Technique for allowing aquatic invertebrate larvae and parents to harmlessly carry Vibrio phages

An invertebrate, phage technology, applied in virus/phage, microorganism-based methods, microorganisms, etc., can solve problems such as inapplicability, and achieve the effect of meeting production needs

Inactive Publication Date: 2012-07-11
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The above research and invention patents, in vertebrates, long-circulating phages strengthen the protective effect and prolong the time, but for aquatic animals, it is not applicable

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 Taking the newly hatched larvae of Litopenaeus vannamei (hereinafter referred to as "prawn larvae") as an example, the technology of the present invention is implemented according to the following steps.

[0034] 1. Isolate Vibrio strains and their phages routinely from the aquaculture water body to obtain a phage lysate.

[0035] 2. The phage lysate obtained in step 1 was recovered by centrifugation, filtered through a 0.22 μm filter membrane, and the titer was measured by the double-plate method, and the obtained concentration was 1.1×10 11 pfu / ml phage solution, stored at 4°C, as a storage solution, for later use; specific method: add freshly prepared phage stock solution from a single phage spot to the logarithmic growth phase bacterial solution cultured by a single colony, and put it into a shaker for vigorous shaking After shaking, the obtained lysate was centrifuged to remove bacterial fragments, and trypanase DNAI and trypsinase were added to remove...

Embodiment 2

[0049] Example 2 Taking the parent Litopenaeus vannamei (hereinafter referred to as "parent prawn") with a body length of 16-18 cm as an example, implement the technology of the present invention according to the following steps.

[0050] 1. Isolate Vibrio strains and their phages routinely from the aquaculture water body to obtain a phage lysate.

[0051] 2. The phage lysate obtained in step 1 was recovered by centrifugation, filtered through a 0.22 μm filter membrane, and the titer was measured by the double-plate method to obtain a concentration of 8.5×10 10 pfu / ml phage solution, stored at 4°C, as a storage solution, for later use; specific method: add freshly prepared phage stock solution from a single phage spot to the logarithmic growth phase bacterial solution cultured by a single colony, and put it into a shaker for vigorous shaking After shaking, the obtained lysate was centrifuged to remove bacterial fragments, and trypanase DNAI and trypsinase were added to remove ...

Embodiment 3

[0065] Example 3 Vibrio parahaemolyticus and its bacteriophage were obtained from a local farm on Donghai Island, Zhanjiang. In clean seawater, the final concentration was 4×10 6 pfu / mL selected Vibrio parahaemolyticus phage, soak the newly hatched Litopenaeus vannamei larvae in it for 1.0h, and continue to cultivate them for 20 days according to the conventional seedling method. No host bacteria were found in the larvae, and the survival rate was 65%. The host bacteria in the larvae of the control group were as high as 4×10 4 cfu / mL, the survival rate is 5%.

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PUM

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Abstract

The invention discloses a technique for allowing aquatic invertebrate larvae and parents to harmlessly carry Vibrio phages. The technique of the invention utilizes the feature that invertebrates do not display specific antibody response, and comprises soaking aquatic invertebrate larvae and parents with phages, collecting culture samples from the aquatic invertebrate larvae and parents, isolating phages from invertebrates, soaking again, repeating the soaking steps for a plurality of times successively, and culturing and selecting phages to obtain phages which can survive in aquatic invertebrates for a long period of time. Phages obtained by use of the technique of the invention are not virulent or harmful, can retain the lysis activity for host bacteria during a long period of time, and can meet the requirements of production of aquatic invertebrate larvae and parents. The invention provides a new technique for preventing and treating vibriosis of aquatic invertebrates.

Description

technical field [0001] The invention relates to a technology for enabling aquatic invertebrate larvae and their parents to carry bacteria, in particular to a technology for enabling aquatic invertebrate larvae and parents to harmlessly carry Vibrio phages, and belongs to the field of aquatic microorganisms. Background technique [0002] In human medicine, phages and their lytic enzymes and specific proteins are used as one of the tools to control drug-resistant strains. As a powerful tool against pathogenic host bacteria, bacteriophages have been applied in aquatic products. They can be put into water, or smeared on wounds, taken orally, or even soaked. In aquatic applications, in addition to using water to effectively reduce the concentration of pathogenic bacteria in the host, the prevention and treatment in aquatic animals mainly rely on soaking and oral administration, which have a good inhibitory effect on intestinal pathogenic bacteria, but Less effective against path...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K61/00C12N7/00C12R1/92
CPCY02A40/81
Inventor 邱德全邱明生杨世平王成桂申玉春
Owner GUANGDONG OCEAN UNIVERSITY
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