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Salmonella phage preparation and preparation method thereof

A Salmonella, phage technology, applied in the field of bioengineering

Active Publication Date: 2015-08-19
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the optimal method of fermentation preparation for phage maturation at home and abroad.

Method used

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  • Salmonella phage preparation and preparation method thereof
  • Salmonella phage preparation and preparation method thereof
  • Salmonella phage preparation and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1: Salmonella phage STP4-a optimum liquid propagation condition optimization

[0056] (1) Determination of optimal culture time for liquid proliferation of Salmonella phage STP4-a

[0057] Add 2 mL of overnight cultured ST14028 (bacteria count 10) to 200 mL NB 9 cfu / mL) and 2mL phage STP4-a (titer 10 9 pfu / mL), set up three parallel groups, shake culture at 37°C 150r / min, take a sample every 2h, centrifuge to remove bacterial sediment, measure its potency by double-layer plate method, the results are shown in figure 1 .

[0058] Depend on figure 1 It can be seen that when phage STP4-a was cultured for 4 hours, its titer had reached 10 9 pfu / mL, especially when the culture time reaches 8h, the titer has basically stabilized at 4×10 9 pfu / mL, by analysis of variance, the titer of phage STP4-a had no significant difference at 8h and 12h (P>0.05), and was significantly higher than that at 0, 2, 4, and 6h (P<0.05). Therefore, the optimal culture time of phag...

Embodiment 2

[0081] Embodiment 2 Salmonella phage STP4-a fermenter test

[0082] (1) Fermentation preparation: Taking the 5L fermentation system as an example, the prepared CaCl 2 After the NB is fully stirred and mixed, it is poured into the tank of the fermenter with a liquid volume of 3L (the maximum liquid volume of the fermenter), and then an appropriate amount of defoamer is added to eliminate the bubbles generated during the fermentation process. Place the body of the fermenter with the culture medium in a high-pressure steam sterilizer at 121°C for 15 minutes, and cool it down before use. The host bacterium ST14028 (bacteria solution concentration is about 10 9 cfu / mL) 30mL (the inoculation volume is 1% of the fermentation volume) and 30mL of bacteriophage STP4-a (the inoculation volume is 1% of the fermentation volume) were successively added into the fermenter. The flame inoculation method is used for inoculation. A certain amount of alcohol is added to the alcohol groove aroun...

Embodiment 3

[0089] The in vitro antibacterial application effect of embodiment 3 phage STP4-a

[0090]The standard strain of Salmonella typhimurium ATCC 14028 preserved in the laboratory and the single colony of Salmonella isolated from sick chickens in chicken farms were inoculated into the nutrient broth medium, and after overnight incubation at 37 °C, the bacterial liquid was dipped with sterile cotton swabs. Then evenly spread it onto a nutrient agar plate, drop 10 μL of bacteriophage STP4-a on each plate respectively, and then place it upside down in a 37°C incubator to cultivate overnight, and observe whether a transparent circle is produced, thus judging the lysis of the bacteriophage. to determine the cleavage profile of phage STP4-a.

[0091] Table 4 shows the bacteriostatic effect of bacteriophage STP4-a on bacteria isolated from sick chickens. The table shows that bacteriophage STP4-a has lytic activity on all 10 isolated Salmonella strains, while on Escherichia coli, Pneumococ...

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Abstract

The present invention relates to a phage preparation, especially a Salmonella phage preparation and a preparation method thereof, and belongs to the field of bioengineering. According to the present invention, phage with a preservation number of CCTCCNO: M 2014145 adopts Salmonella as host bacteria to ferment so as to prepare the Salmonella phage preparation; the optimal Salmonella phage STP4-a liquid proliferation conditions such as the culture temperature, the time, the pH value, the ion concentration, the rotation speed, the inoculation amount and the like are explored so as to determine the optimal culture conditions, and the progressive enlargement culture is performed to determine the optimal phage preparation conditions; and the preparation can be separately used or used in a compounding manner, can specifically inactivate a variety of salmonella, and provides a safe and no toxic-side effect phage product source for current prevention and control of Salmonella contamination.

Description

technical field [0001] The invention relates to a bacteriophage preparation, in particular to a Salmonella bacteriophage preparation and a preparation method thereof, belonging to the field of bioengineering. Background technique [0002] Salmonella (Salmonella) is a Gram-negative straight bacterium, which is an important zoonotic pathogen. Domestic and foreign studies have shown that food-borne diseases caused by Salmonella frequently take the top spot. With the large-scale and long-term use of antibiotics, the increasingly serious drug-resistant Salmonella in food and the environment has affected the treatment of human Salmonella infections. Whether new biological bacteriostatic agents can be developed to replace chemical antibiotics is currently a challenge for researchers. a challenge. Bacteriophages are a class of bacterial viruses. After a virulent phage infects bacteria, it can rapidly lyse the bacteria and cause the death of the host bacteria. Salmonella phages ca...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04
CPCY02A50/30
Inventor 王静雪林洪李梦哲李萌
Owner OCEAN UNIV OF CHINA
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