WO2018030730A1 - 락토바실러스 살리바리우스 cjls1511, 상기 균 또는 이의 사균체를 포함하는 동물 사료 첨가제 조성물, 및 상기 사균체의 제조 방법 - Google Patents
락토바실러스 살리바리우스 cjls1511, 상기 균 또는 이의 사균체를 포함하는 동물 사료 첨가제 조성물, 및 상기 사균체의 제조 방법 Download PDFInfo
- Publication number
- WO2018030730A1 WO2018030730A1 PCT/KR2017/008487 KR2017008487W WO2018030730A1 WO 2018030730 A1 WO2018030730 A1 WO 2018030730A1 KR 2017008487 W KR2017008487 W KR 2017008487W WO 2018030730 A1 WO2018030730 A1 WO 2018030730A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- lactobacillus salivarius
- kccm11829p
- animal feed
- cjls1511
- salivarius cjls1511
- Prior art date
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Images
Classifications
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- 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
- C12N1/205—Bacterial isolates
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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
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/181—Salivarius
-
- 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
- C12N2523/00—Culture process characterised by temperature
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Definitions
- the present invention relates to a novel Lactobacillus salivarius CJLS1511, an animal feed additive composition comprising the bacterium or its microorganism, and a method for producing the microorganism.
- Lactobacillus sp are lactic acid bacilli that are homozygous or heterozygous and are commonly found in the intestines of animals including humans and in the fermentation of dairy products or vegetables.
- Microorganisms of the genus Lactobacillus are lactic acid-producing bacteria found in the intestinal tract of animals, and are subjected to homo-fermentation or hetero-fermentation using dairy products or vegetables as its substrate. Lactobacillus microorganisms keep the intestinal pH acidic, inhibiting the reproduction of harmful bacteria such as E. coli and Clostridium, improving diarrhea and constipation, as well as vitamin synthesis, anticancer activity, serum It is known to play a role in lowering cholesterol.
- a novel Lactobacillus genus that can improve the weight gain of animals when used as feed for livestock, improve the immunity or anti-history of animals, and inhibit the growth of harmful bacteria in the intestine of animals. sp.) to provide microorganisms.
- One aspect of the present invention relates to Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganisms.
- Another aspect of the invention relates to an animal feed additive composition
- an animal feed additive composition comprising Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganism.
- Another aspect of the invention relates to an animal feed comprising the animal feed additive composition described herein.
- Culture medium was prepared by incubating Lactobacillus salivarius CJLS1511 (KCCM11829P),
- the culture is heated at a temperature of 70 °C to 160 °C,
- the heated culture is cooled to a temperature in the range of 10 °C to 60 °C,
- It relates to a method for producing Lactobacillus salivarius CJLS1511 (KCCM11829P) microorganism, comprising separating the Lactobacillus salivarius CJLS1511 (KCCM11829P) microorganism from the cooled culture.
- Lactobacillus salivarius CJLS1511 or its microorganism according to an aspect of the present invention is excellent in triglyceride degrading ability, endotoxin adsorption capacity, pathogenic bacterial growth inhibitory activity and digestive enzyme activity.
- Animal feed additive composition or animal feed comprising Lactobacillus salivarius CJLS1511 or its microorganisms according to one aspect of the present invention may improve the weight gain of the animal and improve immunity to anti-history.
- Figure 1 shows the electron micrograph of Lactobacillus salivarius ( Lactobacillus salivarius ) CJLS1511 strain or its microorganism.
- One aspect of the invention relates to Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganisms.
- KCCM11829P the mycelium of Lactobacillus salivarius CJLS1511) (KCCM11829P) is, for example, inactive cells of Lactobacillus salivarius CJLS1511) (KCCM11829P), specifically, inactivated by heat. May be cells.
- the present inventors collected the small intestine end of broiler, washed with sterile distilled water, and then plated on MRS medium to which 0.001% bromphenicol purple (BCP) was added and incubated anaerobicly at 37 ° C.
- BCP bromphenicol purple
- 50 strains of lactic acid bacteria-producing strains were selected and subcultured, followed by secondary isolation of 24 species of bacilli in morphological separation using these strains.
- Ten isolates were isolated by comparing the antimicrobial activity and digestive enzyme activity of 24 species, and the bile, acid and animal barrier cell adsorption of the lactic acid bacteria-producing strains were measured.
- One strain with the best enzyme activity and triglyceride resolution was isolated.
