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Phage for staphylococcus aureus and application thereof

A staphylococcus and bacteriophage technology, applied in the directions of virus/bacteriophage, application, microorganism-based method, etc., can solve the problem of low inhibitory effect of Staphylococcus aureus strains, and achieve excellent antibacterial effect.

Active Publication Date: 2012-04-18
SHANGHAI XINNONG FEED
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although lysozyme has a good killing effect on some bacteria, its inhibitory effect on most strains of Staphylococcus aureus is very low

Method used

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  • Phage for staphylococcus aureus and application thereof
  • Phage for staphylococcus aureus and application thereof
  • Phage for staphylococcus aureus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Isolation and propagation of phage

[0018] Phage JS01 was isolated from sewage from food processing plants. Its separation and multiplication process are as follows:

[0019] (1) First prepare the required medium: LB liquid medium: add 10g peptone, 5g yeast powder, 10g NaCl to 1L water; 2×LB liquid medium: add 20g peptone, 10g yeast powder, 20g NaCl to 1L water; bottom culture Base: add 1.5% agar powder to LB liquid medium; upper medium: add 0.7% agar powder to LB liquid medium;

[0020] (2) Staphylococcus aureus ATCC25923 (from American Type Culture Collection ATCC) was inoculated into LB liquid medium, and cultured with shaking at 37°C for 10 h to obtain Staphylococcus aureus culture medium.

[0021] (3) After the sewage was sterilized by filtration with a 0.22 μm filter membrane, it was mixed with an equal volume of 2×LB liquid medium, transferred to the culture medium of Staphylococcus aureus, and cultured with shaking at 37°C for 10 hours to prolifera...

Embodiment 2

[0028] Example 2: Inhibition process of bacteriophage JS01 on Staphylococcus aureus ATCC25923

[0029] Inoculate Staphylococcus aureus ATCC25923 into 5 mL of LB liquid medium, cultivate overnight at 37°C and 200 rpm, inoculate 2% into 50 mL of LB liquid medium, and cultivate to OD at 37°C and 200 rpm 600nm is 1.2, that is, the bacterial concentration in the culture medium is 5×10 8 For cfu / mL, add 10 8 1mL of pfu / mL phage solution, after 6 hours, the OD of the culture medium 600nm decreased to 0.152, and the number of viable bacteria decreased to 4.3×10 4 cfu / mL, see results figure 1 shown;

[0030] Contrast test: no phage JS01 was added, the culture medium was cultivated at 37°C and 200rpm, OD 600nm rose to 4.414, and the number of live bacteria reached 3.8×10 9 cfu / mL. Compared with the comparative test in Example 2, when the phage was acted for 6 hours, the number of viable bacteria was reduced by 5.0 log units, and when the phage was acted for 3 hours, the numbe...

Embodiment 3

[0031] Example 3: Antibacterial effect of phage JS01 on Staphylococcus aureus ATCC25923 at different concentrations

[0032] Inoculate Staphylococcus aureus ATCC25923 into 5 mL of LB liquid medium, cultivate overnight at 37°C and 200 rpm, inoculate 1% into 50 mL of LB liquid medium, and cultivate into the culture medium at 37°C and 200 rpm Bacteria solution is 1×10 8 For cfu / mL, add 0, 10 1 、10 2 、10 3 、10 4 、10 5 、10 6 、10 7 、10 8 、10 9 、10 10 pfu of phage JS01 was lysed for 1 hour, and the number of viable bacteria in the liquid was determined. Results When the added amount of phage JS01 was at 10 6 -10 10 When pfu, that is, when MOI is 0.0002~2, Staphylococcus aureus is effectively inhibited, and the number of viable bacteria is reduced by 4.8~7.7 log units. see results figure 2 shown.

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Abstract

A phage for staphylococcus aureus, of which the preservation number is CCTCC M2011409, has an effective splitting effect on the staphylococcus aureus; the storage condition of the phage is that after being filtered and degermed, phage culture solution can be stored for more than two months under 4 DEG C, and can be stored for more than two years in a glycerin tube under 70 DEG C below zero; the concentration of the stored phage culture solution is 1011pfu / mL; and in the application of the phage for the staphylococcus aureus, the phage culture solution is directly sprayed on the surface of solid food or mixed in bacteria-containing food to inhibit the staphylococcus aureus in the food. The dosage of the phage in bacteria-containing material is 0.0001pfu / cfu to 100pfu / cfu. The phage can effectively control the contamination of the staphylococcus aureus in the food system, and relative to antibiotics and chemical preservatives, the phage has the characteristics of high specificity, no residues and safety.

Description

technical field [0001] The invention relates to a phage strain and application thereof, in particular to a phage capable of inhibiting Staphylococcus aureus. Background technique [0002] Staphylococcus aureus is a Gram-positive pathogen that is easily contaminated in food, especially instant food and cold fresh food. It is also a pathogen that is easily contaminated in hospitals. At present, the antibacterial reagents used to inhibit Staphylococcus aureus in food, feed, medical and other fields mainly include antibiotics and chemical preservatives, but due to the overuse of antibiotics, more and more Staphylococcus aureus Antibiotics have developed resistance, and even multidrug-resistant strains and vancomycin-resistant strains have emerged, and antibiotic residues are not easy to be quickly degraded. Chemical preservatives are difficult to be decomposed by the human body, and may have potential toxic side effects after accumulating to a certain amount, so there are poten...

Claims

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Application Information

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IPC IPC(8): C12N7/00A23L3/3571C12R1/445C12R1/92
Inventor 史锋贾静静李永富李烨
Owner SHANGHAI XINNONG FEED
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