Novel compound preparation and application thereof in bacterial diseases

A compound preparation and bacterial technology, which is applied to a new compound preparation and its application in bacterial diseases, can solve the problems that the compatibility of bacteriophage and probiotics is difficult to achieve the expectation of comprehensive prevention and control of bacterial diseases.

Inactive Publication Date: 2021-09-14
PHAGELUX (NANJING) BIO-TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] None of the above-mentioned technical solutions has carried out in-depth research on the use of compatibility of phages and probiotics. It is difficult to achieve the expectation of comprehensive prevention and control of bacterial diseases by using a single combination of phages or probiotics. Technology blank

Method used

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  • Novel compound preparation and application thereof in bacterial diseases
  • Novel compound preparation and application thereof in bacterial diseases
  • Novel compound preparation and application thereof in bacterial diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Preparation of Vibrio parahaemolyticus Phage

[0041] Seed culture: Inoculate the activated Vibrio parahaemolyticus host bacteria into TSB medium with an inoculation ratio of 2%, 220 rpm, and culture at 37° C. for 4 hours to obtain the seed liquid of the Vibrio parahaemolyticus host bacteria.

[0042] Enrichment culture: insert Vibrio parahaemolyticus phage and host bacteria into TSB medium, and the MOI value is 5×10 -6 , the host bacteria inoculation ratio is 5%, 190 rpm, 37 ° C for 5 hours.

[0043] Sterilization and purification: After the fermentation, the fermented liquid was centrifuged at 7000rpm for 10 minutes to collect the supernatant, and the supernatant was sterilized and purified with a 0.22 μm filter membrane to obtain Vibrio parahaemolyticus phage water. The titers of VP46, VP48 and VP7 were respectively 5.7×10 10 PFU / mL, 4.8×10 10 PFU / mL and 4.5×10 10 PFU / mL.

[0044] Drying: add protective agent according to the ratio of 3:1, the ratio of protectiv...

Embodiment 2

[0046] Preparation of Vibrio alginolyticus Phage

[0047] Seed cultivation: Inoculate the activated Vibrio alginolyticus host bacteria into TSB medium with an inoculation ratio of 2%, 220 rpm, and culture at 37° C. for 4 hours to obtain a seed liquid of the Vibrio alginolyticus host bacteria.

[0048] Enrichment culture: insert Vibrio alginolyticus phage and host bacteria into TSB medium, and the MOI value is 5×10 -6 , the host bacteria inoculation ratio is 5%, 190 rpm, 37 ° C for 5 hours.

[0049] Sterilization and purification: After the fermentation, the fermented liquid was centrifuged at 7000rpm for 10 minutes to collect the supernatant, and the supernatant was sterilized and purified with a 0.22 μm filter membrane to obtain Vibrio alginolyticus phage water, the titers of VAP7, VAP9 and VAP21 were 3.7 ×10 10 PFU / mL, 2.8×10 10 PFU / mL and 2.5×10 10 PFU / mL.

[0050] Drying: add protective agent according to the ratio of 3:1, the ratio of protective agent is: 15 parts of...

Embodiment 3

[0052] Preparation of coliphage

[0053] Seed cultivation: Inoculate the activated E. coli host bacteria into TSB medium with an inoculation ratio of 2%, culture at 220 rpm, and 37° C. for 4 hours to obtain E. coli host bacteria seed liquid.

[0054] Enrichment culture: insert Vibrio parahaemolyticus phage and host bacteria into TSB medium, and the MOI value is 5×10 -6 , the host bacteria inoculation ratio is 5%, 190 rpm, 37 ° C for 5 hours.

[0055] Sterilization and purification: After the fermentation, the fermented liquid was centrifuged at 7000rpm for 10 minutes to collect the supernatant, and the supernatant was sterilized and purified with a 0.22 μm filter membrane to obtain E. coli phage water. 10 PFU / mL, 3.4×10 10 PFU / mL and 3.7×10 10 PFU / mL.

[0056] Drying: add protective agent according to the ratio of 3:1, the ratio of protective agent is: 15 parts of skimmed milk powder, 3 parts of glycerin, 4 parts of sodium glutamate, 5 parts of sucrose, mix and dry, crush ...

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Abstract

The invention discloses a novel compound preparation and application thereof in bacterial diseases, and belongs to the technical field of bacterial disease control. The novel compound preparation is a bacteriophage and probiotic compatible preparation, and the component ratio of the bacteriophage to the probiotics is (1-5): (5-1). The invention requests to protect application of the novel compound preparation against bacterial diseases such as vibrio disease, colibacillosis, salmonellosis, staphylococcus aureus disease, canker caused by infection of Xanthomonas axonopodis and the like; and the compound preparation prepared by compatibility of the bacteriophage and the probiotics can effectively control the number of vibrio, escherichia coli, salmonella, staphylococcus aureus and xanthomonas carpet grass, and the effect of the compound preparation is better than that of single application of the bacteriophage or the probiotics.

Description

technical field [0001] The invention relates to the field of prevention and treatment of bacterial diseases, more specifically, it relates to a novel composite preparation and its application in bacterial diseases. Background technique [0002] In recent years, bacterial diseases have caused very large economic losses in the fields of aquaculture, fur animal breeding, poultry and livestock breeding, and crop cultivation. For the problem of bacterial diseases, researchers are constantly exploring new solutions. Bacteria-specific phages and green non-stress probiotics are gradually favored by researchers, such as: [0003] Patent CN201710953780.5 discloses the use of a mixture of 2 strains of Vibrio parahaemolyticus phage and 3 strains of Vibrio alginolyticus phage to control Vibrio in aquaculture; patent CN200510009788.3 discloses the use of Lactobacillus paracasei HD1-7 The natural metabolites of peptides are used as preservatives in the prevention and treatment of Escheric...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N63/40A01N63/20A01N63/27A01N63/22A01N63/23A01N63/25A01N63/32A01N63/30A01P1/00A61K35/76A61P31/04A61P17/00A61K35/744A61K35/742A61K35/747
CPCA01N63/40A01N63/20A01N63/27A01N63/22A01N63/23A01N63/25A01N63/32A01N63/30A61K35/76A61K35/744A61K35/742A61K35/747A61K9/0014A61P31/04A61P17/00A61K2300/00Y02A50/30
Inventor 何四龙谢晓莉丛郁徐旭凌肖逍丁良于浩樊小九许文建徐天舜许小康
Owner PHAGELUX (NANJING) BIO-TECH CO LTD
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