- the isolated lactic acid bacteria producing strain was named Lactobacillus salivarius CJLS1511 ( Lactobacillus salivarius CJLS1511), and was deposited on April 12, 2016 at the Korea Microorganism Conservation Center (Accession Number: KCCM11829P).
- Morphological characteristics (morphology when grown in MRS solid medium) -Shape of the cell Bacillus Cell polymorphism none Mobility none -Formation of apo none 2.
- Physiological Features Gram staining Gram positive Catalase voice Oxidase voice -Growth temperature and time 37 ° C, 18-48 hours -Oxygen demand Anaerobic
- Lactobacillus salivarius CJLS1511 has a rod shape as shown in FIG. 1 and does not form spores.
- the difference between live bacteria and dead bacteria was confirmed by glycoprotein activity on the cell wall surface.
- Lactobacillus Salivarius CJLS1511 No. Our kind Lactobacillus salivarius CJLS1511 0 Temoin - One Glycerol - 2 Erythritol - 3 D-arabinose - 4 L-arabinose - 5 D-ribose - 6 D-xylose - 7 L-xylose - 8 D-adonito - 9 Methyl-aD-xylopyranozide + 10 D-galactose + 11 D-glucose + 12 D-fructose + 13 D-mannose - 14 L-sorbose - 15 L-ramnose - 16 D-ulcitol - 17 Inositol + 18 D-mannitol + 19 D-sorbitol + 20 Methyl-aD-manopyranozide - 21 Methyl-aD-glucopyranozide - 22 N-acetylglucosamine +
- Lactobacillus salivarius CJLS1511 (KCCM11829P) is excellent in acid resistance, bile resistance, antimicrobial activity, digestive enzyme activity and triglyceride degrading ability, and when used in animal feed or feed composition, it can effectively improve feed efficiency and animal growth. There is an advantage.
- the Lactobacillus salivarius CJLS1511 (KCCM11829P) microorganisms exert competitive adhesion with harmful intestinal bacteria and contribute to the formation of intestinal flora, and the cell wall component of the cell wall eluted as the cell walls of the microorganisms were destroyed in the small intestine. Lipotecholic acid can interfere with intestinal harmful bacteria in the gut membrane.
- the Lactobacillus salivarius CJLS1511 (KCCM11829P) microorganisms have a high hydrophobicity and cohesiveness compared to the live bacteria has the advantage that can be attached to pathogenic microorganisms and endotoxins to improve the anti-pathology.
- the Lactobacillus salivarius CJLS1511 (KCCM11829P) is administered to the feed to feed the animal feed can increase both the weight gain and feed rate.
- Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganism may further comprise a protective agent.
- the protective agent may be one or more of sugars such as yeast, yeast extract, monosaccharides, polysaccharides, starch, raw sugar and refined sugar. Specifically, at least one substance selected from the group consisting of yeast extract, dextrose and raw sugar can be mentioned.
- the use of a protective agent protects Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganisms from the external environment to prevent contamination, and has an effect of extending its shelf life.
- Another embodiment of the present invention relates to an animal feed additive composition
- an animal feed additive composition comprising the Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganism.
- the animal feed additive composition may include Lactobacillus salivarius CJLS1511 (KCCM11829P) microorganisms.
- the animal feed additive composition may further comprise an excipient.
- the excipient is not particularly limited, and excipients commonly used in the art may be used. Excipients include, for example, sugars (e.g. lactose, D-mannitol, D-sorbitol, sucrose, etc.), gums (e.g., xanthan gum, guar gum, gum arabic), starch (e.g. corn starch, potato starch, etc.), inorganic Salts such as calcium phosphate, calcium sulfate, precipitated calcium carbonate, and the like.
- sugars e.g. lactose, D-mannitol, D-sorbitol, sucrose, etc.
- gums e.g., xanthan gum, guar gum, gum arabic
- starch e.g. corn starch, potato starch, etc.
- inorganic Salts such as calcium phosphate, calcium sulfate, precipitated calcium carbonate, and the
- the animal feed additive composition may include the Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganism per 1.0 g of the composition as 1.0 ⁇ 10 8 cfu to 1.0 ⁇ 10 10 cfu. Specifically, it may be included as 1.0x10 8 cfu to 3.0x10 9 cfu, in one example may be included as 5x10 8 cfu.
- KCCM11829P Lactobacillus salivarius CJLS1511
- the animal feed additive composition may include the Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganism per 1.0 g of the composition as 1.0 ⁇ 10 8 cfu to 1.0 ⁇ 10 10 cfu. Specifically, it may be included as 1.0x10 8 cfu to 3.0x10 9 cfu, in one example may be included as 5x10 8 cfu.
