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Lytic salmonella phage and separation method thereof

A Salmonella, separation method technology, applied in the field of microbiology, can solve the problem of no drug available, and achieve the effect of high potency and good thermal stability

Inactive Publication Date: 2018-06-05
QINGDAO RUNDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But with the growing problem of bacterial resistance, people are already facing a dangerous situation where no medicine is available

Method used

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  • Lytic salmonella phage and separation method thereof
  • Lytic salmonella phage and separation method thereof
  • Lytic salmonella phage and separation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 The separation method of lytic Salmonella phage, the steps are as follows:

[0033] (1) Preparation of host bacteria: Under sterile conditions, use an inoculation needle to take samples from the internal organs of sick and dead poultry, streak on SS agar medium, and screen out colonies with black spots or no black spots in the transparent center, and then Purify and identify the isolated colonies;

[0034] (2) Isolation of bacteriophage: Soak poultry feces in sterile normal saline overnight, then centrifuge the supernatant and pass it through a 0.22nm sterile filter membrane, take 1mL of supernatant and 0.2mL of bacterial suspension and add 5mL of LB meat In the soup, under the condition of 37°C, shake at 200r / min overnight. Centrifuge the suspension after overnight culture, take 0.1mL supernatant and 0.2mL host bacteria, mix it, let it stand at 37°C for 10min, then add it to LB nutrient agar at 40°C, mix well, and immediately introduce the solidified Mak...

Embodiment 2

[0037] Embodiment 2 Determination method of phage titer: do 10-fold ratio dilution with sterile physiological saline to the phage stock solution after proliferation, take 10 -5 、10 -6 、10 -7 Lay double plates, incubate upside down at 37°C for 4-6 hours, and count the plaques after they grow out. The titer of the phage was determined to be ≥10 9 pfu / mL.

Embodiment 3

[0038] Example 3 Electron microscope observation experiment of phage: specifically: take 20 μL of liquid containing crude phage particles and drop it on a copper grid, let it settle naturally for 15 minutes, and use filter paper to absorb excess liquid from the side, add a drop of 2% phosphotungstic acid (PTA ) to stain the phage on the copper grid for 10 min, then suck the staining solution from the side with filter paper, observe the shape of the phage with an electron microscope after the sample is dry, and observe through the transmission electron microscope, the shape of the phage is tadpole-shaped, and the head is a three-dimensional icosahedron Symmetrical, with retractable tail.

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Abstract

The invention relates to a lytic salmonella phage and a separation method thereof. The phage has a tadpole-shaped form, has the head in an icosahedral cubical symmetry manner and has a telescopic tail. The phage is subjected to biological characteristic study. After multiplication of the phage separated in the invention, the fluid titer of the phage is high, the titer of the phage in the inventionis more than or equal to 10<9>pfu / mL, the phages have a certain activities under different infection multiplicity conditions, and the titer of the phage at a temperature of 30-60 DEG C is basically invariable. The biological characteristic study on the phage separated in the invention provides theoretical guiding significance for further research and development of novel antibacterial drugs.

Description

technical field [0001] The invention belongs to the field of microorganisms, in particular to a strain of lytic Salmonella phage and an isolation method thereof. Background technique [0002] Salmonella belongs to Enterobacteriaceae, a Gram-negative bacterium with more than 2,500 serotypes, and 292 serotypes have been reported in my country. Salmonella infection of animals often manifests clinically as sepsis, gastroenteritis and local inflammation of other tissues. Salmonella avian origin can cause pullorum, chicken typhoid and avian paratyphoid. It has the characteristics of vertical transmission and horizontal transmission. The prevalent dominant strains of pathogenic Salmonella are different at different times [3] . At present, in order to reduce the incidence of salmonellosis from chickens, promote the growth and development of chickens, and increase the egg production rate, antimicrobial drugs are commonly used in farms as growth promoters, preventive agents and cli...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12R1/92
CPCC12N7/00C12N2795/00021C12N2795/00051
Inventor 杜新永刘军伟李先胜马如霞张得彦
Owner QINGDAO RUNDA BIOTECH
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