WO2022154692A1 - Complexe de bifido-bactéries et souches entrant dans la composition du complexe - Google Patents

Complexe de bifido-bactéries et souches entrant dans la composition du complexe Download PDF

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WO2022154692A1
WO2022154692A1 PCT/RU2021/000582 RU2021000582W WO2022154692A1 WO 2022154692 A1 WO2022154692 A1 WO 2022154692A1 RU 2021000582 W RU2021000582 W RU 2021000582W WO 2022154692 A1 WO2022154692 A1 WO 2022154692A1
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strain
consortium
bifidobacteria
bifidobacterium
strains
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PCT/RU2021/000582
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Владимир Владимирович БЕСПАЛОВ
Ирина Валерьевна КАЗАКОВА
Людмила Владимировна МЕРКУЛОВА
Алексей Александрович РОГОВ
Вера Васильевна ЧЕШЕВА
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Общество с ограниченной ответственностью "Научно-производственное объединение Бифилайф"
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745—Bifidobacteria
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor

Definitions

  • the group of inventions connected by a single inventive concept relates to biotechnology, food industry, pharmacology, cosmetology and can be used to obtain bifid-containing products.
  • Bifidobacteria are the most important symbionts of the human gastrointestinal tract. In children by the age of 3 months, their share in the microbiome is more than 90%, which indicates the leading role of this group of bacteria in the formation and development of human homeostasis systems from an early age. They have a morphokinetic effect, form enzymes involved in the metabolism of proteins, carbohydrates, lipids and nucleic acids, produce biologically active compounds, in particular vitamins PP, B1, B6, B9, B 12.
  • Bifidobacteria perform immunogenic and antimutagenic functions, and also participate in detoxification of exogenous and endogenous toxic agents, which is becoming especially relevant in modern conditions of massive anthropogenic impact on the biosphere, in which not only nature suffers, but also man himself.
  • a multifaceted positive effect on the human body allows us to consider bifidobacteria as an effective basis for designing bifid-containing products for various purposes based on promising strains of bifidobacteria.
  • the biocenosis is dominated by the species Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium adolescentis, Bifidobacterium infantis (Bifidobacterium longum spp. infantis) and Bifidobacterium breve. It is these five species of bifidobacteria that are dominant and, in various combinations, create conditions formation of the composition and normal state of the microbiota of a healthy person. (Kozlova E.P., Kurnosova N.A., Feklisova L.V.
  • the bifidoflora has a certain structure, according to which only a consortium of bifidobacteria has a dominant influence on the formation of the microbiota characteristic of a healthy intestine (proven in 88% of the cases studied), while the presence of one or two strains always leads to dysbiosis ( proven in 42% of cases of type 1-2 dysbiosis and in 86% of type 3 dysbiosis).
  • Ivanova E.V The role of bifidoflora in the associative symbiosis of the human intestinal microbiota. Abstract of the thesis ... Doctor of Medical Sciences. Chelyabinsk 2009, p 13).
  • Bifidobacterium longum spp.infantis strains 73-15 and 79-43 Bifidobacterium bifidum strains 1 and 791, 791 BAG, LVA-3, RO71, 8-3; Bifidobacterium breve strain 79-88, 79-119, RO70; Bifidobacterium animalis spp.animalis strain DN-173010 (Danone/Dannon); Bifidobacterium animalis spp. lactis Bbl 2 (Chr. Hansen), HN019 (DR10) (Danisco); Bifidobacterium adolescentis strain G7513, GO-13, MC-42.
  • Bifidobacterium animalis subsp. lactis in various variations of the name, isolated from the rumen of an animal, has the smallest genome size. This is due to the fact that it lives in an environment with a constant and simple composition of the nutrient substrate, in conditions that do not require constant adaptation of the microbial cell. This is confirmed by the presence of only two complete copies of rRNA operons, while other species have up to four to five rRNA operons. Reduced adaptive properties of strains of B. animalis subsp. lactis do not lead to the formation of sustainable eubiosis (Bukharin O.V., Perumova N.B., Ivanova E.V. Bioflora in human associative symbiosis. - Ekaterinburg: Ural Branch of the Russian Academy of Sciences, 2014. - p. eleven)
  • the closest analogue (prototype) of the claimed consortium in terms of species composition is a consortium of five types of breeding strains of bifidobacteria: Bifidobacterium bifidum 791-MB, Bifidobacterium longum B 379M-MB, Bifidobacterium adolescentis G 7513-MB, Bifidobacterium infantis 73-15-MB, Bifidobacterium breve 79 -119- MB used for the preparation of sour-milk, non-fermented products, dietary supplements, bifid-containing preparations, cosmetic and hygiene products (RU 2261909 C1, publ. 10.10.2005).
  • the strains that make up the consortium have similar cultural and biological properties; when co-cultivated, the consortium actively reproduces on nutrient media with the accumulation of production biomass in a short cultivation period, and has specific activity.
  • the finished product contains 10 9 living cells of bifidobacteria in 1 ml.
  • the disadvantage of this consortium is the difficulty of obtaining selection strains, which involves 50 cycles of sequential lyophilized drying and recovery on a thioglycol medium and defatted milk, while the maximum titer of bifidobacteria is only 10 9 living cells of bacteria in 1 ml of the finished product.
  • the technical problem solved as a result of the creation of the present invention is the development of a consortium that includes selection strains selected in composition not only taking into account biocompatibility, but also giving the consortium resistance to a wide range of antibiotics, damaging factors that ensure the process of digestion in the gastrointestinal tract, expressed antagonistic activity with the presence of a high titer of bifidobacteria 10 p CFU per 1 g of dry biomass and the production of bifidus-containing products with high therapeutic and prophylactic properties on its basis, as well as simplification of obtaining a consortium.
