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Screening and application of probiotic bacillus

A Bacillus licheniformis, separation and screening technology is applied in the application field of agricultural microorganisms, which can solve problems such as unseen and unseen separation and screening, and achieve the effects of high safety, strong stability and strong reproductive ability.

Active Publication Date: 2020-12-01
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, no strains with strong activity of degrading cellulose and lignin have been isolated and screened from the rumen of buffaloes, nor has a strain of Bacillus paralicheniformis that has the ability to degrade cellulose and lignin and has a significant growth-promoting effect on beef cattle report

Method used

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  • Screening and application of probiotic bacillus
  • Screening and application of probiotic bacillus
  • Screening and application of probiotic bacillus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Isolation and Identification of Bacillus paralicheniformis SN-6 Strain

[0035] 1. Isolation of strains

[0036] The samples were taken from the rumen of healthy buffalo with rumen fistula in Hubei Jinniu Animal Husbandry Co., Ltd., Shayang County, Jingmen City. The rumen contents were filtered through gauze, and the filtrate was collected in an anaerobic bag and placed in a small incubator on-board at 39°C. Take 1mL of collected samples for 10 -1 -10 -6 Gradual dilution (100 μL culture solution was added to 900 μL sterilized water for gradient dilution), take 10 -4 、10 -5 、10 -6 Coat CMC-Na sole carbon source plate with three dilutions of culture solution, place the plate in an anaerobic box at 39°C for 3 days, pick a single colony, spot on CMC-Na sole carbon source plate, place in 39°C anaerobic chamber Cultivate in the box for 3 days, then stain with 0.1% Congo red staining solution for 10-15min, then wash twice with 1.0mol / L NaCl solution, observe wh...

Embodiment 2

[0062] Example 2: Detection of probiotic properties of Bacillus paralicheniformis SN-6 strain

[0063] 1. Enzyme production test

[0064] (1) Detection of amylase activity

[0065] Spot the Bacillus paralicheniformis SN-6 strain on an LB plate containing 1% soluble starch, put it in a 37°C incubator and cultivate it for 48 hours; add 2mL of iodine solution to the reacted starch medium, and swirl until the iodine solution is uniform Cover the plate and let it stand for 10 minutes. The starch turns blue when it encounters iodine. If amylase is produced, the starch around the colony will decompose, and a faded circle will appear on the blue-purple plate stained with iodine solution. The staining result is as follows: Figure 5 As shown, the detection results of the amylase activity produced by the Bacillus paralicheniformis SN-6 strain are shown in Table 2.

[0066] Table 2 Detection results of amylase produced by Bacillus paralicheniformis SN-6 strain

[0067]

[0068] (2)...

Embodiment 3

[0081] Example 3: Safety Evaluation of Bacillus paralicheniformis SN-6 Strain

[0082] 1. Drug susceptibility test

[0083] 15 kinds of drug-sensitive discs (purchased from Hangzhou Microbiology Reagent Co., Ltd.) including penicillins, cephalosporins, quinolones, aminoglycosides, tetracyclines, and sulfonamides were selected for drug sensitivity test. The test judgment criteria are based on the latest version of NCCLS standards provided by the World Health Organization (WHO) (2018 version). Specific steps are as follows:

[0084] (1) Add 1% of the bacterial solution into LB liquid medium, culture overnight at 37° C. on a shaker at 200 rpm / min.

[0085] (2) With the white paper with black characters as the background, adjust the turbidity to 0.5 McFarland standard turbidimetric tube turbidity. If the concentration of the bacterial solution is too thick, it can be diluted with normal saline.

[0086] (3) Dip the bacterial solution with a sterilized cotton swab, squeeze out ...

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Abstract

The invention belongs to the field of agricultural microorganism application, and particularly relates to screening and application of probiotic bacillus. The invention relates to screening and application of probiotic additives related to ruminant rumen digestion. According to the invention, a bacillus paralicheniformis strain with a remarkable growth-promoting effect on beef cattle is obtained by screening, and 16S rRNA gene identification, probiotic performance detection, safety evaluation and stress resistance identification prove application as a feed additive for beef calves to promote growth. The bacillus paralicheniformis strain SN-6 disclosed by the invention is preserved in the China Center for Type Culture Collection, and the preservation number of the bacillus paralicheniformisstrain SN-6 is CCTCC NO: M2020136. The strain is high in growth speed, strong in stress resistance, safe and capable of promoting growth, and can be used as a micro-ecological additive for livestockand poultry feed.

Description

technical field [0001] The invention belongs to the application field of agricultural microorganisms, and in particular relates to the screening and application of a strain of probiotic bacillus. The invention relates to the screening and application of prebiotic additives related to rumen digestion of ruminants. The present invention screens and obtains a strain of Bacillus paralicheniformis that has significant growth-promoting effects on beef cattle. After isolation and identification, probiotic performance testing, safety evaluation and stress-resistance performance identification, it shows that the present invention can be used as a feed additive for meat calves to promote growth application. Background technique [0002] A key step in the global carbon cycle is the hydrolysis of cellulose in plant cell walls, the most abundant carbon source on land (Malhi 2003, David 2011). Rational use of cellulose resources can alleviate pollution, solve global energy crises and an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K50/10A23K50/60A23K10/18A23K50/75C12R1/07
CPCA23K50/10A23K50/60A23K10/18A23K50/75C12R2001/07C12N1/205
Inventor 石德时杨书敏罗吉周进肖运才李翔王喜亮李筱雯周祖涛
Owner HUAZHONG AGRI UNIV
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