- the animal feed additive composition may be in the form of a powder, pallet, or granular form.
- the content of Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganisms in the animal feed additive composition may range from 0.1 wt% to 10 wt%, specifically 0.1 wt% to 7 wt%.
- the composition includes the strain or the mycelium in the above range, there is an advantage that the triglyceride degrading ability, endotoxin adsorption capacity, pathogenic bacterial growth inhibitory ability and digestive enzyme activity of the animal feed additive composition can be maximized.
- an animal feed prepared using the animal feed additive composition is provided.
- the animal feed may be prepared by mixing an animal feed additive composition according to one aspect of the present invention with a conventional animal feed.
- the amount of the animal feed additive composition herein in the animal feed may be 0.1% to 1% by weight based on the animal feed weight.
- the animal feed is not particularly limited and may be a feed for a domestic animal.
- "Farming animal” is meant to include animals and pets for producing livestock and aquatic products useful to humans, such as milk, meat, eggs, fur, leather, feathers.
- it may be for feeding to a breeding animal such as dog, cow, chicken, pig, horse. More specifically, it may be for feeding poultry such as chickens, and more specifically for broilers.
- Animal feed may include ingredients such as carbohydrates, proteins, fats, vitamins, minerals, etc., which are essential for animal growth.
- the protein may be an animal protein or a vegetable protein, and may be, for example, meat meal, poultry meal, fish meal, soy protein, milk protein, gluten, or the like.
- the carbohydrates include cereals or beans, such as corn, rice, wheat, barley, oats, soybeans or mixtures thereof.
- the lipid may be animal lipid, vegetable lipid or meat. Or, in addition to the above ingredients, may further include other ingredients that add functionality to the animal feed. Alternatively, sugars, salts, spices, seasonings, flavors and the like can be incorporated.
- Another embodiment of the present invention by incubating Lactobacillus salivarius CJLS1511 (KCCM11829P) to prepare a culture solution,
- the culture is heated at a temperature of 70 °C to 160 °C,
- the heated culture is cooled to a temperature in the range of 10 °C to 60 °C,
- It relates to a method for producing Lactobacillus salivarius CJLS1511 (KCCM11829P) microorganism, comprising separating the Lactobacillus salivarius CJLS1511 (KCCM11829P) microorganism from the cooled culture.
- the step of preparing the culture by culturing the Lactobacillus salivarius CJLS1511 may be agar medium, specifically MRS medium.
- the culture is cultured for 5 hours to 48 hours at 25 °C to 40 °C, specifically, 12 hours to 36 hours culture at 30 °C to 40 °C, more specifically 20 to 30 hours culture at 35 °C to 40 °C culture Can be prepared.
- the culture is then heated to a temperature of 70 ° C. to 160 ° C.
- the heating may use direct heating means or indirect heating means, but may be through an indirect heating means such as a heat exchanger.
- the heating may be specifically carried out at a temperature range of 80 °C to 150 °C, more specifically 90 °C to 120 °C.
- the heating may inactivate or sterilize Lactobacillus salivarius CJLS1511.
- the heated culture may then be cooled to a temperature of 10 to 60 °C, specifically 20 to 50 °C, or in one example can be rapidly cooled to 4 °C.
- the cooling may be cooled at a rate of 10 ° C / min to 60 ° C / min, specifically 20 ° C / min to 50 ° C / min, more specifically 30 ° C / min to 40 ° C / min.
- Lactobacillus salivarius CJLS1511 (KCCM11829P) microorganisms can be obtained by separating the microorganisms from the cooled culture.
- the manufacturing method of the Lactobacillus salivarius CJLS1511 (KCCM11829P) microorganism may further comprise mixing a protective agent to the isolated microorganism.
- it may further comprise powdering a mixture of the microorganism and the protective agent.
- the protective agent is not particularly limited, but may be, for example, at least one of sugars such as yeast, yeast extract, monosaccharides, polysaccharides, starches, raw sugars, and refined sugars. Specifically, one or more substances selected from the group consisting of yeast extract, dextrose and raw sugar can be used.
- the powdering may be lyophilization, spray drying or spray flocculation, and more specifically, spray drying may be used.
- Spray drying is also called spray drying, and it is a method of spraying liquid at once in a hot air stream, and obtaining a liquid dried thing instantaneously.
- As a method of making a liquid into a spray shape there exist centrifugal spray by a rotating disk and pressurized spray by a pressure nozzle.
