Clostridium butyricum used for feeding, and application thereof

A technology of Clostridium butyricum and bacteria agent, applied in the direction of application, bacteria, animal feed, etc., to achieve broad application prospects, strong inhibitory effect, and the effect of enhancing animal disease resistance

Inactive Publication Date: 2012-01-18
北京金泰得生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

None of the Clostridium butyricum microecological preparations disclosed in the above patent documents involves the process of anaerobic li...

Method used

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  • Clostridium butyricum used for feeding, and application thereof
  • Clostridium butyricum used for feeding, and application thereof
  • Clostridium butyricum used for feeding, and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1. Isolation, identification and preservation of Clostridium butyricum B1 CGMCC No. 4845

[0030] 1. Isolation of Clostridium butyricum B1 CGMCC No. 4845

[0031] Clostridium proliferation medium (RCM medium): yeast extract 3g, beef extract 10g, tryptone 10g, glucose 5g, soluble starch 1g, sodium chloride 5g, sodium acetate trihydrate 3g, cysteine ​​hydrochloride 0.5g, 0.5% methylene blue 0.2mL, distilled water 1000mL; adjust the pH to 7.1±0.1 before use, cover the culture medium with 5mm liquid paraffin, and sterilize it for use at 115°C for 20 minutes.

[0032] Clostridium selective medium (TSN medium): tryptone 15g, yeast extract 10g, sodium sulfite 1g, iron citrate 0.5g, novobiocin 0.02g, polymyxin 0.05g, agar 0.5g (with solid Used for culture medium), pH 7.2, 115°C, 20min sterilization for later use.

[0033] Multi-point and mixed collection of cellar mud from Hubei Jinpai Wine Co., Ltd. Liuren Bazhen base wine base wine cellar, Hubei Zhijiang Wine Co., Ltd. wine ...

Embodiment 2

[0042] Example 2: Analysis of acid resistance properties of Clostridium butyricum B1 CGMCC No. 4845 spores

[0043] Strain activation, inoculate Clostridium butyricum B1 CGMCC No. 4845 into a test tube containing 9mL RCM medium at an inoculum of 1% (volume / volume percentage), and culture at 37°C for 48h under anaerobic static. Centrifuge and discard the supernatant, wash and suspend the bacteria. Take 0.2mL centrifugal washing bacteria suspension and inoculate respectively 2mL pH 1.0, pH 2.0, pH 3.0 sterile phosphate buffer solution (A solution: phosphoric acid solution, B solution: disodium hydrogen phosphate solution; by adjusting A solution, B solution Proportion, the pH was adjusted to 1.0, 2.0, 3.0) in an anaerobic test tube, placed in a 37°C incubator, and counted viable bacteria at 0h, 1h, 2h, and 3h to calculate the survival rate.

[0044] Results The spore survival rate was 60% after the phosphate buffer with pH value of 1 for 1 hour, and 32.7% after the phosphate buffer ...

Embodiment 3

[0045] Example 3 Analysis of the bile salt tolerance properties of Clostridium butyricum B1 CGMCC No. 4845

[0046] The strain is activated, and Clostridium butyricum B1 CGMCC No. 4845 is inoculated into a test tube containing 9mL RCM medium at an inoculum of 1% (volume / volume percentage), and cultured at 37°C for 48h under anaerobic static. Centrifuge to remove the supernatant, wash and suspend the bacteria. Take 0.2mL centrifugal washing bacteria suspension and inoculate it in 2mL RCM liquid medium containing 0.1%, 0.3%, 0.5% (mass percentage concentration) of porcine bile salt. Placed in a 37°C incubator, counted viable bacteria at 0h, 1h, 2h, 3h, and 20h, and calculated the survival rate.

[0047] The test results showed that the survival rate of the strain was 76.7% after 1h in a bile salt solution of 0.5% (mass percentage concentration) and 24.3% after 20h; in a bile salt solution of less than 0.3% (mass percentage concentration) for 20h The post-survival rate was 66.5%, an...

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Abstract

The invention relates to a clostridium butyricum strain used for feeding, and an application thereof. The provided clostridium butyricum B1 CGMCC No. 4845 is a Gram-positive rod. When observed by using an optical microscope, the sizes of the strains are (0.6-1.2)mum*(3.0-7.0)mum; the ends of the strains are round; the middle parts of the strains are slightly bulged; the strains are single or in couples; the strains are short-chained; filamentous thallus with peritrichous is occurred; the strains are movable; the spores are egg-shaped, and are grew decentrically or on the minor ends; the strains form white or cream-colored small-scaled round colonies on an anaerobic agarose plate; the strains grow under an anaerobic condition, with a growth temperature range of 20 DEG C to 60 DEG C, and preferably 30 DEG C to 42 DEG C; the growth hydrogen power range of the strains is pH 2-10, and preferably pH 7.0. Microbial manure prepared with the strains brings no toxin and harm, and is gastric juice resistant and bile salt resistant. The microbial manure provides strong inhibiting effects against various harmful bacteria. Therefore, the strains can be widely applied in poultry and livestock cultivation industries. With the strain, animal disease resistance can be improved. The strain has potential to be applied as a substitution of antibiotics used for feeding.

Description

Technical field [0001] The invention belongs to microbial strains, and particularly relates to a strain of Clostridium butyricum and its application in preparing feed and feed additives. Background technique [0002] Clostridium butyricum (Clostridium butyricum), also known as Clostridium butyricum or Clostridium butyricum, is an anaerobic beneficial bacteria that exists in the intestines of humans and livestock. Clostridium butyricum can form endophytic spores, which are resistant to high temperature, gastric acid, bile salt, and some antibiotics (such as gentamicin, kanamycin, etc.). As a feed additive or veterinary medicine, Clostridium butyricum preparations have significant advantages compared with non-spore live bacteria preparations. [0003] At present, the research and application of Clostridium butyricum is still in its infancy. Patent document CN 101851596A discloses a high-efficiency phosphate-solubilizing Clostridium butyricum and its preparation method and applicatio...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K1/16A61K35/74A61P31/00C12R1/145A23K10/18A61K35/742
Inventor 张东晓梁运祥赵述淼唐泽华谢树贵雷红升付生慧游伟方莉玉
Owner 北京金泰得生物科技股份有限公司
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