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An enzyme-producing, halo-loving bacillus and its application in the control of pathogenic bacteria in saltwater aquaculture

A bacillus, aquaculture technology, applied in the direction of application, medical preparations containing active ingredients, bacteria, etc., can solve the problems of lack of strain resources and limited research, and achieve great application potential, strong growth adaptability, good inhibition The effect of bacteria

Active Publication Date: 2021-10-15
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the relevant research is still very limited, especially the high-efficiency salt-loving saltwater aquaculture antagonistic probiotics are rarely reported, and the strain resources are very scarce

Method used

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  • An enzyme-producing, halo-loving bacillus and its application in the control of pathogenic bacteria in saltwater aquaculture
  • An enzyme-producing, halo-loving bacillus and its application in the control of pathogenic bacteria in saltwater aquaculture
  • An enzyme-producing, halo-loving bacillus and its application in the control of pathogenic bacteria in saltwater aquaculture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Enrichment culture, isolation, purification and preservation of Bacillus amyloliquefaciens CYZF1-2

[0029] (1) Enrichment culture

[0030] Enrichment medium: NH 4 Cl 0.4g / L, NaNO 2 0.25 g / L, KH 2 PO 4 2.0 g / L, MgSO 4 ·7H 2 O 0.2g / L, Na 2 CO 3 0.4 g / L, the solvent is water; the preparation method is: mix the above ingredients evenly, and sterilize at 121°C for 20 minutes under high temperature and high pressure. Pack into 250mL Erlenmeyer flasks, 90mL per bottle.

[0031] Take 10 g of fresh pig manure refrigerated at 4°C after collection, put it into the Erlenmeyer flask filled with the above medium, culture at 30°C, 180 rpm, for 48 hours, and carry out enrichment culture. Then transfer 10 mL to a new Erlenmeyer flask containing the above-mentioned enrichment medium, culture at 30° C., 180 rpm, for 48 hours, and perform secondary enrichment to obtain a secondary enrichment culture.

[0032] (2) Isolation, purification and preservation of Bacillus CYZ...

Embodiment 2

[0035] The 16S rDNA identification of embodiment 2 Bacillus amyloliquefaciens CYZF1-2

[0036] Genomic DNA of strain CYZF1-2 was extracted, and PCR products were amplified with bacterial 16S rDNA gene amplification universal primers 27F / 1492R (5'-AGAGTTTGATCCTGGCTCAG-3' and 5'-TACGACTTAACCCCAATCGC-3') and sent to Shanghai Meiji Biomedical Technology Co., Ltd. (Guangzhou Branch) performed sequence sequencing, and the length of the 16S rRNA sequence obtained after sequencing was 1410bp, and the sequence is shown in SEQ ID NO.1. The sequencing results were compared with the 16SrDNA sequences in the NCBI and EzBioCloud databases for homology analysis, and then the strain CYZF1-2 and similar strains were selected to build a phylogenetic tree using MEGA 6.0, Kimura2-parameter model, and NJ algorithm (bootstrap repeated 1000 times ), the obtained phylogenetic tree results are shown in Image 6 .

[0037] The analysis of the results showed that it had the highest similarity (99.48%)...

Embodiment 3

[0039] Example 3 Evaluation of the Enzyme Production Ability of Bacillus CYZF1-2

[0040] Enzyme production ability was evaluated by plate hydrolysis circle detection method:

[0041] The main media and reagents used are as follows:

[0042] Protein culture medium: 40.0g skimmed milk powder, 10.0g soluble starch, 3.0g yeast extract, 2.0g potato flour, 15.0g agar, add distilled water to 1000mL, pH 7.0-7.2. Sterilize at 121℃ for 20min at high temperature and high pressure, pour plate .

[0043] Starch medium: 5.0g soluble starch, 5.0g peptone, 5.0g beef extract, 5.0g sodium chloride, 15.0g agar, add distilled water to 1000mL, pH 7.0-7.2. Sterilize under high temperature and high pressure at 121℃ for 20min, pour plate .

[0044] Fat medium (tributyrin): use nutrient broth as the basal medium, add 2mL fat source per 100mL basal medium, agar 15.0g / L, add distilled water to 1000mL, pH 7.0-7.2; The source is obtained by mixing 2% polyvinyl alcohol solution with tributyrin at a ra...

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Abstract

The invention discloses an enzyme-producing, halo-loving bacillus and its application in preventing and controlling pathogenic bacteria in saltwater aquaculture. The bacillus is named: Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) CYZF1‑2, and the preservation number is GDMCC NO: 60904. The Bacillus amyloliquefaciens CYZF1-2 has good enzyme production ability: it has good hydrolysis effects on protein, starch and fat; in the medium with a salt concentration of 4%, the growth of the strain is the best, and the bacterial liquid is cultured for 48 hours OD 600 The value is close to 1.0; the strain CYZF1‑2 has high inhibitory ability to common aquatic pathogens, and the antibacterial substances are mainly protein polypeptide substances, which will not produce drug resistance and no secondary pollution for the prevention and treatment of pathogenic bacteria. It has great application potential in prevention and treatment.

Description

technical field [0001] The invention relates to the technical field of application of aquatic microorganisms, in particular to an enzyme-producing, halo-loving bacillus and its application in preventing and controlling pathogenic bacteria in saltwater aquaculture. Background technique [0002] my country is the only country in the world where mariculture production exceeds marine catches. According to the "World Fisheries and Aquaculture Status" report released by FAO, since 2013, my country's mariculture production has accounted for more than 80% of the world's mariculture production. Marine or brackish water aquaculture occupies an important position in aquaculture in my country. However, in recent years, the aquaculture industry has developed rapidly in the direction of scale and intensification. While promoting the development of the agricultural economy and improving the quality of life of the people, it has also brought many environmental problems and ecological crises...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01K61/13A61K35/742A61P31/04C12R1/07
CPCA61K35/742A01K61/13A61P31/04C12N1/20C12N1/205C12R2001/07Y02A40/81
Inventor 朱红惠陈猛杨敏志李安章张明霞谢小林陈美标
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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