- the well-known spray drying apparatus which employ
- a mixture is prepared by mixing the microorganisms and a protecting agent, and then the mixture is suspended in water, and then the obtained suspension is spray-dried, that is, the inlet temperature of the hot air is 120 to 200 ⁇ , preferably It is preferable to carry out spray drying at 130-170 degreeC and outlet temperature 30-150 degreeC, Preferably it is 50-100 degreeC.
- the amount of the protective agent added may be 0.1 part by weight to 300 parts by weight, specifically 0.1 part by weight to 200 parts by weight based on 100 parts by weight of the dead microorganism.
- the addition amount of the yeast or yeast extract is 0.04 to 50 parts by weight, preferably 0.1 to 10 parts by weight, while the addition amount of the monosaccharide is 1 to 100 parts by weight, preferably 10 to 50 parts by weight,
- the addition amount is 0.2-50 weight part, Preferably you may be 0.4-10 weight part.
- the amount of the inorganic substance in the additive composition is 1 to 99% by weight, specifically 1 to 90% by weight, more specifically 1 to 10% by weight.
- the additive composition comprises from 0.05 to 50% by weight, preferably from 0.5 to 20% by weight, more preferably from 0.5 to 15% by weight, and from 5 to 80% by weight of inorganic material, preferably 5 It contains-50%, More preferably, it is 5-20%.
- the microorganisms in the additive composition may include the Lactobacillus salivarius CJLS1511 (KCCM11829P) or its microorganisms in an amount of 1.0 ⁇ 10 8 cfu to 1.0 ⁇ 10 10 cfu per 1 g of the composition. Specifically, it may be included as 1.0x10 8 cfu to 3.0x10 9 cfu, in one example may be included as 5x10 8 cfu.
- Lactobacillus salivarius KCCM 40210 As a comparative strain, the safety, acid resistance, bile resistance, antimicrobial activity, digestive enzyme activity and triglyceride degrading activity of the Lactobacillus salivarius CJLS1511 (KCCM11829P) strain of the present invention are as follows. It evaluated together.
- Lactobacillus salivarius CJLS1511 (KCCM11829P) was negative in all the gelatin liquefaction test, phenalanlan deamin test and ammonia production test, and appeared to be safe in hemolysis test.
- Lactobacillus salivarius CJLS1511 was less acidic than the comparison strain Lactobacillus salivarius KCCM 40210 standard strain at pH 2 to 4, the acid resistance was excellent.
- Lactobacillus salivarius CJLS1511 reduced the number of viable bacteria in both pH 4, 0.3% and 1% solution of bile acid (Oxgall), but the degree of reduction of the number of bacteria compared to Lactobacillus salivarius KCCM 40210 standard strain Much less was confirmed that even if the bile acid is secreted in the body of the administered strain can grow.
- E. coli K88, E. coli ATCC 25922, Salmonella typhimurium KCCM 25922, Salmonella cholerasuis KCCM 10709 Three pathogens ( E. coli K88, E. coli ATCC 25922, Salmonella typhimurium KCCM 25922, Salmonella cholerasuis KCCM 10709) that were cultured in 24-hour liquid culture in TSB (Tryptic soy broth) medium (BD, USA) were homogenized with sterile swabs. Smeared at 10 5-6 cfu / ml.
- the lactobacillus salivarius CJLS1511 and Lactobacillus salivarius KCCM 40210 standard strains were diluted in 10 9 cfu / mL in PBS buffer, respectively, and 50 ⁇ l were dispensed onto a 4 mm diameter paper disc.
- the mixture was left at room temperature until the dilution penetrated into the paper disc and incubated at 37 ° C. for 18 hours.
- the diluent is centrifuged (3,000 x g , 10 minutes) for the supernatant after the cultivation, and then used only the cells used after washing three times with PBS buffer.
- the size of the inhibitory ring was measured by the difference between the diameter of the entire transparent ring and the diameter of the agar groove. The results are shown in Table 6.
- E. coli K88 E. coli ATCC 25922 Salmonella typhimurium KCCM 25922 Salmonella cholerasuis KCCM 10709 Lactobacillus salivarius CJLS1511 ++ ++ ++ +++ Lactobacillus salivarius KCCM 40210 + ++ + ++ 10-15 mm: +, 15-20 mm: ++, 21-25 mm: +++
- Lactobacillus salivarius CJLS1511 and Lactobacillus salivarius KCCM 40210 standard strains showed proliferation inhibitory action against pathogenic microorganisms
- the Lactobacillus salivarius CJLS1511 strain was lactobacillus salivarius KCCM 40210 standard. It was confirmed that the antibacterial activity is higher than the strain.
