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Bacillus pumilus and application thereof to prevention and control of bombyx mori nuclear polyhedrosis virus infection

A technology of bacillus pumilus and virus infection, applied in the direction of application, antiviral agents, medical preparations containing active ingredients, etc.

Active Publication Date: 2018-05-18
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Bacillus pumilus is a type of bacteria widely distributed in the environment. Most species are friendly to the environment and organisms. So far, there have been some application reports in degrading environmentally harmful substances and promoting the growth of animals and plants, but they can be used as intestinal probiotics There is no report on the prevention and treatment of enteroviral infection at home and abroad. The present invention provides a new way and new technology to prevent and treat viral infection from the intestinal tract by using microorganisms

Method used

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  • Bacillus pumilus and application thereof to prevention and control of bombyx mori nuclear polyhedrosis virus infection
  • Bacillus pumilus and application thereof to prevention and control of bombyx mori nuclear polyhedrosis virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment one, bacillus pumilus SW41 ( Bacillus pumilus SW41) isolation and identification

[0017] 1. Strain isolation: medium: 0.3% beef extract; 1% peptone; 0.5% NaCl; 1.8% agar; distilled water 100ML; pH7.0. The silkworms were fed with mulberry leaves and Zhe leaves, respectively, and the disease models susceptible to and non-susceptible to the silkworm nuclear polyhedrosis virus were established. The possible target bacteria were isolated and cultured at a constant temperature at 30°C.

[0018] 2. Purification: beef extract peptone medium: 0.3% beef extract; 1% peptone; 0.5% NaCl; 2% agar; 1% turmeric dry powder; distilled water; pH7.0. Czapek's modified medium: 0.2% NaNO 3 or NaNO 2 ;0.1%K 2 HPO 4 ;0.05%KCl;0.05%MgSO 4 ;0.001%FeSO 4 ; 2% sucrose; 2% agar; 1% turmeric dry powder; distilled water; pH7.0. Gause No. 1 modified medium: 0.1% KNO 3 ;0.05%K 2 HPO 4 ;0.05%NaCl;0.05%MgSO 4 ;0.001%FeSO 4 ; 2% soluble starch; 2% agar; distilled water; pH7.0. ...

Embodiment 2

[0020] Example 2: Cytological identification of antiviral infection

[0021] 1. Liquid medium: beef extract 3g, peptone 10g, NaCl 5g, distilled water 1000mL;

[0022] 2. Strain fermentation: pick SW41 colonies and inoculate them in beef extract peptone liquid medium, ferment and cultivate for 72 hours at 25~28°C, 200r / min, centrifuge the fermentation broth at 10000rpm / min for 10min, take the supernatant and filter to sterilize, and put Store the filtered supernatant in a sterile centrifuge tube;

[0023] 3. Cell infection test: Use the fermentation supernatant to treat virus particles BV and ODV, incubate at 28°C for 1 hour, then centrifuge at 15000g / min for 30min at 5~10°C, discard the supernatant, add 1mLTC100 cell culture medium, then 15000g / min Centrifuge for 30 min, discard the supernatant, add 1 mL of TC100 cell culture medium, scrape the precipitate, vortex for 10 min, and treat with sterile water as a control. The BV / ODV treated with SW41 fermentation supernatant and...

Embodiment 3

[0024] Embodiment three, SW41 bacterial strain is applied to silkworm rearing antiviral infection

[0025] 1. The results of silkworm rearing after the application of bacterial strain fermentation broth to process the virus

[0026] will be 1.8×10 8 polygons or 1.8×10 9 The polyhedron was centrifuged at 5000rpm / min for 10min to collect the precipitate, added 1mL of lysate, shaken well, incubated at 28°C for 30min, carefully collected the supernatant, and divided into 6 tubes. Centrifuge the supernatant at 12000g / min for 30min, collect the precipitate, add SW41 supernatant (the control is sterile water), and incubate at 28°C for 1h. After the incubation, centrifuge at 12000g / min for 30min to collect the precipitate, add 1mL of sterile water to resuspend. Each group of silkworms (30 heads) took 1 piece of mulberry leaf, applied the treated ODV suspension on the back of the mulberry leaf, dried it, and fed it to the third and fourth instar silkworms. The treatment group had 3 ...

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Abstract

The invention provides bacillus pumilus. The bacillus pumilus has a preservation number of CCTCC (China Center for Type Culture Collection) M 2018057 and is preserved in China Center for Type CultureCollection. The bacillus pumilus provided by the invention has a remarkable anti-infection effect on a bombyx mori nuclear polyhedrosis virus (BmNPV); a cytological infection test proves that the virus infection prevention and control effect reaches 100 percent. It is found out that a bombyx mori intestinal microorganism bacillus pumilus (Bacillus pumilus SW41) has remarkable virus infection resisting activity for the first time at home and abroad; the bacillus pumilus has a wide application prospect of preventing and controlling virus diseases of bombyx mori in a bombyx mori raising industry.

Description

technical field [0001] The invention relates to the technical field of microbial application, in particular to a kind of Bacillus pumilus SW41 ( Bacillus pumilus SW41). Background technique [0002] Bombyx mori nuclear polyhedrosis virus disease is mainly a kind of contagious silkworm disease caused by virus (BmNPV) passing through the intestinal tract of silkworms and causing severe disease in silkworms. It is currently one of the most serious viral diseases harmful to sericulture. The industry has brought huge economic losses. At present, the prevention and treatment of the disease is mainly through the chemical disinfection of the sericulture environment and the selection and use of disease-resistant varieties. [0003] Bacillus pumilus is a type of bacteria widely distributed in the environment. Most species are friendly to the environment and organisms. So far, there have been some application reports in degrading environmentally harmful substances and promoting the...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P1/00A61K35/742A61P31/20A23K10/18A23K50/90C12R1/07
CPCA01N63/00A61K2039/522A23K10/18A23K50/90C12N1/205C12R2001/07
Inventor 万永继周围刘仁华王文慧向庭婷刘晓媛卢延
Owner SOUTHWEST UNIVERSITY
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