Bacillus subtilis and use thereof

A technology of Bacillus subtilis and seeds, which is applied in the direction of bacteria, applications, and microorganisms, can solve the problems of insufficient antibacterial effect, poor resistance to the physiological environment of the digestive tract and feed processing, and unreliable safety. Harmful microorganisms, promote intestinal microecological balance, and improve immunity

Active Publication Date: 2014-03-05
北京昕大洋科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, some feed companies are actively looking for and developing new green and environmentally friendly microbial feed additives, but such additives on the market have poor stress resistance (for example, the ability to resist the physiological environment of the digestive tract and the ability to resist feed processing), and inhibit Bacteria effect is not significant enough, safety is unreliable and other defects

Method used

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  • Bacillus subtilis and use thereof
  • Bacillus subtilis and use thereof
  • Bacillus subtilis and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The separation and purification of embodiment 1 bacterial classification

[0036]Slaughter healthy adult chickens (Beijing Xin Dayang Science and Technology Development Co., Ltd. farm), take about 1 g of the cecum content and place it in a triangular flask filled with 45 mL of sterile distilled water, and shake it well. Place the Erlenmeyer flask in a 75°C water bath for 10 minutes, take 10mL of bacterial liquid and add it to 100mL broth liquid medium, and incubate in a constant temperature shaker at 37°C at 180rpm for 24 hours.

[0037] After 24 hours of culture, the bacterial solution was diluted to 10 -9 After doubling, spread it on the broth solid medium, pick typical colonies with different shapes and inoculate them on the plate of the same medium for pure culture. Pick a small amount of purely cultured single colony, spread it evenly on the glass slide, carry out simple crystal violet staining and microscopic examination, the single colony with spores is detected...

Embodiment 2

[0038] Embodiment 2 antibacterial effect screening

[0039] Preparation of Bacillus fermentation broth: Inoculate each bacterial strain in the above-mentioned Example 1 into 50 mL of complete bacterial culture medium, and culture at 37° C. on a shaker (180 rpm) for 24 hours. Then, it was centrifuged at 5000rpm for 20 minutes, and the supernatant was taken aseptically for antibacterial test.

[0040] Preparation of indicator bacteria plate: Directly use common animal pathogens Escherichia coli (Escherichia coli), Staphylococcus aureus (Staphylococcus aureus) and Salmonella enteritidis (Salmonella enteritidis) as indicator bacteria, and use 5 mL sterile Wash the slanted Escherichia coli, Staphylococcus aureus, and Salmonella enteritidis with water to make a bacterial suspension, mix it with LB solid medium at about 40°C, and pour it into a sterilized plate. After the medium cools down, punch holes in the medium with a 0.5 cm diameter puncher. Add an appropriate amount of super...

Embodiment 3

[0044] Example 3 Screening against digestive tract physiological environment

[0045] bile salt tolerance test

[0046] Add 0.3% porcine bile salt (analytical pure) to the liquid medium (beef extract 3g, peptone 10g, sodium chloride 5g, distilled water 1L, pH 7.2), and autoclave at 121°C for 20 minutes. The 5 strains of bacillus screened out in Example 2 were inoculated into the above-mentioned medium with 10% inoculum respectively, 37 DEG C, shaker (180rpm) culture, in 0, 2, 4 and 24 hours sampling carries out plate colony technique , to calculate the survival rate.

[0047] The specific results are shown in Table 2.

[0048] Strain survival rate (%) = N t / N 0 ×100

[0049] Among them, N t Indicates the number of viable bacteria at different sampling times; N 0 Indicates the number of viable bacteria at 0 hours.

[0050] Table 2

[0051]

[0052]

[0053] The content of bile salts in the small intestine of humans and animals fluctuates between 0.03% and 0.3%...

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Abstract

The invention provides bacillus subtilis K018 which is strong in stress resistance, excellent in probiotics function and safe and reliable, a biological feed additive containing the bacillus subtilis K018 and use thereof. The bacillus subtilis K018 provided by the invention can resist artificial gastric acid, cholate, artificial intestinal juice and high temperature, has stronger inhibiting effects to pathogenic staphylococcus aureus, escherichia coli, salmonella enteritidis and the like in the intestinal tract, is stronger in amylase and cellulase generating capacity, and can degrade starch and cellulose.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Bacillus subtilis and its use. Background technique [0002] my country is a large producer of livestock and poultry products. In recent years, with the scale and intensification of livestock and poultry breeding, there have been some problems that seriously restrict the further development of the breeding industry, such as the pollution of the environment by the stench produced in the manure of livestock and poultry, antibiotic residues, and high breeding costs. In particular, negative effects such as drug residues and drug resistance of antibiotics have become a threat to human health, and more and more countries have begun to ban antibiotics. In my country, the rate of drug-resistant strains is increasing, the difficulty of treating bacterial infectious diseases is becoming more and more difficult, and people's opposition to feed antibiotics is becoming stronger and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K1/16C12R1/125
Inventor 李光智
Owner 北京昕大洋科技发展有限公司
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