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Bacillus amyloliquefaciens and application thereof in pomfret culture

A technology for dissolving amyloid spores and bacilli, applied in the field of microbial screening, can solve the problems of not being able to adapt to the intestinal tract of silver pomfret, difficult to control the content of active ingredients, and the effect of sprinkling probiotics is weak, so as to delay the entry of drugs into the circulatory system and improve the Host digestive enzyme activity, effect on disease risk reduction

Active Publication Date: 2020-09-08
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of silver pomfret breeding, the average daily water change is more than 100%, and the effect of splashing probiotics is very little, and a stable flora cannot be formed. It is necessary to ingest probiotics into the silver pomfret.
But on the one hand, the existing probiotics cannot adapt well to the intestinal environment of silver pomfret due to the difference in the isolation source.
On the other hand, because the feed of silver pomfret is artificial viscous feed, the feed dissolution rate is higher than that of mature pellet feed, and it is difficult to control the content of active ingredients only by adding probiotics to the feed
However, the existing aquaculture technology cannot provide a reasonable solution to the above problems, and innovative technologies must be used to solve the problems of silver pomfret probiotic screening and feeding

Method used

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  • Bacillus amyloliquefaciens and application thereof in pomfret culture
  • Bacillus amyloliquefaciens and application thereof in pomfret culture
  • Bacillus amyloliquefaciens and application thereof in pomfret culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Isolation of Bacillus amyloliquefaciens

[0026] (1) Experimental materials:

[0027] The silver pomfret used in the experiment was obtained from Gangwan Aquatic Seed Co., Ltd., Xiangshan County, Ningbo City, Zhejiang Province. Healthy juvenile fish that were not infected with flatulence and vibriosis during the breeding process. The initial body weight was 70.02±10.94g, the body length was 15.59±2.16cm, the water temperature was 18°C±1°C, the pH was 8.21±0.3, the salinity was 20.3±0.8, and day and night alternated for 12 hours. 6 silver pomfrets were randomly selected in the breeding pond, and 3 were a mixed sample.

[0028] (2) Sample collection:

[0029] After the silver pomfret was anesthetized with an anesthetic, it was quickly dissected, and the intestinal tract was separated, and the fat and blood on the outer wall of the intestinal tract were washed away with 1× phosphate buffer saline (PBS). Use tweezers to gently squeeze out the contents of the ...

Embodiment 2

[0046] Embodiment 2: the safety detection of bacillus amyloliquefaciens

[0047] (1) Selective media screening:

[0048] The special medium MYP and sheep blood TSA medium were used for screening. The purified bacteria were inoculated on the sheep blood TSA medium, cultured at 28°C for 6-12 hours, and the single colony used was inoculated on the sheep blood TSA medium by puncture method, and the size of the hemolysis circle was compared. Observe the hemolysis and record the traits (full hemolysis (β hemolysis); partial hemolysis (α hemolysis); no hemolysis (γ hemolysis)). There are yellow colonies on the MYP medium, and there is no precipitation ring, indicating that no egg phospholipase is produced. It showed gamma hemolysis and no hemolysis ring on the blood plate, and it was initially identified as a non-pathogenic bacteria.

[0049] (2) Animal safety testing

[0050] Activation of strains: Transfer the strains stored at -80°C to LB slant medium; after culturing at 30°C ...

Embodiment 3

[0052] Example 3: Detection of physical and chemical properties of Bacillus amyloliquefaciens

[0053] The strain was identified with a micro biochemical identification tube, and the results showed that the catalase was positive, the nitrate reduction reaction was positive, the starch was hydrolyzed, the gelatin was liquefied, the indole test was negative, the V-P reaction was negative, it could use mannose and arabinose for fermentation, and it could use glycerol , Methyl red reaction is negative, does not produce hydrogen sulfide, and can grow in 7% NaCl.

[0054]

[0055]

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Abstract

The invention provides a bacillus amyloliquefaciens ZM506 strain which is preserved in China General Microbiological Culture Collection Center (CGMCC) on October 21, 2019, wherein the number is CGMCCNO: 18714. The address is No.3, Yard 1, Beichen west road, Chaoyang District, Beijing. The bacillus amyloliquefaciens ZM506 strain provided by the invention is used for preparing a probiotic product for pomfret culture; and the product is a soft capsule and comprises a wall material part and a core material part. The bacillus amyloliquefaciens provided by the invention can enhance the natural barrier of the intestinal tract of a host, improve the composition of intestinal flora of pomfret, reduce the relative abundance of vibrio, reduce the risk of flatulence and vibriosis, and improve the digestive enzyme activity of the host. The feeding mode of the prepared oral soft capsules for the pomfret can delay medicines from entering a circulation system, and ensures that effective components ofprobiotics cannot escape in water in quantity.

Description

technical field [0001] The invention belongs to the technical field of microbial screening, and in particular relates to a bacillus amyloliquefaciens and its application in silver pomfret breeding. Background technique [0002] Silver pomfret (Pampus argenteus) belongs to Perciformes (Perciformes), Pomfretaceae (Stromateidae), and Pomfret (Pampus), which is delicious, nutritious, and widely distributed. It is an important seawater economic fish in my country. In recent years, under the dual requirements of protecting fishery resources and meeting market demand, we have carried out the work of artificial breeding of silver pomfret and gradually realized industrialization. However, the disease problems exposed during the breeding process, especially in the juvenile fish stage, should not be underestimated. On the one hand, the increase of intestinal gas-producing bacteria leads to "flatulence disease", and on the other hand, the outbreak of vibriosis during high temperature in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/742A61K9/48A61K47/46A61K47/42A61P1/00A61P31/04C12R1/07
CPCC12N1/20A61K35/742A61K9/4825A61K9/4816A61P1/00A61P31/04C12R2001/07C12N1/205
Inventor 张曼王亚军胡佳宝顾玮玮孙弋博李亚亚张友仪杨阳王冠林刘哲耀
Owner NINGBO UNIV
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