Strain of microorganism lactobacillus plantarum p4, strain of microorganism lactobacillus buchneri p0 and probiotic preparation based on thereof for correction of dysbacteriosis of different etiology in humans and animals

FIELD: biotechnology, medicine, microbiology, agriculture, food industry.

SUBSTANCE: the strain Lactobacillus plantarum P4 deposited in the collection of NIIM MO RF at number 39 and the strain Lactobacillus buchneri P0 deposited in the collection NIIM MO RF at number 38 form in common a symbiotic system characterizing by syntrophic synergistic type of relationship. The probiotic preparation "Bilakt" based on thereof comprises the dried mixture of strains Lactobacillus plantarum P4 and Lactobacillus buchneri P0 in the ratio of microorganisms from 4:1 to 1:4 and products of their vital activity. This mixture promotes to sanitation of nonsterile cavities of body and correction of dysbacteriosis of different etiology in sick humans and animals at the content of live cells 1 x 108/1 ml of rehydrated preparation, not less. The preparation shows high effectiveness in some human diseases - digestive and urogenital tracts, different cutaneous diseases of different etiology (suppurative and burn injuries of skin) and agricultural animals and poultries.

EFFECT: valuable medicinal properties of strain and probiotic.

4 cl, 3 ex

 

The invention relates to biotechnology, namely strains for the preparation of therapeutic and preventive drugs and technologies of their production in an industrial environment. The invention can be used in food industry, medicine, veterinary medicine and related fields.

Known bacterial preparations for medical purposes, such as colibacterin, Lactobacterin, bifidobakterii (Guide to vaccine and serum case. Ed. by Acad. The Academy of medical Sciences of the USSR Energysave. Moscow, “Medicine”, 1978), containing one or two strains of micro-organisms and inhibit the development of pathogenic bacteria.

Famous promising strains used in the creation of products and milk products, contributing to the regulation of intestinal and nonspecific immunostimulation with dysbacteriosis of various etiologies (Vmichael "the Problem of regulation of the intestinal microflora". Journal of Microbiology, epidemiology and immunology. M., 1995, N 3, p.48-52).

Known Lactobacillus acidophilus VKM B-2020 D-antibiotic-antagonist agents of intestinal infections. The strain is recommended as the basis for the development of antagonistic biological means of prevention and treatment of intestinal diseases caused by pathogenic and conditionally pathogenic microflora. Can be used to create bacterially preparations for the correction microfunctional disorders of the gastrointestinal tract (RF Patent No. 12063436, 12 N 1/20).

A known strain of lactobacilli Lactobacillus acidophilus-13 (Agaric) positively affect the regenerative processes in the intestine for the treatment of gastrointestinal diseases, used for correction of dysbacteriosis of different types and nature of neonates and adults. The strain used in the production of nutritious diet food suppressing the growth of pathogenic forms of microorganisms and normalization of the intestinal microflora due to the high ultimate pH strain, 400° T (RF Patent No. 2103354, With 12 N 1/20).

Known bacterial strain Lactobacillus acidophilus-92 n. v. (Ani)used for the correction of dysbacteriosis in gastrointestinal diseases and in patients with colon cancer (Patent RF №2103353, With 12 N 1/20).

A known strain Lactobacillus acidophilus n. v. 317/402 of "Narine" inmi-9602 (A.S. USSR №163357, With 12 N 1/20), which has a slow acid-forming ability (maximum education - 360° T). The strain produces a significant amount of harmless for children and adults antibiotic substances that inhibit the growth and development of both gram-positive and gram-negative bacteria - pathogens of acute forms of gastrointestinal diseases.

Known drug Lactobacterin, which is based on the strain RA-3 Lactobacillus plantarum.

The disadvantage of this drug and the corresponding strain which is insufficient therapeutic efficacy. Long-term observations showed that at high antagonistic activity and acid-forming ability of the cells of this strain have a low adhesive activity. Some side effects when using Lactobacterin caused by the ability of lactobacilli strains R-3 to produce hydrogen peroxide.

