Bacillus pumilus strain and micro-ecological preparation and feed thereof

A technology of Bacillus pumilus and micro-ecological preparations, applied in the directions of microorganisms, microorganisms, animal feed, etc., can solve the problems of restricting the export and sales of finished tilapia products, restricting the development of tilapia industry, and overusing antibiotic residues, etc. Risk, resistance-enhancing, growth-inhibiting effects

Active Publication Date: 2017-05-17
TIANJIN CHANGNONG TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, antibiotics are mostly used to control the occurrence and infection of Streptococcus tilapia. This farming model has led to the abuse of antibiotics and excessive residues in the farming process, which seriously restricts the export and sales of tilapia finished products, and has become a restricted tilapia. The bottleneck of industrial development

Method used

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  • Bacillus pumilus strain and micro-ecological preparation and feed thereof
  • Bacillus pumilus strain and micro-ecological preparation and feed thereof
  • Bacillus pumilus strain and micro-ecological preparation and feed thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The antibacterial test method of embodiment 1 bacillus pumilus CD6

[0022] 1) Culture method of Streptococcus agalactiae

[0023] Liquid medium: peptone 10.0g / L, dehydrated calf brain extract powder 12.5g / L, dehydrated beef heart extract powder 5.0g / L, sodium chloride 5.0g / L, glucose 2.0g / L, disodium hydrogen phosphate 2.5 g / L, pH7.5.

[0024] Solid medium: peptone 10.0g / L, dehydrated calf brain extract powder 12.5g / L, dehydrated beef heart extract powder 5.0g / L, sodium chloride 5.0g / L, glucose 2.0g / L, disodium hydrogen phosphate 2.5 g / L, pH7.5, agar powder 1.5%.

[0025] 2) Screening plate production

[0026] A single colony of Streptococcus agalactiae on the plate was inoculated into the liquid medium, and cultured at 25°C for 16h. Pour the cultured fermented liquid into the sterilized solid medium at a ratio of 10% under sterile conditions and cool to 40-45°C, mix well, pour it into a sterile plate, 30ml / plate, cool and solidify for later use .

[0027] 3) Scr...

Embodiment 2

[0030] The mensuration of embodiment 2 bacillus pumilus CD6 fermentation supernatant minimum inhibitory concentration

[0031] 1) Preparation method of bacillus pumilus CD6 fermentation broth supernatant

[0032] Inoculate CD6 into 100ml triangular samples containing 25ml LB medium, incubate at 37°C for 24 hours, take 10ml of fermentation broth, centrifuge at 5000r / min for 15min, and collect the supernatant without cells.

[0033] 2) Preparation of different concentrations of CD6 fermentation broth supernatant

[0034] Take 5 glass test tubes, marked as 0, 1, 2, 3, 4 respectively, add 1ml of the supernatant after centrifugation without bacteria to each test tube, add 1ml of pure water to test tube 1, add 2ml of water to test tube 2 , add 3ml of pure water to test tube 3, add 4ml of water to test tube 4. The concentrations of the supernatants of test tubes 1, 2, 3 and 4 were diluted 2 times, 3 times, 4 times and 5 times respectively.

[0035] 3) Determination of minimum inhi...

Embodiment 3

[0040] The preparation of embodiment 3 Bacillus pumilus high-density fermented liquid

[0041] 1) Plate culture rejuvenation: Inoculate the Bacillus pumilus strain on the LB plate medium, culture at 30°C for 24 hours, rejuvenate the Bacillus pumilus, and form a single colony, pick a single colony on the inoculation medium, and culture at 37°C 24h;

[0042]2) Preparation of first-class seeds: transfer the Bacillus pumilus cultured in step 1) to the LB slant medium of eggplant bottle, culture at 37°C for 24 hours, make it in the late logarithmic period, and obtain the first-class seeds;

[0043] 3) Preparation of secondary seeds: make bacterial suspension from the primary seeds prepared in step 2) with sterile water, and inoculate them into a 100L seed tank equipped with 60L LB seed medium at a temperature of 30°C and a rotational speed of 200r / min , tank pressure 0.05MPA, ventilation ratio: 1:0.6 and cultivate for 14 hours to obtain secondary seed liquid.

[0044] 4) Preparat...

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Abstract

The invention belongs to the technical field of biologics, and discloses bacillus pumilus capable of inhibiting streptococcus agalactiae and application thereof. The bacillus pumilus has the advantages that the screened bacillus pumilus metabolite can be used for obviously inhibiting the metabolism and growth of the streptococcus agalactiae, the number of streptococcus agalactiae in the tlapia mossambica culture environment is reduced, and the occurrence of streptococcicosis is inhibited; the bacillus pumilus is subject to multiple times of strain selection and breeding, and can be performed with liquid deep fermenting, so as to prepare water preparation products and powder preparation products; the powder preparation products are prepared into a tlapia mossambica particle feed according to a certain ratio, and then the tlapia mossambica particle feed can be applied into aquaculture.

Description

technical field [0001] The invention belongs to the field of biotechnology. A strain of Bacillus pumilus capable of inhibiting Streptococcus tilapia is screened out, optimized for fermentation, and prepared into water and powder products, which can be used in aquatic feed. Background technique [0002] As one of the most important aquaculture species in the world's aquaculture industry, Tilapia has the advantages of fast growth, miscellaneous food habits, good meat quality, high nutritional value, and strong reproductive ability, and is widely distributed in tropical and subtropical brackish waters. my country's coastal areas, especially Hainan, Guangdong, Fujian, Guangxi and other provinces, account for more than half of the world's tilapia production. However, in recent years, with the increase of tilapia culture area and culture density, tilapia streptococcosis caused by Streptococcus agalactiae infection has become the main obstacle to the healthy and sustainable develop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/747A61P31/04A23K10/18C12R1/07
CPCA61K2035/115C12N1/20C12N1/205C12R2001/07
Inventor 付维来易敢峰卢俊王蕊张聪颖孙强王宁朱传忠
Owner TIANJIN CHANGNONG TECH
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