  • the technical result lies in the fact that the proposed consortium of bifidobacteria used for the preparation of bifid-containing products, and the strains included in its composition, are able to actively accumulate microbial biomass on various nutrient media to a high concentration, have high antagonistic activity in relation to the main test cultures, which is the cause of food poisoning, and, in addition, they have high superoxide dismutase activity with pronounced antioxidant activity, while superoxide dismutase activity is not lower than 37 U/mg of microbial mass protein.
  • strains included in the consortium can be easily identified based on the proposed nucleotide sequences of their DNA fragments.
  • the Bifidobacterium longum B379M-BL strain used to obtain the consortium was deposited in the VKPM Federal State Unitary Enterprise "GosNIIGenetika" under registration No. Ac-2037, and the nucleotide sequence of the genome fragment of this strain SEQ ID NO: 2.
  • the known production strains of five species of bifidobacteria are initially selected without the use of genetic modification methods as a result of 30 cycles of sequential lyophilized drying and recovery on the Bifilife M medium, and then a consortium is obtained by co-cultivation of strains.
  • the method used for selecting breeding strains took into account the study of their antibiotic resistance, cultural, antagonistic, morphological, physiological and biochemical properties, molecular genetic identification and biocompatibility in the co-cultivation of strains, taking into account the growth kinetics of each of them. This made it possible to create a consortium capable of correcting microecological disturbances more efficiently and safely.
  • figure 1 shows a phylogenetic tree for a strain of Bifidobacterium bifidum 791-BL
  • figure 2 shows a phylogenetic tree for a strain of Bifidobacterium longum In 379M-BL
  • fig.3 shows the phylogenetic tree for the Bifidobacterium adolescentis G 7513-BL strain
  • figure 4 shows the phylogenetic tree for the strain Bifidobacterium infantis 73-15-BL
  • figure 5 shows the phylogenetic tree for the strain Bifidobacterium breve 79-119-BL
  • figure 6 presents a graph of the content of bifidobacteria from the time of cultivation.
  • VZ (calcium pantothenate) - 0.001 g
  • the primary inoculum of the production strain Bifidobacterium bifidum 791-MB in a volume of 5% of the volume of the cultivation medium was introduced into the above medium and cultivated at a temperature of 37-38°C for 24 hours. Then 30 cycles of drying-recovery were carried out on the nutrient medium "Bifilife M".
  • strain Bifidobacterium bifidum 791-BL are rod-shaped variable cells with V-shaped bifurcations at one or two ends, immobile, form clusters in the smear, sometimes chains, gram-positive. Facultative anaerobe, the optimum cultivation temperature is 37-38°C, pH 7.0 ⁇ 0.2.
  • Ferment milk with the formation of a stable clot by 16-18 hours. Gas-forming ability, catalase-forming ability, gelatin liquefaction are absent. Acid-forming activity: by 24 h - 70 ° T, by 72 h - 90-100 ° T (on Blaurock's hepatic medium), on hydrolysate-milk medium - up to 100 ° T and 180 ° T, respectively.
  • the maximum titer of bifidobacteria in a hydrolysate-milk medium after 24 hours of cultivation is 1x10 10 CFU/ml; the maximum titer of bifidobacteria in sterile skimmed milk without growth factors after 24 hours of cultivation is 1-3x10 10 cfu/ml.
  • the strain grows on nutrient media used in production with biomass accumulation of at least 1g 9.8-10.2 microbial number for 16-18 hours of cultivation.
  • the claimed strain has a pronounced antagonistic activity against pathogenic and conditionally pathogenic microorganisms: Shigella flexneri, Escherichia coli, Salmonella typhimurium, Shigella sonnei, Candida albicans, Proteus vulgaris, Staphylococcus aureus, Klebsiella pneumonia, Citrobacter freundii, Bacillus subtilis.
  • Table 1 shows the antagonistic activity of the strain against some of the aforementioned microorganisms.
  • the antagonistic activity of a monostrain of bifidobacteria was determined by the zone of growth inhibition of test cultures of pathogenic and conditionally pathogenic microorganisms by the method of perpendicular strokes.
  • a production-useful feature of the B. bifidum 791-BL strain which distinguishes it from the B. bifidum 791-MB strain, is that the culture grown under stationary conditions on the Bifilife M medium in a volume of 300 cm 3 has an increased superoxide dismutase activity. .
  • the SOD activity of the B. bifidum 791-BL strain is 27.7 U/mg of the protein contained in the bacterial mass.
  • the revealed SOD activity of the culture of bifidobacteria indicates that the strain has a pronounced probiotic effect, which can manifest itself both in vivo and in vitro.
  • the high activity of acid formation, the resistance of the strain to major antibiotics contributes to the effective restoration of normal intestinal microflora against the background of antibiotic therapy.
  • Freshly prepared strain B. bifidum 791-BL is stored on a nutrient medium with the addition of a protective medium containing sucrose, gelatin, sodium citrate, and freeze-dried in ampoules. The strain is well stored at a temperature of 6-8 °C without loss of activity during the year.
  • the studied strain of Bifidobacterium bifidum 791-BL belongs to the following systematic group: Bacteria; Actinobacteria; Actinobacteridae; Bifidobacteriales; Bifidobacteriaceae; Bifidobacterium, and the homology with the species Bifidobacterium bifidum is 97%. Analysis in the RDP II 16S rRNA database showed homology with the same bacterial species Bifidobacterium bifidum.
  • the tested Bifidobacterium bifidum 791-BL strain can be attributed to the Bifidobacterium bifidum species (97%).
  • Pavlicek A et al "Free-Tree-freeware program for construction of phylogenetic trees on the basis of distance data and bootstrap ⁇ ackknife analysis of tree robustness".
  • the strain is not genetically modified and does not contain genes from other organisms; transferred resistance genes; genetic changes associated with the use of genetic engineering techniques; the strain is not zoopathogenic and phytopathogenic.