Closest to the claimed invention (the prototype) is the drug “Biosan” (U.S. Pat. 2102075 Russia, MKI6A 61 K 35/74), which is based on the strain plantarum (Kirov-1) and Lactobacillus buchneri (Kirov-4).

Common signs of drug “Biosan” and claimed the drug are as follows:

the basis of the drugs is the Association of microorganisms of two species of lactobacilli L.plantarum and L.buchneri;

lactobacilli derived from natural habitats with the first degree of purity;

lactobacilli have the ability to prejustice on the mucosa;

lactobacilli exhibit antagonistic properties against pathogenic microbes causing endometritis in cows;

Association of lactobacilli used in the treatment of endometritis in cattle.

A comparative juxtaposition of cultural-morphological and biochemical properties of cells, as well as antagonistic activity against the test strains and pathogenic microflora indicate significant differences is tommow L.buchneri P0 and L.buchneri (Kirov-4):

optical properties, color, surface and size of the colonies;

the continuing increase in oxygen atmosphere on a dense nutrient medium MPC-4 at a temperature of 36... 38° C;

the ability to ferment mannitol (mannitol and lactose;

spectrum antimicrobial action, in particular against pathogens of dysentery and salmonellosis.

Data of the authors of the patent (Varganov A.I. Prevention of symptomatic and artificially acquired infertility in cows and heifers: abstract of thesis... SOIC. Dr. vet. Sciences /Institute of Nazareth. animal diseases - Voronezh, 1988. - 31 S.; Varganov A.I., Konoplev I.G. Main drugs for the treatment and prevention of obstetric-gynecologic diseases of animals (Method. the decree. for students of higher and secondary educational institutions on special. “Veterinary medicine” and the wind. specialists). - Kirov, 1995 - 32 C.)) for species typing of strain L.buchneri (Kirov-4) does not allow us to classify it in mind L.buchneri, and the ability to ferment mannitol and the lack of ability to ferment lactose classification Rogosa and Sharpe is more characteristic of L.brevis and L.colinoides. Thus, a comparative juxtaposition of cultural-morphological and biochemical properties of cells L.buchneri P0 and L.buchneri (Kirov-4) gives the basis to consider them if not representatives of different species of lactobacilli, then RA is related strains L.buchneri.

The disadvantages of the drug “Biosan” and its technology are that:

strains of lactobacilli, which constitute its basis, exhibit antagonistic activity only against certain pathogenic microorganisms, causing endometritis in cows;

the drug is characterized by limited duration of therapeutic action;

the method of its preparation of ethnological as for the maintenance of the reference crops, and production of the final form of the drug;

operational characteristics of the drug “Biosan” low due to the fact that the finished form of the drug is a mixture of cultural liquids.

These circumstances reduce the possibilities of use of the drug and do not preclude accumulation of phenotypic and genotypic changes in the cells of the original reference cultures during storage and displaced multiple transfers, inevitably leading to the reduction or complete loss of the required beneficial probiotic properties. Features of delivery and storage of liquid forms of the drug in real conditions of livestock farms do not allow to observe and control the injected dose at the time of application.

Therefore, the task before us was to create a standard with the required performance characteristics of the drug probiotics the first steps on the basis of a combination of lactobacilli strains, who would be in a sustainable ecological balance and not only maintained their properties when used together, but also enhanced therapeutic efficacy of the probiotic.

This problem is solved:

selection (selection) of lactobacilli strains with the desired beneficial probiotic properties;

establishing the optimal ratio of selected strains of lactobacilli in the Association of bacteria;

creation on the basis of the symbiotic Association in relation to each other bacteria of the finished form of the drug, providing standardization and stability of probiotic properties;

the development of technology for a long time to keep useful probiotic properties in the reference culture in the process of its maintenance, the accumulation of biomass of microbial cultures and after transfer of the culture fluid in a dehydrated state.

