Bacillus subtilis PA105 and preparation method and application thereof

A technology of Bacillus subtilis and Escherichia coli, which is applied to the preparation of Bacillus subtilis PA105, and the application field of Bacillus subtilis PA105 in animal feed additives to enhance animal immunity and anti-stress ability, inhibit growth and reproduction, and reduce environmental protection. The effect of pollution

Inactive Publication Date: 2013-03-06
INST OF SUBTROPICAL AGRI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, combining the antibacterial activity and arginine-producing activity of Bacillus subtilis, the application of Bacillus subtilis PA105 in the drinking water and feed of farmed animals has not been reported yet.

Method used

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  • Bacillus subtilis PA105 and preparation method and application thereof
  • Bacillus subtilis PA105 and preparation method and application thereof
  • Bacillus subtilis PA105 and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A preparation method of bacillus subtilis (Bacillus subtilis) PA105, the steps are:

[0034] A, preparation of Bacillus subtilis (Bacillus subtilis) PA105 fermented liquid: get Bacillus subtilis (Bacillus subtilis) PA105 (preservation number: CCTCC M 2011328) 2ml (viable bacteria concentration is 10 10 CFU / ml), inoculated in 100ml medium for shake flask fermentation culture, the fermentation temperature is 37°C, the pH value is 7.2, the rotation speed is 200r / min, and the fermentation time is 24h. The shake flask fermentation medium is LB medium, the ingredients are: tryptone 10g, yeast extract 5g, sodium chloride 10g, distilled water 1000ml, and the pH value is 7.2.

[0035] B. After the shake flask fermentation, carry out the pilot test of the fermenter: take 100ml shake flask fermentation seed liquid and inoculate it into a 10L fermenter, the liquid volume is 5L, the fermentation temperature is 37°C, the pH value is 7.2, and the stirring speed is 300r / min , fermente...

Embodiment 2

[0038] Bacillus subtilis (Bacillus subtilis) PA105 antibacterial activity verification:

[0039] A. Take 2 LB nutrient agar plates, take 20 μL of activated E. coli bacteria liquid, and evenly spread it on one LB solid plate, take 20 μL of activated Salmonella bacteria liquid, evenly spread it on another LB solid plate, place Incubate at 37°C for 10 minutes in an incubator;

[0040] B. Utilize a puncher to punch holes in the culture dish, and the hole diameter is 5 mm;

[0041] C. After centrifuging the Bacillus subtilis PA105 fermentation broth at 12,000 r / m for 1 min, take 20 μL of the supernatant and add it to the well of the culture dish, and incubate in an incubator at 37° C. for 24 h;

[0042] D, the results show that the diameters of the inhibition zone of Bacillus subtilis PA105 to Escherichia coli and Salmonella are 1.1cm and 1.0cm respectively.

Embodiment 3

[0044] Analysis of arginine produced by Bacillus subtilis PA105;

[0045] Take 5ml of Bacillus subtilis (Bacillus subtilis) PA105 fermentation tank fermentation broth, centrifuge at 12000r / min for 10min, take 1ml of supernatant, add 2.5ml of 7.5% trichloroacetic acid per ml, mix well, centrifuge at 18000r / min at 4°C for 15min, draw 1ml of the supernatant was transferred to an amino acid analysis vial, and the contents of arginine and other amino acids were determined with a Hitachi L-8800 automatic amino acid analyzer. The results showed that the content of arginine in the fermentation broth of Bacillus subtilis PA105 fermenter was 1 g / L.

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Abstract

The invention relates to a bacillus subtilis PA105 and a preparation method and application thereof, and the bacillus subtilis is collected in China Center for Type Culture Collection with the collection number of CCTCCM2011328. The preparation method includes the following steps: (a) the bacillus subtilis PA105 receive streak culture on Luria-Bertani (LB) solid medium in a constant-temperature biochemical incubator, and the culture temperature is 30 DEG C to 37 DEG C; (b) a single clone strain of the bacillus subtilis is picked from the LB solid medium in step a, put into a conical flask loaded with fluid nutrient medium and cultured in a constant-temperature biochemical shaker; (c) bacillus subtilis fluid fermentation broth obtained in step b of the experiment is inoculated into a 10L fermentation tank according to 3 to 5 percent of inoculation amount. The bacillus subtilis PA105 can be applied in animal-raising feed additives. The bacillus subtilis PA105 has remarkable activity in inhibiting the growth of pathogenic bacteria, and can effectively inhibit the growth and reproduction of colibacillus, salmonella and other pathogenic bacteria. The bacillus subtilis PA105 can promote the growth of intestinal lactobacillus and inhibit the reproduction of intestinal colibacillus, the growth of animals can be increased by 5 to 10 percent, the economic benefit can be increased, and thereby the bacillus subtilis PA105 has a bright application prospect.

Description

technical field [0001] The invention relates to the field of feed additives, more specifically to a Bacillus subtilis PA105, a preparation method of the Bacillus subtilis PA105, and an application of the Bacillus subtilis PA105 in feed additives for breeding animals. Background technique [0002] In the process of intensive farming, animals are prone to various stresses due to changes in environmental conditions, such as weaning, sudden changes in the weather, etc., which will cause a decrease in animal immunity. Some pathogenic bacteria, especially intestinal conditional pathogenic bacteria, can multiply rapidly and cause corresponding diseases, such as Escherichia coli and Salmonella. At present, the main method is to control and reduce stress by adding antibiotics in the feed. Antibiotic supplementation can inhibit the growth and reproduction of intestinal pathogenic bacteria in piglets and facilitate the growth of beneficial bacteria, thereby maintaining the normal micr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K1/16A61K35/74A61P31/04C12R1/125A61K35/742
CPCY02A50/30
Inventor 王升平印遇龙李丽立姚康
Owner INST OF SUBTROPICAL AGRI CHINESE ACAD OF SCI
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