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Clostridium butyricum used for feeding, and application thereof

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

Inactive Publication Date: 2014-04-16
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

AI Technical Summary

Problems solved by technology

None of the Clostridium butyricum microecological preparations disclosed in the above patent documents involves the process of anaerobic liquid fermentation of Clostridium butyricum, followed by adsorption with solid materials (bran, soybean meal, rice husk, etc.), and finally drying.

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
Comparison scheme
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; before use, adjust the pH to 7.1±0.1, cover the culture medium with 5mm liquid paraffin, and sterilize at 115°C for 20min for later use.

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

[0033] The cellar mud was collected from multiple points and mixed from the base wine cellar of Liurenba Town of Hubei Jinpai Wine Co., Ltd., the wine...

Embodiment 2

[0042] Embodiment 2, the acid resistance property analysis of clostridium butyricum (Clostridium butyricum) B1 CGMCC No.4845 spore

[0043] For bacterial strain activation, Clostridium butyricum (Clostridium butyricum) B1 CGMCC No.4845 was inoculated into a test tube containing 9 mL of RCM medium at an inoculum size of 1% (volume / volume percentage), and cultured anaerobically at 37° C. for 48 hours, Discard the supernatant by centrifugation, wash and suspend the bacteria. Take 0.2mL of centrifuged and washed bacterial suspension and inoculate them into 2mL of sterile phosphate buffer solution containing pH 1.0, pH 2.0, and pH 3.0 respectively (solution A: phosphoric acid solution, solution B: disodium hydrogen phosphate solution; by adjusting solution A, solution B Ratio, the pH was adjusted to 1.0, 2.0, 3.0) in anaerobic test tubes, placed in a 37°C incubator, and the viable bacteria were counted at 0h, 1h, 2h, and 3h respectively, and the survival rate was calculated.

[00...

Embodiment 3

[0045] Example 3, analysis of bile salt resistance properties of Clostridium butyricum (Clostridium butyricum) B1 CGMCC No.4845

[0046] For bacterial strain activation, Clostridium butyricum (Clostridium butyricum) B1 CGMCC No.4845 was inoculated into a test tube containing 9 mL of RCM medium at an inoculum size of 1% (volume / volume percentage), and cultured anaerobically at 37° C. for 48 hours, Centrifuge to remove the supernatant, wash and suspend the cells. Get 0.2mL of centrifuged washing bacterial suspension and inoculate in 2mL RCM liquid medium containing 0.1%, 0.3%, 0.5% (mass percentage concentration) pig bile salt. Put them in an incubator at 37°C, count the viable bacteria at 0h, 1h, 2h, 3h, and 20h, and calculate the survival rate.

[0047] Detecting result shows, this bacterial strain is in the bile salt solution of 0.5% (mass percentage concentration) 1h and the survival rate is 76.7%, and the survival rate after 20h is 24.3%; In the bile salt solution below 0....

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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, in particular to a Clostridium butyricum strain and its application in the preparation of feed and feed additives. Background technique [0002] Clostridium butyricum, also known as Clostridium butyricum or Clostridium butyricum, is an anaerobic beneficial bacterium that exists in the intestines of humans and livestock. Clostridium butyricum can form endophytic spores, and has the characteristics of high temperature resistance, gastric acid resistance, bile salt resistance, and resistance to some antibiotics (such as gentamicin, kanamycin, etc.). Clostridium butyricum preparations, as feed additives or veterinary medicines, have significant advantages over 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-dissolving Clostridium butyricum and its preparati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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