The selection criteria of lactobacilli was the following characteristics:

the ability to actively colonize the mucous membranes conditionally sterile cavities of humans and animals (respiratory, gastrointestinal and urogenital tracts) and to be able to rather long persistence in the host;

save and/or activate the normal biocenosis mucous membranes conditionally sterile cavities of humans and animals (on the respiratory tract, gastrointestinal and urogenital tracts);

the ability to actively suppress the growth and development of gram-positive and gram-negative pathogens, and harmful for the production of microorganisms;

the ability to suppress the growth of microorganisms that cause pulmonary, gastrointestinal, gynecological, skin diseases of different etiology (table 1).

Lactobacillus plantarum P4 was obtained by selection of lactobacilli of the microflora isolated from natural habitat - the vagina of healthy women of reproductive age in 1996 in Kirov there. Culture isolated from monocolonal and identified by the identifier Bergeys (1997) and classification Rogosa and Sharpe.

The strain is deposited in the collection of scientifically MO of the Russian Federation under No. 39.

The strain is characterized by the following features.

Cultural morphological traits

When grown in liquid medium MPC-1 for 48 h at a temperature of 36... 38° With lactobacilli grow as a homogeneous white sediment on the bottom of the tube, while the culture medium is transparent.

On the surface of solid agar medium small 4 after 48 h incubation at a temperature of 36... 38° formed With colony size 1,0... 2,0 mm white, opaque, having the form of hemispheres with smooth sharp edges, convex, shiny, smooth surface profile of the colonies to flavigny, consistency soft, slimy, stringy.

Pigment does not form.

The morphology of cells

Gram long curved sticks with straight or slightly rounded poles. The length of the cells is 1.5 to 5.0 µm in diameter 0.6 μm.

Physiological and biochemical characteristics of strain

Optional gone anaerobic, catalytically, zithromaxadministering. Ammonia from arginine does not form. Nitrates are not getting. Gelatino not thins.

Caroliina activity.

Sprayway: glucose, cellobiose, esculin, fructose, gluconate, galactose, lactose, maltose, mannitol, mannose, melibiose, salicin, D-raffinose, sorbitol, sucrose, trehalose, D-arabinose, ribose, melezitose.

Not fermented: rhamnose, xylose.

The final pH value on formatiruem sugars (4,8± 0,2) pH in the environment MPC-1. The type of fermentation - fermentation homofermentative with the formation of lactic acid without gas. Milk roll with the formation of the clot, the maximum titratable acidity which is (246± 15)° So

The temperature optimum of 36... 38° C. Temperature limit growth in air 15... 45° C. Limit value pH environment in which there is a growth - 5,0... 8,0 pH unit. The optimum value of pH - 6.3 pH unit.

In the process of life accumulates glycine, glutamine, isoleucine, and tyrosine.

Sensitivity to antibiotics

The strain is resistant to metronidazole, kanamycin, nalidixic acid, moderately resistant to fluoroquinolones, sensitive to amoxicillin, doxycycline, clarithromycin (table 2).

Adhesive properties

High adhesive strain. The index of the adhesive - the average number of lactobacilli attached to a single erythrocyte, when calculating the 25... 50 erythrocytes (no more than 5 red blood cells per field) - is 8.2± 1,14 (table 3).

Antagonistic activity.

The strain is characterized by a high antagonistic activity against the suppression of the growth and development of gram-positive and gram-negative pathogens, and harmful for the production of microorganisms (table 4).

Hydrogen peroxide does not produce, lysozyme, hemolytic, plasmalogens and gelatinase activity is not.

Plasmid composition.

The strain has four plasmids with molecular masses of 5.2 MT 5,9 MD, 8,2 MD 40 MD.

The method, conditions and medium composition for storage of cultures of strain.

Culture of the strain stored under oil on medium sheep, with slices of liver in test tubes at a temperature of 4... 6° with periodic reseeding 1 every 6... 12 months or in lyophilized form in a sealed ampoule (storage duration 2 years or more). Protective environment during drying of 1.5% poliglyukin.

