Bacteriophage composition, bacteriostatic agent and application thereof

A phage and composition technology, applied in the field of bioengineering, can solve problems such as ineffective prevention and control of animal intestinal diseases

Active Publication Date: 2021-07-02
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no information about the inhibition of Clostridium perfringens, Salmonella, E. records and reports, an...

Method used

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  • Bacteriophage composition, bacteriostatic agent and application thereof
  • Bacteriophage composition, bacteriostatic agent and application thereof
  • Bacteriophage composition, bacteriostatic agent and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Isolation, preparation and purification of phage

[0037] 1. Phage isolation

[0038]Add peptone solution to the feces samples collected from poultry farms, shake at room temperature for 2-4 hours, and take the supernatant at 5000×g min -1 Centrifuge for 30 min, and filter the supernatant with a 0.22 μm filter membrane.

[0039] Take 1ml of the supernatant and add it to 50ml of TY medium, then add 1ml of the overnight culture of ATCC 13124 Clostridium perfringens strain (TY overnight culture), wherein the overnight culture of ATCC 13124 Clostridium perfringens strain produces perfringens The concentration of Clostridium was 1 x 10 8 cfu / ml~1×10 10 cfu / ml.

[0040] Get 1ml supernatant and add in 50ml LB culture medium respectively, then add the overnight culture of 1ml Salmonella ATCC 13076, avian Escherichia coli E-0503 (isolated from a chicken farm in Zhengzhou, Henan) and Shigella flexneri CMCC 51572 respectively, Among them, the concentration of corresponding ba...

Embodiment 2

[0048] Phage lysis in vitro

[0049] Mix Clostridium perfringens, Salmonella, Escherichia coli and Shigella cultured overnight at a concentration ratio of 1:1:1:1, and the final concentration of the mixed bacteria is about 10 9 cfu / ml, centrifuged to remove medium, adjusted to OD with 0.85% saline 600 About 0.5. After mixing the 4 kinds of phages according to the titer ratio of 1:1:1:1, the final concentration of the mixed phages is about 10 10 pfu / ml, added to the above mixed bacterial solution (MOI is 0.1 and 1 respectively), cultivated at 37°C, and set up a bacterial control group at the same time, that is, no phage composition was added to the bacterial solution; After 10 hours, take 0.2ml to detect OD 600 value, compare the OD after adding the phage composition 600 Variety.

[0050] The result is as figure 2 As shown, starting OD in MOI 0.1 and 1 groups 600 The values ​​are 0.51 and 0.52, respectively. After acting for 2 hours, compared with the control group, th...

Embodiment 3

[0052] Bacteriostasis Effect of Phage Composition in Poultry Breeding

[0053] Forty 1-day-old chicks were selected and randomly divided into 4 groups, 10 in each group. Among them, group A was fed normally as the control group, and group B was fed normally with phage composition at the same time, about 1×10 8 pfu / 0.1ml / cat, fed continuously for 7 days, group C was fed normally and added ampicillin drinking water (0.1% W / V), drank water continuously for 7 days, group D was the blank control group. On the 14th to 21st day, groups B and C were fed with phage composition and ampicillin respectively; groups A, B, and C were respectively fed with Clostridium perfringens, Salmonella, Escherichia coli and Shigella flexneri on day 22 Bacteria composition (approximately 10 8 ~10 9 cfu / 0.1ml / bird, the ratio is 10:1:1:1) to infect, the chickens were weighed on the 25th day, and the intestines were taken to analyze the occurrence of necrotic enteritis and scored (0 was normal small int...

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Abstract

The invention provides a bacteriophage composition, a bacteriostatic agent and application thereof, and belongs to the technical field of bioengineering. The bacteriophage composition comprises a clostridium perfringens bacteriophage (CCTCC NO: M2020047), a salmonella bacteriophage (CCTCC NO: M2020710), an escherichia coli bacteriophage (CCTCC NO: M2020711) and a shigella flexneri bacteriophage (CCTCC NO: M209029). The bacteriophage composition has a strong lysis effect on clostridium perfringens, salmonella, escherichia coli and shigella flexneri, and can be used for inhibiting and/or eliminating clostridium perfringens, salmonella, escherichia coli and shigella flexneri; by inhibiting the overgrowth of clostridium perfringens, salmonella, escherichia coli and shigella flexneri, the occurrence of intestinal diseases of animals is effectively prevented and controlled; a safe bacteriophage antibacterial product source without toxic and side effects is provided for preventing and controlling animal intestinal diseases caused by clostridium perfringens, salmonella, escherichia coli and shigella flexneri.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a phage composition, a bacteriostatic agent comprising the phage composition and applications thereof. Background technique [0002] The intestinal flora is a complex group composed of hundreds of microorganisms. The interaction between microorganisms and the metabolites produced by microorganisms play a role in the health of the host, and have the functions of immune regulation and nutrient digestion. However, there are many pathogenic bacteria in the intestinal flora, such as Clostridium perfringens, Salmonella, Escherichia coli and Shigella, which will colonize the chicken intestines, and the excessive growth will cause intestinal flora imbalance and systemic infection. Seriously affect production performance, even cause death, bring huge losses to the breeding industry. In addition, due to the excessive use of growth-promoting agents such as antibiotics in...

Claims

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Application Information

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IPC IPC(8): C12N7/00A01N63/40A01P1/00A61K35/76A61P1/00A61P31/04C12R1/92
CPCC12N7/00A01N63/40A61K35/76A61P31/04A61P1/00Y02A50/30C12N2795/00032
Inventor 张辉包红朵周艳王冉
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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