Screening and application of buffalo rumen-derived bacillus amyloliquefaciens
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A technology for dissolving starch spores and water buffalo, applied in the field of agricultural microorganism application, can solve problems such as insufficient cellulose degradation
Active Publication Date: 2020-11-27
HUAZHONG AGRI UNIV
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[0006]
The purpose of the present invention is to solve the problem of insufficient cellulose degradation in the existing silage fermentation. Inspired by the buffalo's resistance to roughage and the buffalo rumen's stronger degradation of lignocellulose, the present invention screens the cellulose degrading ability from the buffalo rumen. Bacillus amyloliquefaciens (Bacillus am
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Embodiment 1
[0037] Example 1: Isolation and identification of bacterial strains
[0038] 1. The preparation of culture medium:
[0039] Table 1 Buffer A
[0040]
[0041] NOTE: This buffer is used to prepare enrichment media.
[0042] Table 2 Buffer B
[0043]
[0044] NOTE: This buffer is used to prepare enrichment media.
[0045] Table 3 enrichment medium
[0046]
[0047]
[0048] Note: Resazurin is blue, and the color of the medium will turn pink after sterilization, and the solution is colorless when the medium does not contain oxygen.
[0049] Prepare the antifungal agent according to the formula in Table 4. After the preparation is completed, use a 0.45 μm filter to filter and sterilize, aliquot into 1.0 mL, and store at 4°C.
[0050] The configuration of table 4 antifungal agent
[0051]
[0052] Note: Cycloheximide is a highly toxic drug that inhibits yeast, mold and protozoa; the solvent used for nystatin is DMSO, dissolve a small amount of DMSO and then mix...
Embodiment 2
[0109] Example 2: Safety Evaluation of Bacillus amyloliquefaciens SN-C strain
[0111] The gene primers of non-hemolytic enterotoxin NHE (nheA, nheB, nheC), hemolysin Hbl (hblA, hblC, hblD), cytotoxin CytK, enterotoxin T (BceT) and enterotoxin FM (EntFM) were used to determine the virulence Amplify the gene-positive Bacillus cereus, detect the virulence genes contained in it, and use 1% agarose gel electrophoresis (120V, 90mA), and the positive identification of the virulence gene is that the given Bacillus cereus contains nheA, nheB, and nheC , EntFM, hblA, hblC, hblD, CytK, BceT a total of 9 virulence genes; gene primer sequences are shown in Table 11, amplification results are shown in Figure 7 .
[0112] Table 11 Virulence gene PCR primer sequence
[0113]
[0114] (2) Amplification of virulence genes of Bacillus amyloliquefaciens SN-C strain
[0115] Bacterial Genomic DN...
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Abstract
The invention belongs to the technical field of agricultural microorganism application, and relates to screening and application of buffalo rumen-derived bacillus amyloliquefaciens. The invention relates to the field of feed additive preparation and the field of ruminant rumen digestion, development and utilization. The invention relates to stain separation and identification of the bacillus amyloliquefaciens with a degradation effect on cellulose in a feed, safety evaluation, identification of the fiber degradation capacity of silage, and development and application of the bacillus amyloliquefaciens as an additive for promoting silage fermentation of the feed. The strain disclosed by the invention is a Bacillus amyloliquefaciens SN-C strain, and the preservation number is CCTCC NO:M2020137. The strain is high in growth speed, can remarkably improve the degradation rate of cellulose in corn silage, can reduce the cellulose content by 3.10% and the hemicelluloses content by 1.75% compared with a control group, is high in stress resistance and safe, and can be used for preparing an additive for promoting silage fermentation.
Description
technical field [0001] The invention belongs to the technical field of agricultural microorganism application, and in particular relates to the screening and application of buffalo rumen-derived Bacillus amyloliquefaciens. The field of the present invention is related to the field of preparation of feed additive preparations and the field of rumen digestion biological additives for ruminants. The present invention provides the isolation, identification, safety assessment and treatment of a strain of Bacillus amyloliquefaciens that can degrade cellulose in feed. The identification of silage fiber degradation ability can be used as an additive to promote the development and utilization of feed silage fermentation. Background technique [0002] my country is a large agricultural country, with an annual output of about 630 million tons of agricultural by-product straw, but the amount of straw used as feed for herbivores only accounts for 15% to 24% of the total output each year,...
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