours on sterile skimmed milk and shifted in a ratio of 1:11 with a protective medium. Protective » medium during lyophilization in a 90:2095 aqueous solution of sucrose 10

30% aqueous solution of sodium citric acid 10% gelatose solution (dissolve 1 part of gelatose in b parts of distilled water). 30

Information on pathogenicity (for humans, animals or plants). The strain is non-pathogenic for warm-blooded animals. Auxotrophicity, resistance to antibiotics, phages, etc. were not studied.

Next, the claimed invention is confirmed by examples of specific implementation for the dry bac concentrate "Biocor-1" and the biologically active supplement "Biocorectin". Example 1. For the production of dry concentrate

"Biokor-1" was taken: strain Viidobasiegisht riyyacht 1MV B-7019, strain Viidorasiegisht yuopdit 1MV B-7020, strain I asiorasiyiv asidorpyiiz MV B-7021, strain Zigeriosossiv (pegtorpiyiz 1MV B-7026, liver medium (Blauroka) for cultivation of bifidobacteria, drinking water according to GOST 2874, magnesium sulfate 7-hydrate according to GOST 4523, 70 sodium hydroxide according to GOST 4328, sodium citric acid according to GOST 22280, dry skimmed milk according to GOST 10970, skimmed milk with an acidity of no more than 197T, obtained from milk according to DSTU 3662.

To accumulate the biomass of microorganisms, a culture medium was prepared, the basis of which is 7/5 reconstituted skimmed milk, the proteins of which are hydrolyzed by protosubtilin with the addition of lactose, sodium citric acid, iron sulfate, and growth stimulants. Seed material, which was prepared in this way, was added to the prepared nutrient medium. For the production of seed material (inoculum) bifidobacteria 590 day culture, which was grown in Blaurok's medium, is added to the prepared cultivation medium for biomass accumulation. Cultivation was carried out during Zbhod. at a temperature of 35:-176.

The seed dose of bifidobacteria in the fermenter was 7905. Inoculum of lactic acid bacteria Zigeriosossiv

ISheptornuins were prepared on sterilized skimmed milk with a seed dose of 195 at a temperature of 37170 for 1 hour. The inoculum dose of thermophilic streptococcus in the fermenter was 395. The inoculum I asiobacilliis acidorpiinz was prepared on sterilized skimmed milk with an inoculum dose of 0.795 at a temperature of 38-17 for 1 hour. The seed dose of Iasiorasiyshz acidorpis in the fermenter was 295. The accumulated biomass of the microorganisms was separated from the culture medium on a supercentrifuge, weighed and mixed with a sterile aqueous solution of the protective medium containing 595 skimmed milk powder, 89o sucrose, 4,590 sodium citric acid. The suspension obtained in this way was frozen in a freezer at a temperature of minus (35-45)"C and dried in a freeze-dryer. The "Biokor-1" buck concentrate was obtained, which had the following properties. A loose powder of light brown color. The taste and smell are impersonal, without extraneous flavors and odors. The mass fraction of moisture is 595. The duration of fermentation of milk (with the addition of 1 g per 1 liter) at 377C was 4 hours. The ratio of the number of bifidobacteria and lactobacilli, in 90: o "- bifidobacteria 40 lactobacilli 60 "

From Example 2. Bac concentrate "Biocor-1" was prepared as described in example 1, except that the ratio of bifidobacteria and lactobacilli inocula was slightly changed. Dry milk concentrate was obtained with the following ratio of the number of bifidobacteria and lactobacteria: bifidobacteria 80 h s 40 lactobacteria 20 Z "» The milk concentrate had a cream color, the mass fraction of moisture was 4.995. The duration of milk fermentation when 1 g of milk concentrate was added to 1 liter of milk was 5 hours. at 3770.

Example 3. Bac concentrate "Biokor-1" was prepared as described in example 1, except that the ratio of inoculums of bifidobacteria and lactobacilli was slightly changed. We obtained a dry yeast concentrate with the following (22) ratio of the number of bifidobacteria and lactobacilli, i.e.: bifidobacteria 60 - lactobacilli 40 AND

"Biokor-1" dry baking concentrate is a cream-colored loose powder without extraneous flavors and odors. (Che) Mass fraction of moisture - 4.895. Duration of milk fermentation - 15 hours.

