Strain bifidobacterium longum 379-in used for preparing bacterial preparations, biologically active supplements for food, ferments, fermented-dairy and nonfermented dairy foodstuffs, hygienic and cosmetic agents
FIELD: biotechnology, food and medicinal industry, microbiology.
SUBSTANCE: the strain Bifidobacterium longum 379-IN is obtained by selection without using methods of genetic modification of the strain Bifidobacterium longum B379M and distinct by ability to utilize insulin. The strain is deposited in GKNM GU "MNIIEM named for G. N. Gabrichevskiy Russia Ministry of Public Health" at № 172. The strain shows high technological effectiveness, accumulates biomass with substrates of vegetable origin and artificial nutrient media for short periods with concentration of bifidobacteria, it elicits acid-forming and antagonistic properties with respect to pathogenic and putrid microflora. This allows its using in manufacturing bacterial preparations, biologically active supplements for food, fermented-dairy and nonfermented-dairy foodstuffs, ferments, hygienic and cosmetic agents providing probiotic effect and normalization of microbiocenosis in human body, among them in gastroenteric and urogenital tracts, cutaneous and mucosa integuments. Invention can be used in manufacturing bacterial preparations, biologically active supplements for food, fermented-dairy and nonfermented-dairy foodstuffs, hygienic and cosmetic agents.
EFFECT: valuable properties of strain, expanded assortment of similar agents.
The invention relates to biotechnology, food processing and medical industry and can be used in the production of bacterial preparations, biologically active additives to food, fermented milk fermented and non-fermented foods, hygienic and cosmetic products.
Known strain of Bifidobacterium longum B379M, which is the closest analogue to the claimed strain (Patent SU 849779). The strain isolated from the intestine of healthy adult deposited in Vniigenetika in CMPM under the number In 2000 and has the following features:
1. Cultural and morphological characteristics.
Gram polymorphic still sticks with bifurcation and thickening at one or two ends. Incubation with 37,5-38°C.
In liquid nutrient media (liver, hydrolysate-dairy, casein-yeast) with the addition of 0.75-1% agar grow throughout the volume, except for the zone of aerobiosis. Logarithmic growth phase ends by 24 hours. Individual colonies are of the form a compact, dense grains.
2. Physiological and biochemical characteristics.
Obligate gone anaerobic. The enzyme catalase does not form, gelatino not thins, glucose decomposes to acetic acid and L+lactic acid without the formation of gas. Ferments lactose, arabinose, xylose, ribose, weak and fickle - cellobiose and inulin; weak, in pain who s late - mannose, does not decompose mannitol, inulin, sorbitol, Inositol, salicin.
Energetic kislotoobrazoutei. Skachivat milk to 18-20 hours of cultivation with the formation of a clot of soft consistency.
Acid-forming activity in the hepatic environment: 24 h - 100-110°T, 72 - 150-180°T; on GM medium: up to 120°T 240°T, respectively.
3. Antagonistic properties of the strain manifests against Shigella sonnei, Flexner, enteropathogenic Escherichia coli, Staphylococcus.
In the human colon, the main nutrient substrate for bifidobacteria are the carbohydrates of plant origin. However, as can be seen from the data presented, strain Bifidobacterium longum B379M adapted to the substrates on the basis of animal protein and carbohydrate, inulin decomposes weak and unstable.
Consequently, when injected into the human body lack of viability, sustainability and colonizing activity when passing through the digestive tract.
The objective of the invention is to improve the digestion of carbohydrates due to the utilization of inulin from the culture medium is valuable from a medical point of view of the carbohydrate with a wide range of indications therapeutic and prophylactic properties, including with prebiotic activity.
To obtain Bifidobacterium longum 379-IN was conducted breeding piece is MMA Bifidobacterium longum B379M. Selection was carried out without using the techniques of genetic modification by multiple passages through a synthetic nutrient medium of the following composition:
to 1 liter of distilled water:
Epidermal-0 10 g
inulin 10 g
Cysteine, 0.5 g
Sodium chloride 2.5 g
Magnesium sulfate 0.5 g
Sodium acetate 0.3 g
Agar-agar 0.75 g
After dissolving the above components in the environment brought the pH to 7.2 to 7.4, was poured into the flasks and sterilized at 0.5 psi for 30 minutes.
Primary seed material Bifidobacterium longum B379M in the amount of 5% was introduced into the above medium and cultivated at 37°With 24 hours. It was further held 35 passages grew up in this environment culture through the medium of the same composition, after each of which made the selection of clones on the grounds of maximum accumulation of biomass during the cultivation cycle. If first passages got microbial yield no more than 1051 ml, then at the 30th passage, the number of CFU of bifidobacteria reached 109and remained at this level in subsequent subcultures. This is due to the increased production of inulinase in the long passage on the medium containing inulin.