Pic is b, conditions and medium composition for growth of strain

The strain is grown at 36... 38° in liquid medium MPC-1, solid medium Mrs-4 and liquid casein planting environment.

The composition of the medium MPC-1, g 1 l:

peptone - 10,0;

distilled water - up to 500,0;

the enzymatic hydrolysate meat - 100,0;

MnS4·4H2O - 0, 05;

MgS4·4H2O - 0, 02;

cysteine hydrochloric acid - 0,2;

glucose - 20,0;

lactose - 2,0;

To2NRA4- 2,0;

KN2RHO4- 2,0;

sodium acetate - 5,0;

ammonium acetate - 2,0;

tween-80 - 1,0;

liver extract - 100,0;

milk gidrolizovannogo - up 1000,0;

the concentration of hydrogen ions was adjusted to 6.2... pH of 6.4 and sterilized fluid ferry 20 min at 100° C.

For solid medium (Mrs-4) injected 15 g of agar.

The sowing medium g in 1 l of:

peptone - 10,0;

distilled water - up to 500,0;

yeast extract - 50,0;

hydrolyzed casein - 100,0;

MnSO4·4H2O - 0,05;

MgSO4·4H2O - 0, 02;

cysteine hydrochloric acid - 0,2;

glucose - 20,0;

To2NRA4- 3,0;

KN2RHO4- 3,0;

tween-80 - 1,0;

milk gidrolizovannogo up to 500,0;

the concentration of hydrogen ions was adjusted to 6.2... pH of 6.4 and sterilized fluid ferry 20 min at 100° C.

Lactobacillus buchneri P0 was obtained by selection lacto is atill of microflora, isolated from the natural environment of the vagina of healthy women of reproductive age in 1996 in Kirov there. Culture isolated from monocolonal and identified by the identifier Bergeys (1997) and classification Rogosa and Sharpe.

The strain is deposited in the collection of scientifically MO of the Russian Federation under No. 38.

The strain is characterized by the following features.

Cultural morphological traits

When grown in liquid medium MPC-1 for 72 h at a temperature of 36... 38° With lactobacilli grow as a homogeneous white sediment on the bottom of the tube, while the culture medium is transparent.

On the surface of solid agar medium small 4 through 96...120 h incubation at a temperature of 36... 38° formed colonies with a diameter of 0,5... 1,0 mm white and brownish color with a yellow tinge. Form colonies round with scalloped edges are wavy, uneven, but clear. Colony structure is coarse-grained. The surface of the colony is smooth, raised, bumpy and shiny. Profile colonies curved. The center colonies conical, gradually turns into a flattened peripheral portion. The texture is dense, viscous, difficult to be removed from the environment.

Pigment does not form.

The morphology of cells

Gram-positive short cuckoobananas sticks with clear contours and rounded poles length is 1.0... 1.5 mm and a diameter of 0.4-0.6 μm.

Phys the I-biochemical characteristics of the strain

Optional gone anaerobic, catalytically, zithromaxadministering. Ammonia from arginine does not form. Nitrates are not getting. Gelatino not thins.

Caroliina activity.

Sprayway: glucose, D-esculin, fructose, D-galactose, D-lactose, maltose, melibiose, sucrose, D-raffinose, D-xylose, gluconate, melibiose, ribose, arabinose.

Not fermented: rhamnose, sorbitol, mannitol, salicin, mannose, cellobiose, trehalose.

The final pH value on formatiruem sugars - (4,8± 0,2) pH unit in the base environment MPC-1. The type of fermentation - fermentation heterofermentative with the formation of lactic and acetic acid, ethanol and carbon dioxide. Milk roll with the formation of the clot, the maximum titratable acidity which is (246± 15)° So

The temperature optimum of 36... 38° C. Temperature limit growth in air 15... 45° C. Limit value pH environment in which there is a growth - 5,0... 8,0 pH units. The optimum value of pH - 6.3 pH unit.