  • strain Bifidobacterium longum B 379M-BL the selection of the strain Bifidobacterium longum B 379M-MB was carried out, which was deposited on July 15, 2004 in the VKPM Federal State Unitary Enterprise GosNIIgenetika of the Ministry of Education and Science under No. Ac-1689.
  • the original strain is protected by RF patent No. genetic modification through multiple cycles of lyophilized drying and recovery on the previously mentioned nutrient medium "Bifilife M".
  • the primary inoculum of the production strain Bifidobacterium longum B 379M-MB in a volume of 5% of the volume of the cultivation medium was introduced into the above medium and cultivated at a temperature of 37-38°C for 24 hours. Then 30 cycles of drying-recovery were carried out on the nutrient medium "Bifilife M".
  • Facultative anaerobe optimum temperature 37 - 38°C, pH 7.0 ⁇ 0.2.
  • Acid-forming activity when cultivated on sterile skimmed milk, titratable acidity reaches 100° T by 18 hours, 120° T by 20 hours, and 145° T by 24 hours.
  • the maximum titer of bifidobacteria in the milk clot reaches 18 hours of cultivation and is 10 9 CFU/ml.
  • the strain grows on nutrient media used in production with biomass accumulation of at least 1g 9-10 microbial number for 16-18 hours of cultivation.
  • the strain has a pronounced antagonistic activity against pathogenic and conditionally pathogenic microorganisms: Shigella sonnei, Shigella flexneri, Salmonella typhimurium, Escherichia coli, Candida albicans, Proteus vulgaris., Staphylococcus aureus, Klebsiella pneumonia, Citrobacter freundii, Bacillus subtilis.
  • Table 4 shows the antagonistic activity against some of the aforementioned microorganisms.
  • the antagonistic activity of a monostrain of bifidobacteria was determined by the zone of growth inhibition of test cultures of pathogenic and conditionally pathogenic microorganisms by the method of perpendicular strokes.
  • Determination of the level of resistance of the strain to antibiotics was carried out by the method of serial dilutions in a liquid nutrient medium - sterile skimmed milk. Various concentrations of sterile solutions of antibiotics and a 16-hour culture of bifidobacteria in an amount of 5% were introduced into the nutrient medium. Stability was determined by the change in titratable acidity in the samples after 48 h of incubation at a temperature of 37-38°C.
  • a production-useful feature of the B. longum B379M-BL strain which distinguishes it from the B. longum B379M-MB strain, is that the culture grown under stationary conditions on a nutrient medium "Bifilife M" in a volume of 300 cm 3 has a pronounced superoxide dismutase activity.
  • SOD - the activity of the strain B. longum B379M-BL is 32.0 units/mg of the protein contained in the bacterial mass.
  • the revealed SOD activity of the culture of bifidobacteria indicates that the strain has a pronounced probiotic effect, which can manifest itself both in vivo and in vitro.
  • the high activity of acid formation, the resistance of the strain to major antibiotics contributes to the effective restoration of normal intestinal microflora against the background of antibiotic therapy.
  • the strain is well stored at a temperature of 6-8°C without loss of activity during the year.
  • the studied strain of Bifidobacterium longum V379M-BL belongs to the following systematic group: Bacteria; Actinobacteria; Actinobacteridae; Bifidobacteriales; Bifidobacteriaceae; Bifidobacterium.
  • Analysis of the no RDP II 16S pRNA database showed the best homology with the bacterial species Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium suis, Bifidobacterium subtile, Bifidobacterium pseudoIongum, Bifidobacterium pullorum.
  • a phylogenetic tree with homologous strains was constructed (Fig. 2).
  • the strain Bifidobacterium longum V379M-BL can be attributed to the species Bifidobacterium longum (99%).
  • Pavlicek A et al "Free-Tree-freeware program for construction of phylogenetic trees on the basis of distance data and bootstrap ⁇ ackknife analysis of tree robustness.”
  • the strain is not genetically modified and does not contain genes from other organisms; transferred resistance genes; genetic changes associated with the use of genetic engineering techniques; the strain is not zoopathogenic and phytopathogenic.
  • strain Bifidobacterium adolescentis G 7513-BL the selection of the strain Bifidobacterium adolescentis G 7513-MB was carried out, which was deposited on July 15, 2004 in the VKPM FSUE GosNIIgenetika of the Ministry of Education and Science under No. Ac-1690.
  • the original strain of Bifidobacterium adolescentis G 7513-MB is protected by RF patent No.
  • the selection was carried out without the use of genetic modification methods by means of multiple cycles of lyophilized drying and recovery on the previously mentioned nutrient medium "Bifilife M".
  • the primary inoculum of the production strain Bifidobacterium adolescentis G 7513-MB in a volume of 5% of the volume of the cultivation medium was introduced into the above medium and cultivated at a temperature of 37-38°C for 24 hours. Then 30 cycles of drying-recovery were carried out on the nutrient medium "Bifilife M".
  • Ferment sterile milk with the formation of a stable dense clot by 22-24 hours. Gas-forming ability, catalase-forming ability, gelatin liquefaction are absent.
  • Acid-forming activity when cultivated in sterile skimmed milk, titratable acidity reaches 70 ° T by 24 hours, and accumulates the microbial mass of bifidobacteria in 1 ml of 1g 9.0-9.4 microbial cells.
  • Acid-forming activity by 24 hours - 70 ° T, by 72 hours - 90-100 ° T (on a liver medium), on a hydrolyzate-milk medium - up to 100 ° T and 180 ° T, respectively.
  • the maximum titer of bifidobacteria in a hydrolysate-milk medium after 24 hours of cultivation is 1x10 10 CFU/ml.
  • the strain grows on nutrient media used in production with biomass accumulation of at least 1g 9.8-10.0 microbial number for 18-20 hours of cultivation.
