Bacillus subtilis as well as extraction method and application thereof

Abstract

The invention discloses bacillus subtilis as well as an extraction method and application thereof, and relates to the field of bacterial preparation, the preservation number of the bacillus subtilis in the China Center for Type Culture Collection (CCTCC) is CCTCC M 20242567, the name is bacillus atrophaeus Y10, and the preservation date is November 15, 2024. The extraction method of the bacillus subtilis comprises the following steps: S1, adopting contents of ileum and cecum of a slaughtered goose, and performing sterilization treatment to obtain different types of single colonies; s2, respectively performing purification culture and gram staining on the obtained single colonies to obtain different colonies; and S3, carrying out single colony propagation and identification on each obtained bacillus strain to obtain the required bacillus subtilis. The bacillus subtilis can remove peculiar smell, decompose harmful microorganisms in sewage, purify goose farm sewage and promote growth of hydroponic crops, has better tolerance compared with traditional bacillus subtilis, and can better adapt to the environment of the goose farm sewage.

Description

Technical Field

[0001] The present invention relates to the field of bacterial preparation, and particularly to a Bacillus subtilis and an extraction method and application thereof. Background Art

[0002] Bacillus subtilis has been widely used in sewage treatment. In practical applications, the components of different sewage vary widely, so the actual decontamination effects of Bacillus subtilis are also significantly different. For example, in the treatment of goose farm sewage, since the sewage from goose farms mainly comes from excreta such as goose feces, the internal bacteria, pH value, etc. are more complex. When the Bacillus subtilis obtained by traditional methods is applied therein, there are obvious problems of poor tolerance and unsatisfactory decontamination effect. Summary of the Invention

[0003] To solve the above technical problems, the present invention provides a Bacillus subtilis and an extraction method and application thereof, which can remove odors, decompose harmful microorganisms in sewage, purify goose farm sewage, promote the growth of hydroponic crops, have better tolerance than traditional Bacillus subtilis, and can better adapt to the environment of goose farm sewage.

[0004] The technical solution adopted by the present invention to solve its technical problems is: a Bacillus atrophaeus Y10, whose preservation number in the China Center for Type Culture Collection is: CCTCC No: M 20242567, and the preservation date is: November 15, 2024.

[0005] An extraction method of a Bacillus subtilis includes the following steps:

[0006] S1, using the ileum and cecum contents of slaughtered geese, and performing sterilization treatment to obtain different types of single colonies;

[0007] S2, respectively purifying and culturing the obtained single colonies, and performing Gram staining to obtain different colonies;

[0008] S3, performing single colony propagation and identification on each of the obtained bacillus strains to obtain the required Bacillus subtilis.

[0009] Preferably, in step S3, the preliminary identification is as follows: extracting DNA from the bacterial strain sample, performing amplification sequencing using the bacterial 16S universal primer, and performing comparison in the NCBI database using the sequencing results.

[0010] Preferably, in step S3, after amplifying and sequencing this sequence and comparing it with the known sequences in GenBank, the bacterial species is determined and classified into a genus or species.

[0011] Preferably, the sequencing results are assembled using ContigExpress, and the inaccurate parts at both ends are removed; the assembled sequences are aligned in the NCBI database, and the species alignment result with the highest homology is selected as Bacillus subtilis.

[0012] An application of Bacillus subtilis for sewage decontamination in a goose farm.

[0013] Advantages of the present invention:

[0014] The Bacillus subtilis in this solution has high tolerance to the sewage environment of a goose farm, better decontamination effect, can remove odors, decompose harmful microorganisms in the sewage, purify the sewage of the goose farm, and promote the growth of hydroponic crops compared with traditional Bacillus subtilis. Specific implementation manner

[0015] To deepen the understanding of the present invention, the following will further describe the present invention in detail in conjunction with embodiments. The embodiments are only used to explain the present invention and do not limit the protection scope of the present invention.

[0016] Embodiment

[0017] A Bacillus atrophaeus Y10, its preservation number in the China Center for Type Culture Collection is: CCTCC No: M 20242567, and the preservation date is: November 15, 2024.

[0018] An extraction method of Bacillus subtilis, comprising the following steps:

[0019] S1, using the ileum and cecal contents of slaughtered geese, and performing sterilization treatment to obtain different types of single colonies;

[0020] S2, respectively purifying and culturing the obtained single colonies, and performing Gram staining to obtain different colonies;

[0021] S3, performing single colony propagation and identification on each obtained bacillus strain to obtain the required Bacillus subtilis.

[0022] In step S3, the preliminary identification is as follows. DNA is extracted from the bacterial strain sample, amplified and sequenced using universal bacterial 16S primers, and the sequencing results are aligned in the NCBI database.

[0023] In step S3, this sequence is amplified by PCR, sequenced, and compared with the known sequences in GenBank, and then the bacterial species is determined and classified into a genus or species.

[0024] Use ContigExpress to splice the sequencing results and remove the inaccurate parts at both ends; compare the spliced sequences in the NCBI database and select the species with the highest homology as Bacillus subtilis.

[0025] An application of Bacillus subtilis for sewage decontamination in a goose farm.

[0026] The beneficial effects of the present invention:

[0027] The Bacillus subtilis in this solution has high tolerance to the sewage environment in the goose farm, better decontamination effect, can remove odors, decompose harmful microorganisms in the sewage, purify the sewage in the goose farm, and promote the growth of hydroponic crops compared with the traditional Bacillus subtilis.

[0028] The technical features of the above embodiments can be combined arbitrarily. For the sake of concise description, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as the scope described in this specification.

[0029] The above-described embodiments only represent several implementation manners of the present application. The description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present application, several modifications and improvements can still be made, and these all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims.

Claims

1. A Bacillus subtilis, characterized in that Its preservation number in the China Center for Type Culture Collection is: CCTCC M 20242567, its name is: Bacillus atrophaeus Y10, and its preservation date is: November 15, 2024.

2. A method for extracting Bacillus subtilis, characterized in that: The steps include: S1, the ileum and cecum contents of slaughtered geese were sterilized to obtain different types of single colonies; S2, purifying and culturing the obtained single colonies respectively, and performing Gram staining to obtain different colonies; S3, performing single colony propagation and identification on each obtained Bacillus strain to obtain the required Bacillus subtilis.

3. The method for extracting Bacillus subtilis according to claim 1, characterized in that: In step S3, preliminary identification is as follows: DNA is extracted from bacterial samples, amplified and sequenced using bacterial 16S universal primers, and the sequencing results are compared in the NCBI database.

4. The method for extracting Bacillus subtilis according to claim 3, characterized in that: In step S3, the sequence is amplified and sequenced by PCR, and after being compared with known sequences in GenBank, the bacterial species is determined and classified into genus or species.

5. The method for extracting Bacillus subtilis according to claim 4, characterized in that: The sequencing results were spliced ​​using ContigExpress, and the inaccurate parts at both ends were removed; the spliced ​​sequences were compared in the NCBI database, and the species with the highest homology was selected, and the comparison result was Bacillus subtilis.

6. The use of Bacillus subtilis according to claim 1, characterized in that: Used for sewage decontamination in goose farms.