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Ox-bile-tolerant bezoar transformation bacterium screening culture medium, preparation method and application

A technology for transforming Calculus Bovis and selecting culture medium, which is applied in the directions of bacteria, resistance to vector-borne diseases, fermentation, etc. It can solve the problems of difficulty in screening out strains, survival, affecting the transformation efficiency of Calculus Bovis, etc., and achieve the effect of high transformation efficiency of Calculus Bovis.

Active Publication Date: 2022-07-05
SHANDONG UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, bovine bile has a broad-spectrum antibacterial effect. Most gallbladder bacteria are intolerant to bile and cannot survive in bile for a long time. In particular, some high-efficiency bezoar-transformed bacteria are difficult to survive in bile for more than 48 hours, seriously affecting Calculus conversion efficiency
At present, it is difficult to screen out the strains that can survive in ox bile for a long time in the various media (LB, TSA, BHI, etc.) in vitro cultured bezoar

Method used

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  • Ox-bile-tolerant bezoar transformation bacterium screening culture medium, preparation method and application
  • Ox-bile-tolerant bezoar transformation bacterium screening culture medium, preparation method and application
  • Ox-bile-tolerant bezoar transformation bacterium screening culture medium, preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Preparation of bovine intestine infusion: take fresh bovine small intestine, cut open to remove the contents, rinse clean, cut into small pieces, homogenize into meat puree, add 3 times the volume of normal saline, continue to homogenize, filter with gauze, filtrate at 4000rpm After centrifugation for 15 min, the supernatant was lyophilized to prepare lyophilized powder of bovine intestine infusion. Take an appropriate amount of beef intestine infusion freeze-dried powder and prepare 15% beef intestine infusion with deionized water.

[0028] The preparation method of bovine liver extract is as follows: cut fresh bovine liver into 1-2 cm 3 Add 3 times the volume of 15% beef intestine infusion to the beef liver puree, mix well, adjust the pH of the mixture to 7, stir and keep at 37°C for 6h, 121°C High-pressure cooking for 1 hour, after cooling, put the top layer of oil at a low temperature of 15 °C to condense the top layer of oil into a layer, remove the condensed oil ...

Embodiment 2

[0033] Preparation of bovine intestine infusion: take fresh bovine small intestine, cut it open to remove the contents, rinse clean, cut into small pieces, homogenize into meat puree, add 3 times the volume of deionized water, distilled water or normal saline, and continue to homogenize , filtered with gauze, the filtrate was centrifuged at 4000 rpm for 30 min, the supernatant was taken and freeze-dried to prepare beef intestine infusion freeze-dried powder. Take an appropriate amount of beef intestine infusion freeze-dried powder and prepare 10% beef intestine infusion with deionized water or distilled water.

[0034] The preparation method of bovine liver extract is as follows: cut fresh bovine liver into 1-2 cm 3 Add 3 times the volume of 10% beef intestine infusion to the beef liver puree, mix evenly, adjust the pH of the mixture to 7.5, stir and keep at 37°C for 6h, High-pressure cooking at 121°C for 1 h, after cooling, place the top layer of oil at a low temperature of ...

Embodiment 3

[0040] Preparation of bovine intestine infusion: take fresh bovine small intestine, cut it open to remove the contents, rinse clean, cut into small pieces, homogenize into meat puree, add 3 times the volume of deionized water, distilled water or normal saline, and continue to homogenize , filtered with gauze, the filtrate was centrifuged at 4000 rpm for 30 min, the supernatant was taken and freeze-dried to prepare beef intestine infusion freeze-dried powder. Take an appropriate amount of beef intestine infusion freeze-dried powder and prepare 20% beef intestine infusion with deionized water or distilled water.

[0041] The preparation method of bovine liver extract is as follows: cut fresh bovine liver into 1-2 cm 3 Add 3 times the volume of 20% beef intestine infusion to the beef liver meat puree, mix well, adjust the pH of the mixture to 7.2, stir and keep at 37°C for 6h, High-pressure cooking at 121°C for 1 h, after cooling, place the top layer of oil at a low temperature ...

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Abstract

The invention relates to a culture medium for screening microorganisms, in particular to a culture medium for separating efficient bezoar transforming bacteria resistant to oxgall, a preparation method and application. The formula of the culture medium comprises the following components: relative to the amount of 1000.0 ml of a solvent, the culture medium further comprises 100.0 to 300.0 g of a bovine liver extract, 100.0 to 300.0 g of a bovine kidney extract, 5.0 to 15.0 g of peptone, 3.0 to 5.0 g of glucose, 3.0 to 5.0 g of NaCl, 3.0 to 5.0 g of disodium hydrogen phosphate and 100.0 to 600.0 ml of ox bile, 10.0 to 30.0 g of agar is added on the basis of the solid culture medium, and the pH value is adjusted to 6.8 to 7.5. The strain separated and screened by using the culture medium disclosed by the invention has the characteristics of being capable of tolerating ox bile for a long time and high in bezoar conversion efficiency.

Description

technical field [0001] The invention relates to a culture medium for microbial screening, in particular to a culture medium, preparation method and application for isolating high-efficiency oxbile-tolerant bacteria transformed from bovine bile. Background technique [0002] Bezoar is a bovine bovine ( Bostaurus domestlcus Gmel) or buffalo ( Bubalus bubalisl ) is a dry product of stones formed in the gallbladder or bile duct under pathological conditions. About 650 kinds of 4,500 kinds of proprietary Chinese medicines in my country contain bezoar. The occurrence rate of natural bezoar in Chinese cattle is only 0.21%, and the natural output is very rare. In the 1990s, Professor Cai Hongjiao first developed the in vitro cultivation of Bezoar (CN1022668C, CN1044402A, CN1041105A), which was included in the Chinese Pharmacopoeia (2020 Edition). Compound bilirubin calcium, etc. At present, only Wuhan Jianmin Dapeng Pharmaceutical Co., Ltd. exclusively produces bezoar cultivat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12P1/04
CPCC12N1/20C12N1/02C12P1/04Y02A50/30
Inventor 王厚伟窦彦玲王康诚张珂璠徐凌川李依展
Owner SHANDONG UNIV OF TRADITIONAL CHINESE MEDICINE
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