  • the strain has a pronounced antagonistic activity against pathogenic and conditionally pathogenic microorganisms: Shigella sonnei, Shigella flexneri, Escherichia coli, Proteus vulgaris, Staphylococcus aureus, Candida albicans, Klebsiella pneumonia, Citrobacter freundii, Bacillus subtilis.
  • Table 7 shows the antagonistic activity against some of the aforementioned microorganisms.
  • the antagonistic activity of a monostrain of bifidobacteria was determined by the zone of growth retardation of test cultures of pathogenic and conditionally pathogenic microorganisms by the method of perpendicular strokes.
  • Determination of the level of resistance of the strain to antibiotics was carried out by the method of serial dilutions in a liquid nutrient medium - sterile skimmed milk. Various concentrations of sterile solutions of antibiotics and a 16-hour culture of bifidobacteria in an amount of 5% were introduced into the nutrient medium. Stability was determined by the change in titratable acidity in the samples after 48 hours of incubation at a temperature of 37-38°C.
  • a production-useful feature of the strain B. adolescentis G 7513 - BL which distinguishes it from the strain B. adolescentis G 7513-MB, is that the culture grown under stationary conditions on a nutrient medium "Bifilife M" in a volume of 300 cm 3 has pronounced superoxide dismutase activity.
  • SOD - the activity of the strain B. adolescentis G 7513-BL is 25.0 units/mg of the protein contained in the bacterial mass.
  • the revealed SOD activity of the culture of bifidobacteria indicates that the strain has a pronounced probiotic effect, which can manifest itself both in vivo and in vitro.
  • the high activity of acid formation, the resistance of the strain to major antibiotics contributes to the effective restoration of normal intestinal microflora against the background of antibiotic therapy.
  • Freshly prepared strain B. adolescentis G 7513 - BL is stored on a nutrient medium with the addition of a protective medium containing sucrose, gelatose, sodium citrate, dried by sublimation in ampoules. The strain is well kept at a temperature of 6-8 °C without loss of activity during the year.
  • the studied strain of Bifidobacterium adolescentis G7513-BL belongs to the following systematic group: Bacteria; Actinobacteria; Actinobacteridae; Bifidobacteriales; Bifidobacteriaceae; Bifidobacterium, and the homology with the species Bifidobacterium adolescentis is 97%.
  • the tested Bifidobacterium adolescentis G7513-BL strain can be attributed to the Bifidobacterium adolescentis species (97%).
  • Pavlicek A et al "Free-Tree-freeware program for construction of phylogenetic trees on the basis of distance data and bootstrap ⁇ ackknife analysis of tree robustness.”
  • the strain is not genetically modified and does not contain genes from other organisms; transferred resistance genes; genetic changes associated with the use of genetic engineering techniques; the strain is not zoopathogenic and phytopathogenic.
  • strain Bifidobacterium infantis 73-15-BL selection was carried out strain Bifidobacterium infantis 73-15-MB, which was deposited on July 15, 2004 in VKPM Federal State Unitary Enterprise GosNIIgenetika of the Ministry of Education and Science under No. Ac-1692.
  • the original strain Bifidobacterium infantis 73-15-MB is protected by RF patent No. 2261906. lyophilized drying and recovery on the previously mentioned nutrient medium "Bifilife M".
  • the primary inoculum of the production strain Bifidobacterium infantis 73-15-MB in a volume of 5% of the volume of the cultivation medium was introduced into the above medium and cultivated at a temperature of 37-38 °C for 24 hours. Then 30 cycles of drying-recovery were carried out on the nutrient medium "Bifilife M".
  • Acid-forming activity when cultivated on sterile skimmed milk, titratable acidity reaches 24 h - 80 ° T and the strain accumulates a microbial mass of bifidobacteria in 1 ml of 1 g of 10.2-10.6 microbial cells.
  • the strain grows on nutrient media used in production with biomass accumulation of at least 1g 9-10 microbial number for 18-22 hours of cultivation.
  • the strain has a pronounced antagonistic activity against pathogenic and conditionally pathogenic microorganisms: Shigella sonnei, Shigella flexneri, Escherichia coli, Proteus vulgaris, Staphylococcus aureus, Candida albicans, Proteus mirabilis, Klebsiella pneumoniae, Citrobacter freundii, Bacillus subtilis.
  • Table 10 shows the antagonistic activity against some of the aforementioned microorganisms.
  • the antagonistic activity of a monostrain of bifidobacteria was determined by the zone of growth inhibition of test cultures of pathogenic and conditionally pathogenic microorganisms. by the method of perpendicular strokes.
  • Determination of the level of resistance of the strain to antibiotics was carried out by the method of serial dilutions in a liquid nutrient medium - sterile skimmed milk. Various concentrations of sterile solutions of antibiotics and a 16-hour culture of bifidobacteria in an amount of 5% were introduced into the nutrient medium. Stability was determined by the change in titratable acidity in the samples after 48 h of incubation at a temperature of 37-38°C.
  • a production-useful feature of the B. infantis 73-15-BL strain which distinguishes it from the B. infantis 73-15-MB strain, is that it is grown under stationary conditions on a nutrient medium "Bifilife M" in a volume of 300 cm 3 culture has a pronounced superoxide dismutase activity.
  • SOD - the activity of the strain B. infantis 73-15-BL is 29 units/mg of protein contained in the bacterial mass.
  • the revealed SOD activity of the culture of bifidobacteria indicates that the strain has a pronounced probiotic effect, which can manifest itself both in vivo and in vitro.
  • the high activity of acid formation, the resistance of the strain to major antibiotics contributes to the effective restoration of normal intestinal microflora against the background of antibiotic therapy.
  • a freshly prepared strain of B. infantis 73-15-BL is stored on a nutrient medium with the addition of a protective medium containing sucrose, gelatin, sodium citrate, and freeze-dried in ampoules.
