CN103283643A - Method for detecting cryptocaryon irritans brown chemotaxis - Google Patents
Method for detecting cryptocaryon irritans brown chemotaxis Download PDFInfo
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- CN103283643A CN103283643A CN2013101721652A CN201310172165A CN103283643A CN 103283643 A CN103283643 A CN 103283643A CN 2013101721652 A CN2013101721652 A CN 2013101721652A CN 201310172165 A CN201310172165 A CN 201310172165A CN 103283643 A CN103283643 A CN 103283643A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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Abstract
The invention discloses a method for detecting cryptocaryon irritans brown chemotaxis. The method includes (1) taking a clean glass slide as a carrier, dropping 40-60 microliter of seawater droplets containing cryptocaryon irritans brown larvae on one side of the glass slide, dropping test solution with the same volume on the other side of the glass slide, and the space between the seawater droplets and the test solution is 1-2cm; (2) using a clean toothpick to draw lines gently from the seawater droplets to the test solution to make the droplets of two sides to be communicated; (3) standing for 20-40 minutes at the temperature of 25-28 DEG C, and observing the amount of the cryptocaryon irritans brown larvae moved to the test solution through a microscope and calculating the chemotaxis. Since the glass slide is adopted to detect the cryptocaryon irritans brown chemotaxis, the method is more convenient to operate as compared with the internationally conventional petri dish punching method and 96-well plate method. By the aid of the technology of detecting the cryptocaryon irritans brown chemotaxis through the glass slide, large amount of time, reagents and consumable items are saved.
Description
Technical field
The present invention relates to a kind of aquaculture field, particularly relate to the chemotactic assay method of a kind of stimulation cryptonucleus insect.
Background technology
Along with the continuous deterioration of breeding environment, the lifting gradually of cultivation density, very huge to the economic loss that the aquaculture seawater fish causes by " ichthyophthirius " that stimulate cryptonucleus insect to cause.And " ichthyophthirius " have the infection rate height, involve characteristics such as scope is wide, lethality height, and nowadays also do not have good treatment way.The chemotaxis research that stimulates cryptonucleus insect is helped deep discussion to the mechanism problem that stimulates the cryptonucleus insect infection host, advance from filtering out anthelmintic drug for the control of " ichthyophthirius ".Abroad research has methods such as plate punch method, 96 orifice plates to the eliminating fish parasites chemotaxis at present, and these methods exist complex operation, are not easy to dynamic observe defectives such as counting.Domestic also do not have a chemotactic research method of special detection eliminating fish parasites.
Summary of the invention
The object of the present invention is to provide a kind of easy and simple to handle, chemotactic assay method of stimulation cryptonucleus insect of saving cost.
For achieving the above object, technical solution of the present invention is:
The present invention is the chemotactic assay method of a kind of stimulation cryptonucleus insect, it may further comprise the steps: (1) is carrier with clean slide, side at slide drips the sea water drops that 40-60 μ l contains cryptocaryon irritans larva, at the test solution of opposite side dropping equal volume, distance is 1-2 cm between sea water drops and the test solution; (2) rule lightly with clean toothpick to the test solution direction from sea water drops, the drop of both sides is communicated with; (3) under the environment of 28 ℃ of temperature 25-, after leaving standstill 20-40 minutes, examine under a microscope the quantity that moves to test solution moderate stimulation cryptonucleus insect larva and calculate the chemotactic rate, chemotactic rate (IC)=move to and contain larva number/larva sum * 100% in the test substances one side drop.Be reference with the clean sea water, namely test solution is clean sea water, measures its chemotactic rate, and then judges that test solution is to the chemotaxis of cryptocaryon irritans larva.
Cryptocaryon irritans larva is 3.4% ± 1.6% to the chemotactic rate of clean sea water, has namely regarded as chemotaxis so cryptocaryon irritans larva is higher than 5% to the chemotactic rate of test solution, does not have chemotaxis otherwise regard as.
Described test solution is a kind of in egg-shaped pompano serum, egg-shaped pompano mucus, fructose soln, large yellow Crocker serum or the glutamic acid solution.
After adopting such scheme, because the present invention uses the chemotaxis that slide detects stimulates cryptonucleus insect, more easy and simple to handle than the plate punch method of abroad commonly using, 96 well plate method.Adopting slide to detect stimulates cryptonucleus insect chemotaxis technology, has saved a large amount of time and reagent, consumptive material.