Buck concentrate "Biokor-1" is used in the food industry for the production of preventive lactic acid products and biologically active additives.

In addition, the bacterial concentrate "Biokor-1" has a high biological activity, promotes the active colonization of bifidobacteria and lactobacilli strains in the human digestive system, and exhibits a high level of » antagonistic activity towards the unwanted microflora of the mucous membranes. "Biokor-1" is a therapeutic and preventive product, normalizes the microflora of mucous membranes, restores the functions of the intestinal tract, activates the synthesis of vitamin-enzyme complexes, increases immunity, promotes the disappearance of allergic manifestations on the skin. According to the data of clinical studies, the therapeutic effect in patients with exacerbations of pathology and dysbiosis occurred on the 2nd-4th day of treatment in 5095 cases, on the 4th-6th day in another 3095 cases, making a total of 8095. For patients with chronic intestinal diseases, the therapeutic effect was manifested on 6-12 days in 8095 patients.

"Biokor-1" baking concentrate is recommended to be used in a liquid or lyophilized state, as a leavening agent for the preparation of home and industrial curative-prophylactic fermented milk products, b5 cocktails, etc.

The biologically active additive "Biocorectin" (hereinafter, biocorectin) was produced using the following raw materials and materials: the "Biocor-1" bac concentrate 7, described in examples 1-3, as well as apple powder, pear powder, grape powder, citrus powder, pineapple powder, carrots, beetroot powder, garlic powder, pumpkin powder, watermelon powder, melon powder, cabbage powder, parsley powder, dill powder, spinach powder, Jerusalem artichoke powder, milk sugar, vitamins C, B, B, B, PP, A, 0, E, calcium stearate, hard gelatin capsules.

Example 4. For the production of biocorrectin, the following was taken per 1 kg of the finished product: "Biocor-1" bac concentrate 120 g; 70 milk sugar 265g, calcium stearate 15g, carrier (apple powder) god;

The component parts of the composition were mixed to form a homogeneous powder. Then, after checking the properties, the obtained biocorrectin powder was sent for packaging or for the preparation of gelatin capsules, packaging, labeling and storage of finished products.

Biocorrectin is a highly dispersed powder, homogeneous throughout the mass with a pleasant taste and smell characteristic of apple powder, without extraneous tastes and smells. 100 g of biocorrectin contains 0.4 g of proteins, 9 g of carbohydrates, and the energy value is 45 kcal. According to microbiological indicators, biocorrectin meets the following requirements: the total number of viable microorganisms is at least 1-10 CFU/g (absence of bacteria of the group of Escherichia coli (coli forms) and pathogenic microorganisms, including bacteria of the genus Salmonella, 51. .

Examples 5-8. Biocorrectin was made as described in example 4, except that the ratio of components was changed. The composition of the obtained biocorrectin is given in examples 5-8 of table 10.

Organoleptic and microbiological indicators were practically the same as for example 4.

Examples 9-11. Biocorrectin was made as described in example 4, except that vitamins were additionally added to the composition of 29. The composition of biocorrectin with added vitamins is given in examples 9-11 of Table 10.

Examples 12-28. Biocorrectin was produced as described in Example 4, except that the carrier was a powder selected from the following series: apple powder, pear powder, grape powder, citrus powder, pineapple powder, carrot powder, beetroot powder, garlic powder, pumpkin powder, F watermelon powder, melon powder, cabbage powder, parsley powder, dill powder, spinach powder, M Jerusalem artichoke powder. These carriers increase the histaadhesive activity of the strains included in biocorrectin, and also expand the range of organoleptic and therapeutic properties of biocorrectin. Examples 12-28 -- table 11 show some features of the properties of biocorrectin with different carriers, which had the following composition: 60 - s Therefore, biocorrectin is a source of pure cultures of symbiotic bifidobacteria in the amount of 1.107-1.1012 CFU/g, lactobacilli in the amount of 1.107-1.1012 CFU/g, as well as vitamins and minerals of natural origin. 5 of them "Biokor-1", CSU/g of product

