Freeze-drying protective additive of pseudoalteromonas bacteria and freeze-drying method

A technology of pseudoalteromonas and freeze-drying protective agent, which is applied in the field of bacterial freeze-drying and storage, can solve the problems of different freeze-drying protective agent and freeze-drying method, and achieves the effect of good protection effect and long storage time.

Active Publication Date: 2013-12-18
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the large differences in the habitat conditions, nutritional requirements, and cultivation methods of various microbial strains, the freeze-drying protective agents and freeze-drying methods of different microbial strains are also different.
Before the present invention was made, the lyoprotectants and lyophilization methods involving marine bacteria were seldom reported

Method used

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  • Freeze-drying protective additive of pseudoalteromonas bacteria and freeze-drying method
  • Freeze-drying protective additive of pseudoalteromonas bacteria and freeze-drying method

Examples

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Embodiment 1

[0018] In November 2011, "rotting skin syndrome" occurred in japonicus seedlings in a japonicus seedling protection farm in Qingdao City, Shandong Province. A japonicus seedlings with typical symptoms were selected from the breeding pond and brought back to the laboratory of the Yellow Sea Fisheries Research Institute for pathogen isolation. and nurture. Under sterile conditions, pick 3 to 5 diseased sea cucumber seedlings and put them in a sterile petri dish, gently drip wash the lesion several times with sterilized 1.5% NaCl solution, and then scrape it with a sterile scalpel. Take a small amount of tissue from each diseased ginseng lesion in the same new sterile petri dish, cut up and mix these tissues by aseptic operation, and use an inoculation loop to pick a small amount of crushed tissue on tryptone soybean broth agar medium Bacteria were isolated by streaking. Place the streaked medium in a constant temperature incubator at 28°C for 30 hours to obtain a dominant bacte...

Embodiment 2

[0027]In August 2012, "ascites disease" occurred in a semi-smooth tongue sole breeding factory in Weifang City, Shandong Province. The diseased semi-smooth tongue sole fish with obvious symptoms were selected from the breeding pond, packed and oxygenated, and brought back to the laboratory of the Yellow Sea Fisheries Research Institute for pathogen isolation. In the laboratory, aseptically draw the "ascites" of 3 to 5 diseased fish with a disposable syringe into a sterile centrifuge tube, add an equal volume of sterile NaCl solution with a concentration of 1.5%, and oscillate evenly . Use a pipette to draw 100 μL of the mixture and add it to a tryptone soybean broth agar medium, and spread the culture. After the coated medium was placed in a constant temperature incubator at 28° C. for 36 hours, three dominant bacterial colonies with different forms were obtained. Single colonies of these three bacteria were picked and transferred to a new tryptone soybean broth agar medium f...

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Abstract

The invention provides a freeze-drying protective additive of pseudoalteromonas bacteria and a freeze-drying method, and relates to the field of freeze vacuum drying protection method of ocean bacteria. The freeze-drying protective additive is liquid and comprises dried skim milk, milk sugar, cane sugar, vitamin C, gelatin, NaC1 and water. Pseudoalteromonas bacterium viable bacterium suspension and the freeze-drying protective additive are fully mixed uniformly according to the volume ratio of 1:4-1:1, are freezed quickly for 3-4 hours at the temperature of minus 80 DEG C after being subpackaged, and then are placed in a vacuum freezing drying machine to be dried in a freezing mode for 20 hours with a vacuum degree of 0.1 mbar in a freeze-drying bin at the temperature of minus 54 DEG C, high-purity drying nitrogen is injected in the freeze-drying bin, so that standard atmospheric pressure is achieved, and the freeze-drying bin is sealed through a pressing cover. The average survival rate of the pseudoalteromonas bacteria after being dried in a freezing mode exceeds 30%, and the freeze-drying protective additive can be stored for a long time at a normal temperature.

Description

technical field [0001] The invention belongs to the technical field of bacterial freeze-vacuum drying preservation, and in particular relates to a freeze-drying protective agent and a freeze-drying method suitable for bacteria of the genus Pseudoalteromonas. Background technique [0002] Pseudoalteromonas (Pseudoalteromonas spp.) is a very common type of bacteria in the marine environment, which is widely distributed in the seawater environment, the substrate, and the body or surface of marine organisms (Torben L. Skovhus, et al. 2004). Pseudoalteromonas has low nutritional requirements and can survive for a long time in a relatively nutrient-poor marine environment in most cases. But it is also an opportunistic pathogen at the same time, especially in the case of high-density farming, the massive reproduction of Pseudoalteromonas often infects marine animals and plants and causes disease, causing rapid spread and prevalence of diseases, making Mariculture produces serious ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/04C12R1/01
Inventor 王印庚张正于永翔李彬荣小军廖梅杰史秀清王岚
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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