The present invention is further illustrated below in conjunction with specific embodiment.
Embodiment
Embodiment 1: under 28 ℃ of conditions, get clean slide, in the worm liquid of a side Dropwise 50 μ l cryptocaryon irritans larva of wave carrier piece, at opposite side Dropwise 50 μ l egg-shaped pompano serum, both sides make passage of its formation at a distance of 2 cm with clean toothpick line.Behind 30 min, microscopically is observed counting, it is 35 that discovery moves to the larva number that contains egg-shaped pompano serum one side, and the sum of larva is 55, and calculating the chemotactic rate is 64 ﹪ (being that egg-shaped pompano serum has strong chemotaxis to cryptocaryon irritans larva).
Embodiment 2: under 26 ℃ of conditions, get clean slide, in the worm liquid of a side Dropwise 50 μ l cryptocaryon irritans larva of wave carrier piece, in the egg-shaped pompano mucus of opposite side Dropwise 50 μ l, both allow it form a passage at a distance of 1 cm with clean toothpick line.Behind 30 min, microscopically is observed counting, it is 21 that discovery moves to the larva number that contains egg-shaped pompano mucus one side, and the sum of larva is 60, and calculating the chemotactic rate is 35 ﹪ (being that egg-shaped pompano mucus has certain chemotaxis to cryptocaryon irritans larva).
Embodiment 3: under 28 ℃ of conditions, get clean slide, drip the worm liquid of 40 μ l cryptocaryon irritans larvas in a side of wave carrier piece, drip the fructose soln of 40 μ l at opposite side, both sides are at a distance of 2 cm, allow passage of its formation with clean toothpick line.Behind 30 min, microscopically is observed counting, find that moving to the larva number that contains fructose soln one side is 1, and the sum of larva is 50, and calculating the chemotactic rate is 2 ﹪ (being that fructose soln does not have chemotaxis to cryptocaryon irritans larva).
Embodiment 4: under 25 ℃ of conditions, get clean slide, drip the worm liquid of 40 μ l cryptocaryon irritans larvas in a side of wave carrier piece, drip the large yellow Crocker serum of 40 μ l at opposite side, both are at a distance of 1 cm, allow passage of its formation with clean toothpick line.Behind 40 min, microscopically is observed counting, it is 29 that discovery moves to the larva number that contains egg-shaped pompano mucus one side, and the sum of larva is 55, and calculating the chemotactic rate is 70 ﹪ (being that large yellow Crocker serum has very strong chemotaxis to cryptocaryon irritans larva).
Embodiment 5: under 25 ℃ of conditions, get clean slide, drip the worm liquid that 40 μ l stimulate latent nuclear larva worm in a side of wave carrier piece, drip 40 μ l glutamic acid solutions at opposite side, both sides make passage of its formation at a distance of 1 cm with clean toothpick line.Behind 40 min, microscopically is observed counting, find that moving to the larva number that contains glutamic acid solution one side is 3, and the sum of larva is 97, and calculating the chemotactic rate is 3 ﹪ (being that glutamic acid solution does not have chemotaxis to cryptocaryon irritans larva).
Can find that by above five embodiment the serum of various seawater fishs (even some freshwater fishes) and mucus have chemotaxis to stimulating cryptonucleus insect, and some small-molecule substances (carbohydrate, amino acid etc.) there is not chemotaxis to stimulating cryptonucleus insect.
The chemotactic time: 20-40 min.
28 ℃ of chemotactic temperature: 25-(stimulating the cryptonucleus insect suitable growth temperature).
Chemotactic rate computing formula: chemotactic rate (IC)=move to and contain larva number/larva sum * 100% in the test substances one side drop.
At present, with criterion and the foundation of chemotactic rate as chemotaxis.Be reference with the clean sea water, the chemotactic rate of general clean sea water is lower than 5%.If the chemotactic rate is higher than 5%, just thinking has chemotaxis, and the chemotactic rate is more high, just illustrates that chemotaxis is more strong.
Emphasis of the present invention just is: be to set up the detection method that cryptocaryon irritans larva drives voltinism, and by determining concrete parameter and the scope of application of this detection method.
The above, only for preferred embodiment of the present invention, so can not limit scope of the invention process with this, i.e. the equivalence of doing according to the present patent application claim and description changes and modifies, and all should still belong in the scope that patent of the present invention contains.