- a medium to which 0.5 (w / v)% of phytate calcium salt (Ca-phytate) was added was prepared. The strains isolated on each medium prepared above were streaking and observed after culturing for 24 hours.
- A-amylase and cellulase activity was treated with 2% congo red (Sigma, USA) reagent in culture medium for 24 hours, washed with 1M sodium chloride (Nacl), and then washed by color. It was confirmed whether or not the enzyme. In addition, the determination of protease and phytase activity was confirmed by the presence of a transparent ring, and the results are shown in Table 7 below.
- Protease activity Phytase activity Lipase activity Cellulose activity Amylase activity Lactobacillus salivarius CJLS1511 +++ ++ + +++ +++ Lactobacillus salivarius KCCM 40210 + + ++ ++ ++ 1-10 mm: +, 11-20 mm: ++, 21-30 mm: +++
- Lactobacillus salivarius CJLS1511 was found to have a higher digestive enzyme activity such as protease activity, phytase activity, cellulose degradation activity, amylase activity and the like compared to Lactobacillus salivarius KCCM 40210 standard strain. .
- BSH Bile salt hydrolase
- Lactobacillus salivarius CJLS1511 has a high bile resistance and converts it into a form capable of degrading triglycerides by the production of BSH, which can affect the weight gain when applied to animals It was confirmed that the strain.
- Lactobacillus salivarius CJLS1511 and Lactobacillus salivarius KCCM 40210 standard strains were respectively plated in MRS agar medium in a loop and incubated at 37 ° C. for 48 hours to prepare a culture solution.
- the culture solution was indirectly heated to a temperature of 100 ° C. using a heat exchanger, and it was rapidly cooled to 10 ° C. at a rate of 30-100 L / min.
- Lactobacillus salivarius CJLS1511 bacterium and Lactobacillus salivarius KCCM 40210 were prepared by separating the bacterium from the rapidly cooled culture medium through a centrifuge (3,000 ⁇ g , 10 minutes).
- Lactobacillus salivarius CJLS1511 and Lactobacillus salivarius KCCM 40210 standard strains were respectively plated in MRS agar medium in a loop and incubated at 37 ° C. for 48 hours to prepare a culture solution.
- Lactobacillus salivarius CJLS1511 and Lactobacillus salivarius KCCM 40210 were prepared by separating centrifugal cells from a rapidly cooled culture medium through a centrifuge (3,000 x g , 10 minutes), wherein yeast extract, Dextrose and raw sugar were mixed respectively. After the mixture was suspended in water, the suspension obtained was spray dried to obtain a powder. That is, spray-drying was carried out by making the inlet temperature of hot air 150 degreeC, and outlet temperature 100 degreeC.
- the addition amount of the protective agent was based on 100 parts by weight of the dead microorganisms, the addition amount of yeast extract 20 parts by weight, the addition amount of dextrose 30 parts by weight, the addition amount of raw sugar was 5 parts by weight.
- microorganisms prepared in Preparation Example 1 hydrophobicity and cohesiveness were tested by the method described below, and the weight gain and feed rate of the basic feed including Lactobacillus salivarius CJLS1511 microorganisms Compared.
- the autocoagulation reaction was vortexed for 90 seconds by adding 1 mL of toluene to 3 mL of Lactobacillus diluted to be OD 600 0.5 against Lactobacillus salivarius CJLS1511 bacteria and dead bacteria. After leaving for 1 hour in a 37 °C water bath to remove the toluene and measured the OD 600 of the aqueous layer.
- pathogenic microorganisms E. coli K88, Salmonella typhimurium KCCM 25922, Salmonella cholerasuis KCCM 10709
- Hydrophobicity (%) is 100 x - was calculated as (initial OD 600 1 sigan OD 600) / initial OD 600.
- Lactobacillus salivarius CJLS1511 microorganism-administered group of the present invention showed superior effects in both weight gain and feed requirement compared to the control group not administered to it.
