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 the problems of less inhibitory effect, achieve the effects of improving product quality, high biological safety, and wide application prospects

Active Publication Date: 2013-06-19
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) Isolation of intestinal adherent bacteria

[0029] Five batches of healthy Litopenaeus vannamei shrimp were purchased at different times and in different markets, with 30 tails in each batch. From them, 5 tails with large individual, bright body color, smooth body, no damage and brown liver were selected, with an average body length of 12cm. Disinfect the body surface of prawns with alcohol with a volume fraction of 75%, and then wash them with sterile seawater. Rinse the intestinal tract 2-3 times until there is no residual content in the intestinal tract with the naked eye. Add sterile saline to a sterile homogenizer to homogenize the intestinal tract. Take 0.5 mL homogenate and dilute it 10 times serially with sterile physiological saline (1% NaCl) to obtain the intestinal adhesion bacteria suspension.

[0030] The intestinal adhesion bacteria suspension was coated on the shrimp feed medium....

Embodiment 2

[0042] Example 2: Vibrio inhibition spectrum analysis and hemolytic analysis of Bacillus pumilus LV149

[0043] The inhibition spectrum of Bacillus pumilus LV149 was analyzed using 13 Vibrio strains from 9 different species. The above-mentioned flat filter paper diffusion method was also used. The strength of the antibacterial ability was determined 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; ++: Moderate antibacterial activity; +: Has antibacterial activity.

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

[0048] The h...

Embodiment 3

[0049] Embodiment 3: the preparation of bacillus pumilus probiotic preparation

[0050] First-class seed liquid: LB is used as the medium, and Bacillus pumilus LV149 cryopreserved is added to the LB liquid medium at a ratio of 1% (volume percentage), and shake cultured at 30° C. for 24 hours to obtain the first-class seed liquid.

[0051] Secondary seed liquid: with 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-state 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 multimineral 10g (Guangzhou Guoxin Feed Technology Co., Ltd., product name For: Jiangfeng multi-mineral, product number: JC1000), total 10kg, dissolved in 11kg tap water, the ratio of material to water is 1:11, mix evenly.

[0053]Sterili...

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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 in particular relates to a bacillus pumilus with high enzyme activity and vibrio-inhibiting activity, a probiotic preparation using it as an active ingredient, a preparation method and application thereof. Background technique: [0002] With the rapid development of large-scale and intensive aquaculture, aquaculture diseases occur frequently. In order to control diseases, some antibiotics are widely used and abused in aquaculture, which not only causes drug residues in the environment and increases bacterial resistance, Moreover, due to the fluidity of the water environment, drug-resistant genes are easier to transfer horizontally, resulting in the emergence of more drug-resistant strains. In order to cope with drug resistance, the amount of drugs used in farming has to be increased and the types of drugs are constantly replaced, thus causing a vicious circle. On the other hand, due to the exte...

Claims

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Application Information

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Patent Type & Authority Patents(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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