  • the strain is well stored at temperature of 6-8 °C without loss of activity during the year.
  • the studied strain of Bifidobacterium infantis 73-15-BL belongs to the following systematic group: Bacteria; Actinobacteria; Actinobacteridae; Bifidobacteriales; Bifidobacteriaceae; Bifidobacterium. Analysis of the no RDP II 16S rRNA database showed the best homology with the bacterial species Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium suis, Bifidobacterium pseudoIongum.
  • the studied strain of Bifidobacterium infantis 73-15-BL belongs to the species Bifidobacterium infantis.
  • the species Bifidobacterium infantis is currently referred to as Bifidobacterium longum subsp. infantis.
  • the tested strain Bifidobacterium infantis 73-15-BL can be assigned to the species Bifidobacterium longum subsp. infantis (97%).
  • Pavlicek A et al "Free-Tree-freeware program for construction of phylogenetic trees on the basis of distance data and bootstrap ⁇ ackknife analysis of tree robustness.”
  • the strain is not genetically modified and does not contain genes from other organisms; transferred genes resistance; genetic changes associated with the use of genetic engineering techniques; the strain is not zoopathogenic and phytopathogenic.
  • strain Bifidobacterium breve 79-119-BL the selection of the strain Bifidobacterium breve 79-119-MB was carried out, which was deposited on July 15, 2004 in the VKPM Federal State Unitary Enterprise GosNIIgenetika of the Ministry of Education and Science under registration No. Ac-1691. patent of the Russian Federation No. 2261905. Selection was carried out without the use of genetic modification methods by means of multiple cycles of lyophilized drying and recovery on the previously mentioned nutrient medium "Bifilife M".
  • the primary inoculum of the production strain Bifidobacterium breve 79-119-MB in a volume of 5% of the volume of the cultivation medium was introduced into the above medium and cultivated at 37-38°C for 24 hours. Then 30 drying-recovery cycles were carried out on the nutrient medium "Bifilife M".
  • Ferment milk with the formation of a dense clot by 16-18 hours. Gas-forming ability, catalase-forming ability, gelatin liquefaction are absent.
  • Acid-forming activity when cultivated on sterile skimmed milk, titratable acidity reaches 120° T by 18 hours, 140° T by 24 hours. .
  • the strain grows on nutrient media used in production with biomass accumulation of at least 1g 10.2 microbial number for 16-18 hours of cultivation.
  • the strain has a pronounced antagonistic activity against pathogenic and conditionally pathogenic microorganisms: Shigella sonnei, Shigella flexneri, Escherichia coli, Staphylococcus aureus, Proteus vulgaris, Candida albicans, Klebsiella pneumoniae, Citrobacter freundii, Bacillus subtilis.
  • Table 13 shows the antagonistic activity against some of the aforementioned microorganisms.
  • the antagonistic activity of a monostrain of bifidobacteria was determined by the zone of growth retardation of test cultures of pathogenic and conditionally pathogenic microorganisms by the method of perpendicular strokes.
  • Determination of the level of resistance of the strain to antibiotics was carried out by the method of serial dilutions in a liquid nutrient medium - sterile skimmed milk. Various concentrations of sterile solutions of antibiotics and a 16-hour culture of bifidobacteria in an amount of 5% were introduced into the nutrient medium. Stability was determined by the change in titratable acidity in the samples after 48 hours of incubation at a temperature of 37-38°C.
  • a production-useful feature of the B. breve 79-119-BL strain which distinguishes it from the B. breve 79-119-MB strain, is that it is grown under stationary conditions on a nutrient medium "Bifilife M" in a volume of 300 cm 3 culture has a pronounced superoxide dismutase activity.
  • the detected SOD activity of the B. breve 79-119-BL strain indicates that the strain has a pronounced probiotic effect, which can manifest itself both in vivo and in vitro.
  • the high activity of acid formation, the resistance of the strain to major antibiotics contributes to the effective restoration of normal intestinal microflora against the background of antibiotic therapy.
  • the claimed strain has a pronounced antioxidant activity: superoxide dismutase the activity of the strain is 27.0 units/mg of microbial mass protein.
  • Freshly prepared strain B. breve 79-119-BL is stored on a nutrient medium with the addition of a protective medium containing sucrose, gelatin, sodium citrate, and freeze-dried in ampoules.
  • the strain is well stored at a temperature of 6-8 °C without loss of activity during the year.
  • the studied strain Bifidobacterium breve 79-119-BL belongs to the following systematic group: Bacteria; Actinobacteria; Actinobacteridae; Bifidobacteriales; Bifidobacteriaceae; Bifidobacterium. Analysis of the no RDP II 16S pRNA database showed homology with the same bacterial species Bifidobacterium breve.
  • the tested strain Bifidobacterium breve 79-119-BL can be attributed to the species Bifidobacterium breve (97%).
  • Pavlicek A et al "Free-Tree-freeware program for construction of phylogenetic trees on the basis of distance data and bootstrap ⁇ ackknife analysis of tree robustness.”
  • Information about the safety of using the strain the strain is not genetically modified and does not contain genes from other organisms; transferred resistance genes; genetic changes associated with the use of genetic engineering techniques; the strain is not zoopathogenic and phytopathogenic.
  • 3rd passage - Sowing of inoculums of monocultures into the fermenter is carried out in stages in the amount of 3-5% of each strain of the volume of the nutrient medium.
  • the process of co-cultivation lasts 10-12 hours, the pH of the finished biomass is 4, 6-5, 2.
  • Protective media (sucrose-gelatin, skim milk) are added to the resulting biomass, the suspension is frozen at a temperature not higher than minus 45 ° C, lyophilized, a dry concentrate is obtained with the content of bifidobacteria not less than 10 11 CFU/g.