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Abstract
Description
1. 형태학적 특징(MRS 고체배지에서 성장시켰을 때의 형태) | |
- 세포의 모양 | 간균 |
- 세포의 다형성(多形性) | 없음 |
- 운동성 | 없음 |
- 아포 형성 | 없음 |
2. 생리학적 특징 | |
- 그람 염색(Gram staining) | 그람 양성(Gram positive) |
- 카탈라아제(Catalase) | 음성 |
- 옥시다아제(Oxidase) | 음성 |
- 생육 온도 및 시간 | 37°C, 18~48시간 |
- 산소 요구도 | 통성혐기성 |
No. | 당종류 | 락토바실러스 살리바리우스 CJLS1511 |
0 | Temoin | - |
1 | Glycerol | - |
2 | Erythritol | - |
3 | D-arabinose | - |
4 | L-arabinose | - |
5 | D-ribose | - |
6 | D-xylose | - |
7 | L-xylose | - |
8 | D-adonito | - |
9 | Methyl-aD-xylopyranozide | + |
10 | D-galactose | + |
11 | D-glucose | + |
12 | D-fructose | + |
13 | D-mannose | - |
14 | L-sorbose | - |
15 | L-ramnose | - |
16 | D-ulcitol | - |
17 | Inositol | + |
18 | D-mannitol | + |
19 | D-sorbitol | + |
20 | Methyl-aD-manopyranozide | - |
21 | Methyl-aD-glucopyranozide | - |
22 | N-acetylglucosamine | + |
23 | Amygdalin | - |
24 | Arbutin | + |
25 | Asculin | - |
26 | Salicin | + |
27 | D-celobiose | - |
28 | D-maltose | + |
29 | D-lactose | + |
30 | D-mellibiose | + |
31 | D-saccharose | + |
32 | D-trehalose | + |
33 | Inulin | - |
34 | D-melesitose | - |
35 | D-rafinose | + |
36 | Amidon | - |
37 | Glycogen | - |
38 | Xylitol | - |
39 | Genthiobiose | - |
40 | D-turanose | - |
41 | D-lyxose | - |
42 | D-tagatose | - |
43 | D-fucose | - |
44 | L-fucose | - |
45 | D-arabitol | - |
46 | L-arabitol | - |
47 | Gluconate | - |
48 | 2-ketogluconate | - |
49 | 5-ketogluconate | - |
균주 | 항목 | |||
젤라틴 액화 반응 검사 | 페닐알라닌 탈아민 검사 | 용혈 현상 검사 | 암모니아 생성 | |
락토바실러스 살리바리우스 CJLS1511 | 음성 | 음성 | a-용혈, 안전함 | 음성 |
Strains | MRS broth (pH7) | MRS broth (pH2) | MRS broth (pH3) | ||||||
생균수 (CFU/mL) | 생균수 (CFU/mL) | 생균수 (CFU/mL) | |||||||
0 h | 1 h | 3 h | 0 h | 1 h | 3 h | 0 h | 1 h | 3 h | |
락토바실러스 살리바리우스 CJLS1511 | 5.8 x 108 | 6.0 x 108 | 6.2 x 108 | 5.8 x 108 | 2.3 x 105 | 2.0 x 105 | 5.8 x 108 | 3.5 x 108 | 2.0 x 108 |
락토바실러스 살리바리우스 KCCM 40210 | 6.4 x 108 | 6.8 x 108 | 8.6 x 108 | 6.4 x 108 | 2.0 x 102 | 2.0 x 102 | 6.4 x 108 | 1.0 x 108 | 5.0 x 106 |
Strains | MRS broth (pH4) | MRS broth(pH4 and Oxgall 0.3%) | MRS broth(pH4 and Oxgall 1%) | ||||||
생균수 (CFU/mL) | 생균수 (CFU/mL) | 생균수 (CFU/mL) | |||||||
0 h | 1 h | 3 h | 0 h | 1 h | 3 h | 0 h | 1 h | 3 h | |
락토바실러스 살리바리우스 CJLS1511 | 2.4 x 108 | 4 x 108 | 5.6 x 108 | 2.4 x 108 | 4.0 x 106 | 2.0 x 104 | 2.4 x 108 | 5.6 x 105 | 5.2 x 103 |
락토바실러스 살리바리우스 KCCM 40210 | 2.0 x 108 | 2.0 x 108 | 2.2 x 108 | 2.0 x 108 | 2.6 x 105 | 2.0 x 102 | 2.0 x 108 | 2.0 x 102 | 2.0 x 102 |
Strains | E. coli K88 | E. coliATCC 25922 | Salmonella typhimuriumKCCM 25922 | Salmonella cholerasuisKCCM 10709 |
