Bacillus pumilus, probiotics preparation and preparation method and application thereof

A technology of Bacillus pumilus and probiotic preparations, applied in the field of microorganisms, can solve problems such as low inhibition effect, and achieve the effects of improving commodity quality, inhibiting the growth of shrimp enteric pathogenic Vibrio, and reducing the bait coefficient

Active Publication Date: 2012-07-18
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The inhibitory effect of Bacillus on these Gram-negative bacteria from aquatic animals has been rarely reported

Method used

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  • Bacillus pumilus, probiotics preparation and preparation method and application thereof
  • Bacillus pumilus, probiotics preparation and preparation method and application thereof
  • Bacillus pumilus, probiotics preparation and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Screening and isolation of Bacillus pumilus LV149

[0028] (1) Intestinal adhesion bacteria separation

[0029] Five batches of healthy Litopenaeus vannamei were purchased at different times and in different markets, each with 30 tails, and 5 large, bright, smooth, undamaged, liver-brown individuals were selected from them, with an average body length of 12 cm. Disinfect the surface of the prawns with 75% alcohol, then rinse with sterile seawater. After dissection, place the intestines on sterile tin foil. Use a syringe to absorb the sterilized seawater to quickly flush out the feces in the intestines. Wash the tract 2-3 times, and observe the intestinal tract with naked eyes until there is no residual content. Add sterile normal saline in a sterile homogenizer to homogenize the intestine. Take 0.5 mL of the homogenate and dilute 10-fold serially with sterile normal saline (1% NaCl) to form the intestinal adhesion bacterial suspension.

[0030] Coat the shrimp fe...

Embodiment 2

[0042] Example 2: Analysis of Vibrio Inhibitory Spectrum and Analysis of Hemolysis of Bacillus pumilus LV149

[0043] 13 strains of Vibrio from 9 different species were used to analyze the inhibitory Vibrio spectrum of Bacillus pumilus LV149. The above-mentioned flat filter paper diffusion method is also used. The antibacterial ability is judged according to the size of the inhibition zone, and the results of the antibacterial spectrum analysis are shown in Table 1:

[0044] Table 1: Analysis of Vibrio inhibitory activity of Bacillus pumilus LV149

[0045]

[0046] Note: +++: strong antibacterial activity; ++: medium antibacterial activity; +: antibacterial activity.

[0047] It can be seen from Table 1 that from the analysis of dozens of common pathogenic Vibrio species, Bacillus pumilus LV149 has broad-spectrum antibacterial activity against Vibrio. The typical inhibition zone formed by Bacillus pumilus LV149 against Vibrio image 3 .

[0048] The hemolytic activity of Bacillus pum...

Embodiment 3

[0049] Example 3: Preparation of Bacillus pumilus probiotic preparation

[0050] First-level seed liquid: Using LB as the medium, add the ultra-low-temperature stored Bacillus pumilus LV149 to the LB liquid medium at a ratio of 1% (volume percentage), and culture with shaking at 30°C for 24 hours to obtain the first-level seed liquid.

[0051] Secondary seed liquid: Using LB as the medium, the primary seed liquid was added to the LB liquid medium at a ratio of 2% (volume percentage), and cultured with shaking at 30°C for 24 hours to obtain the secondary seed liquid.

[0052] Solid fermentation medium: rice bran 1000g, bran 2500g, soybean meal 3000g, corn flour 3000g, skimmed milk powder 200g, sucrose 200g, yeast extract 50g, NaCl 40g, feed multi-mineral 10g (Guangzhou Guoxin Feed Technology Co., Ltd., product name It is: Jiangfeng Multi-Mining, the article number is: JC1000), total 10kg, dissolved in 11kg tap water, the ratio of material to water is 1:11, and the mixture is uniform. ...

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Abstract

The invention discloses bacillus pumilus, a probiotics preparation and a preparation method and application thereof. The Bacillus pumilus LV149 is preserved in China center for type culture collection (CCTCC) in Nov. 23th, 2011, and the preservation number is CCTCC NO: M 2011411. The bacillus pumilus LV149 has strong extracellular protease, lipase and amylase activities, has wide rejection capability to vibrio and has no hemolytic activity. The Bacillus pumilus LV149 serves as fermenting bacterial strains and is performed with solid fermentation, drying and smashing so as to prepare bacillus pumilus probiotics preparation which uses Bacillus pumilus LV149 as the active ingredients. The bacillus pumilus probiotics preparation can be added into prawn feeds for feeding prawns, so that growth of prawn intestinal pathogenic vibrio can be restrained, prawn vibriosis can be reduced, simultaneously prawn growth is promoted, fish bait coefficient is reduced, quality of commodity is improved, culture cycle is shortened, accordingly culture risk and culture cost are reduced, biological safety is high and the bacillus pumilus, the probiotics preparation and the preparation method and application thereof have wide application prospect in aquaculture.

Description

Technical field [0001] The invention belongs to the field of microorganisms, and specifically relates to a Bacillus pumilus with high enzyme activity and vibriostatic activity, and a probiotic preparation using it as an active ingredient, and a preparation method and application thereof. Background technique: [0002] With the rapid development of large-scale and intensive aquaculture industry, aquaculture diseases frequently occur. In order to control diseases, some antibiotic drugs are widely used and abused in aquaculture, which not only causes drug residues in the environment and increases bacterial resistance, but also Moreover, due to the fluidity of the water environment, resistance genes are easier to transfer horizontally, resulting in the emergence of more resistant strains. In order to deal with drug resistance, the aquaculture has to increase the dosage and constantly change the types of drugs, which will cause a vicious circle. On the other hand, the large-scale use...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/74A61P31/04A23K1/16C12R1/07A61K35/742
Inventor 罗鹏江海英胡超群王艳红任春华
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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