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Lactobacillus johnsonii, microbial inoculum, application and premix thereof

A Lactobacillus johnsonii, premix technology, applied in the application, bacteria, animal feed and other directions, to achieve strong gastric acid tolerance, improve economic benefits, and improve the effect of production performance

Active Publication Date: 2014-07-09
BEIJING DABEINONG TECH GRP CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The joint application of Lactobacillus johnsonii, Lactobacillus acidophilus and Bacillus pumilus to prepare microecological preparations or premixes has not been reported at home and abroad

Method used

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  • Lactobacillus johnsonii, microbial inoculum, application and premix thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Verification of Stress Resistance of Lactobacillus johnsonii CGMCC No.4926

[0022] Inoculate 1 mL of Lactobacillus johnsoni CGMCC No.4926 bacteria solution into 10 mL of MRS medium, and incubate at 37°C for 20 hours, then take 3 mL of the bacterial suspension, centrifuge at 12,000 rpm for 5 minutes, discard the supernatant, and add 10ml pH 2.0, 1% pepsin artificial gastric juice, stand at 37°C for 2h; centrifuge at 12,000rpm for 5min, discard the supernatant, add the bacterial pellet to 10ml artificial bile salt with a concentration of 0.3%, let stand at 37°C for 3h . After oscillating and mixing, take 1 mL of the bacterial suspension and dilute it with sterile physiological saline gradient to coat the MRS medium plate. After anaerobic culture at constant temperature at 37°C for 20 h, colonies were purified. The strains obtained from the primary screening were further screened.

[0023] (1) Tolerance of artificial gastric juice

[0024] Inoculate 1 mL of ...

Embodiment 2

[0031] Example 2 Verification of Lactobacillus johnsonii CGMCC No.4926 antibacterial ability

[0032] Melt the LB solid medium and cool to 45°C. Add E.coli K88, K99 and Staphylococcus aureus cultured overnight (add 1 microliter of bacterial solution per milliliter of culture medium), shake and mix well, pour into a sterile plate, solidify horizontally, and place gently on each agar plate 4 Oxford cups should be equally spaced. Add 50 μL of Lactobacillus johnsonii bacteria solution to each Oxford cup, cover the dish cover, carefully move it to a 37°C incubator, and place the dish for static culture. After cultivating for 20 hours, open the lid of the dish, remove the Oxford cup, and measure the diameter of the inhibition zone with a caliper (results are shown in Table 1).

[0033] Table 1 Determination of inhibition zone

[0034]

example 3

[0035] The preparation of example 3 Lactobacillus johnsonii CGMCC No.4926 live bacteria preparation

[0036] Preparation of Lactobacillus johnsonii seed solution: Inoculate the strains preserved on the slant of the test tube into an anaerobic tube containing 20 mL of MRS medium, and culture at a constant temperature for 20 hours at 37°C until the number of viable bacteria reaches 10 7 CFU / mL, inoculate at 1% in a 250mL Erlenmeyer flask containing 100mL seed medium, culture anaerobically at 37°C for 20h, until the number of viable bacteria reaches 10 7 CFU / mL or more, as a seed solution for later use.

[0037] Seed liquid MRS medium is: peptone, 10.0g / L; beef extract, 8.0g / L; yeast extract, 4.0g / L; glucose, 20.0g / L; Tween 80, 1mL / L; dipotassium hydrogen phosphate, 2.0g / L; Sodium acetate trihydrate, 5.0g / L; Triammonium citrate, 2.0g / L; Magnesium sulfate heptahydrate, 0.2g / L; Manganese sulfate, 0.05g / L; Add distilled water to 1000mL, pH 6.2 ±0.2. liquid submerged fermentation ...

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Abstract

The invention belongs to the technical field of microecological preparations, and particularly relates to lactobacillus johnsonii, a microbial inoculum, application and a premix thereof. The lactobacillus johnsonii, lactobacillus acidophilus and bacillus pumilus has a good health-benefiting function and high stress resistance; the three bacteria are prepared into a bacterial agent first and then into a feed additive to be added into the premix, and after animals are fed with the microbial inoculum and the feed additive, the productivity of weaned pigs can be improved obviously, the diarrhea rate of the pigs can be reduced, and the economic benefit can be improved. The lactobacillus johnsonii has a popularization and application prospect, and provides a theoretical and practice foundation for seeking antibiotic substitutes.

Description

technical field [0001] The invention belongs to the technical field of microecological preparations, and in particular relates to Lactobacillus johnsonii and its bacterial preparation, application and premix. Background technique [0002] As people's awareness of the negative effects of antibiotic feed additives continues to increase, feed microecological preparations, as a new type of antibiotic substitute, have many advantages such as no toxic side effects, no drug resistance, no residue, and significant effects. welcomed by the farming industry. [0003] As a kind of microecological preparation, compound live bacteria preparation has been widely used in animal husbandry. It is mainly composed of two or more different genera of probiotics, focusing on the complementarity between different genera, and generally exerting a positive combination effect. Probiotics with different biological functions are combined into a compound live bacterial preparation to maintain the stab...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K1/16C12R1/225C12R1/23C12R1/07
Inventor 胡婷刘滢赵莉莉刘婷
Owner BEIJING DABEINONG TECH GRP CO LTD
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