락토바실러스 살리바리우스 CJLS1511 | ++ | ++ | ++ | +++ |
락토바실러스 살리바리우스 KCCM 40210 | + | ++ | + | ++ |
10~15 mm: +, 15~20 mm: ++, 21~25 mm: +++ |
Strains | Protease activity | Phytase activity | Lipase activity | Cellulose activity | Amylase activity |
락토바실러스 살리바리우스 CJLS1511 | +++ | ++ | + | +++ | +++ |
락토바실러스 살리바리우스 KCCM 40210 | + | + | ++ | ++ | ++ |
1~10 mm: +, 11~20 mm: ++, 21~30 mm: +++ |
복합 담즙산/균주명 | 락토바실러스 살리바리우스 CJLS1511 | 락토바실러스 살리바리우스 KCCM 40210 |
TDCA | O | O |
GDCA | + | - |
항목/병원성 균주 | 락토바실러스 살리바리우스 CJLS1511 | ||
생균 | 사균체 | ||
자가응집반응 | 11% | 23% | |
동시응집반응 | E. coli K88 | 54% | 64% |
Salmonella typhimurium KCCM 25922 | 45% | 65% | |
Salmonella cholerasuis KCCM 10709 | 30% | 45% |
구분 | 대조군 | 락토바실러스 살리바리우스 CJLS1511 사균체 투여군 |
시험 마리수 | 150 | 150 |
개시 평균 체중(g) | 38.48 | 38.47 |
종료 평균 체중(g) | 1582.00a | 1690.24b |
일당 증체량(g/d) | 53.23a | 56.96b |
일당 사료 섭취량(g/d) | 78.76 | 79.48 |
사료 요구율 | 1.48a | 1.40b |
Claims (15)
- 락토바실러스 살리바리우스 CJLS1511 (Lactobacillus salivarius CJLS1511)( KCCM11829P) 또는 이의 사균체.
- 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 또는 이의 사균체를 포함하는, 동물 사료 첨가제 조성물.
- 제2항에 있어서, 상기 동물 사료 첨가제 조성물이 상기 사균체를 포함하는, 동물 사료 첨가제 조성물.
- 제2항에 있어서, 상기 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 또는 이의 사균체가 동물 사료 첨가제 조성물 1g 당 1.0x108 cfu 내지 1.0x1010 cfu로 포함되는, 동물 사료 첨가제 조성물.
- 제2항 내지 제4항 중 어느 하나의 항에 있어서, 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 또는 이의 사균체의 보호제를 추가로 포함하는, 동물 사료 첨가제 조성물.
- 제5항에 있어서, 상기 보호제가 효모, 효모추출물, 단당류, 다당류, 전분류, 설탕류 중 1종 이상인, 동물 사료 첨가 첨가제 조성물.
- 제2항 내지 제4항 중 어느 하나의 항에 있어서, 상기 동물이 육계인, 동물 사료 첨가제 조성물.
- 제2항 내지 제4항 중 어느 하나의 항에 있어서, 상기 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 또는 이의 사균체가 동물 사료의 중량 기준으로 0.1 중량% 내지 1 중량%인, 동물 사료 첨가제 조성물.
- 제8항에 따른 동물 사료 첨가제 조성물을 포함하는 동물 사료.
- 락토바실러스 살리바리우스 CJLS1511(KCCM11829P)를 배양하여 배양액을 제조하고,상기 배양액을 70℃ 내지 160℃의 온도에서 가열하고,상기 가열된 배양액을 10℃ 내지 60℃의 범위의 온도로 냉각하고,상기 냉각된 배양액에서 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 사균체를 분리하는 것을 포함하는, 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 사균체의 제조 방법.
- 제10항에 있어서, 상기 가열이 열교환기를 통해 간접 가열되는, 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 사균체의 제조 방법.
- 제10항에 있어서, 상기 냉각이 10℃/min ~ 60℃/min의 속도로 냉각되는, 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 사균체의 제조 방법.
- 제10항 내지 제12항 중 어느 하나의 항에 있어서, 상기 분리된 사균체에 보호제를 혼합하는 것을 추가로 포함하는, 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 사균체의 제조 방법.
- 제13항에 있어서, 상기 혼합된 사균체를 분말화하는 것을 추가로 포함하는, 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 사균체의 제조 방법.
- 제13항에 있어서, 상기 보호제가 효모, 효모추출물, 단당류, 다당류, 전분류, 설탕류 중 1종 이상인, 락토바실러스 살리바리우스 CJLS1511(KCCM11829P) 사균체의 제조 방법.
