Bdellovibrio bacteriovorus microcapsule and preparation method thereof

A technology of phagocytic Bdellovibrio and microcapsules, which is applied in the field of microbial additives for aquatic feed, can solve the problems of rapid decline in bacterial count, inconvenient transportation, short storage period, etc., achieve good dispersibility and solubility, shorten fermentation time, The effect of simple production process

Inactive Publication Date: 2014-04-16
SHANGHAI LYUAO BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the vast majority of Bdellovibrio phage preparations in my country are liquid preparations and freeze-dried powders, but liquid preparations generally have rapid bacterial count decline, short storage period, inconvenient transportation, and are easily affected by the high acid and high enzyme environment in the digestive tract. Many defects such as death, the production cost of freeze-dried powder is high, and it is difficult to meet the requirements of the aquaculture industry
[0004] The Chinese invention patent with the application number 201110088162.1 and the name of a powdery Bdellovibrio preparation and its preparation method discloses a powdery Bdellovibrio preparation, but this preparation is also susceptible to high acid in the digestive tract, The effect of high enzyme environment affects the anti-infective effect

Method used

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  • Bdellovibrio bacteriovorus microcapsule and preparation method thereof
  • Bdellovibrio bacteriovorus microcapsule and preparation method thereof
  • Bdellovibrio bacteriovorus microcapsule and preparation method thereof

Examples

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preparation example Construction

[0054] Preparation of Escherichia coli DH5α suspension

[0055] Under sterile conditions, Escherichia coli DH5α preserved in glycerol at -70°C was inoculated in common nutrient broth at pH 7.2, cultured at 30°C, shaken at 200r / min for 18h, and then centrifuged at 6000r / min for 20min. Wash 3 times with saline to make 2.0 × 10 10 Escherichia coli DH5α suspension in pfu / ml.

[0056] Preparation of culture medium of bacteriophage Bdellovibrio F16

[0057] The final concentration of glycerol preservation at -70°C was 2.0×10 4 Pfu / ml Bdellovibrio F16 was inoculated in 100ml of 10-fold diluted common nutrient broth with pH 7.2, and 100μl / 100ml of Escherichia coli DH5α suspension was added at the same time, shaken at 30°C and 200r / min After shaking culture for 48 hours, the live bacteria content of Bdellovibrio F16 was 4.0×10 5 pfu / ml, stored in a refrigerator at 4°C.

Embodiment 1

[0059] The effect of gelatin concentration on the effective bacterial content of Bdellovibrio phage F16 microcapsules: aseptically take 1 mL of Bdellovibrio phage F16 culture solution and add the final concentrations of 1%, 2%, 3%, 4%, 5% respectively In sterile gelatin solution, then fully mix evenly on a magnetic stirrer at 30°C, 100r / min, and under the conditions of air inlet temperature 130°C, feeding speed 10ml / min, air flow rate 700L / h, needle passing frequency 10s, etc. spray-dried under different gelatin concentrations, respectively, to determine the effective bacteria content of Bdellovibrio F16 microcapsules prepared under different gelatin concentrations. See the experimental results figure 1 .

[0060] figure 1 The histogram, the ordinate is the logarithmic value of the bacteria content, and the abscissa is the concentration of the gelatin solution. It can be seen from the histogram that when the gelatin concentration is 2%, the effective bacterial content of th...

Embodiment 2

[0062] Effect of air flow rate on the effective bacterial content of Bdellovibrio phage F16 microcapsules: Aseptically take 1 mL of Bdellovibrio phage F16 culture solution and add it to a sterile gelatin solution with a final concentration of 2%, and then place it on a magnetic stirrer. Mix well at 30°C and 100r / min, and carry out under the conditions of air flow rate of 600, 650, 700, 750, 800L / h, inlet air temperature of 130°C, feed rate of 10ml / min, needle frequency of 10s, etc. After spray drying, the effective bacteria content of Bdellovibrio F16 microcapsules prepared under different air flow rates were determined. See the experimental results figure 2 .

[0063] figure 2 The histogram, the vertical axis is the logarithmic value of the bacteria content, and the horizontal axis is the air inlet temperature. It can be seen from the histogram that when the air flow rate is 750L / h, the effective bacterial content of the Bdellovibrio microcapsules is the highest. Specif...

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Abstract

The invention discloses a bdellovibrio bacteriovorus microcapsule, the wall of the microcapsule is gelatin, the particle is in a ball shape, the surface has recess without holes and cracks, the particle size of the particle is 3-12 mum, the distribution is basically uniform, wherein the content of bdellovibrio bacteriovorus is 3.69*10<7>pfu / g-3.70*10<7>pfu / g, and the content of escherichia coli DH5 alpha is 8.6*10<9>pfu / g-2.10*10<10>pfu / g. The invention provides a preparation method of the microcapsule, which comprises the following steps: 1) preparing an escherichia coli DH5 alpha bacterium suspension; 2) preparing a bdellovibrio bacteriovorus nutrient solution; and 3)uniformly mixing the escherichia coli DH5 alpha bacterium suspension obtained in the step 1) and the bdellovibrio bacteriovorus nutrient solution obtained in the step 2) with an aseptic gelatin solution, performing spray drying, and finally preparing the microcapsule. The microcapsule obviously increases the anti-infection effect in aquatic product breeding industry.

Description

technical field [0001] The invention belongs to the technical field of microbial additives for aquatic feeding, and in particular relates to a microcapsule of bacteriophage Bdellovibrio and a preparation method thereof. Background technique [0002] In recent years, the rapid development of intensive aquaculture industry has led to the excessive abuse of antibiotics as therapeutic agents and growth promoters for aquatic animal diseases, which has increasingly challenged the safety of aquatic products and caused a series of environmental, health and social problems. At present, developed countries such as Europe, the United States, and Japan have banned antibiotics as feed additives, and are actively looking for alternatives to antibiotics. Therefore, the aquaculture industry is in urgent need of new feed additives that can meet the pollution-free requirements and enable aquatic animals to grow rapidly and healthily. Numerous studies have shown that feeding aquaculture anima...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K1/00
Inventor 曹海鹏何珊
Owner SHANGHAI LYUAO BIOTECH CO LTD
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