Salmonella phage capable of reducing vertical propagation of salmonella pullorum and application of salmonella phage

A Salmonella, vertical transmission technology, applied in the direction of bacteriophage, virus/bacteriophage, resistance to vector-borne diseases, etc., can solve problems such as large losses, affecting feeding efficiency, and decreasing egg production of laying hens, achieving good purification effect, reducing The effect of vertical transmission and high fermentation efficiency

Active Publication Date: 2021-08-24
武汉格瑞农生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And it will also cause the disease and death of the breeding chickens, and the egg production of laying hens will decline, which will hinder the healthy development of the chicken industry while affectin...

Method used

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  • Salmonella phage capable of reducing vertical propagation of salmonella pullorum and application of salmonella phage
  • Salmonella phage capable of reducing vertical propagation of salmonella pullorum and application of salmonella phage
  • Salmonella phage capable of reducing vertical propagation of salmonella pullorum and application of salmonella phage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Isolation and Purification of Salmonella Phage GSMP2233

[0019] Collect 3 parts of water samples near a chicken farm in Hubei, each 30mL, centrifuge at 5000rpm for 10min, take 15mL supernatant to filter and sterilize, and the filtrate is mixed with 15mL 2×TSB liquid culture medium and 1mL Salmonella bacteria liquid in logarithmic phase (10 8 cfu / mL) were evenly mixed, and cultured overnight at 180 rpm at 37°C to enrich the phage. The sample enrichment solution was centrifuged at 5000 rpm for 10 min, and the supernatant was sterilized through a 0.22 μm microporous membrane to obtain a filtrate containing phage. Take 100uL of the filtrate and mix it evenly with 300uL of the host Salmonella bacteria liquid, and let it stand for 15min to fully bind with the receptors on the surface of the bacteria. Add the above mixed solution to 4mL TSB semi-solid agar medium cooled to 50°C, mix well and spread it on the solidified TSA plate immediately, after the agar is solidified, inc...

Embodiment 2

[0022] Determination of titer of Salmonella phage GSMP2233 under different multiplicity of infection and different infection time

[0023] Pick a single bacterium colony of host Salmonella (Salmonella Pullorum), inoculate it into a test tube filled with 3ml TSB culture solution, and shake it at 180rpm in a shaker at 37°C for 12h to obtain a host bacterium suspension. The bacterial suspension was transferred to 10ml TSB culture medium at a ratio of 1:100, and cultured with shaking at 180rpm at 37°C to the pre-logarithmic phase. The phage GSMP2233 and its host Salmonella were diluted to a certain concentration and counted, and mixed cultured according to different multiplicity of infection ratios (MOI=number of phages / number of bacteria), and TSB liquid medium was added to make the total volume of each tube the same. Shake culture at 180rpm in a shaker at 37°C for 12h. Take out part of the culture solution every 2h and carry out 10000g centrifugation for 10min and collect the s...

Embodiment 3

[0029] pH Stability Test of Salmonella Phage GSMP2233

[0030] Take sterile bacterial bottles and add 9 mL of TSB medium with different pH (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) respectively, and then place the above bacterial bottles in a constant temperature water bath at 25°C After the temperature was balanced, 1mL of pure phage culture solution was added, and allowed to stand at 25°C for 240min. The samples were taken at 1h, 2h and 4h respectively, and after appropriate dilution, the phage titer was determined by the double-layer plate method. Each point was cultured in duplicate tubes to obtain the average value, and the experiment was repeated 3 times.

[0031] The results are shown in Table 2. Salmonella phage GSMP2233 has no significant change in titer when treated at pH 3-10 for 1 hour, and still has relatively high titer after being treated at pH 4-10 for 4 hours.

[0032] The pH value stability (initial titer: 5.2x10 8 PFU / mL)

[0033]

[0034] Note: ND: not det...

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Abstract

The invention belongs to the field of livestock and poultry breeding salmonella prevention and control in biotechnology, and discloses a salmonella bacteriophage capable of reducing vertical propagation of salmonella pullorum and application of the salmonella bacteriophage, and the preservation number of the bacteriophage is CCTCC NO:M 2020661. The bacteriophage GSMP2233 is high in fermentation efficiency, when MOI is equal to 0.01, the titer reaches 8.9*10<10> PFU/ml after the bacteriophage GSMP2233 is cultured for 6 h, the bacteriophage GSMP2233 has a good purification effect on salmonella pullorum, vertical propagation of the salmonella pullorum can be effectively reduced, and a biological prevention and control measure which is safe, free of toxic and side effects and capable of achieving large-scale fermentation production is provided for salmonella purification.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Salmonella phage capable of reducing the vertical transmission of Salmonella pullorum and its application. Background technique [0002] Salmonella (Salmonella) is an important zoonotic pathogen, mainly living in the intestines of humans and animals. Studies have shown that food poisoning caused by Salmonella is the highest proportion and most harmful of all bacterial food poisoning. . Most of the ubiquitous Salmonella serotypes are pathogenic in both animals and humans, and they can cause zoonotic diseases. The World Health Organization (WHO) survey shows that there are hundreds of millions of cases of human gastroenteritis caused by Salmonella infection in the world every year, and the death cases are as high as 3 million. Poultry is considered a major reservoir of Salmonella. In recent years, due to the vertical transmission of Salmonella pullorum, the eggs are contaminated a...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04C12R1/92
CPCC12N7/00A61K35/76A61P31/04C12N2795/00021C12N2795/00032Y02A50/30
Inventor 王喜亮黄金梅徐岳张秀玲胡雄张晓东田甲
Owner 武汉格瑞农生物科技有限公司
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