In the process of life accumulates glycine, glutamine, isoleucine, and tyrosine.

Sensitivity to antibiotics

The strain is resistant to metronidazole, kanamycin, nalidixic acid, moderately resistant to fluoroquinolones; sensitive to amoxicillin, doxycycline, clarithromycin (table 2).

Adhesive properties

High adhesive strain. The index of the adhesive - the average number of lactobacilli attached to a single erythrocyte, when calculating the 25... 50 erythrocytes (no more than 5 red blood cells per field) - amounted to 9.1± 1,18 (table 3).

Antagonistic activity.

The strain is characterized by a high antagonistic activity against the suppression of the growth and development of gram-positive and gram-negative pathogens, and harmful for the production of microorganisms (table 4).

Hydrogen peroxide does not produce, lysozyme, hemolytic, plasmalogens and gelatinase activity is not.

Plasmid composition

Strain the plasmid has not.

The method, conditions and medium composition for storage of cultures of strain

Culture of the strain stored under oil on medium sheep, with slices of liver in test tubes at a temperature of 4... 6° with periodic reseeding 1 every 6... 12 months or in lyophilized form in a sealed ampoule (storage duration 2 years or more). Protective environment during drying of 1.5% poliglyukin.

The method, conditions and medium composition for growth of strain

The strain is grown at 36... 38° in liquid medium MPC-1, a dense Mrs-4 and liquid casein planting environment

The composition of the medium MPC-1, g 1 l:

peptone - 10,0;

distilled water - up to 500,0;

the enzymatic hydrolysate meat - 100,0;

MnSO ·4H2O - 0,05;

MgSO4·4H2On - 0,02;

cysteine hydrochloric acid - 0,2;

glucose - 20,0;

lactose - 2,0;

To2NRA4- 2,0;

KN2RHO4- 2,0;

sodium acetate - 5,0;

ammonium acetate - 2,0;

tween-80 - 1,0;

liver extract - 100,0;

milk gidrolizovannogo - up 1000,0;

the concentration of hydrogen ions was adjusted to 6.2... pH of 6.4 and sterilized fluid ferry 20 min at 100° C.

For solid medium (Mrs-4) injected 15 g of agar.

The sowing medium g in 1 l of:

peptone - 10,0;

distilled water - up to 500,0;

yeast extract - 50,0;

hydrolyzed casein - 100,0;

MnSO4·4H2O - 0,05;

MgSO4·4H2O - 0, 02;

cysteine hydrochloric acid - 0,2;

glucose - 20,0;

To2NRA4- 3,0;

KN2RHO4- 3,0;

tween-80 - 1,0;

milk gidrolizovannogo up to 500,0;

the concentration of hydrogen ions was adjusted to 6.2... pH of 6.4 and sterilized fluid ferry 20 min at 100° C.

Lactobacillus plantarum P4 and Lactobacillus buchneri P0 similar on a wide range of characteristics.

The main difference between strains are antagonistic properties: Lactobacillus buchneri P0 has a high antagonistic activity of Lactobacillus plantarum P4 has a high antagonistic activity.

Overall positive effect when the joint is used and both strains is the ability to form a stable symbiotic system, characterized citrinum and synergistic type of relationship and are able to exert a therapeutic effect on the human body, exposed to the impact of negative external factors.

This system exists when the ratio in the mixed cell culture strains No. 39 and No. 38 from 1:4 to 4:1, while maintaining the specified properties.

The basic properties of the strains and their mixtures in different proportions of microorganisms are given in tables 5 and 6.

The experiments showed that the resulting symbiotic system efficiency higher than monoculture of microorganisms increases the size of the zone of inhibition of growth of cultures of E.coli 0157, Sh.flexneri 170, Sh.sonnei 5063, Pr.vulgaris 177, St.aureus 209, Pr.mirabilis (table 7).

