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Probiotic for inhibiting clostridium perfringens, and screening method and application thereof

A Clostridium perfringens and screening method technology, applied in the biological field, can solve problems affecting growth and development, shock and even death, and achieve the effect of improving weight loss, improving the structure of intestinal flora, and reducing the diarrhea rate of mice

Active Publication Date: 2021-08-27
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If sick animals are not treated correctly and in a timely manner, it is easy to cause shock or even death. Even if young livestock and poultry infected with Clostridium perfringens can survive after treatment, it will affect their growth and development

Method used

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  • Probiotic for inhibiting clostridium perfringens, and screening method and application thereof
  • Probiotic for inhibiting clostridium perfringens, and screening method and application thereof
  • Probiotic for inhibiting clostridium perfringens, and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 strain screening:

[0030] (1) Preliminary screening of Bacillus with antibacterial ability from pig-derived intestinal contents and feces

[0031] Take 1g of jejunum, ileum intestinal content and feces samples from 180-day-old Jinhua pigs, dilute them with sterilized normal saline at a ratio of 1:10, bathe in 75°C water for 15 minutes, and then dilute them in a 10-fold gradient with sterile water. -3 、10 -4 、10 -5 、10 -6 For four dilution gradients, draw 100 μL of the bacterial suspension and spread it on the LB plate, at least 3 plates for each dilution gradient, cool the upper layer of the LB semi-solid medium to about 50°C, add 5% of the activated product Clostridium perfringens (ATCC 13124), after mixing, pour it into the upper plate immediately and spread it, place it in a constant temperature incubator at 37°C for 12 hours, make at least 3 plates for each dilution gradient and each pathogenic bacteria, and screen Bacillus having the ability to inhib...

Embodiment 2

[0055] Example 2: Identification of strain BA40

[0056] The 16S 16S rDNA of the strain BA40 of the present invention was sequenced, and compared with the NCBI database by Blast, it was shown that the homology between BA40 and the type strain Bacillus amyloliquefaciens AB301002.1 was 99.72%, and the homology with the type strain Bacillus amyloliquefaciens KX137853.1 was 99.38% %, so it was determined to be Bacillus amyloliquefaciens. Strain BA40 was deposited in China Center for Type Culture Collection, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province on May 17, 2021, and the preservation number is CCTCC NO: M2021535.

Embodiment 3

[0057] Example 3 Application of Bacillus amyloliquefaciens BA40 to prevent Clostridium perfringens infection

[0058] 1) Preparation of probiotic bacteria liquid

[0059] Inoculate the Bacillus amyloliquefaciens BA40 described in claim 6 in the LB liquid medium, and culture it on a shaking table at 37° C. for 12 hours, so that the concentration of the bacterial solution reaches 1.0×10 8 ~1.0×10 9 CFU / mL, prepare Bacillus amyloliquefaciens BA40 bacterial liquid; follow the same method, prepare bacterial liquid for commercially available Bacillus subtilis PB6.

[0060] 2) Application of probiotic liquid

[0061] 24 6-week-old C57B / L mice were selected and randomly divided into four groups, namely control group, Clostridium perfringens infection group, BA40+Clostridium perfringens infection group, PB6+Clostridium perfringens infection group, and PB6+Clostridium perfringens infection group. infection group. The BA40+Clostridium perfringens infection group was orally administe...

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Abstract

The invention discloses a probiotic for inhibiting clostridium perfringens, and a screening method and application thereof. The probiotic is bacillus amyloliquefaciens BA40 and is preserved in the China Center for Type Culture Collection with the preservation number of CCTCC NO: M2021535. The screening source of the probiotic is pig source intestinal content and excrement sample. Probiotic strains are subjected to primary screening through a double-layer agar method and an agar diffusion experiment, and then two probiotics, namely bacillus amyloliquefaciens BA40, with remarkable inhibition capacity on clostridium perfringens are obtained through acid-resistant, cholate-resistant, pancreatin-resistant and other experiments. When the strain is used for animal experiments for preventing the infection of the clostridium perfringens in mice, the problems of weight loss, intestinal morphological injury, intestinal inflammatory response and the like caused by the infection of the clostridium perfringens can be remarkably improved, meanwhile, the content of IgA and IgG in serum of the mice is increased by the probiotic treatment group, the intestinal flora structure of the mice is remarkably improved, the diarrhea rate of the mice is reduced, and a new way is provided for replacing antibiotics.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to the field of development and application of functional bacillus, in particular to probiotics inhibiting Clostridium perfringens and its screening method and application. Background technique [0002] Clostridium perfringens can cause a variety of diseases in humans and other animals, including human food poisoning, lamb dysentery, piglet diarrhea, chicken necrotic enteritis and other diseases. If the sick animals are not treated correctly and in time, it will easily lead to shock or even death. Even if young livestock and poultry infected with Clostridium perfringens can survive after treatment, their growth and development will be affected. Clostridium perfringens is the main pathogenic bacteria of zoonotic diseases. Once the pathogenic conditions are triggered, it will bring serious economic losses to human health and animal husbandry and other industries. [0003] Probiot...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A61K35/742A61P31/04C12R1/07
CPCC12N1/20C12N1/02A61K35/742A61P31/04
Inventor 汪以真姜子鹏李文涛文超越苏维发路则庆靳明亮王凤芹
Owner ZHEJIANG UNIV
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