Priority Applications (9)
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EP17839748.5A EP3498820A4 (en) | 2016-08-09 | 2017-08-07 | LACTOBACILLUS SALIVARIUS CJLS1511, ANIMAL FOOD ADDITIVE COMPOSITION COMPRISING THE SAME OR DEAD CELLS THEREOF AND PROCESS FOR PRODUCING SAID DEAD CELLS |
MYPI2019000683A MY193328A (en) | 2016-08-09 | 2017-08-07 | Lactobacillus salivarius cjls1511, animal feed additive composition comprising same bacterium or dead cells thereof, and method for producing same dead cells |
AU2017309095A AU2017309095B2 (en) | 2016-08-09 | 2017-08-07 | Lactobacillus salivarius CJLS1511, animal feed additive composition comprising same bacterium or dead cells thereof, and method for producing same dead cells |
RU2019104984A RU2721127C1 (ru) | 2016-08-09 | 2017-08-07 | Lactobacillus salivarius cjls1511, композиция кормовой добавки для животных, содержащая указанную бактерию или ее мертвые клетки, и способ получения указанных мертвых клеток |
JP2019506700A JP6715386B2 (ja) | 2016-08-09 | 2017-08-07 | ラクトバチルスサリバリウスcjls1511、該菌またはその死菌体を含む動物飼料添加剤組成物、及び該死菌体の製造方法 |
BR112019002545-6A BR112019002545B1 (pt) | 2016-08-09 | 2017-08-07 | Composição para aditivos de ração animal, ração animal e método para preparar uma composição para aditivos de ração animal |
US16/324,462 US11382940B2 (en) | 2016-08-09 | 2017-08-07 | Lactobacillus salivarius CJLS1511, animal feed additive composition comprising same bacterium or dead cells thereof, and method for producing same dead cells |
CN201780049252.5A CN109563473B (zh) | 2016-08-09 | 2017-08-07 | 唾液乳杆菌cjls1511,含该菌或死细胞的动物饲料添加组合物及生产死细胞的方法 |
PH12019500264A PH12019500264A1 (en) | 2016-08-09 | 2019-02-07 | Lactobacillus salivarius cjls1511, animal feed additive composition comprising same bacterium or dead cells thereof, and method for producing same dead cells |
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KR1020160101355A KR101873899B1 (ko) | 2016-08-09 | 2016-08-09 | 락토바실러스 살리바리우스 cjls1511, 상기 균 또는 이의 사균체를 포함하는 동물 사료 첨가제 조성물, 및 상기 사균체의 제조 방법 |
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EP (1) | EP3498820A4 (ko) |
JP (1) | JP6715386B2 (ko) |
KR (1) | KR101873899B1 (ko) |
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BR (1) | BR112019002545B1 (ko) |
MY (1) | MY193328A (ko) |
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KR102130646B1 (ko) * | 2018-10-15 | 2020-07-06 | 주식회사 제일바이오 | 신규한 락토바실러스 살리바리우스 및 이를 유효성분으로 포함하는 양식 어류 및 갑각류용 사료첨가제 |
KR102251294B1 (ko) | 2018-11-06 | 2021-05-12 | 주식회사 메디뉴트롤 | 락토바실러스 살리바리우스 v133 균주의 사균체를 유효성분으로 함유하는 알코올성 위염의 예방, 개선 또는 치료용 조성물 |
KR102251295B1 (ko) | 2020-04-02 | 2021-05-12 | 주식회사 메디뉴트롤 | 락토바실러스 살리바리우스 v133 균주의 사균체를 유효성분으로 함유하는 숙취 예방 또는 해소용 조성물 |
KR20220011293A (ko) | 2020-07-21 | 2022-01-28 | 주식회사 종근당바이오 | 대사성 질환의 예방 또는 치료 활성을 갖는 락토바실러스 플란타룸 사균체 |
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JP6715386B2 (ja) | 2020-07-01 |
CN109563473B (zh) | 2022-05-27 |
JP2019524129A (ja) | 2019-09-05 |
CN109563473A (zh) | 2019-04-02 |
MY193328A (en) | 2022-10-05 |
AU2017309095A1 (en) | 2019-02-28 |
BR112019002545B1 (pt) | 2023-01-24 |
BR112019002545A2 (pt) | 2019-05-21 |
PH12019500264A1 (en) | 2019-06-17 |
KR20180017489A (ko) | 2018-02-21 |
US20190216867A1 (en) | 2019-07-18 |
EP3498820A1 (en) | 2019-06-19 |
EP3498820A4 (en) | 2020-01-22 |
AU2017309095B2 (en) | 2020-10-01 |
RU2721127C1 (ru) | 2020-05-18 |
US11382940B2 (en) | 2022-07-12 |
KR101873899B1 (ko) | 2018-07-03 |
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