Salmonella sp.phage JNwz02 and application thereof

A Salmonella and phage technology, applied in the direction of phage, virus/phage, application, etc., to achieve the effect of pollution prevention and control, good temperature and pH tolerance

Pending Publication Date: 2022-03-18
JIANGXI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The above-mentioned technical scheme has the following defects: the research on Salmonella phage mainly focuses on the cracking and application of different serotypes of Salmonella, and the research on the cracking of other genera or species of food-borne pathogenic bacteria and its application on Salmonella phage is rare to report

Method used

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  • Salmonella sp.phage JNwz02 and application thereof
  • Salmonella sp.phage JNwz02 and application thereof
  • Salmonella sp.phage JNwz02 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Isolation and identification of phage

[0041] Isolation of a phage

[0042] (1) Take 500mL water sample from the lake near Jiangxi Agricultural University’s farmer’s market, add 0.055g calcium chloride to mix, and then let it settle for 2h; Concentrate the supernatant, and centrifuge the concentrate at 6000g, 4°C for 10min; The supernatant was filtered through a 0.22 μm membrane filter. Take 10 mL of the above filtrate and mix with 10 mL of TSB liquid medium, and then add 600 μL of logarithmic phase host bacteria (Salmonella Stanleyville RSE39) (OD 600 =0.5–0.6) in the above-mentioned TSB liquid medium, cultivated at 37°C, 220rmp for 12h, collected the bacterial solution and centrifuged at 12000g, 4°C for 15min, and filtered the supernatant with a 0.22μm filter membrane. Repeat the above steps three times, and store the filtrate at 4°C, which is the phage stock solution.

[0043] (2) Take 100 μL log phase host bacteria (OD 600 =0.5–0.6) on the plate of L...

Embodiment 2

[0054] Example 2: Determination of cleavage profile of bacteriophage JNwz02

[0055] The experimental selection included 14 strains of bacteria such as Salmonella and Escherichia coli, and the lysis profile of phage JNwz02 was determined.

[0056] Among them, 14 strains of bacteria are:

[0057] 1) 1 strain of Salmonella Stanleyville RSE39;

[0058] 2) 1 strain of Salmonella Hillingdon N1529-D3;

[0059] 3) 2 strains of Salmonella typhi ATCC BAA-664, NCTC 8271;

[0060] 4) 1 strain of Salmonella Arizona LHICA_AZ23;

[0061] 5) 1 strain of Salmonella Bareilly CFSAN000189;

[0062] 6) 1 strain of Salmonella Heidelberg AR-0404;

[0063] 7) 1 strain of Salmonella Corvallis AR-0406;

[0064] 8) 1 strain of Salmonella Albany CVM N18S2238;

[0065] 9) 1 strain of Salmonella Kentucky 161365;

[0066] 10) 1 strain of Salmonella enteritidis BNCC336875;

[0067] 11) 1 strain of Salmonella typhimurium BNCC185946;

[0068] 12) 1 strain of Escherichia coli ATCC 25922;

[0069] 13...

Embodiment 3

[0075]Example 3: Determination of the optimal multiplicity of infection (MOI) and one-step growth curve of phage JNwz02

[0076] MOI determination:

[0077] Mix 100 μL phage (MOI: 100, 10, 1, 0.1, 0.01, 0.001, 0.0001, respectively) with 100 μL log-phase host bacterial solution (OD 600 =0.5–0.6) and mix well, incubate at 37°C for 15 min, add to 10 mL TSB liquid medium, and incubate at 37°C with shaking at 220 rpm for 5 h. Centrifuge at 12000g at 4°C for 15min, and take the supernatant and filter it with a 0.22μm membrane filter to sterilize. The titer of phage under different multiplicity of infection was measured by the double-layer agar plate method, and the infection ratio with the highest titer was the best MOI;

[0078] The best MOI results are shown in Table 2. The best MOI between phage JNwz02 and host Salmonella Stanleyville RSE39 was 0.01, and the best MOI between phage JNwz02 and EHEC O157:H7 AV4997 was 10.

[0079] One-step growth curve determination:

[0080] Ac...

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Abstract

The invention discloses a salmonella phage (salmonella sp. Phage) JNwz02 and application of the salmonella phage JNwz02. The strain is preserved in Guangdong Microbial Culture Collection Center on May 12, 2021, and the preservation number is GDMCC No: 61662-B1. The bacteriophage is a virulent bacteriophage, can be used for cracking salmonella of eight serotypes such as stetank vil, hirgania and typhoid, including multiple strains of drug-resistant salmonella, and also can be used for cracking enterohemorrhagic Escherichia coli O157: H7. The invention also discloses an application of the bacteriophage JNwz02 as a bacteriostatic agent in food storage.

Description

technical field [0001] The invention relates to the field of microbial strains, in particular to a Salmonella phage (salmonellasp.phage) JNwz02 and its application. Background technique [0002] Salmonella and Enterohemorrhagic Escherichia coli (EHEC) O157:H7 are two important zoonotic foodborne pathogens causing significant economic losses worldwide and seriously threaten public health security. Since Fleming discovered penicillin in the last century, a series of antibiotics have been discovered and synthesized and used in clinical practice and the prevention and treatment of livestock and poultry diseases, and foodborne pathogenic bacteria infections have been effectively controlled. However, in recent years, the abuse of antibiotics has led to the continuous emergence of a large number of drug-resistant strains, and there is a serious development trend of high-level drug resistance, multi-drug resistance and cross-drug resistance. Therefore, finding bioinhibitory agents...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A23L3/3571A23B4/22
CPCC12N7/00A23L3/3571A23B4/22C12N2795/10321C12N2795/10331A23V2002/00A23V2200/10
Inventor 吴国平舒梅张慧珍钟婵
Owner JIANGXI AGRICULTURAL UNIVERSITY
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