Copper-resistant cholate-resistant bacterial strain, probiotic agent and application of copper-resistant cholate-resistant bacterial strain and probiotic agent

A technology of bile salt resistance and antibacterial agents, applied in the field of microorganisms, can solve the problems of short shelf life, poor tolerance, and small number of viable bacteria in products, and achieve good probiotic effects

Active Publication Date: 2022-02-01
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, probiotics have poor tolerance to copper. When copper is contained in animal feed, it is easy to affect the probiotics in animal feed, so that the probiotics cannot perform their own functions.
[0005] Therefore, due to the poor tolerance of the current probiotics to bile salts and copper, the number of live bacteria in the product is small, the shelf life is short, and it is difficult to achieve the probiotic effect in the host body

Method used

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  • Copper-resistant cholate-resistant bacterial strain, probiotic agent and application of copper-resistant cholate-resistant bacterial strain and probiotic agent
  • Copper-resistant cholate-resistant bacterial strain, probiotic agent and application of copper-resistant cholate-resistant bacterial strain and probiotic agent
  • Copper-resistant cholate-resistant bacterial strain, probiotic agent and application of copper-resistant cholate-resistant bacterial strain and probiotic agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Screening of copper-resistant and bile-resistant bacterial strains

[0046] (1) Take the feces of domestic livestock and poultry from a farmer in Qingdao City, Shandong, dip the tip of a pipette into 1 mL of sterilized water, shake and mix to obtain a suspension, and inoculate the suspension with a 5% inoculum Copper broth liquid culture medium (wherein, the formula of broth liquid culture medium containing copper is: add 1g tryptone, 0.5g yeast extract powder, 0.2g dipotassium hydrogen phosphate, 0.2g diammonium citrate per 100mL distilled water , 2g glucose, 0.058g magnesium sulfate heptahydrate, 0.025g manganese sulfate, 0.5g sodium acetate, 0.1ml Tween 80, 0.1g copper sulfate pentahydrate and 0.5ml glacial acetic acid), at 37°C, 220r / min Under the condition of shaking and culturing for 24 hours, the bacterial liquid was obtained.

[0047] (2) Inoculate the bacterium liquid with an inoculum amount of 5% in a copper-containing broth liquid medium and subcul...

Embodiment 2

[0050] The identification of embodiment 2 bacterial strains

[0051] (1) Identification of the morphological characteristics of the culture medium

[0052] The bacillus NGTNDY.D8 obtained in Example 1 was streaked on an LB medium plate, cultured at 28° C. for 24 hours, and characteristics such as colony shape, size, and color were observed. Wherein, LB culture medium comprises the component of following concentration:

[0053] Add 1g of tryptone, 0.5g of yeast extract powder, 1g of sodium chloride, and 1.5g of agar powder per 100ml of distilled water.

[0054] Observation results such as figure 1 As shown, the colony of Bacillus NGTNDY.D8 is round, white, with rough edges and glossy.

[0055] (2) Colony PCR amplification and 16S rDNA sequencing

[0056] Extract the genomic DNA of the copper-resistant and bile-resistant bacterial strain D8 obtained in Example 1: extract the probiotic genome obtained according to the instructions of the bacterial genome extraction kit purcha...

Embodiment 3

[0057] The copper-resistant bile salt test of embodiment 3 bacterial strains

[0058] (1) The bacillus NGTNDY.D8 obtained by the above screening was inoculated respectively in broth solid medium containing 0.3% porcine bile salt and in broth medium containing 0.3% copper sulfate and cultured for 1 day to observe its growth (set 3 parallel control group), and tested their survival rate, the results are shown in Table 1.

[0059] Table 1 Test results of copper resistance and bile salt resistance

[0060] Survival rate under 0.3% pig bile salt Survival rate under 0.3% copper sulfate Bacillus NGTNDY.D8 99％～99.8％ 98.5％～99.7％

[0061] Therefore, Bacillus NGTNDY.D8 can grow well in the medium with 0.3% high copper and 0.3% bile salt, and it has good tolerance to copper and bile salt.

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Abstract

The invention discloses a copper-resistant and cholate-resistant strain, a probiotic agent and an application of the copper-resistant and cholate-resistant strain and the probiotic agent, and relates to the technical field of microorganisms. The copper-resistant and cholate-resistant strain is bacillus (Bacillus sp.) NGTNDY.D8, the preservation number of the strain is CCTCC NO: M 2021477, and the preservation time of the strain is April 28, 2021. The copper-resistant and cholate-resistant strain provided by the invention has tolerance to copper and cholate, the survival rate of bacillus NGTNDY.D8 is 99-99.8% under the condition that the mass fraction of copper is 0.3%, the survival rate of bacillus NGTNDY.D8 is 98.5-99.7% under the condition that the mass fraction of cholate is 0.3%, the good probiotic effect can be fully exerted, and beneficial strain resources are provided for the field of animal feed.

Description

technical field [0001] The invention relates to the technical field of microorganism technology, in particular to a copper-resistant and bile-salt-resistant bacterial strain, a probiotic bacterial agent and applications thereof. Background technique [0002] Probiotics are a type of active microorganisms that are beneficial to the host by colonizing the animal body and changing the flora composition of a certain part of the animal. They are often added to animal feed. Probiotics can promote nutrient absorption and maintain intestinal health by regulating the immune function of the host mucosa and system or by regulating the balance of intestinal flora, thereby producing single microorganisms or mixed microorganisms with a defined composition that are beneficial to animal health. [0003] Bile salts secreted by animal liver into the duodenum can inhibit the growth and activity of most probiotics. However, many probiotics can only regulate the balance of intestinal flora after...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K10/18A23K20/20C12R1/07
CPCC12N1/20A23K10/18A23K20/30
Inventor 张西锋马浩海李万芬李兰沈伟
Owner QINGDAO AGRI UNIV
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