As a result of breeding the resulting strain Bifidobacterium longum 379-IN which is deposited in the Public collections of microorganisms-performance, the representatives of normal microflora of man SE “mniam them. Hingamisega Ministry of health of Russia” No. 172.
The inventive strain can be used in the production of bacterial preparations in various forms, including capsules, suppositories, ointments; biologically active additives to food, sourdough, fermented milk fermented and non-fermented foods, juices, drinks, kvass, etc. Also strain Bifidobacterium longum 379-IN can be used in the production of hygienic and cosmetic products, for example, creams, gels, nourishing masks.
The technical result of the claimed invention to provide a bacterial preparations, biologically active additives to food, sourdough, fermented milk fermented and non-fermented foods, hygienic and cosmetic products, which as of the applicable commencement probiotic orientation contain Bifidobacterium longum 379-IN, which has a higher biological activity and resistance to external influences, which provides a more effective colonization of the intestine and the recovery of the resident microflora, and will expand the Arsenal of technical means of preventive and curative orientation. In addition, the absorption of a wider range of carbohydrates increases as the biological activity and stability of the proposed strain to external influence is, and its adaptability: it accumulates biomass not only on conventional production environments, skachivat milk is getting sour-milk product with valuable biological and organoleptic properties, but also cultivated on nutrient substrates from cheap natural plant materials.
The inventive strain has the following properties.
Cultural-morphological: gram-positive homogeneous sticks, evenly painted methylene blue, located mainly chain.
Physiological-biochemical: ferments lactose, inulin, cellobiose with the formation of acetic and lactic acid and traces of formic and propionic acids.
The arabinose, xylose, mannose, mannitol, sorbitol, salicin not fermented. All taken in the production of nutrient media grows with the accumulation of biomass of bifidobacteria not less than lg 8-9 microbial numbers for 16-20 hours. Possesses a high activity of acid generating, skvashivaya milk for 16-18 hours with the formation of a clot of soft consistency and good organoleptic properties. Under cultivation in the medium with inulin biomass yield exceeds the prototype 4 of the order.
The data presented in table 1.
The number of CFU in 1 ml and the active key is latest in° T after 16-18 hours of cultivation
|Nutrient medium||C. longum||WM||C. longum 379-IN|
|Epidermal with inulin||1,5x105||40°T||1,1x109||78°T|
|An aqueous extract of the tubers of Jerusalem artichoke||1,2x105||35°T||5,5x108||70°T|
Antagonistic properties of the inventive strain manifested against Shigella sonnei, Flexner, Salmonella, enteropathogenic Escherichia coli, Staphylococcus, Proteus.
Thus, these data indicate that a new strain of Bifidobacterium longum 379 IN high technology, is expanding biomass of bifidobacteria not only on production environments and milk, and vegetable substrates containing carbohydrates of vegetable origin, in particular inulin. This paragraph is more than functional value products and preparations, prepared on the basis of such a strain. When injected into the human body a culture of this strain utilizes more plant substrates, resulting in more intensive colonization of the mucous membrane of the intestine, the displacement of pathogenic and putrefactive microflora, more effectively recovers the resident microflora.
Example 1. Getting dried bacterial preparation.
To a suspension of 22 to 24-hour culture of Bifidobacterium Bifidobacterium longum 379-IN adds a protective environment drying (sucrose 80% and gelatin 8% in the volume of 10-12%) and poured into 5 ml penicillin vials, frozen at 40-45°C and freeze-dried from the frozen state in a vacuum drying apparatus. The resulting preparation contains 1 g of dry biomass of not less than 108living cells of bifidobacteria.
Example 2. The use of Bifidobacterium longum 379-IN as a starter for the preparation of a medicinal product is a functional food.
Suspension 14-16 hour culture of Bifidobacterium longum 379 IN making sterile milk in an amount of 5% by volume of the milk, mix and cultivate 16-18 hours prior to the formation of a clot, then the product is cooled and drained. As a result of receiving treatment-and-prophylactic fermented milk product containing Bifidobacterium not less than 108in 1 ml.
Example 3. Getting the liquid form of biologists who Eski supplements.
The inoculum culture of Bifidobacterium longum 379-IN in the amount of 5-10% contribute in a fermenter with a nutrient medium of the following composition per 1 liter of product: 5-%aqueous extract of Jerusalem artichoke, 2% of hydrolyzed collagen, 0.5% of sodium chloride, 0.5% rosehip extract, 1% pectin, 0.1% agar-agar and cultured for 6-8 hours at a 37.5-38°and poured into sterile containers. The result is a biologically active food Supplement containing bifidobacteria not less than 1081 ml of the Obtained additive has a strong immunocorrective action.
