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Bacillus subtilis fermentation broth of Chinese herbal medicine as well as preparation and application of bacillus subtilis fermentation broth

A technology of Bacillus subtilis and Chinese herbal medicine, applied in the fields of application, botany equipment and methods, medical preparations containing active ingredients, etc., can solve the problems of double infection, endogenous infection, secondary pollution, etc., and reduce the incidence of disease The effect of rate and degree of harm

Inactive Publication Date: 2015-02-18
HUNAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

However, the extensive use of antibiotics has caused a series of serious problems such as endogenous infections and superinfections, the emergence of drug-resistant bacterial strains, the decline of livestock and poultry immunity, and secondary pollution caused by residues in livestock products and the environment. Many countries and regions such as Antibiotics have been banned in both the EU and South Korea

Method used

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  • Bacillus subtilis fermentation broth of Chinese herbal medicine as well as preparation and application of bacillus subtilis fermentation broth
  • Bacillus subtilis fermentation broth of Chinese herbal medicine as well as preparation and application of bacillus subtilis fermentation broth
  • Bacillus subtilis fermentation broth of Chinese herbal medicine as well as preparation and application of bacillus subtilis fermentation broth

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Experimental program
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Effect test

Embodiment 1

[0029] Preparation of Bacillus subtilis Fermentation Broth of Chinese Herb Galla chinensis

[0030] A. Resurrection: Bacillus subtilis AP139 (preservation number: CCTCC NO: M 2014053) was cultured by streaking in a constant temperature biochemical incubator on LB solid medium, with a culture temperature of 30-37°C and a culture time of 18-24h; LB The composition and mass percentage of the solid medium are: 1% of bacteriological peptone, 0.5% of yeast extract, 0.5% of sodium chloride, 1.5% of agar powder, and the pH value is 6.8-7.2.

[0031] B, then from the LB solid medium, pick the revived Bacillus subtilis AP139 single clone and inoculate it in the 500ml shake flask that 100ml medium is housed and cultivate, the fermentation temperature is 35 ℃, the rotating speed is 200r / min, and the cultivation time is 24h. The composition and mass percentage of the shake flask medium are: 1% of bacteriological peptone, 0.5% of yeast extract, 0.5% of sodium chloride, and the pH value is ...

Embodiment 2

[0034] Inhibitory effect of Bacillus subtilis AP139 fermented without gallnut components

[0035] Take 2 LB nutrient agar plates, respectively take 10 μL of activated Staphylococcus aureus liquid (1×10 8 CFU / ml) and Pasteurella porcine toxin (1×10 8CFU / ml), spread evenly on the LB solid plate, place in an incubator at 35-37°C and incubate for 10 minutes, use a puncher to punch holes in the culture dish with a diameter of 5 mm; gallnuts are not added, and the rest follow the After centrifuging the Bacillus subtilis fermentation broth prepared by the method of Example 1 at 12,000 r / m for 3-5 minutes, take 80-100 μL of the supernatant of the fermentation broth and add it to the well of the culture dish, and incubate in an incubator at 35-37° C. for 24-48 hours. Observe the size and clarity of the Bacillus inhibition zone to judge the antibacterial ability of the test bacteria against the indicator bacteria. The results showed that the diameters of the inhibition zones of the Ba...

Embodiment 3

[0037] Verification of the antibacterial activity of Chinese herbal medicine Galla chinensis:

[0038] A, the preparation of Chinese herbal medicine Galla gall fermentation solution not containing Bacillus subtilis AP139

[0039] Weigh 3 g gallnut powder with an electronic balance, and carry out shake flask culture in 100 ml medium, the fermentation temperature is 35° C., the rotational speed is 200 r / min, and the fermentation time is 24 hours. The composition and mass percentage of the shake flask fermentation medium are: 1% of bacteriological peptone, 0.5% of sodium chloride, 1% of glucose, 1% of magnesium sulfate heptahydrate, and the pH value is natural.

[0040] B. Take 2 LB nutrient agar plates, take 10 μL of activated Pasteurella multocida bacterial liquid, and evenly spread it on one LB solid plate, take 10 μL of activated Staphylococcus aureus bacterial liquid, and spread it evenly on the second Place on an LB solid plate and incubate at 37°C for 10 minutes in an inc...

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Abstract

The invention relates to bacillus subtilis fermentation broth of a Chinese herbal medicine as well as preparation and an application of the bacillus subtilis fermentation broth. The Chinese herbal medicine refers to Chinese gall; according to pasteurella multocida and staphylococcus aureus inhibiting screening tests, the Chinese gall can inhibit growth of the pasteurella multocida and the staphylococcus aureus, bacillus subtilis AP139 is used for fermenting the Chinese gall, spraying is performed in an animal breeding shed, the fermented Chinese herbal medicine remarkably inhibits the growth of the pasteurella multocida and the staphylococcus aureus in the air, the effect is superior to those of a bacillus subtilis AP139 separate fermentation group and a Chinese herbal medicine control group, and the fermentation broth has the broader application prospect.

Description

technical field [0001] The invention belongs to the field of microbial fermentation bactericides, and in particular relates to a Bacillus subtilis fermented liquid of Chinese herbal gall gall and its preparation method, and its application for inhibiting Pasteurella multocida and Staphylococcus aureus. Background technique [0002] Pasteurella multocida and Staphylococcus aureus are important commensal pathogens of livestock, poultry, wild animals and humans, and are mainly transmitted through the respiratory tract. Pasteurella multocida can cause fowl cholera, swine pneumonia and bovine hemorrhagic sepsis, etc. Toxigenic Pasteurella multocida can also cause atrophic rhinitis in pigs, and Staphylococcus aureus can cause mastitis in cows, food contamination, wounds, etc. suppuration etc. Staphylococcus aureus and Pasteurella multocida are spread throughout the air, water, and human and animal excreta. Pasteurella multocida and Staphylococcus aureus are common in livestock an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K1/16A01N63/02A01P1/00A61K35/742A61P31/04A61K35/64
Inventor 舒燕王升平刘惠知郭照辉张德元邬理洋周映华胡新旭高书锋周小玲缪东曾发姣
Owner HUNAN INST OF MICROBIOLOGY
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