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Nocardia anfenacea bacteriophage P69 and application thereof

A Nocardia and bacteriophage technology, applied in the direction of phage, virus/bacteriophage, special treatment targets, etc., can solve the problems of disease risk, turbidity of effluent, and reduce the ability of activated sludge to treat pollutants, so as to improve the universality and effectiveness, expansion of host spectrum, and rapid and efficient lysis

Pending Publication Date: 2022-03-11
UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When Nocardia overgrows, a large amount of stable foam floats on the surface of the aeration tank, resulting in poor sludge settling performance and turbid effluent
And when the bubbles burst, Nocardia will spread to the air in the form of aerosols, posing a risk of disease
Traditional physical methods (such as salvage, water spray defoaming, etc.) and chemical methods (such as adding aluminum salts, iron salts, or disinfectants, etc.) have not fundamentally solved the problem of Nocardia overgrowth. , will also affect the normal flora in the activated sludge system and reduce the ability of activated sludge to treat pollutants

Method used

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  • Nocardia anfenacea bacteriophage P69 and application thereof
  • Nocardia anfenacea bacteriophage P69 and application thereof
  • Nocardia anfenacea bacteriophage P69 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The separation and purification of embodiment 1 phage

[0021] Preparation of PYCa liquid medium:

[0022] at 1L ddH 2 Add 1 g of glucose, 1 g of yeast extract, and 15 g of tryptone to O, mix well and then autoclave at 121°C for 20 min. After cooling, add 4.5 mL of calcium chloride solution with a concentration of 1 mol / L.

[0023] Preparation of PYCa solid medium:

[0024] at 1L ddH 2 Add 1g of glucose, 1g of yeast extract, 15g of tryptone, and 15g of agar powder to O, mix well and then autoclave at 121°C for 20min. Add 4.5mL calcium chloride solution with a concentration of 1mol / L in the uncoagulated state.

[0025] Preparation of PYCa semi-solid medium:

[0026] at 1L ddH 2 Add 1 g of glucose, 1 g of yeast extract, 15 g of tryptone, and 7.5 g of agar powder into O, mix well, and then sterilize under high pressure at 121 ° C for 20 min. Add 4.5mL calcium chloride solution with a concentration of 1mol / L in the uncoagulated state.

[0027] The sewage sample use...

Embodiment 2

[0030] Morphological observation of embodiment 2 bacteriophage

[0031] Take 20 μL of the phage suspension and drop it on the copper grid, wait for its natural precipitation for 10 minutes, blot it dry from the side with dry filter paper, let it air for about 1 minute, add 1 drop of 1% uranyl acetate on the copper grid, stain for 2 minutes, and then use it carefully to dry Blot the excess dye from the side of the filter paper, let it dry naturally in the dark for 30 minutes, and observe it with a transmission electron microscope (JEM-1400).

[0032] The results of transmission electron microscopy are shown in figure 2 , the results showed that phage P69 had a spherical head and a long tail, the diameter of the head was about 68.47nm, the length of the tail was about 239.25nm, and the diameter of the tail was about 13.12nm. According to the eighth report on the virus classification of the International Organization for Taxonomy of Viruses (ICTV), this strain of phage P69 can ...

Embodiment 3

[0033] Example 3 Phage Genome Analysis and Identification

[0034] Phage whole genome sequencing and analysis: IlluminaNextSeq500 was used for PE 2×150 to sequence the DNA of the sample. Then, the optimized sequence was spliced ​​using spades v.3.11.1 splicing software, and the optimal assembly result was obtained. Use the biological software PHASTER to predict and analyze the open reading frame of the genome, and use NCBI Blastp to complete the preliminary annotation of functional genes, and use tRNAscan-SE (http: / / lowelab.ucsc.edu / / tRNAscan-SE / ) to predict tRNA online, Use CGView Server software (http: / / cgview.ca / ) to complete the drawing of the whole genome circle map (see image 3 ), a phylogenetic tree was constructed using MEGA X software based on the whole genome data.

[0035]The results showed that the genome size of phage P69 was 68913bp, and the G+C content was 65.56mol%. According to the results of Blast comparison, it seems to be a highly novel phage, with the ...

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Abstract

The invention discloses a strain of Nocardia anfenacea bacteriophage P69 and application thereof, and belongs to the technical field of biology. The preservation number of the nocardia orientalis bacteriophage P69 disclosed by the invention is CGMCC (China General Microbiological Culture Collection Center) No.23088. The nocardia orientalis bacteriophage P69 provides a selectable basic material for prevention and control of a sludge foaming event caused by nocardia orientalis, and enriches a bacteriophage seed bank; the functional gene and protein of the bacteriophage P69 are further deeply excavated through separation identification and whole genome sequencing of the bacteriophage P69, and one or more enzymes and the like with a splitting effect on Nocardia soufeniana are provided for bacteriophage treatment and biological prevention and control, so that the host spectrum is expanded, and the application universality and effectiveness of the bacteriophage are improved; the bacteriophage P69 can be used for rapidly and efficiently cracking Nocardia soufeniformis and can be used for sludge foaming events caused by excessive propagation of Nocardia in a sewage treatment system.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a strain of Nocardia phage P69 and its application. Background technique [0002] Nocardia africa is a rare human pathogen. Like other Nocardia species, this bacterium can cause illness when inhaled or inoculated through the skin. South African Nocardia infection occurs more often in immunocompromised patients, such as organ transplant patients or AIDS patients. Mortality rates in patients infected with Nocardia aeruginosa can be as high as 75%, and when the central nervous system is involved, the mortality rate is 100%. Studies in laboratory animals have shown that the organism is more virulent than Nocardia brasiliensis (but less than Nocardia stellatus). In addition, Nocardia africa has some resistance to aminoglycosides, including amikacin. At present, trimethoprim-sulfamethoxazole (TMP-SMX) is considered to be the most effective drug for the treatment of Noca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C02F3/34C12R1/92
CPCC12N7/00C02F3/34C12N2795/10321C12N2795/10331C02F2303/04
Inventor 刘新春熊文斌卢晗
Owner UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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