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Bioculture method for entomopathogenic nematodes

An entomopathogenic nematode and culture method technology, applied in the field of biological reproduction and culture, can solve the problems of slow inoculation and low efficiency, and achieve the effects of simple and feasible steps, low cost and improved survival rate

Inactive Publication Date: 2019-03-22
李小龙
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved in the present invention is to provide a method for in vivo culture of entomopathogenic nematodes aiming at the shortcomings of slow inoculation and low efficiency in the cultivation of entomopathogenic nematodes in the prior art

Method used

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  • Bioculture method for entomopathogenic nematodes
  • Bioculture method for entomopathogenic nematodes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] S1. Preparation of inoculum: according to the required concentration, prepare an entomopathogenic nematode inoculum with a concentration of 10,000 tails / ml;

[0032] S2. Inoculation: Soak the larvae of the greater wax moth into the entomopathogenic nematode inoculum obtained in step S1 at 20°C; the ratio of the number of the greater wax moth larvae to the entomopathogenic nematode in the inoculum is 1:10000, and when the soaking time is 5 minutes, 75% The greater wax moth larvae can be taken out when the greater wax moth is suspended in motion; the room temperature for inoculation is 20°C;

[0033] S3. Cultivation: Move the soaked live S. mellonella larvae obtained in step S2 onto a stackable tray covered with sterilized cloth, and continuously feed it with 0.1% under the conditions of 20° C. and 93% humidity. The saturated water vapor obtained by atomizing the sodium hydroxide solution was incubated at constant temperature and humidity;

[0034] S4. Detection: On the ...

Embodiment 2

[0037] S1. Preparation of inoculum: according to the required concentration, prepare an entomopathogenic nematode inoculum with a concentration of 20,000 tails / ml;

[0038] S2. Inoculation: Soak the larvae of the greater wax moth into the entomopathogenic nematode inoculum obtained in step S1 at 22°C; the ratio of the number of the greater wax moth larvae to the entomopathogenic nematode in the inoculum is 1:20000, and when the soaking time is 7 minutes, 80 The larvae of the greater wax moth can be taken out when % of the greater wax moth is feigned dead; the room temperature for inoculation is 22°C;

[0039] S3. Cultivation: move the soaked live S. mellonella larvae obtained in step S2 onto a stackable tray covered with sterilized cloth, and continuously inject 0.3% under the conditions of 25° C. and 95% humidity. The saturated water vapor obtained by atomizing the sodium hydroxide solution was incubated at constant temperature and humidity;

[0040] S4. Detection: On the 7t...

Embodiment 3

[0043] S1. Preparation of inoculum: according to the required concentration, prepare an entomopathogenic nematode inoculum with a concentration of 50,000 tails / ml;

[0044] S2. Inoculation: soak the larvae of the greater wax moth into the entomopathogenic nematode inoculum obtained in step S1 at 25°C; the ratio of the number of the larvae of the greater wax moth to the entomopathogenic nematode in the inoculum is 1:30000, and when the soaking time is 7 minutes, 80 The larvae of the greater wax moth can be taken out when % of the greater wax moth is feigned dead; the room temperature for inoculation is 25°C;

[0045] S3. Cultivation: move the soaked live S. mellonella larvae obtained in step S2 onto a stackable tray covered with sterilized cloth, and continuously pass through 0.6% under the conditions of 28° C. and 95% humidity. The saturated water vapor obtained by atomizing the sodium hydroxide solution was incubated at constant temperature and humidity;

[0046] S4. Detecti...

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Abstract

The invention discloses a bioculture method for entomopathogenic nematodes. A large amount of the entomopathogenic nematodes can be obtained through the steps of preparation of an inoculation solution, immersed inoculation with the entomopathogenic nematodes, collection of inoculated last instar larvae of large wax moths, and aerated culturing of the entomopathogenic nematodes through a self-madedrawer type incubator. The large-scale bioculture of the entomopathogenic nematodes is realized through the bioculture of the last instar larvae of large wax moths, a creative immersed inoculation process and aerated drawer type incubator culture. The mass propagation of parasitic entomopathogenic nematodes in the larvae of large wax moths is realized at a suitable temperature and a suitable humidity. The method realizes the intensive inoculation and culture of the large wax moths during the bioculture of entomopathogenic nematodes, and solves the contradiction of large labor force required intraditional bioculture methods.

Description

technical field [0001] The invention relates to the technical field of biological propagation and cultivation, and more specifically, to a method for in vivo cultivation of entomopathogenic nematodes. Background technique [0002] Entomopathogenic nematodes are a type of pathogenic nematodes that use insects as hosts. They survive in the soil during the infestation stage. The nematodes do not feed during the infestation stage and live freely independently of insects, looking for or invading insect hosts. Nematode infection pests mainly go through three stages: infecting the host, latently releasing symbiotic bacteria and toxins, and killing the pest. After entering the insect body, the intestinal symbiotic bacteria will multiply in the blood cavity of the insect, produce toxins to kill the insect, and decompose the insect. tissue as a food source for nematodes. This symbiotic system is beneficial for biological control of pests. [0003] Entomopathogenic nematodes have a s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/033
CPCA01K67/033
Inventor 李小龙张凤琴何农跃
Owner 李小龙
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