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Lactobacillus johnsonii and application thereof in degradation of deoxynivalenol and inhibition of pathogenic bacteria

A technology for deoxynivalenol and Lactobacillus johnsonii, which is applied in the field of microorganisms and achieves the effects of being safe, reliable, easy to use and effective.

Active Publication Date: 2022-08-09
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the application of Lactobacillus johnsonii in the preparation of animal feed has been studied at present, there are few reports on its application in the degradation ability of deoxynivalenol and the ability to inhibit pathogenic bacteria

Method used

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  • Lactobacillus johnsonii and application thereof in degradation of deoxynivalenol and inhibition of pathogenic bacteria
  • Lactobacillus johnsonii and application thereof in degradation of deoxynivalenol and inhibition of pathogenic bacteria
  • Lactobacillus johnsonii and application thereof in degradation of deoxynivalenol and inhibition of pathogenic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 - Isolation, screening and identification of Lactobacillus johnsonii NJD412

[0047] The present embodiment adopts the following method to obtain Lactobacillus johnsonii NJD412:

[0048] (1) Isolation and screening of lactic acid bacteria

[0049] A total of 10 different dog feces samples were collected in Xuanwu District, Nanjing, and 0.1 g of feces were weighed and diluted to 10 with sterile normal saline. -4 times, spread on MRS plate (purchased from Qingdao Haibo Biological Co., Ltd., HB0384), cultured at 37°C for 48h, and the plate colonies were preliminarily observed. Aseptically pick a suitable single colony and inoculate it in MRS liquid medium (purchased from Qingdao Haibo Biological Co., Ltd., HB0384-1), and after standing at 37°C for 48 hours, take 10 μL of bacterial liquid MRS plate for streaking. This step was repeated three times to obtain a purified culture strain.

[0050] The DNA of the purified strain was extracted from the purified strain...

Embodiment 2

[0061] Example 2 - Growth curve, in vitro acid production, acid resistance and bile salt resistance test of Lactobacillus johnsonii NJD412

[0062] (1) Determination of growth curve

[0063]Lactobacillus johnsonii NJD412 with deoxynivalenol enol degradation ability in Example 1 was activated, and the activated bacterial solution was passaged into 100 mL MRS liquid medium at 5%, placed in a 37°C constant temperature incubator, and allowed to stand. Incubate for 24h, draw 1mL of bacterial liquid every 3h for continuous 10-fold dilution, and adjust the concentration to 10 -1 to 10 -7 , respectively draw 100 μL of the dilution solution and spread it on the MRS plate medium, place it in a constant temperature incubator at 37°C for 24 hours and count it. Calculate the bacterial concentration and record, repeat 3 times to get the average value. The growth curve of Lactobacillus johnsonii NJD412 was drawn with the culture time (hours) as the abscissa and the bacterial concentration...

Embodiment 3

[0076] Embodiment 3-Inhibition test of Lactobacillus johnsonii NJD412 to pathogenic bacteria

[0077] In this example, the test of the bacteriostatic ability of Lactobacillus johnsonii NJD412 was carried out by taking enterotoxogenic Escherichia coli and Salmonella typhimurium as examples.

[0078] Lactobacillus johnsonii NJD412 was inoculated into MRS liquid medium at a proportion of 5%, and cultured at 37°C for 24h, and the concentration of the bacterial solution was adjusted to 1×10 8 CFU / mL. After the bacterial liquid was centrifuged at 4000 rpm for 10 min, and filtered with a bacterial filter with a diameter of 0.22 μm, the fermentation broth of Lactobacillus johnsonii NJD412 without bacterial cells was obtained.

[0079] Activated enterotoxogenic Escherichia coli and Salmonella typhimurium were evenly inoculated on LB plates (purchased from Qingdao Haibo Biological Co., Ltd., HB0129), inverted at 37°C for 1 hour, and then taken out. Circular holes were punched on the p...

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Abstract

The invention discloses lactobacillus johnsonii and application of the lactobacillus johnsonii in degradation of deoxynivalenol and inhibition of pathogenic bacteria, the lactobacillus johnsonii is NJD412, the lactobacillus johnsonii is classified and named as Lactobacillus johnsonii, and the preservation number of the lactobacillus johnsonii is CGMCC No.23553. The lactobacillus johnsonii is named as Lactobacillus johnsonii, and the lactobacillus johnsonii is named as Lactobacillus johnsonii. The lactobacillus johnsonii NJD412 is separated and purified from dog feces, and an MRS plate bacterial colony is semitransparent and circular, has the diameter of about 1-2 mm and is irregular in edge. Tests prove that the lactobacillus johnsonii NJD412 disclosed by the invention can effectively degrade deoxynivalenol in vivo and in vitro, and meanwhile, the lactobacillus johnsonii NJD412 has a remarkable inhibition effect on enterotoxigenic escherichia coli and salmonella typhimurium. The lactobacillus johnsonii NJD412 and the fermentation liquor thereof can be used for relieving animal poisoning caused by deoxynivalenol pollution and preventing and treating animal diseases induced by other pathogenic bacteria, are safe to use and have a good market application prospect.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Lactobacillus johnsonii and its application in degrading deoxynivalenol and inhibiting pathogenic bacteria. Background technique [0002] At present, animal husbandry is facing many challenges, especially environmental pollution, feed pollution and pathogenic microorganisms, which seriously affect the economic interests of animal husbandry and hinder the sustainable development of animal husbandry. Among them, the feed involves many links such as raw material supply, processing process, transportation and storage, etc., and is easily polluted by environmental factors. The main manifestations are the pollution of feed raw materials, improper processing, and poor transportation and storage conditions. Feed is affected by fungi and bacteria in the environment. As well as its various toxic metabolite pollution, it causes or aggravates the diseases of the digestive system, im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L5/20A61K35/747A61P31/04C12R1/225
CPCC12N1/20A23L5/28A61K35/747A61P31/04Y02A50/30
Inventor 黄克和葛雷郭俊妍尹刘雯甘芳
Owner NANJING AGRICULTURAL UNIVERSITY
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