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A kind of culture medium and culture method for long-term and continuous culture of deuterostomum

A culture medium and long-term technology, applied in fish farming, application, animal husbandry and other directions, can solve the problems of endangering the healthy and sustainable development of the sea cucumber industry, affecting the growth and development of sea cucumbers, and the death of sea cucumbers. Application, stable and reliable source of raw materials, low cost of cultivation

Active Publication Date: 2021-12-03
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In parasitic diseases, the infection of a deuterostomum parasitized on the respiratory tree of sea cucumbers can lead to decreased adhesion, reduced food intake, slow growth, and even visceral excretion reactions, which seriously affect the growth and development of sea cucumbers, and cause Its immunity is reduced, and it is susceptible to secondary infection by other microorganisms, leading to individual or even group death, which seriously endangers the healthy and sustainable development of the sea cucumber industry.
In 2020, the outbreak of japonicus japonicus disease in many coastal provinces led to mass deaths of aquacultured japonicus in many main breeding areas and heavy economic losses
So far, there is no relevant industrial culture method that can realize the pure culture of deuterostomum

Method used

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  • A kind of culture medium and culture method for long-term and continuous culture of deuterostomum
  • A kind of culture medium and culture method for long-term and continuous culture of deuterostomum
  • A kind of culture medium and culture method for long-term and continuous culture of deuterostomum

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: Comparison of the survival status of deuterostomes under different medium conditions

[0021] The specific steps are as follows: filter seawater, boil and sterilize, cool to 8-10°C, add yeast fermentation liquid concentrate to a final concentration of 2.5 mg / L, mix well and add Lactobacillus paracasei CSDN-6, photosynthetic bacteria, Bacillus to a final concentration of 10 5 CFU / ml, wherein group 1 is Lactobacillus paracasei CSDN-6 + yeast fermentation broth concentrate, group 2 is photosynthetic bacteria + yeast fermentation broth concentrate, and group 3 is Bacillus + yeast fermentation broth concentrate. The above-mentioned liquids are three culture media for the pure culture of Deuterostomum in vitro. The comparison experiment was carried out with the culture medium added with 1:500 body cavity fluid and respiratory tree tissue fragments in sterilized seawater as the control.

[0022] In November 2020, an ostomy disease occurred in sea cucumbers in a ...

Embodiment 2

[0026] Embodiment 2: the selection of the addition amount of yeast fermented liquid concentrate

[0027] The specific steps are as follows: After filtering the seawater, boil it for 5-10 minutes, cool it to 8-10°C, add 0.1mg / L, 1.0mg / L, 2.5mg / L, 5.0mg / L, 10.0mg / L, 25mg / L The yeast fermented liquid concentrate, mix well and add Lactobacillus paracasei CSDN-6 to the final concentration of 10 5 CFU / ml, the liquid is the culture medium for deuterostomum pure culture in vitro.

[0028] In December 2020, the sea cucumbers suffering from deuterostomum disease were transported back to the laboratory, and the respiratory trees were dissected to make water-soaked slices. Microscopic observation showed that a large number of deuterostomum parasitized in the respiratory trees. The deuterostomum isolated from the respiratory tree of the diseased sea cucumber is counted under the microscope after removing large tissue fragments and other organisms; that is, according to the result of count...

Embodiment 3

[0032] Embodiment 3: culture salinity control

[0033] The specific steps are as follows: adjust the salinity of the medium to 0‰, 2.5‰, 5‰, 10‰, 15‰, 20‰, 25‰, 30‰, 35‰, 40‰ by adding fresh water and adding sodium chloride to seawater ‰, 45‰, add 2.5mg / L yeast fermentation liquid concentrate and 10 5 The Lactobacillus paracasei CSDN-6 of CFU / ml, this liquid is the culture medium of deuterostomum pure culture in vitro.

[0034] In December 2020, the sea cucumbers suffering from deuterostomum disease were transported back to the laboratory and dissected, and the respiratory trees were taken to make water-soaked slices. Microscopic observation showed that a large number of deuterostomum parasitized in the respiratory trees. The deuterostomum isolated from the respiratory tree of the diseased sea cucumber is counted under the microscope after removing large tissue fragments and other organisms; that is, according to the result of counting under the microscope, the deuterostomum ...

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Abstract

The invention relates to a culture medium and a culture method for long-term and continuous culture of Deuterostomum, belonging to the field of biotechnology. The culture medium is liquid and contains a yeast fermentation liquid concentrate with a final concentration of 1 to 3 mg / L, and the final concentration is 1 ×10 5 CFU / ml Lactobacillus paracasei CSDN‑6 (Lactobacillus paracasei), and the remaining ingredients are filtered and boiled natural seawater with a salinity of 28-32‰ and a pH of 7.8-8.5. The present invention also provides a method for long-term and continuous cultivation of Deuterostomum using the above-mentioned culture medium. During the culture process, the culture medium of Deuterostomum can be maintained in a clear state for a long time without corruption and peculiar smell, and can be continuously and stably cultivated for a long time. Deuterostomum provides technical support for subsequent research on basic biology and drug screening of parasites.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a long-term and continuous culture medium and a culture method of Deuterostomum. Background technique [0002] Apostichopus, also known as Apostichopus japonicus, has become a pillar industry of marine aquaculture in my country since the success of large-scale artificial propagation technology in the 1980s. In 2019, the national sea cucumber breeding area was about 246,745 hectares, the output reached 171,700 tons, the direct output value was more than 30 billion yuan, and the economic output value of the industrial chain was nearly 100 billion yuan. One (2020 Fishery Statistical Yearbook). [0003] With the gradual expansion of the sea cucumber industry, disease problems have become increasingly prominent, causing economic losses of more than 3 billion yuan each year. The reported causes of the disease include bacterial infection, fungal infection, and parasitic infecti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K61/30
CPCA01K61/30
Inventor 王印庚王锦锦廖梅杰李彬畅孟阳尚晓金
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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