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Bacillus pumilus and application thereof to preparation of exopolysaccharides

A technology of Bacillus pumilus and exopolysaccharides, applied in the direction of application, bacteria, and microbial-based methods, can solve the problems of economic losses for shrimp farmers, improve non-specific immunity, strong acid-base resistance, and improve shrimp The effect of resistance to viral infection

Inactive Publication Date: 2015-06-17
威海光洋生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, virus infection is a serious threat affecting shrimp farming, causing severe economic losses to shrimp farmers

Method used

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  • Bacillus pumilus and application thereof to preparation of exopolysaccharides

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Experimental program
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Effect test

Embodiment 1

[0012] Embodiment 1: Screening of Bacillus pumilus (Bacillus pumilus) GY001 bacterial strain

[0013] (1) Separation and purification of bacteria:

[0014] In February 2013, intestinal samples of disease-resistant Penaeus vannamei from ponds where WSSV had occurred were extracted from Jiaonan, Qingdao, and put into 100ml sterile water, and the bacterial suspension was obtained after shaking for 20 minutes. Pipette 1ml respectively, dilute 1000 times, take 30μl and spread on LB plates, dry slightly and incubate at 30°C upside down. After 48 hours, according to the characteristics of the colony shape and color on the plate, select representative colonies and streak on the LB plate for purification. The purified bacterial isolates were inoculated on the slant medium and stored at 4°C until use. Finally, a total of 125 strains were isolated from the intestinal tract of prawns.

[0015] (2) Antibacterial screening

[0016] Bacteriostasis test - double-layer plate, Oxford cup met...

Embodiment 2

[0022] Example 2: Sensitivity test of Bacillus pumilus GY001 to antibiotics in vitro

[0023] 1) Antibiotic selection: 1-2 commonly used and relatively easy-to-obtain antibiotics are selected for each major category of veterinary antibiotics. According to the conventional therapeutic dosage of various antibiotics in the full-price feed, 2 times of the proposed therapeutic dosage is the severe dosage, and 1 / 2 of the therapeutic dosage is the preventive dosage.

[0024] 2) Bacterial solution preparation: Inoculate Bacillus pumilus GY001 into liquid 2216E medium and culture at 28°C for 48 hours, then dilute to 10 with 2216E medium 8 Magnitude.

[0025] 3) Sensitivity test: Take 5mL of the diluted GY001 bacterial liquid, add the antibiotics selected in 1) to it so that the concentration reaches the severe dose, therapeutic dose and preventive dose respectively, place it at 37°C for 4 hours, and use 2216E agar count. Calculate the survival rate of Bacillus pumilus GY001 under di...

Embodiment 3

[0033] Embodiment 3, exopolysaccharide produced by Bacillus pumilus

[0034] (1) Strain activation: take the Bacillus pumilus GY001 strain, culture the strain on the culture medium at 37°C for 2 days, and culture it in the fermentation medium at 37°C and 180rpm for 3 days to obtain liquid strains; the culture medium is: 1L distilled water, 10g glucose, 25g peptone, 5g sodium chloride, 1.0g beef extract, 18g agar; fermentation medium: 10g soluble starch, 15g corn flour, 10g lactose, 2.5g glucose, 0.5g yeast extract, 21g CaCl, MgSO4 7H2O1g, dissolved in deionized water to 1000mL, pH7.6;

[0035] (2) Strain expansion: Inoculate the liquid strain of step (1) into a triangular flask containing fermentation medium with an inoculum of 10 6 The inoculum amount of CFU / mL was inoculated into 100mL LB Erlenmeyer flasks, cultured in shake flasks at 37°C, 180rpm for 5 days, and kept at 4°C for 16h.

[0036] 2. Extraction of Exopolysaccharides

[0037](1) Casein removal by isoelectric po...

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Abstract

The invention provides Bacillus pumilus which is a GY001 strain and has a collection number of CGMCC No.8639. The screened Bacillus pumilus is used for fermenting and producing exopolysaccharides. According to the screened Bacillus pumilus for exopolysaccharides, the acid and alkali resistance of the exopolysaccharides is high, and the unique activity structure has a unique effect of resisting prawn viruses and has an ecological effect of maintaining good prawn digestive tract balance. The nonspecific immunity level of prawn breeding can be obviously improved, the pathogen infection resistance of the bred prawns is greatly improved, particularly multiple biological functions, such as virus infection resistance, of the prawns can be obviously improved, and the Bacillus pumilus is low in cost and easy to transport and store.

Description

technical field [0001] The invention belongs to the technical field of microbial screening applications, and in particular relates to a bacillus pumilus and its application in preparing exopolysaccharides. The exopolysaccharides produced by the bacillus pumilus are used for the prevention and treatment of WSSV in prawns. Background technique [0002] The large-scale development of prawn farming began in the 1970s and 1980s. The production of prawn farming from south to north along the coast of my country is very large, which has played a great role in promoting the development of coastal economy. Currently, virus infection is a serious threat affecting shrimp farming, causing severe economic losses to shrimp farmers. Shrimp white spot syndrome virus (White Spot Syndrome Virus, WSSV) disease is one of the viral diseases that seriously endanger the healthy development of the shrimp farming industry. According to the shrimp disease report of the World Bank, in 1994, shrimp vir...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P19/04A23K1/16C12R1/07
Inventor 刘旭雅元福杰姜英俊
Owner 威海光洋生物科技有限公司
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