Example 4. Obtain a capsulated form of biologically active food supplements.
To a suspension of 22 to 24-hour culture of Bifidobacterium Bifidobacterium longum 379-IN type environment drying in the amount of 20% and spray dried. 1 g of the prepared biomass contains not less than 108viable bifidobacteria.
For encapsulation prepare a mixture, which is in equal parts includes the following ingredients: dried biomass of bifidobacteria, lactulose, pectin, dextrin. To a mixture of calculating the resulting mass was added 1% of magnesium stearate and 1% of ascorbic acid. After thorough mixing spend encapsulation in capsules No. 0 to 0.3, One capsule contains not less than 108viable bifidobacteria.
Example 5. The use of Bifidobacterium longum 379 IN the La receiving cosmetic products.
The dry biomass of bifidobacteria strain containing bifidobacteria not less than 1091 g enter into the composition of the nutrient cream at the rate of 1 g per 100 g of cream.
Example 6. The use of Bifidobacterium bifidum 379-IN to make juice.
The inoculum culture of Bifidobacterium bifidum 379 IN the amount of 5-10% contribute in a fermenter with a nutrient medium of the following composition per 1 liter of product: 5-%aqueous extract of Jerusalem artichoke, 2% of hydrolyzed collagen, 0.5% of sodium chloride, 0.5% rosehip extract, 1% pectin, 0.1% agar-agar and cultured for 6-8 hours at a 37.5-38°and contribute to the juice mix and pour in the mixture. The result is a product with a concentration of live bifidobacteria not less than 106/ml.
Example 7. The use of Bifidobacterium bifidum 379-IN to receive hygiene products.
The dry biomass of bifidobacteria strain containing bifidobacteria not less than 1091 g enter into the composition of toothpaste at a rate of 1 g per 100 g of paste.
Strain Bifidobacterium longum 379 IN GCNM state mniam health Ministry to them. Hingamisega No. 172 used to obtain bacterial preparations, biologically active additives to food, sourdough, fermented milk fermented and non-fermented foods, hygienic and cosmetic products.
FIELD: biotechnology, microbiology.
SUBSTANCE: method for preparing liquid lactobacterin involves regeneration, culturing passages of lyophilized culture and culturing ferment of lactobacilli in liquid lyophilized nutrient medium containing dry defatted milk enzymatic hydrolyzate with the content of amine nitrogen 1 485 mg%, 30.0 ± 3.0 g/l; yeast concentrated autolyzate, 110.0 ± 10 g/l; food agar, 0.8 g/l, and distilled water, up to 1 l. Culturing ferment is carried out up to accumulation of biomass of lactobacilli 109-1010 CFU/ml. Then 10-30% of supernatant liquid is removed from the ready product and replaced it with equal volume of fresh nutrient medium. Invention provides simplifying technology in preparing liquid lactobacterin and to elevate the storage period of viable lactobacilli. Invention can be used in producing probiotic preparations.
EFFECT: improved preparing method.
1 tbl, 3 ex
FIELD: biotechnology, microbiology, veterinary science.
SUBSTANCE: for preparing a preparation cells of microorganism Escherichia coli VKM CR-322D is cultured in nutrient medium containing Hottinger's broth, glucose, yeast extract, manganese sulfate, potassium hydrophosphate, sodium chloride and tap water in the content of amine nitrogen 125-155 mg%. Glucose is added by batch portions in the process of culturing cells that is carried out at temperature 30-31oC at stirring and aeration for 10-12 h. Prepared cultural fluid containing 3 x 109 bacterial cells/ml is mixed with protective sucrose-gelatin medium and subjected for lyophilic drying. Dried mass is stored under nitrogen that enhances safety of viable cells in the preparation. Applying the preparation for prophylaxis and treatment of agricultural and domestic animals and poultries with gastroenteric diseases provides its effectiveness.
EFFECT: improved preparing method, valuable veterinary properties of preparation.
17 cl, 8 ex
FIELD: biotechnology, biochemistry, microbiology.
SUBSTANCE: the strain Bacillus circulans B-65 a producer of cyclodextrin glucanotransferase is isolated and selected form the soil sample by culturing in nutrient medium with amylolytic activity 12.17 U/ml and cyclodextrinogenic activity up to 0.530 U/ml. Cyclodextrin glucanotransferase isolated from B. circulans B-65 shows the high degree for conversion of starch to cyclodextrins and this enzyme is specific for formation of β-cyclodextrin. Invention can be used in food industry for preparing cyclodextrins and cyclodextrin glucanotransferase used in different branches of industry.
