Bacillus subtilis AP139 and fermented microbial inoculum thereof, and application method of bacillus subtilis AP139 and fermented microbial inoculum

A technology of Bacillus subtilis and AP139, applied in botany equipment and methods, microbial-based methods, biochemical equipment and methods, etc., can solve problems such as double infection, endogenous infection, and secondary pollution, and achieve reduction Incidence and Harm Effects

Active Publication Date: 2014-10-15
HUNAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the extensive use of antibiotics has caused a series of serious problems such as endogenous infections and superinfections, the emergence of drug-resistant strains, the decline of livestock and pou

Method used

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  • Bacillus subtilis AP139 and fermented microbial inoculum thereof, and application method of bacillus subtilis AP139 and fermented microbial inoculum
  • Bacillus subtilis AP139 and fermented microbial inoculum thereof, and application method of bacillus subtilis AP139 and fermented microbial inoculum
  • Bacillus subtilis AP139 and fermented microbial inoculum thereof, and application method of bacillus subtilis AP139 and fermented microbial inoculum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Screening of a Bacillus subtilis AP139

[0035]More than 200 strains of bacillus were isolated from the intestinal tract of animals, and these more than 200 strains of bacillus were tested for Escherichia coli, Staphylococcus aureus, Salmonella paratyphi, Pasteurella 439 (low-toxic Pasteurella), and Pasteurella 2010 ( Toxic Pasteurella), Pasteurella 2011 (virulent Pasteurella) bacteriostasis test, the results showed that the Bacillus 139 strain had the best inhibitory effect on Pasteurella, reaching 18-25mm, and the Bacillus 139 strain was tested by biochemistry and molecular biology. It was identified as Bacillus subtilis and named as Bacillus subtilis AP139. It was preserved in China Center for Type Culture Collection as CCTCC M2014053, and its 16S rDNA sequence is shown as SEQ ID NO:1.

[0036] Table 1. The inhibitory effects of some Bacillus strains on Pasteurella and other pathogenic bacteria

[0037]

Embodiment 2

[0039] A preparation method of Bacillus subtilis (Bacillus subtilis) AP139 fermentation agent:

[0040] A. Bacillus subtilis AP139 (preservation number: CCTCC M2014053) was streak cultured on LB solid medium, the culture temperature was 30-37°C, and the culture time was 18-36h;

[0041] B, the preparation of bacillus subtilis (Bacillus subtilis) AP139 fermented liquid: the bacillus subtilis (Bacillus subtilis) AP139 that step A is cultivated is picked single clone, is placed in 10ml EP tube (4ml LB substratum is housed), The culture temperature is 37°C, the rotation speed is 200r / min, and the culture time is 24h. Then use LB liquid medium to adjust the bacterial concentration to 10 10 CFU / ml, take 4ml and inoculate in 100ml medium for shake flask fermentation culture, the fermentation temperature is 37°C, the rotation speed is 200r / min, and the fermentation time is 24h. The shake flask fermentation medium is LB medium, and its composition is: tryptone 10g, yeast extract 5g, ...

Embodiment 3

[0044] Bacillus subtilis (Bacillus subtilis) AP139 antibacterial activity verification:

[0045] A. Take 3 LB nutrient agar plates, take 20 μL of activated Pasteurella 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, take 20 μL of activated Escherichia coli liquid was evenly spread on the third LB solid plate, and placed in an incubator at 37°C for 10 minutes;

[0046] B. Utilize a puncher to punch holes in the culture dish, and the hole diameter is 5 mm;

[0047] C. After centrifuging the Bacillus subtilis AP139 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;

[0048] D, the results show that the diameters of the inhibition zone of Bacillus subtilis (Bacillus subtilis) AP139 to Pasteurella, Staphylococcus aureus, Escherichia coli and Salmonella are 22m...

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Abstract

The invention discloses bacillus subtilis AP139 and fermented microbial inoculum thereof, and an application method of the bacillus subtilis AP139 and the fermented microbial inoculum. The bacillus subtilis is preserved in the China Center For Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: M2014053. The bacterial strain can restrain the growth and the propagation of porcine toxigenic pasteurella multocida in a culture dish experiment by screening porcine toxigenic pasteurella multocida resistance. The bacillus subtilis AP139 is atomized in an animal breeding shed, the bacillus subtilis AP139 has an obvious characteristic of restraining the growth of pasteurella multocida in air, the pasteurella multocida attack rate in a breeding farm is reduced, at the same time, the bacillus subtilis AP139 also has a good effect on restraining escherichia coli, staphylococcus aureus and salmonella, and the bacillus subtilis AP139 has a favorable application prospect.

Description

technical field [0001] The present invention relates to the field of microbial bactericides, more specifically to a kind of Bacillus subtilis AP139 and its fermentation bacterium, and also to their preparation of one or more of the inhibitors of Pasteurella, Escherichia coli, Staphylococcus aureus and Salmonella application in formulations. Background technique [0002] Pasteurella multocida is an important comorbid pathogen of livestock, poultry, wild animals and humans. It is mainly transmitted through the respiratory tract and can cause fowl cholera, swine pneumonia and bovine hemorrhagic sepsis. It is a very common disease in farms. The incidence of respiratory infectious diseases is higher in the absence of antibiotics. Pasteurella exists widely in nature, including air, water, dust, and human and animal excrement. There are many kinds of Pasteurella in the farm environment. A certain titer of Pasteurella in the air can Under the pathogenic. [0003] As the intensifi...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00A23K1/17C12R1/125A23K20/195
Inventor 王升平舒燕刘惠知周映华郭照辉胡新旭高书峰周小玲缪东曾发姣
Owner HUNAN INST OF MICROBIOLOGY
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