On the basis of a symbiotic mixture created a dry product containing along with cells of strains No. 39 and No. 38 in the ratio from 1:4 to 4:1 products of their metabolism. The total content of living organisms is not less than 1× 108cells in 1 ml registrationpage drug.

Dry product has a light brown color with a characteristic sour smell and taste. When dissolved in water dry product forms a homogeneous suspension. When dissolved in 0.85% sodium chloride solution, the pH of the solution is not less than 4.5 pH. The activity of acid generating drug - no less than 200° So

The dry preparation does not change its properties is within one year when stored in a dry dark place at a temperature of 2... 8° C. By the end of the specified term in the drug content of viable bacteria of at least 50% (table 8).

The safety of the drug “Bellakt was examined by studying the factors of pathogenicity in vitro and in vivo. It is established that the drug hemolytic, plasmalogens and gelatinase activities does not, is not toxigenic and toxic to white mice. The drug is specifically safe for Guinea pigs and white mice when administered via the mouth and under the skin. When examining the internal organs of slaughtered white mice pathological changes of the abdominal and thoracic cavity, gastrointestinal tract and parenchymatous organs is not revealed. In bacterial examination of internal organs (liver, spleen, lung, heart) method prints on the surface of nutrient agar (Mrs-4) growth of lactobacilli was not found. The obtained results indicate the absence of acute and chronic toxicities of the studied strains of lactobacilli and drug prepared on their basis (tables 9, 10 and 11).

The impact of introduction of the developed drug detoxifying function of the liver and processes in the CNS, is not revealed. Allergenic effect is absent.

The drug does not cause local reactions, aerogenes. Does not cause macro - and microscopic changes in the internal lanah experimental animals. Microscopic examination of histological preparations morphological changes were localized in nature, as evidenced by intactness reticulo-histiocytomas apparatus outside the lymph nodes draining the injection site.

The results of studies conducted using standard tests for the determination of the effect of study drug on the detoxifying function of the liver, also testified about his safety.

Example 1.

Sublimated Museum culture of lactobacilli strains No. 39 and No. 38 rehydration by diluting the contents of the vials of purified water for their subsequent separate growing in test tubes (MPC-1) and mattresses (s-4).

Received rehydration suspension cultures of strains of lactobacilli contribute 0.5... 1.0 ml separately in tubes with nutrient liquid MPC-1 (sowing dose of not less than 1× 106cells in 1 ml).

Tubes with culture is incubated at a temperature of 36... 38° C for 48 hours

In mattresses with beveled dense nutrient medium small 4 separately contribute to 4.0... 5.0 ml seed culture of lactobacilli strains from test tubes. Crops incubated at a temperature of 36... 38° C for 48 hours

The process of obtaining biomass for each strain includes separate introduction of the suspension agar cultures of lactobacilli strains in AIDS-cultiva the ora, containing a liquid nutrient medium MPC-1, 50 ml cultures in 1 l of liquid culture medium, and cultivation for periodic mechanical stirring speed of 60 rpm./min-1and a temperature of 36... 38° C for 48 hours

Culture grown in apparatus of the cultivators must meet the following requirements:

the concentration of lactobacilli, billion· ml-1- not less than 5.0;

pH, pH - 3,5... 4,0;

extraneous microflora is not allowed.

Separately grown in apparatus of the cultivators culture strains No. 39 and No. 38 decanted into a bottle in a desired ratio, stirred for 15 min and enter the protective environment of the calculation of 80 ml per 1000 ml of microbial suspension. After stirring the microbial suspension is poured into 2.0 ml in sterile penicillin vials that load in the dryer installation.

