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Low temperature storage method for cryptocaryon irritans trophont and cyst

A technology that stimulates Cryptonucleosis and cryopreservation, applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as interruption of passage of worm strains, economic and time waste, aging, etc.

Inactive Publication Date: 2008-09-10
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in order to preserve the worm strain on the host fish, the subculture process cannot be interrupted. Even when the parasite is not needed, the subculture work must continue without interruption, which will cause a huge waste of economy and time
In addition, Burgesset et al. (1994) found that the vitality of stimulating Cryptocaryon worms would gradually weaken with the increase of the number of passages, that is, "aging phenomenon", and the "aging" of the insect body would eventually lead to the interruption of the passage of the strains.
Houghton et al. (1986) and Noe et al. (1995) found that the subculture of Ichthyophthirius multifiliis strains also has aging phenomenon

Method used

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  • Low temperature storage method for cryptocaryon irritans trophont and cyst
  • Low temperature storage method for cryptocaryon irritans trophont and cyst

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1 stimulates the preservation method of the trophozoite and cyst of Cryptocaryon

[0020] Collect the trophozoites and cysts of Cryptocaryon stimuli, put them into 24-well tissue culture plates (50 parasites per well) and 5cm petri dishes (1000 parasites per petri dish), respectively, at 8-15°C Store in an artificial climate box with a relative humidity of 90% and 12hr light / 12hr light. In order to ensure consistent storage conditions, put all 24-well tissue culture plates and petri dishes containing parasites into a 10L beaker, add 5L sterilized seawater, keep a small amount of gas, and change the sterilized seawater every 5d. The water volume is 2L. The seawater for cultivating parasites was first filtered, then sterilized at 120° C. for 30 minutes, and 100 IU / ml of penicillin and 100 ul / ml of streptomycin were added before use.

Embodiment 2

[0021] Example 2. Validity test of cryopreservation stimulating trophozoites and encapsulation methods of Cryptocaryonia

[0022] Collect trophozoites and cysts of Cryptocaryon stimuli, store them at 8-15°C according to the above method, and test the hatching rate of trophozoites and cysts at 27.0°C and the infectivity of hatched larvae every one month. It was found that with the increase of storage time, the hatching rate of trophozoites and cysts and the infection rate of larvae would gradually decrease, and the trophozoites and cysts stored for 3 months at 12°C could still recover. Infectious larvae hatch. Larvae hatched from trophozoites stored at 12°C for 4 months can reproduce a large number of offspring under suitable conditions, and the virulence of the offspring has not decreased, because they are more sensitive to eggs of the same size than before storage. The lethal dose of pomfret was not lowered.

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Abstract

The invention discloses a method for low-temperature preservation of trophonts and cysts of Cryptocaryon irritans, comprising the following steps: firstly, the trophonts and the cysts of the Cryptocaryon irritans are collected and preserved in a manual climatic cabinet at the temperature of 8 to 15 DEG C with a relative humidity of 90 percent according to 12 hour-light: 12 hour-dark cycle; sterilizing seawater is exchanged once at intervals of 5 days and the water exchange quantity is 20 percent; secondly, the trophonts and the cysts of the Cryptocaryon irritans which are preserved at low temperature are detected once every month, and the hatchability of the trophonts and the cysts and the infectivity of hatched larvaes when the temperature is returned to 27.0 DEG C are measured. By adoption of the method for preservation of the trophonts and the cysts of the Cryptocaryon irritans, the trophonts which are preserved for 4 months and the cysts which are preserved for 3 months can still hatch out larvaes with infectivity; the larvaes can propagate a great amount of offsprings under proper conditions, and the virulence of the offsprings is not reduced.

Description

technical field [0001] The invention relates to a method for preserving parasites, in particular to a method for low-temperature preservation to stimulate trophozoites and cysts of Cryptocaryon. Background technique [0002] Cryptocaryon irritans Brown (1951) is an obligate fish parasite, parasitic in the gills and subcutaneous skin of tropical and subtropical marine fish, causing highly lethal "white spot disease". In order to study Cryptocaryon stimuli systematically, we used Pomfret ovata as a model animal to establish a set of efficient passage and reproduction system, which can obtain a large number of samples of the worm, laying the foundation for further research on the worm. However, in order to preserve the worm strain on the host fish body, the subculture process cannot be interrupted. Even when the parasite is not needed, the subculture work must continue without interruption, which will cause a huge waste of economy and time. In addition, Burgesset et al. (1994)...

Claims

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Application Information

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IPC IPC(8): C12N1/10C12R1/90
Inventor 李安兴但学明
Owner SUN YAT SEN UNIV
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