A Salmonella phage that can reduce the vertical transmission of Salmonella pullorum and its application

A Salmonella, vertical transmission technology, applied in the direction of bacteriophage, virus/phage, and disease resistance to vector transmission, can solve the problems of large losses, affecting feeding efficiency, and loss of chicken industry, achieving good purification effect, reducing vertical transmission, Effect without toxic side effects

Active Publication Date: 2022-06-07
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 affecting the feeding benefits, and bring huge losses to the chicken industry. Treatment with various drugs is not ideal
In addition, due to the pollution of the hatchery, more deaths were caused, and the loss was huge.

Method used

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  • A Salmonella phage that can reduce the vertical transmission of Salmonella pullorum and its application
  • A Salmonella phage that can reduce the vertical transmission of Salmonella pullorum and its application
  • A Salmonella phage that can reduce the vertical transmission of Salmonella pullorum and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Isolation and purification of Salmonella phage GSMP2233

[0019] Collect 3 water samples near a chicken farm in Hubei, each 30 mL, centrifuge at 5000 rpm for 10 min, and filter sterilize 15 mL of supernatant. 8 cfu / mL), mixed evenly, and incubated overnight at 37°C at 180 rpm to enrich for phage. The sample enrichment solution was centrifuged at 5000 rpm for 10 min, and the supernatant was taken through a 0.22 μm microporous membrane for sterilization to obtain a filtrate containing bacteriophages. Take 100uL of the filtrate and evenly mix it with 300uL of its host Salmonella bacterial solution, and let it stand for 15min to fully bind to the receptors on the bacterial surface. Add the above mixture to 4 mL of TSB semi-solid agar medium cooled to 50 °C, and immediately spread on the solidified TSA plate after mixing. . On the plaque-forming double-layer plate, pick out large and translucent plaques with a sterile pipette tip, desorb them in 1 mL of SM solution by sha...

Embodiment 2

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

[0023] Pick a single colony of the host Salmonella Pullorum, inoculate it into a test tube containing 3 ml of TSB culture solution, and shake and culture at 180 rpm in a 37°C shaker for 12 hours to obtain a host bacterial suspension. The bacterial suspension was transferred to 10 ml of TSB medium at a ratio of 1:100, and cultured with shaking at 180 rpm at 37 °C to the early logarithmic stage. Bacteriophage GSMP2233 and its host Salmonella were diluted to a certain concentration and counted, mixed and 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. Incubate with shaking at 180 rpm in a shaker at 37°C for 12 h. Every 2h, take out part of the culture liquid and carry out centrifugation at 10000g for 10min and ...

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 place the above bacterial bottles in a constant temperature water bath at 25 °C. , after the temperature was equilibrated, 1 mL of pure phage culture solution was added, and it was allowed to stand at 25 °C for 240 min. The samples were taken at the 1st, 2h and 4h respectively for proper dilution, and then the phage titer was determined by the double-layer plate method. Each point was cultured in duplicate and the average value was obtained, and the experiment was repeated 3 times.

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

[0032] Table 2 pH value stability of Salmonella phage after different reaction times (in...

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Abstract

The invention belongs to the field of prevention and control of Salmonella in livestock and poultry breeding of biotechnology, and discloses a Salmonella phage capable of reducing the vertical transmission of Salmonella pullorum and its application. The preservation number of the phage is CCTCC NO: M 2020661. The bacteriophage GSMP2233 fermentation efficiency in the present invention is high, when MOI=0.01, culture 6h titer reaches 8.9x10 10 PFU / ml has a good purification effect on Salmonella pullorum, can effectively reduce the vertical transmission of Salmonella pullorum, and provides a safe, non-toxic and side-effect-free biological control measure for large-scale fermentation and production of Salmonella purification.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Salmonella phage that can reduce vertical transmission of Salmonella pullorum and application thereof. Background technique [0002] Salmonella is an important zoonotic pathogen that mainly resides in the intestines of humans and animals. Studies have shown that food poisoning caused by Salmonella is the highest proportion and most widespread of all bacterial food poisonings. . Most ubiquitous Salmonella serotypes are pathogenic to 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 every year in the world, and the death cases are as high as 3 million. Poultry is considered a major reservoir of Salmonella. In recent years, there have been frequent cases of high morbidity and high mortality in chicks due to the contamination o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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