The drying process of the preparation includes:

freezing of the material in the vials to a temperature of minus 30... 40° With a speed of 10... 15°× h-1;

two-hour exposure of the frozen material;

one hour exposure of the frozen material under vacuum to 300 microns Hg;

drying of the material in the range from minus 30... 40° 0° at the rate of 1... 2°× h-1, during the next 4... 5 h bring the temperature of the material up to 25... 30° C and d is sosialt it within 7... 10 o'clock

The drug Bellakt” must meet the following requirements:

be a porous or a crystalline mass, in the form of tablets are light brown in color with a yellowish tinge;

this tablet should be dissolved in 5 ml of distilled water under stirring for 3 min;

to have the mass loss on drying not more than 3.5%;

pH after rehydration of the drug in 1.8 ml of 0.9% sodium chloride solution should be not less than 4.5 pH units;

not contain extraneous microflora;

to be harmless;

to contain not less than 1× 108live lactic acid bacteria in 1 ml registrationpage drug;

to possess antagonistic activity zone of inhibition of growth of test strains should be not less than 20 mm;

to be acid generating activity by Turner no less than 200° So

The drug is used in therapeutic and prophylactic purposes:

when intestinal dysfunctions as a result of dysbiosis caused by prolonged antibiotic, hormonal, radiation and other therapies, in stressful situations and stay in extreme conditions;

in acute intestinal infections (in the complex therapy of acute dysentery, salmonellosis, escherichiosis, viral gastroenteritis, etc.), with long-term bowel dysfunction staphylococcal etiology, as well as in the treatment of reconcile the cents after acute intestinal infections in the ongoing bowel dysfunction;

in acute and chronic inflammatory diseases of the colon and small intestine (colitis, enterocolitis), occurring against the background of violations of the microflora with deficiency or absence of lactobacilli;

in the treatment of a dysbacteriosis of a vagina (vaginosis);

in the treatment of purulent wounds, trophic ulcers, complicated by infection.

Method of use

Inside. The drug in the vial is dissolved in 5.0 ml of warm boiled water 25... 30° C. the resulting suspension was diluted in 1/2 Cup of warm water and drink 30...40 min before meals 2 times a day. The course of treatment is 5 days, then made a break for 1 to 2 weeks. If necessary, the course is repeated.

Intrawaginalno. The contents of the vial are dissolved in 5 ml of water. Obtained a suspension of the drug is impregnated with a sterile swab which is injected intrawaginalno and leave for 2... 3 hour treatment starts with 10... 12 days of the menstrual cycle.

Topically. The contents of the vial are dissolved 5 ml of water. Obtained a suspension of the drug is impregnated with a sterile swab which is applied to the damaged surface and stand at least 10 minutes

The stability of the Association of lactobacilli strains to metronidazole allows for comprehensive treatment of gastric ulcer and 12 duodenal ulcer if a test is positive for Helicobacter pylori, acute and hronicheskoj is trichomoniasis in women, combining prescription probiotics and antimicrobial and antiparasitic funds.

Example 2

Therapeutic preparation containing as the basis of two selected strains of lactobacilli and their metabolic products, due to the high specific activity of lactobacilli - colonizing ability of strain No. 38 and strong antimicrobial activity of the strain No. 39 in relation to a wide spectrum of pathogenic and conditionally pathogenic microorganisms - has a strong ability to adjust the dysbacteriosis of various etiologies in patients with humans and animals.

The drugs were used in the complex treatment of gastric ulcer and 12 duodenal ulcer, purulent paraproctitis, chronic nonspecific colitis, extensive or deep festering and rotting wounds with deep traumatic decomposition of soft tissues or deep corruption as a result of burns or frostbite.

During the treatment and observation of these patients were obtained the following results:

when gastric ulcer and 12 duodenal ulcer recovery time was reduced by 30... 50%. The drug is given 2 times a day. The course of treatment is 10 days;

paraproctitis - local application in the form of Turunc resulted in the rapid rejection of pus and wound cleansing to the third - fourth day, then the how without the use of the preparation process was delayed until 8... 9 days. The drug is used topically for the dressings once daily for 7 days;

chronic nonspecific colitis - oral use, which contributed to the normalization of microflora and removal of pain. The drug was applied inside 2 times a day. The course of treatment is 10... 15 days;

purulent-necrotic processes in daily use in the form of applications has been rapid relief of inflammation. The drug is used topically for the dressings once daily for 7 days.

