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Preparation method of bacillus licheniformis and butyric acid bacterium composite bacterial agent

A technology of Bacillus licheniformis and composite bacterial agents, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of unstable growth of the two bacteria, high production costs, and high equipment requirements, and achieve improved intestinal micro-environment, improve production efficiency and reduce production cost

Inactive Publication Date: 2018-01-09
SHANXI DAYU BIOLOGICAL ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Since butyric acid bacteria is a strict anaerobic bacteria, if it is produced alone, it needs to be produced in an anaerobic environment maintained by paraffin oil, nitrogen or carbon dioxide, etc., which requires high equipment and high production costs; while Bacillus licheniformis belongs to aerobic bacteria, The joint culture method often leads to the phenomenon that the growth of both bacteria is unstable or severely degenerated

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Activation and expansion of strains.

[0043] (1) Bacillus licheniformis:

[0044] a) Strain activation: Take the glycerol tube strain of Bacillus licheniformis, inoculate it into the beef extract peptone liquid medium, incubate at 120 r / min, 30°C for 24 hours, and count on the beef extract peptone plate.

[0045] b) Bacteria expansion: select a single colony counted on the beef extract peptone plate, inoculate it into the beef extract peptone liquid medium, and culture at 30°C and 120 r / min for 24 hours with shaking.

[0046] (2) Butyric acid bacteria:

[0047] a) Activation of strains: Inoculate the glycerol tube strains of butyric acid bacteria into the corresponding seed medium, culture anaerobically at 32°C for 12 hours, and count on RCM plates.

[0048] b) Strain expansion: select a single colony on the RCM plate, inoculate it in RCM liquid medium, place it in an anaerobic bag, and culture it anaerobically at 37°C for 24 hours.

[0049] 2. Cultivate strains ...

Embodiment 2

[0071] 1. Activation and expansion of strains.

[0072] (1) Bacillus licheniformis:

[0073] a) Strain activation: take the Bacillus licheniformis glycerol tube strain, inoculate it into the beef extract peptone liquid medium, culture at 140 r / min, 34°C for 24 hours, and count on the beef extract peptone plate.

[0074] b) Bacteria expansion: select a single colony counted on the beef extract peptone plate, inoculate it into the beef extract peptone liquid medium, and incubate at 34°C for 24 hours with shaking at 140 r / min.

[0075] (2) Butyric acid bacteria:

[0076] a) Activation of strains: Inoculate the glycerol tube strains of butyric acid bacteria into the corresponding seed medium, culture anaerobically at 34°C for 15 hours, and count on RCM plates.

[0077] b) Strain expansion: select a single colony on the RCM plate, inoculate it in RCM liquid medium, place it in an anaerobic bag, and culture it anaerobically at 37°C for 24 hours.

[0078] 2. Cultivate strains in s...

Embodiment 3

[0100] 1. Activation and expansion of strains.

[0101] (1) Bacillus licheniformis:

[0102] a) Strain activation: take the glycerol tube strain of Bacillus licheniformis, inoculate it into the beef extract peptone liquid medium, culture at 160 r / min, 37°C for 24 hours, and count on the beef extract peptone plate.

[0103] b) Bacteria expansion: select a single colony counted on the beef extract peptone plate, inoculate it into the beef extract peptone liquid medium, and incubate at 37°C with shaking at 160 r / min for 24 hours.

[0104] (2) Butyric acid bacteria:

[0105] a) Activation of strains: Inoculate the glycerol tube strains of butyric acid bacteria into the corresponding seed medium, culture anaerobically at 37°C for 18 hours, and count on RCM plates.

[0106] b) Strain expansion: select a single colony on the RCM plate, inoculate it in RCM liquid medium, place it in an anaerobic bag, and culture it anaerobically at 37°C for 24 hours.

[0107] 2. Cultivate strains i...

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Abstract

The invention discloses a preparation method of a bacillus licheniformis and butyric acid bacterium composite bacterial agent, and belongs to the technical field of biological fermentation. Accordingto the method, bacillus licheniformis secondary seed liquid is inoculated into a fermentation tank; when the viable count during the microscopic examination is 1.0*10<9> to 3.0*10<9>CFU / ml, the butyric acid bacterium secondary seed liquid is inoculated into a fermentation tank according to the proportion of the weight percentage being 2 to 6 percent; when the butyric acid bacterium is fermented to13 to 15h, a glucose solution starts to be added in a flowing manner; when the butyric acid bacterium is fermented to 14 to 16h, a nitrogen source supplementary material culture medium starts to be added in a flowing manner. The method provided by the invention uses a combined culture mode, the mutualism among microbes is utilized; the production cost is reduced; the production benefits are improved; the maintenance of the animal microecosystem is facilitated; the digestive absorption rate of the feed is improved through various kinds of secreted digestive enzymes and organic acids; the method can be applied to feed additives.

Description

technical field [0001] The invention relates to the technical field of biological fermentation, in particular to a method for preparing a compound bacterial agent of bacillus licheniformis and butyric acid bacteria by fed-batch fermentation. Background technique [0002] As the phenomenon of adding antibiotics to feed is very common at present, for safety reasons, many countries prohibit the use of meat and bone meal and other animal protein raw materials produced from leftovers of livestock products for feed. Therefore, from the perspective of safety and cost, plant-based protein As a substitute for animal protein raw materials has become a trend. However, in order to efficiently utilize plant-derived protein, it is necessary to solve the problems of anti-nutritional factors and palatability that affect digestion and absorption; at the same time, in the prevention and treatment of diarrhea in young poultry and livestock breeding, the use of antibiotics seriously threatens o...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/10C12R1/07
Inventor 胡红伟段明房麻啸涛闫凌鹏杨京娥党亚鹏
Owner SHANXI DAYU BIOLOGICAL ENG CO LTD
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