Claims (2)
1. one kind stimulates the chemotactic assay method of cryptonucleus insect, it is characterized in that: it may further comprise the steps: (1) is carrier with clean slide, side at slide drips the sea water drops that 40-60 μ l contains cryptocaryon irritans larva, at the test solution of opposite side dropping equal volume, distance is 1-2 cm between sea water drops and the test solution; (2) rule lightly with clean toothpick to the test solution direction from sea water drops, the drop of both sides is communicated with; (3) under the environment of 28 ℃ of temperature 25-, leave standstill 20-40 minutes after, examine under a microscope the quantity that moves to test solution moderate stimulation cryptonucleus insect larva and calculate the chemotactic rate, thereby judge that test solution is to the chemotaxis of cryptocaryon irritans larva.
2. the chemotactic assay method of a kind of stimulation cryptonucleus insect according to claim 1 is characterized in that: described test solution is a kind of in egg-shaped pompano serum, egg-shaped pompano mucus, fructose soln, large yellow Crocker serum or the glutamic acid solution.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105660462A (en) * | 2015-12-18 | 2016-06-15 | 中山大学 | Cyst inactivation method for controlling cryptocaryon irritans disease of fishes |
| CN106508754A (en) * | 2016-11-02 | 2017-03-22 | 集美大学 | Biological prevention and treatment method for cryptocaryon irritans disease |
| CN114164078A (en) * | 2021-11-30 | 2022-03-11 | 齐齐哈尔大学 | Bacterial chemotactic substance screening glass slide and application thereof |
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| US4309416A (en) * | 1979-09-20 | 1982-01-05 | Research Corporation | Vaccines from taxonomically similar organisms |
| CN101260367A (en) * | 2008-04-22 | 2008-09-10 | 中山大学 | Low temperature storage method for cryptocaryon irritans trophont and cyst |
| CN202364674U (en) * | 2011-12-06 | 2012-08-08 | 中国水产科学研究院东海水产研究所 | Micropipette for separating and absorbing trophozoite cells of cryptocaryon irritans brown from fish bodies |
| CN102726327A (en) * | 2012-06-20 | 2012-10-17 | 中国水产科学研究院东海水产研究所 | Method for removing residual cryptocryon irritans from culture pond in pollution-free way |
| TW201313247A (en) * | 2011-09-26 | 2013-04-01 | Univ Nat Taiwan | Vaccine produced using optimized immobilization antigen cDNA of Cryptocaryon irritans and producing method and use thereof |
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2013
- 2013-05-10 CN CN2013101721652A patent/CN103283643A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4309416A (en) * | 1979-09-20 | 1982-01-05 | Research Corporation | Vaccines from taxonomically similar organisms |
| CN101260367A (en) * | 2008-04-22 | 2008-09-10 | 中山大学 | Low temperature storage method for cryptocaryon irritans trophont and cyst |
| TW201313247A (en) * | 2011-09-26 | 2013-04-01 | Univ Nat Taiwan | Vaccine produced using optimized immobilization antigen cDNA of Cryptocaryon irritans and producing method and use thereof |
| CN202364674U (en) * | 2011-12-06 | 2012-08-08 | 中国水产科学研究院东海水产研究所 | Micropipette for separating and absorbing trophozoite cells of cryptocaryon irritans brown from fish bodies |
| CN102726327A (en) * | 2012-06-20 | 2012-10-17 | 中国水产科学研究院东海水产研究所 | Method for removing residual cryptocryon irritans from culture pond in pollution-free way |
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| GYÖRGY CSABA ET AL: "Effect of Oxytocin and its Analogues on the Chemotaxis of Tetrahymena:Evolutionary Conclusions", 《ACTA PROTOZOOL》 * |
| 夏春: "《水产动物疾病》", 30 June 2011, 中国农业出版社 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105660462A (en) * | 2015-12-18 | 2016-06-15 | 中山大学 | Cyst inactivation method for controlling cryptocaryon irritans disease of fishes |
| CN106508754A (en) * | 2016-11-02 | 2017-03-22 | 集美大学 | Biological prevention and treatment method for cryptocaryon irritans disease |
| CN114164078A (en) * | 2021-11-30 | 2022-03-11 | 齐齐哈尔大学 | Bacterial chemotactic substance screening glass slide and application thereof |
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Application publication date: 20130911 |