Vijdorasiegisht I asiorasiv | tank sugar stearate | carrier |vitamin vitamin |vitamin vitamin vitamin |vitamin | vitamin | vitamin cancer h Biyait 1MV asidorniish5 concentrate | dairy | of calcium Сg Vi,go |Вв,го |В42, g IRR, go Ag b,g E, g v-7019 1 1MV V-7021 i | "Biokor-1"

Iospdit 1MV Iegtornishv

Ге в-7020 1мМВ в-7026 рю юю 5 рю юю 11th CENTURY OF POMSNNNNY POETS POED NOE NI NO UNONIA NOE UAN NOSY PON KON PANNO PON PON KON NOYA PON z" ryu yuyu 11111 ru yuyu vv i royov yuyu 12651666 10 100080 160050 08020605 ”Vol 1 Yv1YUV BY WILL WILL WIGHT T 65 MO CRAKE CONTRACTS Some features

12 apple powder contains pectins, organic acids, flavonoids, vitamins, organic compounds of iron and phosphorus, tannins, catechins, is used in dyskinesia of the biliary tract, in spastic colitis, in vitamin deficiency, contains tannins and pectins, carotene, vitamins, mineral salts, an astringent (14 grape powder has a tonic, diuretic, laxative and choleretic effect on the body citrus powder improves digestion in hyposecretory gastritis, weak bile secretion, tendency to spastic constipation, has tonic properties 16 pineapple powder contains proteins, carbohydrates, vitamins, enzymes, including the bromelain enzyme, which has unique properties to accelerate the breakdown of fats in the body 17 carrot powder contains p-carotene and other carotenoids, vitamins, pectin substances, macro- and microelements, a valuable 70 treatment tool for vitamin deficiency, as an auxiliary tool for conjunctivitis, keratitis, etc. 18 beetroot powder stimulates hematopoiesis with, stimulates gastric secretion and intestinal peristalsis, promotes cholesterol excretion, exhibits antitumor properties, is effective in spastic colitis, atherosclerosis, thyrotoxicosis, arrhythmias, hypertension, liver diseases, intestinal atony 19 garlic powder stimulates the secretion of gastric juice and bile, increases intestinal peristalsis, contributes to the development of normal intestinal flora, expands blood vessels, lowers blood pressure 19 20 pumpkin powder contains sugars, organic acids, vitamins, minerals, has a diuretic, choleretic and laxative effect contains sugars, organic acids, pectin substances, vitamins, diuretic and choleretic agent 22 melon powder contains pectin substances, vitamins, mineral salts, has moderate laxative, diuretic and choleretic properties 23 cabbage powder contains proteins, sugars, organic acids, minerals, vitamins, including the so-called vitamin B, which provides good therapeutic effect in peptic ulcer disease stomach and duodenum, with enteritis, colitis, etc. 24 parsley powder contains flavonoids, vitamins, proteins, stimulates appetite, improves digestion, inhibits fermentation processes in the intestines, promotes milk secretion in nursing mothers (25 dill powder contains flavonoids, vitamins, minerals, has a multifaceted positive effect on the body 29 26 spinach powder |increases intestinal peristalsis, has a mild laxative effect, has capillary-strengthening, anti-sclerotic and " anti-inflammatory properties, is also used as a means of creating a therapeutic background, and to potentiate the action of medicines for a number of diseases 27 powder contains carbohydrates in the form of polymers (fructooligosaccharides) that are not broken down in the gastric juice because Jerusalem artichoke lacks the inulinase enzyme; as a result of this, fructooligosaccharides reach the intestines, where they are fermented by bifidobacteria, which intensively reproduce and displace pathogenic and opportunistic microorganisms from the intestines. fruit, properties, contributes to the normalization of human intestinal microflora, provides effectiveness in vegetable, therapeutic and preventive nutrition in the presence of diseases of the gastrointestinal tract, which is associated with plantains and dysbacteriosis; with diseases of the stomach and duodenum, liver, metabolic disorders; for citrus crops/treatment of intestinal infections (shigellosis, salmonellosis, staphylococcal enterocolitis and others), organ infections

From urogenital and upper respiratory tract organs; after radio irradiation, transfer of acute infections, re) food poisoning, immunodeficiencies, long-term use of antibiotics and chemotherapeutic drugs