  • the consortium is well stored at a temperature of 6-8 °C without loss of activity within one year.
  • the created consortium of bifidobacteria showed a high compatibility of five strains of bifidobacteria with each other. Co-cultivation of a mixed population of all five strains proceeds much faster compared to separate cultivation of strains.
  • Acid-forming activity by 24 hours - - 150 ° T, by 72 hours - 180 ° T (on the nutrient medium "Bifilife M"), on the hydrolyzate-milk medium - up to 140-160 0 T and 170-190 ° T, respectively.
  • the consortium actively reproduces on media of different composition.
  • the consortium In vitro on standard nutrient media, at a temperature of T 37° C, the consortium accumulates biomass to a high concentration of probiotic microorganisms - 10 11 CFU per 1 g of biomass due to the absence of mutual inhibition of individual strains. (Fig 6)
  • the maximum titer of bifidobacteria of the consortium in sterile skimmed milk without growth factors by 22 hours is 2x10 10 CFU / ml, on corn-lactose medium by 20 hours it is 2x10 10 CFU / ml, on the nutrient medium "Bifilife M" after lyophilization, a dry concentrate is obtained containing bifidobacteria not less than 10 11 CFU/g.
  • Bifidobacteria of the consortium are able to utilize free amino acids in sterile skimmed milk, such as: valine, methionine, alanine, arginine, isoleucine, glycine.
  • the consortium exhibits antagonistic activity against pathogenic and conditionally pathogenic microorganisms: Escherichia coli, Shigella sonnei, Staphylococcus aureus, Shigella fexneri, Candida albicans, Proteus vulgaris, Proteus mirabilis, Klebsiella pneumoniae, Citrobacter freundii, Bacillus subtilis.
  • Table 17 shows antagonistic activity against some of the aforementioned microorganisms.
  • the antagonistic activity of a consortium of bifidobacteria strains was determined by the method of developing mixed populations in co-cultivation with pathogenic microorganisms in the growth retardation zone of test cultures by the method of perpendicular strokes.
  • the synergistic effect of the consortium is also manifested in the greater antagonistic activity of the consortium compared to the prototype.
  • Determination of levels of resistance of strains of bifidobacteria to antibiotics was carried out by the method of serial dilutions in a liquid nutrient medium for each strain separately. Sterile skimmed milk was used as a nutrient medium.
  • Table 18 shows the resistance to antibiotics of individual strains of the proposed consortium.
  • V379M-BL longum V379M-BL - to amikacin, ampicillin, benzylpenicillin, gentamicin, kanamycin, lincomycin, oleandomycin, ofloxacin, streptomycin, tetracycline, sulfadimidine, benzathine benzylpenicillin.
  • the consortium can be cultured on various nutrient media with consistently high CFU/ml of probiotic microorganisms and produced organic acids.
  • the culture results are shown in Table 20.
  • the resulting consortium and the strains included in the consortium can be used as a biologically active principle of probiotic orientation in the production of food products, functional foods, dietary (therapeutic and preventive) foods, biologically active additives (BAA), cosmetics, hygiene products, medical products. and veterinary drugs.
  • BAA biologically active additives
  • the use of the claimed consortium and the strains that make up the consortium in the production of fermented, non-fermented food products, dietary supplements, bifid-containing preparations, cosmetic and hygiene products - creams, wipes, tampon lotions, nourishing masks, feminine pads, soap, cosmetic milk provides a probiotic and bacteriological effect , contributes to the restoration of the normal microflora of the skin and mucous membranes, and is also aimed at expanding the arsenal of similar tools.
  • Example 1 The use of the proposed consortium of strains of bifidobacteria in the composition of the starter for the preparation of fermented milk products.
  • 3rd passage - inoculation of inoculums of monocultures into the fermenter is carried out in stages in the amount of 3-5% of each strain of the volume of the nutrient medium.
  • the process of co-cultivation lasts 10-12 hours.
  • the pH of the finished biomass is 4.6-5.2.
  • the biomass of the consortium of bifidobacteria, obtained as shown above, in a volume of not more than 5% (of the volume of milk) is introduced into sterile milk and cultivated until a clot is formed, which is poured into containers and cooled.
  • Example 2 The use of a consortium of strains of bifidobacteria in the composition of dietary supplements.
  • a dry biologically active additive for this, protective media (sucrose-gelatin, skim milk) are added to the biomass obtained according to Example 1, the suspension is frozen at a temperature not exceeding minus 45 ° C, lyophilized, sealed and a dry concentrate is obtained with a content of bifidobacteria of at least 10 11 CFU/g, which can be used as a biologically active additive by adding it to food raw materials immediately before use.
  • protective media sucrose-gelatin, skim milk
  • Example 3 Use of a consortium of strains of bifidobacteria to obtain a bifid-containing medical preparation.
  • Example 1 In the biomass of the consortium of bifidobacteria obtained according to Example 1, which is a liquid concentrate with a high titer of bifidobacteria, protective media (sucrose gelatin, skim milk) are added. The resulting suspension is frozen at a temperature not exceeding minus 45°C, lyophilized, sealed and a dry concentrate is obtained with a bifidobacteria content of at least 10 11 CFU/g, which can be used as a bifid-containing medical product.
  • protective media sucrose gelatin, skim milk
  • Example 4 Use of a consortium of strains of bifidobacteria to produce kvass or juice.
  • the biomass of the consortium of bifidobacteria obtained according to Example 1, in in a volume of not more than 5% of the volume of the normalized mixture (kvass or juice) is added to bread kvass or juice after pasteurization so that the content of bifidobacteria in 1 ml of kvass or juice is at least 10 7 CFU / ml. Kvass or juice is cooled and consumed without subsequent fermentation.
  • Example 5 Use of a consortium of strains of bifidobacteria to obtain a cosmetic product.