After 2... 3 dressings disappeared tension and pain on the edge of wounds, inflammation was ogranichivalos, necrotic processes stopped, decreased purulent discharge, patients noted improvement in health. Early removal of the drug led to a new exacerbation of the inflammatory process in the wound, so applications used throughout the treatment period, which contributed to the rapid filling of the wound channel granulations and early (day 12) epithelialization of the wound defect. The drug was applied topically at dressing once daily for 10... 15 days.

Example 3.

Based on the Department of Microbiology of the all-Russian research veterinary Institute, poultry RAAS (G.C.-Petersburg) and the laboratory of veterinary immunology, research Institute of agriculture of North-East of them. Rudnicki (Kirov) shows high is th antagonistic activity of the investigated strains of lactobacilli against pathogens, isolated from sick calves, cows, pigs, and poultry of different ages farms Saratov (Mikhailovskaya poultry"), Tula (JSC APF “Ivan Lake) and Kirov ("Path of Lenin", "Izhevsk") areas (E. coli strain P16; Salmonella dublin 12/1; Pasteurella multocida 98/4; Pseudomonos aeruginosa L-4; Bordetella bronchiseptica W-97/4; Staphylococcus al-bus KB 97/2; S.enteritidis and St.aureus).

In experiments based poultry production complex JSC APF “Ivan-Ozero” (Saratov region) and sue Mikhailovskaya poultry” (Tula region) have shown that feeding (100 ml suspension with a cell concentration of 4... 5 billion 15000 heads) and feeding (daily dose of 3... 4 million microbial cells per 1 head) of the probiotic Bract young chicken cross the "T-46" and broiler chickens aged 1 to 7 days increases the safety of the young birds by 3%.

Such a therapeutic effect on the first day of life birds observed in experiments on Local poultry (Kirov region). Received therapeutic effect of the probiotic on the first day of life young was reproduced on the chicken cross “Smena-2” Aranskoy poultry (Kirov region). It is shown that the death of chickens in the control group to 15 days was higher than experienced. Thus the gain of body weight in the experimental group of chickens was higher than that of the chickens of the control group. Increasing the dose of the probiotic twice in the period is the increase in bird deaths from colibacillosis allowed to reduce bird deaths by 10%. At the same time daily increase in the number of surviving birds in the experimental group was significantly higher than in the control group.

1. The bacterial strain Lactobacillus plantarum P4 scientifically MO of the Russian Federation No. 39 symbiont strain of Lactobacillus buchneri P0 scientifically MO of the Russian Federation No. 38, used for preparation of the preparation of the probiotic.

2. The bacterial strain Lactobacillus buchneri P0 scientifically MO of the Russian Federation No. 38 symbiont strain Lactobacillus plantarum P4 scientifically MO of the Russian Federation No. 39, used for preparation of the preparation of the probiotic.

3. Probiotic preparation, facilitating correction of dysbacteriosis of various etiologies in humans and animals based on the biomass of lactobacilli, characterized in that as the biomass it contains dried mixture of bacteria strains Lactobacillus plantarum P4 scientifically MO of the Russian Federation No. 39 and Lactobacillus buchneri P0 scientifically MO of the Russian Federation No. 38 in the ratio of microorganisms from 4:1 to 1:4 when the contents of living cells is not less than 1×108cells per 1 g of the drug.



 

Same patents:

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: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 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: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention proposes a method for preparing exopolysaccharide by culturing microorganisms in nutrient medium containing one or more carbon source assimilated by microorganisms and caruba seeds fraction as nitrogen organic source. Applying the proposed method provides preparing exopolysaccharide eliciting improved organoleptic, sensor and visual properties. Invention can be used in building, paper, textile, cosmetic, food, oil output industry and agriculture.

EFFECT: improved preparing method.

15 cl, 4 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

Up!