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Production method of bacillus subtilis biological agent used for sodium humate-containing feed in medium

A technology of Bacillus subtilis and sodium humate, applied in the direction of microorganism-based methods, microorganisms, animal feed, etc., to achieve the effects of long-term storage resistance, many products, and high bacterial counts

Active Publication Date: 2013-07-03
SHENYANG BOSHAN YINGSHENG BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The design and production of this process is still domestic and international initiative

Method used

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  • Production method of bacillus subtilis biological agent used for sodium humate-containing feed in medium
  • Production method of bacillus subtilis biological agent used for sodium humate-containing feed in medium
  • Production method of bacillus subtilis biological agent used for sodium humate-containing feed in medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] 1) Purification and optimization of a single colony: the slant strain of Bacillus subtilis is spread on a petri dish filled with bacterial culture medium, and cultivated to obtain a single colony seed of Bacillus subtilis.

[0055] 2) First-level solid seed cultivation: the single colony seed obtained in step 1), single colony is picked out and inoculated into the first-level solid seed medium, the solid seed culture bottle is a 200ml eggplant-shaped bottle, and the loading capacity is 50ml solid medium , after cultivation, the first-grade solid seeds are obtained;

[0056] 3) Single colony purification and optimization of primary solid seed medium

[0057] Under laboratory conditions, carry out the optimization test of culture factor, test design is 6 factors 3 levels, test design and test result are as follows table 1:

[0058] Table 1. Bacillus subtilis primary solid seed medium formula test

[0059]

[0060] From the test results in Table 1, it is determined th...

Embodiment 2

[0062] 1) Two-stage liquid seed fermentation culture: adopt the submerged fermentation method to produce liquid seeds, and connect the cultivated first-grade solid seeds to the seed fermentation tank, and the inoculation ratio of the first-grade solid seeds to a 1-ton fermenter (70% liquid content) For 1 bottle: 200 kg. After inoculation, it is cultured by ventilation and fermentation. When the culture is mature, the bacterial liquid has a slightly sour aroma. The Bacillus subtilis cells are robust and free of abnormal phenomena such as contamination by bacteria, and can be used as secondary liquid seeds for solid fermentation materials.

[0063] 2) Optimization test of secondary liquid seed fermentation medium

[0064] Under the conditions of the fermentation factory, carry out the preferred test of fermentor medium culture, the test design is 6 factors and 3 levels, and the test design and test results are as follows in Table 2:

[0065] Table 2. Bacillus subtilis secondary...

Embodiment 3

[0077] 1) Tertiary solid fermentation material fermentation and cultivation: inoculate the liquid seeds into the solid fermentation material, supply oxygen through ventilation, and ferment and cultivate to obtain the cultured solid fermentation material.

[0078] 2) The weight ratio of various raw materials in the solid fermentation material: 2 parts of glucose, 30 parts of sodium humate, 10 parts of soybean meal, 10 parts of corn flour, and 50 parts of bran; the weight ratio of the mixture of secondary liquid seeds and solid fermentation material is 40:100 . Oxygen is supplied by ventilation, and the ventilation rate is 0.5V / V·min. The influence of solid fermentation materials with different water contents on temperature is shown in the following table:

[0079] Table 4. Effect of solid fermentation materials with different water contents on Bacillus fermentation temperature (°C)

[0080]

[0081] In the case of the above solid fermented material formula, the test result...

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Abstract

The invention relates to a microbe biological agent, and concretely relates to a production method of a bacillus subtilis biological agent used for a sodium humate-containing feed in a medium. The bacillus subtilis bacterial strain is subjected to first-stage solid seed culture and second-stage liquid seed fermentation culture to obtain a bacterial classification liquid. The bacterial classification liquid is inoculated in the medium which takes glucose, sodium humate, soybean meal, corn and wheat bran as raw materials for three-stage solid aerobic fermentation, the culture condition is adjusted to form spore, the spore can be treated, dried, and crushed to produce the bacillus subtilis biological agent. The biological agent contains more than or equal to 200 hundreds million / g of acillus subtilis, the pH value is 5.6-6.6, the moisture is less than or equal to 10%, and the granularity is greater than or equal to 100 meshes, the color presents special shallow grey or grey black, and the shelf life is 18 months. The biological agent can be taken as an additive application in aquatic product, and livestock and poultry culture, can increase the production performance and improve the culture environment.

Description

technical field [0001] The invention relates to a microbial biological preparation, in particular to a production method of a high-concentration Bacillus subtilis biological preparation for feed containing sodium humate in a culture medium. Background technique [0002] Bacillus subtilis (Bacillus subtilis), is a kind of Bacillus, single cell 0.7~0.8×2~3 microns, Gram-positive bacteria, no capsule, perinatal flagella, able to move. The spores are large, elliptical to columnar, 0.6-0.9×1.0-1.5 microns, located in the center of the thallus or slightly off, and the thallus does not expand after spore formation. The surface of the colony is rough, stained white or yellowish, and often forms wrinkles when cultured in liquid. Bacillus subtilis has the characteristics of fast reproduction speed, high temperature resistance, acid and alkali resistance, extrusion resistance, good stability, and wide application range. It is often used as a model strain in the scientific research and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K1/16C02F3/34C12R1/125
Inventor 潘军薛德林宋颖张伟强
Owner SHENYANG BOSHAN YINGSHENG BIOLOGICAL TECH CO LTD
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