  • the biomass of the consortium of bifidobacteria obtained according to Example 1 is destroyed by heating in an autoclave or membrane filtration is carried out.
  • the resulting biologically active components are cooled and mixed with a cream base in a ratio of 1:2-1:20, depending on the purpose of the cream being produced.
  • Example 6 Use of a consortium of strains of bifidobacteria for the production of cooked smoked sausages.
  • the biomass of the consortium of bifidobacteria obtained according to Example 1, in a volume of not more than 5% of the minced meat volume, is added to the prepared sausage minced meat after adding salt, spices and sodium nitrite solution. After shaping, sausage sticks are sent to the draft, which is carried out on frames at a temperature of 22 ° C, humidity 90%. Rainfall duration 6 hours. Cooking and smoking are carried out according to standard technology.
  • Example 7 Use of a consortium of strains of bifidobacteria for the production of raw smoked sausages.
  • the consortium of bifidobacteria in dry form obtained according to Example 2, is preliminarily restored in boiled chilled water at a temperature of 37 ° C, or activated in sterile cow's skimmed milk at a temperature of 37 ° C for 8-10 hours to an acidity of 60-65 ° T.
  • the biomass of the consortium of bifidobacteria is introduced into the prepared sausage mince, while the dosage of the introduced reduced consortium is 0.1-0.2% of the mince mass, and the dosage of the activated consortium is 2-5% of the mince mass.
  • enriched sausage raw materials are processed according to the traditional scheme according to the technology for producing raw smoked sausages.
  • Example 8 Use of a consortium of bifidobacteria strains for the production of fortified dry infant formula.
  • a small amount of dry milk mixture is thoroughly mixed with a dry concentrate of a consortium of bifidobacteria obtained according to Example 2 with a bifidobacteria content of at least 10 11 CFU / g based on at least 100 g of concentrate per 1 ton of dry milk formula. Then, the entire remaining batch of the milk mixture is thoroughly mixed with enriched dry milk mixture to achieve a content of bifidobacteria of at least 10 7 CFU / g.
  • Example 9 Use of a consortium of bifidobacteria strains to produce fortified beer.
  • Example 10 Use of a consortium of bifidobacteria strains for the production of ice cream.
  • ice cream is added to milk, the resulting mixture is heated to a temperature of 35 ° C, dry ingredients are added: granulated sugar, a stabilizer, heated to a temperature of 85 ° C, homogenized, cooled to a temperature of 6 ° C, a dry bacterial concentrate of a consortium of bifidobacteria is added, obtained according to Example 2 with a content of bifidobacteria of at least 10 11 CFU / g in the amount of 1-5% by weight of the mixture.
  • Example 11 The use of a consortium of strains of bifidobacteria for the production of feed additives in animal husbandry and poultry farming.
  • Example 12 Use of a consortium of bifidobacteria strains for the production of toothpaste.
  • Example 13 Use of a consortium of bifidobacteria strains for the production of soft rennet cheese.
  • Normalized milk is subjected to heat treatment at a temperature of 74-76 ° C for 1-5 minutes and cooled to a fermentation temperature of 37-38 ° C, a 40% aqueous solution of calcium chloride is added at the rate of 50 ml per 100 kg of milk, 1% dry bacterial concentrate of a consortium of bifidobacteria obtained according to Example 2, with a bifidobacteria content of at least 10 11 CFU/g. The mixture is stirred and a solution of a milk-clotting enzyme is added at the rate of 1.0-1.2 g per 100 kg of milk, then coagulation is carried out. The subsequent production of soft cheese is carried out according to standard technology.
  • Example 14 Use of a consortium of bifidobacteria strains for the production of brine mature cheese.
  • Coagulation of milk is carried out at a temperature of 32-34 ° C for 30-45 minutes, a combined starter is added in the amount of 1.0-2.0%, consisting of salt-tolerant pure cultures of mesophilic lactic streptococci; a consortium of bifidobacteria obtained according to Example 1; mesophilic lactic acid bacilli. Starter cultures of bifidobacteria, streptococci and mesophilic lactobacilli are prepared separately. Streptococci, bifidobacteria and mesophilic lactic acid bacilli are added to the production starter. Then calcium chloride and rennet are added to the resulting mixture to obtain a clot. The subsequent production of brine mature cheese is carried out according to standard technology.
  • consortium based on five breeding strains of bifidobacteria was obtained, which is more resistant to damaging factors of the aggressive environment of the gastrointestinal tract during the passage of bifidobacteria in the gastrointestinal tract, which is expressed in increased colonizing activity.
  • the consortium has a pronounced antioxidant activity and probiotic action in the absence of mutual inhibition of individual strains.
  • the proposed consortium is high-tech, accumulates biomass on nutrient media in a short period of cultivation with a high concentration of probiotic microorganisms - 10 11 CFU / g
  • the method used for selecting breeding strains which involves the study of their antibiotic resistance, cultural, antagonistic, morphological, physiological and biochemical properties and molecular genetic identification, made it possible to create a consortium capable of correcting microecological disorders more effectively and safely and expanding the arsenal of probiotic agents.

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Abstract

Ce groupe d'inventions, liées par un seul concept inventif, concerne les biotechnologies, l'industrie alimentaire, la pharmacologie, la cosmétologie, et peut être utilisé afin de produire des produits contenant du bifidus. Afin de produire ce complexe, on utilise des souches sélectionnées de bifidobactéries: Bifidobacterim bifidum 791-BL, Bifidobacterim longum В 379М-BL, Bifidobacterim adolescentis G 7513-BL, Bifidobacterim infantis 73-15-BL, Bifidobacterim breve 79-119-BL, dont la particularité caractéristique est la capacité à accumuler activement une biomasse sur divers milieux de culture jusqu'à une concentration élevée, une stabilité antibiotique élevée avec une activité antioxydante élevée. L'activité de superoxydes dismutases du complexe est d'au moins 37 unités/mg de protéine de la masse microbienne. Cette activité spécifique du complexe et des souches entrant dans la composition du complexe offre une possibilité de suppression d'une large gamme d'acides aminés, une résistance aux antibiotiques, une stabilité envers les facteurs nocifs de milieux agressifs des voies gastro-intestinales, une grande capacité de colonisation permettant d'augmenter son efficacité à des fins de prévention et de traitement.