EFFECT: improved isolating method, valuable properties of strain and enzymes.
9 cl, 8 tbl, 2 ex
FIELD: biotechnology, microbiology, dairy industry.
SUBSTANCE: the strain of microorganism Lactobacillus lactis VKPM B-8354 is prepared without using mutagens and genetic methods and shows resistance against broad spectrum of lactophages. The strain ferments effectively milk from different trading sorts, with broad range of fatness and different methods of thermal treatment. Individual specific properties of the strain allows its applying as a monostrain ferment. Curd obtained with applying the strain L. lactis VKPM B-8354 shows good organoleptic qualities, nonacid taste and homogenous consistence. The strain is suitable especially for plants with small volume of manufacture but with varied assortment.
EFFECT: valuable properties of strain.
3 tbl, 2 dwg, 5 ex
FIELD: biotechnology, microbiology, agriculture.
SUBSTANCE: the strain Lactobacillus plantarum 578/25 is obtained by method of step-by-step selection and selected by its ability to produce significant amount of crude protein and to accumulate the biomass. The strain is deposited in the VGNKI collection at number VGNKI-03.04.09.-DEP. Invention provides eliminating the pollution of environment in producing the protein fodder, to elevate the protein specific yield, to reduce energy consumptions in preparing protein fodder, to simplify and to accelerate the process of its preparing, to simplify apparatus equipment, to utilize waste in manufacturing using the natural raw.
EFFECT: valuable properties of strain.
2 tbl, 10 ex
FIELD: biotechnology, agriculture, microbiology.
SUBSTANCE: invention relates to a new isolated strain of Lactobacillus acidophilus 1660/08 as a producer of the protein fodder. The strain Lactobacillus acidophilus 1660/08 is obtained by the selection method and selected by its ability to form significant amount of crude protein and to accumulate the biomass. The strain is deposited in the VGNKI collection at number VGNKI-03.04.10.-DEP. Invention provides eliminating the environment pollution in producing the protein fodder, to enhance the specific protein yield, to reduce energy consumptions in preparing protein fodder, to simplify and to accelerate the process in its preparing, to simplify equipment fitting out and to utilize waste in manufactures using natural raw.
EFFECT: valuable properties of strain.
2 tbl, 10 ex
FIELD: biotechnology, microbiology, agriculture.
SUBSTANCE: the strain Lactobacillus buchneri 600 is isolated from barley grains at milkwax stage of ripeness. The strain Lactobacillus buchneri 600 is registered in 05. 08. 2002 year in the All-Russian State collection of microorganism strains used in veterinary science and animal husbandry at number Lactobacillus buchneri VGNKI 02.08.04-DEP and deposited in the collection 000 "Biotrof". Invention provides the more effective multiplication of the strain in maize ensilaging green mass and preserving flattened grains with enhanced formation of lactic acid, inhibition of putrid microflora that allows preparing fodder from vegetable raw with enhanced quality. Invention can be used in fodder production for ensilage maize green mass and preserving flattened grains.
EFFECT: valuable properties of strain.
3 tbl, 2 ex
FIELD: biotechnology, microbiology, medicine.
SUBSTANCE: invention relates to the strain Lactobacillus paracasei CNCM I-2116 used for diarrhea prophylaxis causing by pathogenic microorganisms. Supernatant of this strain culture elicits ability to prevent colonization of intestine with pathogenic microorganisms causing diarrhea also and this strain is designated for preparing agent used for prophylaxis and/or treatment of disorders associated with diarrhea. Agent for oral administration represents therapeutically effective dose of the strain L. paracasei CNCM I-2116 or supernatant of its culture and acceptable foodstuff. Invention provides the enhanced viability of the strain in its applying and effectiveness in prophylaxis of adhesion to intestine cells and invasion to intestine cells of pathogenic microorganisms causing diarrhea.
EFFECT: valuable medicinal properties of strain.
5 cl, 8 dwg, 10 ex
FIELD: biologically active substances.
SUBSTANCE: invention relates to medical desinsection, household chemistry, and plant protection agents, in particular to contact- and intestinal-type insecticidal and acaricidal preparation, which can be used in medical desinsection area, agriculture, personal support farms, and urban domestic life for controlling pest insects and ticks. Insectoacaricidal agent of invention containing an active principle, preferably of pyrethroid series (e.g. cypermetrin), and solvent additionally contains effective amounts of dodecyldimethylammonium bromide molten together with active principle. Amounts of dodecyldimethylammonium bromide and cypermetrin may vary each between 10 and 500 g/L. Solvent can be deodorized kerosene.
EFFECT: achieved stability, efficiency, low price, and toxic safety of claimed agent.
5 cl, 2 tbl, 2 ex