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A kind of forage Bacillus subtilis and its application

A technology of Bacillus subtilis and bacterial agent, which is applied in the field of application of Bacillus subtilis and its bacterial agent as a feed additive for breeding animals, can solve the problems of genotoxicity and tumor-causing toxicity, and achieve improved feed conversion rate, The effect of promoting animal growth and good application prospects

Active Publication Date: 2021-02-19
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, studies have confirmed that zearalenone also has cytotoxicity, genotoxicity, immunotoxicity, and tumorigenic toxicity.

Method used

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  • A kind of forage Bacillus subtilis and its application
  • A kind of forage Bacillus subtilis and its application
  • A kind of forage Bacillus subtilis and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Isolation and Identification of Bacillus subtilis Ma.K

[0027] 1. Isolation of strain Ma.K

[0028] 1. Isolation and cultivation of strains

[0029] Take 1g of pig intestinal digesta sample from the Pig Experimental Base of China Agricultural University in Haidian District, Beijing, and put it into a test tube with 9ml of normal saline, and vortex it to mix well, which is a 1:10 dilution, and then take the dilution Perform 10-fold incremental dilution, and then select 1 ml of each of three appropriate gradient dilutions and spread them on the LB medium. Cultivate at 37°C for 48-72 hours, observe and record the colony shape, pick a single colony with good growth, and separate and purify by streaking.

[0030] 2. UV mutagenesis and screening of strains

[0031]Pour the sterilized LB medium into a petri dish, and after solidification, take the bacterial suspension obtained in step 1 and spread it on a plate. Control the number of colonies in each petri dish t...

Embodiment 2

[0047] Stress resistance detection of embodiment 2 Bacillus subtilis Ma.K

[0048] 1. Heat resistance test

[0049] Put the Bacillus subtilis Ma.K (CGMCC No.17613) bacteria solution in the water bath for 20 minutes, and treat them at 60°C, 80°C, and 90°C respectively, and repeat each treatment three times. After the treatment, use the pouring method to measure its activity. The number of bacteria, the number of live bacteria and the survival rate of spores meet the minimum requirements for the processing characteristics of probiotics in actual production.

[0050] The results show that initially greater than 10 10 After CFU / ml of Bacillus subtilis Ma.K CGMCC No.17613 was treated at 60°C and 80°C for 20 minutes, the number of viable bacteria could be maintained at 10 10 More than CFU / ml; the number of live bacteria of Bacillus subtilis Ma.K after treatment at high temperature 90°C for 1 hour and 2 hours is greater than 10 10 CFU / ml.

[0051] 2. Acid resistance test

[0052...

Embodiment 3

[0065] The growth curve determination of embodiment 3 subtilis Ma.K

[0066]The growth curve represents the dynamic changes in the whole process of bacteria growing and multiplying in a new suitable environment until aging and death. Bacillus subtilis Ma.K CGMCC No.17613 was inoculated into LB medium at an inoculation amount of 10% (v / v), and cultivated at 37° C. for 36 hours. The LB medium without bacterial solution was used as a blank control. Measure the OD600 value every 2-6 hours to calculate the number of viable bacteria. The experiment was repeated three times, the results were averaged, the data were recorded and the growth curve was drawn. like Figure 5 As shown, in 0-14 hours, Bacillus subtilis Ma.K is in the logarithmic growth stage, and the reproduction rate is high. The number of Bacillus subtilis Ma.K tended to be stable in the stationary phase at 14-24 hours. After 24 hours, the growth rate decline phase was entered.

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Abstract

The invention provides a bacillus subtilis for feed and an application thereof, belonging to the technical field of preparation of microorganisms and feed additives. The feed Bacillus subtilis (Bacillus subtilis) provided by the present invention is named Ma.k, and the preservation number is CGMCC NO.17613. It has strong salt capacity and has the effect of degrading mycomycin. It is safe and reliable to feed animals after making Bacillus subtilis Ma. It has a positive effect on promoting growth.

Description

technical field [0001] The invention relates to the field of microbes and feed additives, in particular to the application of a feed bacillus subtilis and its agent as a feed additive for breeding animals. Background technique [0002] Early weaning is implemented in modern large-scale pig raising. Weaned piglets are prone to a series of stress syndromes such as loss of appetite, abnormal digestive function, reduced immunity, high-frequency diarrhea, and growth retardation, which is a major problem in pig production. In the past breeding process, weaning stress of piglets was dealt with by adding antibiotics. Due to problems such as the continuous emergence of drug-resistant strains caused by the long-term use of antibiotics, my country will implement a complete ban on antibiotics in the breeding process by the end of 2020. As an effective substitute for a class of antibiotics, probiotics have received more and more attention. Prebiotics have become the first choice for an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K10/18A23L5/20A61K35/742A61P1/14C12R1/125
CPCA61K35/742A23K10/18A23L5/28A61P1/14C12N1/20C12N1/205C12R2001/125
Inventor 马曦郭娉婷姬琳堡孙美鸽孙敏燕付函君
Owner CHINA AGRI UNIV
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