PCT/RU2021/000582 2021-01-12 2021-12-17 Complexe de bifido-bactéries et souches entrant dans la composition du complexe WO2022154692A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998044090A1 (fr) * 1997-04-01 1998-10-08 Moskovsky Nauchno-Issledovatelsky Institut Epidemiologii I Mikrobiologii Imeni G.N.Gabrichevskogo Association de lignees de bacteries bifidus pour la preparation de lait suri, de produits alimentaires non fermentes et de preparations bacteriennes
RU2261909C1 (ru) * 2004-10-20 2005-10-10 Общество с ограниченной ответственностью "Мир биотехнологий (НПО)" КОНСОРЦИУМ БИФИДОБАКТЕРИЙ Bifidobacterium bifidum 791-МБ, Bifidobacterium longum В 379М-МБ, Bifidobacterium adolescentis Г-7513- МБ, Bifidobacterium infantis 73-15-МБ, Bifidobacterium breve 79-119-МБ, ИСПОЛЬЗУЕМЫЙ ДЛЯ ПРИГОТОВЛЕНИЯ КИСЛО-МОЛОЧНЫХ, НЕФЕРМЕНТИРОВАННЫХ ПРОДУКТОВ, БИОЛОГИЧЕСКИ АКТИВНЫХ ДОБАВОК, БИФИДОСОДЕРЖАЩИХ ПРЕПАРАТОВ, КОСМЕТИЧЕСКИХ И ГИГИЕНИЧЕСКИХ СРЕДСТВ
RU2264450C2 (ru) * 2003-10-08 2005-11-20 Закрытое Акционерное Общество "Биокад" Биопрепарат "бифистим для взрослых" для профилактики и лечения дисбактериозов и инфекционных болезней желудочно-кишечного тракта у людей старше 12 лет, биологически активная добавка к пище " бифистим для взрослых" для профилактики дисбактериозов и восстановления микрофлоры желудочно-кишечного тракта после антибиотикотерапии и других неблагоприятных воздействий у людей старше 12 лет и консорциум бифидобактерий для коррекции микрофлоры желудочно-кишечного тракта у людей старше 12 лет
RU2491331C1 (ru) * 2012-05-14 2013-08-27 Федеральное бюджетное учреждение науки "Московский научно-исследовательский институт эпидемиологии и микробиологии им. Г.Н. Габричевского" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека Консорциум бифидобактерий и лактобацилл, используемый для приготовления бактерийных препаратов и биологически активных добавок к пище, предназначенных для коррекции микрофлоры желудочно-кишечного тракта детей в возрасте до 3-х лет, и способ его получения, биологически активная добавка к пище и бактериальный препарат для лечения дисбиотических состояний желудочно-кишечного тракта детей в возрасте до 3-х лет

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998044090A1 (fr) * 1997-04-01 1998-10-08 Moskovsky Nauchno-Issledovatelsky Institut Epidemiologii I Mikrobiologii Imeni G.N.Gabrichevskogo Association de lignees de bacteries bifidus pour la preparation de lait suri, de produits alimentaires non fermentes et de preparations bacteriennes
RU2264450C2 (ru) * 2003-10-08 2005-11-20 Закрытое Акционерное Общество "Биокад" Биопрепарат "бифистим для взрослых" для профилактики и лечения дисбактериозов и инфекционных болезней желудочно-кишечного тракта у людей старше 12 лет, биологически активная добавка к пище " бифистим для взрослых" для профилактики дисбактериозов и восстановления микрофлоры желудочно-кишечного тракта после антибиотикотерапии и других неблагоприятных воздействий у людей старше 12 лет и консорциум бифидобактерий для коррекции микрофлоры желудочно-кишечного тракта у людей старше 12 лет
RU2261909C1 (ru) * 2004-10-20 2005-10-10 Общество с ограниченной ответственностью "Мир биотехнологий (НПО)" КОНСОРЦИУМ БИФИДОБАКТЕРИЙ Bifidobacterium bifidum 791-МБ, Bifidobacterium longum В 379М-МБ, Bifidobacterium adolescentis Г-7513- МБ, Bifidobacterium infantis 73-15-МБ, Bifidobacterium breve 79-119-МБ, ИСПОЛЬЗУЕМЫЙ ДЛЯ ПРИГОТОВЛЕНИЯ КИСЛО-МОЛОЧНЫХ, НЕФЕРМЕНТИРОВАННЫХ ПРОДУКТОВ, БИОЛОГИЧЕСКИ АКТИВНЫХ ДОБАВОК, БИФИДОСОДЕРЖАЩИХ ПРЕПАРАТОВ, КОСМЕТИЧЕСКИХ И ГИГИЕНИЧЕСКИХ СРЕДСТВ
RU2491331C1 (ru) * 2012-05-14 2013-08-27 Федеральное бюджетное учреждение науки "Московский научно-исследовательский институт эпидемиологии и микробиологии им. Г.Н. Габричевского" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека Консорциум бифидобактерий и лактобацилл, используемый для приготовления бактерийных препаратов и биологически активных добавок к пище, предназначенных для коррекции микрофлоры желудочно-кишечного тракта детей в возрасте до 3-х лет, и способ его получения, биологически активная добавка к пище и бактериальный препарат для лечения дисбиотических состояний желудочно-кишечного тракта детей в возрасте до 3-х лет

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