Bacillus subtilis for feeding and application thereof

A technology of Bacillus subtilis and inoculum, which is applied to the application field of forage Bacillus subtilis and inoculum as feed additives for breeding animals, can solve the problems of tumor-causing toxicity and genotoxicity, and achieves the goal of promoting animal growth and improving Feed conversion ratio, the effect of enhancing immune function

Active Publication Date: 2019-11-15
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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  • Bacillus subtilis for feeding and application thereof
  • Bacillus subtilis for feeding and application thereof
  • Bacillus subtilis for feeding and application thereof

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. Such as 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 is entered.

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Abstract

The invention provides bacillus subtilis for feeding and application thereof, and belongs to the technical field of microorganisms and feed additive preparation. The provided bacillus subtilis is named as Ma. k, and the preservation number is CGMCC NO.17613. The bacillus subtilis can resist the high temperature of 90 DEG C, can grow in the acid environment with the pH value of 2.0 or above, has astrong bile salt tolerance capacity and has a function of degrading moulds and mycin, and after the bacillus subtilis Ma. k is prepared into a bacterial agent, the effect of the bacterial agent for feeding animals is safe and reliable, and the bacterial agent plays positive roles in promoting the digestion and absorption of nutrients, reducing the diarrhea rate, improving the feed conversion efficiency and 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 Applications(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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