A kind of bacillus pumilus strain and its application

A technology of Bacillus pumilus and seeds, applied in the direction of application, bacteria, microorganisms, etc., can solve the problems of unstable effect, low number of viable bacteria, no strains resistant to shrimp virus disease, etc., to reduce virus infection, slow down the value-added rate, The effect of enhancing the non-specific immune function of prawns

Active Publication Date: 2017-12-15
湖北宜水生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] But at present, there are few Bacillus pumilus strains used in the production of prawn farming, and there are deficiencies such as mediocre biological properties, low number of viable bacteria, and unstable effect, especially the application of strains against prawn virus disease.

Method used

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  • A kind of bacillus pumilus strain and its application
  • A kind of bacillus pumilus strain and its application
  • A kind of bacillus pumilus strain and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: Screening of Bacillus pumilus

[0014] The Bacillus pumilus LV012 strain of the present invention is screened from the digestive tract of Litopenaeus vannamei. The collected healthy prawn samples were transported back to the laboratory in inflatable bags filled with seawater. Wipe the body surface of live shrimp with 75% alcohol on the aseptic operation table for disinfection, dissect the live shrimp by aseptic operation, and take out the intestinal tissue of the prawn. After washing the removed intestinal tissue with sterile PBS (pH=7.0) buffer solution for 3-4 times, grind it evenly with a sterile grinding rod. Afterwards, it was appropriately diluted with sterile PBS buffer, and 0.1 ml was taken and spread on the 2216E solid medium, and placed upside down in a constant temperature incubator at 28°C for 16h-20h. Observe the colony shape of the bacteria on the 2216E plate, number the bacteria with the same colony shape, and pick a single colony of the domi...

Embodiment 2

[0022] With the 100mL bacillus pumilus LV012 solution that makes and 10Kg basic prawn feed material (containing fish meal 30%, shrimp meal 10%, soybean meal 25%, peanut meal 25%, flour 3%, corn meal 3%, multivitamin 1% ) mixed to make immune feed containing Bacillus pumilus LV012. Various raw materials were crushed through a 60-mesh sieve, accurately weighed, and mixed well step by step. Sodium alginate was used as a binder, and an appropriate amount of water was added to extrude it with a small meat grinder to make pellet feed, and dried at 40°C. Dried until the moisture content is below 10%, packed in separate bags, and stored in a refrigerator at 4°C for later use.

[0023] There were more than 300 healthy Litopenaeus vannamei prawns, and the initial average body length of the experimental prawns was 6.3 cm. Two treatment groups were randomly divided into the experimental group and the control group. Each treatment group had 3 parallels, and 50 prawns were cultured in each ...

Embodiment 3

[0027] The prawn immunization feed containing 1% (V / W) Bacillus pumilus LV012 solution (total concentration of viable bacteria ≥ 107 CFU / mL) The preparation method is the same as embodiment 1.

[0028] There were more than 300 healthy Litopenaeus vannamei prawns, and the initial average body length of the experimental prawns was 6.3 cm. They were randomly divided into two treatment groups, the experimental group and the control group, and 150 prawns were cultured in each treatment group. The experimental group was fed the immune feed for 4 consecutive days, and the basal feed for 3 days, with a cycle of 7 days until the end of the experiment. The control group was fed the basal feed throughout the experimental period. After 21 days of breeding with immunized feed, the artificial injection infection experiment of WSSV was carried out. Feed was stopped 1 day before infection, and 50 μL of WSSV crude extract (concentration: 0.16 g / mL) was injected into the second abdominal segme...

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Abstract

The object of the present invention is to provide a strain of Bacillus pumilus, the preservation number of which is CCTCC M2013263. The Bacillus pumilus of the present invention can improve the composition and quantity of dominant bacterial flora in the digestive tract of prawns, form and maintain a good micro-ecological balance in the digestive tract, and resist the adhesion and proliferation of viruses in the digestive tract through nutrient competition, space competition or secretion of bacteriocins, etc. . It can activate the expression of anti-pathogenic infection-related molecules such as pro-prawn phenol oxidase, superoxide dismutase, lysozyme, heat shock protein, lipopolysaccharide-glucan binding protein, signal transduction and transcription activator, and slow down the virus in the Value-added rate in prawns. The bacillus pumilus of the present invention can be added to the prawn feed raw materials, and can also be directly applied to the prawn breeding water environment.

Description

technical field [0001] The invention belongs to the technical field of probiotic screening, and in particular relates to a Bacillus pumilus strain and an application thereof. Background technique [0002] Bacillus pumilus belongs to Firmicutes (low G+C Gram-positive bacteria), Bacilli class (Bacilli), Bacillales (Bacillales), Bacillus family ( Bacillaceae), Bacillus (Bacillus). Bacillus pumilus is ubiquitous in soil and on the surface of plants, and is also one of the common symbiotic bacteria in the intestines of humans and animals. Bacillus pumilus is one of the 44 microorganisms that can be directly fed and are generally considered safe as announced by the US Food and Drug Administration (FDA) and the American Feed Standards Association (AAFCO) in 1989. Bacillus pumilus has strong protein secretion ability and low extracellular protease activity, so it has the natural advantages of secreting and expressing foreign proteins, and it is expected to establish an efficient p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K10/18A23K50/80C12R1/07
CPCC12N1/20C12N1/205C12R2001/07
Inventor 宋晓玲李玉宏张庆利许华黄倢
Owner 湖北宜水生物科技有限公司
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