Preparation method and application of compound microbial substrate modifier with aquatic pathogen antagonistic characteristic

A substrate improvement and microbial technology, applied in the field of microorganisms, can solve problems such as limited use conditions, instability, and single strain structure, and achieve the effects of wide growth temperature range, high vitality, and strong degradation ability

Active Publication Date: 2020-01-17
江苏省苏微微生物研究有限公司
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Problems solved by technology

[0008] Therefore, in view of the low-oxygen environment conditions of the pond bottom, an important direction for the creation of microbial preparations for aquaculture pond bottom improvement is to develop dissolved oxygen that does not consume aquaculture. The multifunctional and facultative anaerobic new beneficial bacterial strains with antagonistic effects on aquatic pathogens, through the mixing of several ...

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  • Preparation method and application of compound microbial substrate modifier with aquatic pathogen antagonistic characteristic
  • Preparation method and application of compound microbial substrate modifier with aquatic pathogen antagonistic characteristic
  • Preparation method and application of compound microbial substrate modifier with aquatic pathogen antagonistic characteristic

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Experimental program
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Effect test

Embodiment 1

[0048] Embodiment 1: the screening of strain

[0049] (1) Screening of Lactobacillus gasseri: Allogeneic gibel carp samples were collected in the Wuxi Goose Lake breeding pond, the fish body was dissected to take the intestinal tract, and physiological saline was added to make a homogenate, and a pipette was used to pipette 0.1mL into the MRS Spread on the solid medium, cover with a layer of MRS medium, and culture at 37°C. According to the size of the colony, the well-growing colony was selected and screened repeatedly until a uniform single colony was obtained, which was named 88.

[0050] (2) Screening of Bacillus subtilis: The samples were collected in the herring pond of Wuxi Goose Lake, weighed 10 g of the bottom mud of the pond into a 250 ml Erlenmeyer flask filled with 90 ml of sterile water and a small amount of glass beads, oscillated for 30 min, and stood still . The supernatant was taken, inoculated in the separation medium, and cultured at 37°C for 7 days. Take...

Embodiment 2

[0052] Embodiment 2: strain identification

[0053] (1) Morphological characteristics:

[0054] Strain 88, a gram-positive bacterium, is spherical under the microscope and does not produce spores. This strain belongs to non-strict anaerobic bacteria and has a wide growth temperature range, and can grow at 20°C-45°C. The diameter of the colony on the LB plate is 0.5-1.0 mm, the shape is regular, the surface is smooth, opaque, and pigment-free. A large number of round, smooth, opaque, white colonies can be obtained on the MRS medium.

[0055] Strain AB90008-15, a Gram-positive bacterium, is rod-shaped under the microscope, with endospores of 0.8×1.5-1.8µm in diameter, round or irregular colonies on the agar medium, and the surface is dark, opaque, creamy or brown.

[0056] (2) Biochemical properties:

[0057] strain 88, capable of producing H 2 S, tryptophan deaminase, acetylmethylmethanol, can ferment glucose, mannitol, sucrose, amygdalin, galactose, fructose, N-acetyl-gluc...

Embodiment 3

[0064] Embodiment 3: Bacillus subtilis AB90008-15 bacteriostatic assay

[0065] Place sterile filter paper at a concentration of approximately 10 6 Soak in CFU / mL fresh Bacillus subtilis culture solution for 1.0 h. Take a concentration of about 10 8 Common pathogenic bacteria in aquaculture of CFU / mL Aeromonas hydrophila, Aeromonas sobria, Aeromonas victoria, Aeromonas caviae, Edwardsiella, Vibrio parahaemolyticus, Escherichia coli, Spread 0.1 mL of liquid culture solution of Vibrio alginolyticus on LB agar medium plates respectively, and then paste the filter paper soaked in the bacterial solution on the culture plates, stick 3 pieces on each plate, and make 3 pieces for each plate repeat. The plates were placed in an incubator at 25-30°C for 24 and 48 h, and the size of the inhibition zone was measured. The measurement results are shown in Table 1:

[0066] Table 1 Bacillus subtilis AB90008-15 antibacterial assay result (inhibition zone diameter: mm)

[0067]

[006...

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Abstract

The invention belongs to the technical field of microorganisms and discloses a preparation method and application of a compound microbial substrate modifier with an aquatic pathogen antagonistic characteristic. Two strains namely bacillus subtilis AB-90008-15 and lactobacillus gasseri 88 are provided, and the compound microbial substrate modifier is obtained by strain activation, fermentation culture, centrifugal strain collection and vacuum drying and grinding. The compound microbial substrate modifier has an antagonistic action on aquatic pathogens such as aeromonas hydrophila, aeromonas sobria, aeromonas veronii, aeromonas caviae, Edwardsiella tarda, listonella anguillarum, vibrio parahaemolyticus and vibrio alginolyticus and is capable of degrading carbonaceous organics and nitrogenousorganics in pond sediment, promoting sediment mineralization, reducing contents of total nitrogen, total phosphorus, nitrite, ammonia nitrogen and COD of aquaculture water, promoting virtuous cycle of waternutrients, preventing accumulation of toxic and harmful substances and improving an aquaculture substrate ecological environment.

Description

technical field [0001] The invention relates to a preparation method and application of a composite microbial substrate modifier with antagonistic properties of aquatic pathogens, belonging to the technical field of microorganisms. Background technique [0002] To raise fish, raise the water first, and raise the water first to raise the bottom. Pond culture is based on water, and the quality of water is the main reason affecting the survival rate and output of aquaculture organisms. The condition of the sediment environment has a great influence on the water quality of the water body. Therefore, its health directly affects the economic and ecological benefits of aquaculture. If the bottom mud is too thick, it often brings many potential hazards to the water body. In ponds with a lot of silt, the oxidative decomposition of organic matter will consume the oxygen that is not much at the bottom, causing the bottom to be in a state of anoxic conditions, anaerobic bacteria multi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C02F11/02C12R1/125C12R1/225C02F103/20
CPCC12N1/20C02F3/341C02F3/348C02F11/02C02F2103/20C12R2001/125C12N1/205
Inventor 施大林张维娜高亮于丹孙梅匡群
Owner 江苏省苏微微生物研究有限公司
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