Biocorrectin has general strengthening and adaptogenic properties, contributes to the normalization of human intestinal microflora by suppressing pathogenic and conditionally pathogenic microorganisms and increasing the number of 420 Lactic acid microflora. Clinical studies, which were conducted in hospitals of the city of Kyiv, allow 8 s to recommend biocorrectin with a high effect in preventive and dietary nutrition in case of: и - the use of antibiotics and chemotherapeutic drugs; "" - radiation therapy; - diseases of the gastrointestinal tract associated with dysbacteriosis; - food allergies;

Ge") - intestinal infections; - secondary immunodeficiencies. shk Biocorrectin is recommended to use 2-3 capsules for adults, 1-2 capsules per day for children 30 minutes before - taking food for 1-2 months. The method of application proposed by the authors has passed clinical trials and does not cause objections. i-th Biologically active supplement "Biocorrectin" is approved for use by the Ministry of Health of Ukraine. ie)

Claims (9)

The formula of the invention is what and what about and 1. Vijdobrasiegisht Biydyt 1MV B-7019 strain for the production of bacterial concentrates and food products that correct the microflora of the digestive tract of humans and animals. 2. Bacterial strain Viidobasiegist Iopdit 1MV B-7020 for the production of bacterial concentrates and food products that correct the microflora of the digestive tract of humans and animals. in Z. Bacterial strain Iasiorasiyz acidorpiyzIMV 8-7021 for the production of bacterial concentrates and food products that correct the microflora of the digestive tract of people and animals. 4. Strain of bacteria Bigeriosossivz (Sheptpornpishz 1MV 8-7026 for the production of bacterial concentrates and food products that correct the microflora of the digestive tract of people and animals. 5. Buckwheat concentrate containing a mixture of lyophilized cultures of bifidobacteria and lactobacilli strains, 65 which differs in that it contains the strains of Viidorasiegt bBiidit 1MV B-7019 and Viidobasiegisht Iopdit 1MV B-7020 from bifidobacteria, and as lactobacilli it contains strains of I asioraciiyvs asidorpiche 1MV B- 7021 and zigeriosossisis (pegptorpiyiz 1MV B-7026 with this ratio, because by weight: bifidobacteria 40 - 80 lactobacteria 20-60. 6. A biologically active supplement containing a bac concentrate and a carrier, which differs in that as a bac concentrate it contains the bac concentrate "Biokor-1", which consists of bifidobacteria of the strains Viidorasiegisht Biydit 1MV B-7019 and Viidobasiegicht Iyupdit 1MV B-7020 in the amount of 107 1012 CFU/g and lactobacilli strains I asiobacilliv 70 asidorpytse 1MV B-7021 and Zigeriosossiv (Sheptorpyshv 1MV B-7026 in the amount of 1071-1010 CFU/g, as a carrier it contains fruit, citrus or vegetable powder or powder of pineapple or melon crops, or their mixture, as well as milk sugar and calcium stearate with this ratio of components, 95 wt.: baking concentrate 10-25 milk sugar 20-35 and calcium stearate 1-2 carrier the rest. 7. Biologically active supplement according to point b, which is distinguished by the fact that it additionally contains vitamins per kg of product: vitamins 1.0-20g vitamin B 15 - VOmg vitamin Bb 15-8Omg vitamin B12 1.0-4, Omg vitaminRR o /10-20mg « vitaminA 2.0 - B.Omg vitamin O 10-BOmMg vitamin E 5.0-10g. Gee! , and -. but also U 8. Biologically active supplement according to claim 6, which is characterized by the fact that as a carrier it contains a powder selected from the following series: apple powder, pear powder, grape powder, citrus fruit powder, pineapple powder, "- carrot powder, beetroot powder, garlic powder, pumpkin powder, watermelon powder, melon powder, cabbage powder, parsley powder, dill powder, spinach powder, Jerusalem artichoke powder. " 9. Biologically active supplement according to point b, which differs in that it contains a mixture of fruit, vegetable, melon and citrus powders as a carrier. Official bulletin "Industrial Property". Book 1 "Inventions, useful models, topographies of integrated microcircuits", 2004, M 2, 15.02.2004. State Department of Intellectual Property of the Ministry of Education and Science of Ukraine. Torment t kr o, . and? (o) Cheka" - 1 iChe) 60 b5