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Application of burkholderia ubonensis JD03 and fermentation broth thereof and active compound in preparation of products for preventing and controlling saprolegniasis

An antibacterial and saprolegnia technology, applied in the application field of products, can solve the problems of mammalian and human teratogenicity, malachite green is difficult to degrade, and the residual time is long, and achieves inhibition of mycelial growth, strong antagonistic ability, The effect of reducing morbidity

Active Publication Date: 2019-09-06
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Malachite green used to be the most effective chemical drug against saprolegniasis since the 1930s. However, studies have found that malachite green is difficult to degrade and remains in the water environment and aquatic animals for a long time. It is harmful to both mammals and humans. Teratogenic, carcinogenic, and mutagenic risks

Method used

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  • Application of burkholderia ubonensis JD03 and fermentation broth thereof and active compound in preparation of products for preventing and controlling saprolegniasis
  • Application of burkholderia ubonensis JD03 and fermentation broth thereof and active compound in preparation of products for preventing and controlling saprolegniasis
  • Application of burkholderia ubonensis JD03 and fermentation broth thereof and active compound in preparation of products for preventing and controlling saprolegniasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Identification of antagonistic strain JD03

[0054] (1) Activation of the JD03 strain: The JD03 strain was isolated from the pond sediment samples and kept in the laboratory. Take the preserved JD03 strain and activate it on the beef extract peptone solid medium by streaking on the plate. After culturing at a constant temperature of 28°C for 16 hours, pick a single clone and inoculate it in the beef extract peptone liquid medium. 10h. The bacterial liquid can be used not only as the seed liquid for fermentation culture, but also as the bacterial liquid for the antibacterial test.

[0055] (2) Identification of JD03 strain:

[0056] The morphological, physiological and biochemical indicators of the JD03 strain were determined, and the specific methods were referred to "Berger's Bacterial Identification Handbook (Ninth Edition)" (English version).

[0057] The results showed that the JD03 strain was a Gram-negative bacterium with extremely clustered flagella...

Embodiment 2

[0061] Embodiment 2: the inhibitory effect of antagonistic bacterial strain JD03 to Saprolegnia

[0062] (1) Preparation of Saprolegnia mycelium and Saprolegnia spore suspension: Take the preserved Saprolegnia agar plate, use a sterile puncher with a diameter of 8mm to punch holes on the Saprolegnia agarose plate to obtain Saprolegniasis pieces, and take 4 Saprolegniasis pieces evenly Inoculate it on a potato dextrose agar (PDA) plate (a plate formed by pouring into a petri dish: 200g of peeled potatoes, 20.0g of glucose, 20.0g of agar, 1000mL of deionized water, autoclave at 121°C for 20min, and place at room temperature When the temperature drops to more than 50 degrees, pour the plate), incubate at a constant temperature of 25°C for 3 days, until the Saprolegnia mycelium completely covers the entire surface of the plate, and store it in a refrigerator at 4°C for subsequent experiments. Use a sterile puncher to punch holes on a fresh water mold plate to get water mold pieces...

Embodiment 3

[0070] Embodiment 3: the preventive effect of antagonistic bacterial strain JD03 fermented liquid to grass carp artificial infection saprolegniasis

[0071] (1) Preparation of JD03 strain fermentation broth and Saprolegnia spore suspension: same as in Example 2.

[0072](2) Take a healthy grass carp, wipe off the mucus on the body surface with gauze near the tail behind the dorsal fin, then scratch the surface of the body with a rough object, and use a syringe needle to draw a line about 1.5 cm long and deep on both sides of the back here. About 1mm wound. The prepared saprolegnia spore suspension is added dropwise to the wound and then put into the water added with saprolegnia spores. The experiment was divided into two groups: JD03 experimental group and control group, each with three parallels. 25L of water is put into each barrel, and 15 grass carp after the above-mentioned treatment are put into it. Add 500mL of prepared Saprolegnia spore suspension (2×10 4 spores / mL)...

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Abstract

The invention discloses an application of burkholderia ubonensis JD03 and a fermentation broth thereof and an active compound in preparation of products for preventing and controlling saprolegniasis.The burkholderia ubonensis JD03 is burkholderia ubonensis JD03, and has the preservation number of GDMCC NO:60660. The burkholderia ubonensis JD03 and the fermentation broth thereof have good, safe, effective and stable antagonistic effect against water mould, can be used as a novel biological control product instead of malachite green, avoids environmental pollution and malachite green and metabolite residues thereof in aquatic products, and not only meets the need of preventing and controlling saprolegniasis in fishes, but also ensures the quality and safety of the aquatic products. The active compound 2-pyrrolidone-5-carboxylic acid is separated from the fermentation broth of the burkholderia ubonensis JD03 for the first time. The compound can significantly inhibit the water mould sporegermination and mycelial growth, is environmentally friendly and safe to fishes, and has great potential to develop the novel product for prevention and control of saprolegniasis in fishes.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to the application of a Saprolegnia antagonistic bacteria JD03 and its fermented liquid and the active compound 2-pyrrolidone-5-carboxylic acid in the preparation of products for preventing and controlling Saprolegniasis, especially in the preparation of water for preventing and controlling fish. Mildew application in products. Background technique [0002] Saprolegniasis is a common fungal parasitic disease in freshwater aquaculture. It has the characteristics of wide epidemic range, long duration, and no selectivity to hosts. The breeding industry has caused serious economic losses. Saprolegnia spp. is the main parasitic fungal pathogen that causes Saprolegnia. Malachite green used to be the most effective chemical drug against saprolegniasis since the 1930s. However, studies have found that malachite green is difficult to degrade and remains in the water environment and aquat...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/10A61K35/74A61P31/10C12R1/01
CPCA61K35/74A61P31/10C12P17/10C12N1/205C12R2001/01
Inventor 张其中刘彦甍
Owner JINAN UNIVERSITY
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