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Bacillus subtilis strain and application thereof

A Bacillus subtilis, bacterial liquid technology, applied in the application, bacteria, microorganisms and other directions, can solve the problems of no anti-shrimp virus disease strain, low number of viable bacteria, unstable effect, etc., to improve the anti-stress response ability of prawns , the effect of reducing virus infection and slowing the growth rate

Inactive Publication Date: 2015-01-28
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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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 subtilis strains used in the production of prawns, and there are deficiencies such as mediocre biological properties, low number of viable bacteria, and unstable effects, especially the application of anti-prawn virus disease strains.

Method used

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  • Bacillus subtilis strain and application thereof
  • Bacillus subtilis strain and application thereof
  • Bacillus subtilis strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: Screening of Bacillus subtilis

[0013] The Bacillus subtilis LV023 strain of the present invention is selected from the digestive tract of Litopenaeus vannamei. Put the collected healthy prawn samples in an air bag filled with sea water and transport them back to the laboratory. The surface of the live shrimp is wiped with 75% alcohol on a sterile operating table for disinfection, the live shrimp are dissected aseptically, and the shrimp intestinal tissue is taken out. Wash the removed intestinal tissue with sterile PBS (pH=7.0) buffer solution 3-4 times, and then grind it evenly with a sterile grinding rod. After that, it is appropriately diluted with sterile PBS buffer, and 0.1 ml is applied to 2216E solid medium, and placed in a 28°C constant temperature incubator for 16h-20h. Observe the morphology of the bacterial colonies on the 2216E plate, number the bacteria with the same colony morphology, use an inoculation loop to pick a single colony of the domin...

Embodiment 2

[0021] Combine the prepared 100mL Bacillus subtilis LV023 solution with 10Kg of basic shrimp feed ingredients (containing 30% fish meal, 10% shrimp meal, 25% soybean meal, 25% peanut meal, 3% flour, 3% corn meal, and 1% compound vitamins. ) Mixed to prepare immune feed containing Bacillus subtilis LV023. The various raw materials were crushed through a 60-mesh sieve, accurately weighed, and mixed step by step. Sodium alginate was used as a binder, and a small amount of water was added to squeeze into a pellet feed. The feed was baked at 40°C. Dried until the moisture content is below 10%, packaged in bags, and stored in a refrigerator at 4°C for later use.

[0022] There are more than 300 healthy Litopenaeus vannamei, and the initial average body length of the experimental prawns is 6.3cm. They are randomly divided into two treatment groups, the experimental group and the control group, each treatment group has 3 parallels, and each parallel group breeds 50 prawns. The experimen...

Embodiment 3

[0025] Prawn immune feed containing 1% (V / W) Bacillus subtilis LV023 solution (total concentration of live bacteria ≥ 10 7 The preparation method of CFU / mL) is the same as in Example 2.

[0026] There are more than 300 healthy Litopenaeus vannamei. The initial average body length of the experimental prawns is 6.3cm. They are randomly divided into two treatment groups, the experimental group and the control group, and 150 prawns are cultured in each treatment group. The experimental group was fed with immune feed for 4 consecutive days and basic feed for 3 days, a cycle of 7 days until the end of the experiment. The control group was fed basic feed throughout the experimental period. After 21 days of immunized feed culture, the WSSV artificial injection infection experiment was carried out. Feeding was stopped one day before infection, and 50μL of WSSV crude extract (concentration of 0.16g / mL) was injected at the second abdominal segment of each shrimp on the next day, and PBS so...

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Abstract

The invention aims to provide bacillus subtilis with a collection number of CCTCC M2013264. The bacillus subtilis preparation provided by the invention has a variety of biological functions of improving the composition and number of advantageous floras in alimentary canals of prawns, forming and maintaining good micro-ecological balance in the alimentary canals, resisting adherence and propagation of viruses in the alimentary canals by virtue of nutrient competition, spatial competition or bacteriocin excretion and the like, being able to enhance nonspecific immunity of the prawns to reduce the chance of virus infection, activating expressions of prophenoloxidase, hyperoxide dismutase, lysozyme, heat shock protein, lipopolysaccharide-glucosan conjugated protein, signal conduction and transcription activator and the like of the prawns and antipathogen infection related molecules to slow down the propagation rate of viruses in bodies of the prawns, improving activity of protease, lipase and amylase of the prawns, promoting digestive ability of the prawns and digestion of feed nutrients, improving stress reaction resistance of the prawns, and the like.

Description

Technical field [0001] The invention belongs to the technical field of probiotics screening, and specifically relates to a Bacillus subtilis strain and its application. Background technique [0002] Bacillus subtilis belongs to Firmicutes (low G+C gram-positive bacteria), Bacilli, Bacillales, Bacillus ( Bacillaceae), Bacillus. Bacillus subtilis ubiquitously exists in soil and plant surfaces, and is also one of the common symbiotic bacteria in the intestines of humans and animals. Bacillus subtilis is one of 44 "microorganisms that can be directly fed and generally considered safe" announced by the U.S. Food and Drug Administration (FDA) and the American Feed Association (AAFCO) in 1989. It was also published by the Ministry of Agriculture in December 2008. One of the 16 feed-grade microbial additives that can be directly fed to animals specified in the "Catalogue of Feed Additives" in Announcement No. 1126. Because Bacillus subtilis has the characteristics of non-pathogenicity...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K1/18A23K1/16C12R1/125
CPCC12N1/205C12R2001/125
Inventor 宋晓玲许华孙艳李玉宏黄倢
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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