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A kind of Bacillus subtilis producing aflatoxin b1 degrading enzyme and its application

A Bacillus subtilis, aflatoxin technology, applied in the application, bacteria, microorganism-based methods, etc., can solve the problems of microorganisms and metabolites safety doubts, destruction of feed nutrients, animal body damage, etc., and achieve fast reproduction. , the effect is mild, the effect of promoting healthy growth

Active Publication Date: 2020-02-14
武汉格瑞农生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional physical and chemical detoxification methods all have defects such as destroying the nutritional content of the feed and affecting the quality of the feed.
The biological method provides a new development direction for the detoxification of aflatoxin B1, but the safety of the screened microorganisms and metabolites is questionable, which will cause additional damage to the animal body and is difficult to be widely used in actual production

Method used

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  • A kind of Bacillus subtilis producing aflatoxin b1 degrading enzyme and its application
  • A kind of Bacillus subtilis producing aflatoxin b1 degrading enzyme and its application
  • A kind of Bacillus subtilis producing aflatoxin b1 degrading enzyme and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Bacillus subtilis HDR-02 collection, isolation and primary screening

[0017] 1. Collection and isolation of strains

[0018] Samples were taken from 90-day-old healthy broiler manure and moldy grain. After the sample is collected, put it into a 7mL EP tube filled with 3mL sterile water, and culture it in LB activation medium in a water bath at 80°C for 10 minutes. , and colonies with neat edges and a size of about 1 mm, pick suspected colonies for pure culture.

[0019] The purely cultured colonies are screened by colony morphology observation and Gram staining. The bacteria are in the shape of long rods, and the Gram-positive bacilli arranged in single, pair or chain are all passaged and preserved.

[0020] 2. Primary screening of strains

[0021] The isolated strains were placed in the improved primary screening medium mixed with coumarin solution to ensure that coumarin became the only carbon source and energy source, and placed in a 37°C incubator for...

Embodiment example 2

[0038] Implementation Case 2: Identification of HDR-02 Strain

[0039] 1. Morphological identification and identification of physical and chemical characteristics

[0040] The cell morphology and physicochemical characteristics of the above-mentioned HDR-02 strains are shown in Table 3.

[0041] Table 3 Cell morphology and physicochemical characteristics of HDR-02 strain

[0042] Identification index result Identification index result Colony color white or yellowish glucose acid production + a positive xylose acid production + Cell morphology Rod arabinose acid + spore + Mannitol Acidogenic + Catalase + Propionate — hydrolyzed gelatin + Nitrate reduction + Formation of indole — Hydrolyzed starch + anaerobic growth — 7% NaCl growth +

[0043] 2. Molecular identification

[0044] The strain HDR-02 was identified by the strain 16S rRNA sequence. The primers used for PCR amplification...

Embodiment example 3

[0049] Implementation Case 3: Application of Bacillus subtilis HDR-02 in ducklings

[0050] 1-day-old Cherry Valley meat ducks were pre-fed with the basic diet for 3 days, randomly selected healthy ducklings with similar body weight, and randomly divided them into 4 groups, with 4 replicates in each group and 5 ducklings in each replicate. The specific groups are as follows: blank control group, Bacillus subtilis HDR-02 group, aflatoxin B 1 Challenge group, aflatoxin B 1 Challenge+Bacillus subtilis HDR-02 group. Among them, the challenge dose of aflatoxin B1 is 0.1 mg / kg·BW, and the dose of Bacillus subtilis HDR-02 is 10 per day. 8 CFU / mL, Bacillus subtilis HDR-02 was added by gavage, and the gavage was continued for 1 week.

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Abstract

The invention discloses a bacillus subtilis strain for producing an aflatoxin B1 digestive enzyme. The bacillus subtilis for producing the aflatoxin B1 digestive enzyme provided by the invention is concretely bacillus subtilis HDR-02; the preservation code of the bacillus subtilis in CCTCC (China Center for Type Culture Collection) is CCTCC NO:M3016298; and the nucleotide sequence of the 16S rRNAgene is shown as SEQ ID No.1. Experiments prove that the bacillus subtilis HDR-02 provided by the invention has strong detoxification capability on aflatoxin B1; the action effect is mild; nutrition ingredients in the feed cannot be damaged; no influence is caused on the sensory quality of the feed; meanwhile, the bacillus subtilis HDR-02 has high reproduction speed and strong environmental tolerance; the health growth of animals can be promoted; the production performance of the animals can be improved; and the bacillus subtilis is applicable to animal husbandry production.

Description

technical field [0001] The invention relates to the technical field of aflatoxin, in particular to a bacillus subtilis producing aflatoxin B1 degrading enzyme and application thereof. Background technique [0002] Aflatoxins are mainly secondary metabolites produced by fungi such as Aspergillus flavus and Aspergillus parasiticus. Aflatoxins are the most toxic type of biological toxins found so far in contaminated agricultural products. Among them, the most toxic and harmful one is aflatoxin B1, which is 10 times more toxic than potassium cyanide and 68 times that of arsenic. B1, can cause acute poisoning death in sensitive animals. Aflatoxin is also a strong carcinogen. The carcinogenic ability of aflatoxin B1 is 70 times that of dimethyl nitramine. According to a survey in my country's high-incidence areas of liver cancer, the high incidence of liver cancer is related to eating food contaminated by aflatoxin. [0003] Aflatoxin B1 has a wide range of pollution and strong...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K10/18C12R1/125
Inventor 王喜亮陈翔晏涛肖运才金秀娥周祖涛吴贝
Owner 武汉格瑞农生物科技有限公司
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