CN104823882A - Golden apple snail egg mass scattering method - Google Patents
Golden apple snail egg mass scattering method Download PDFInfo
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- CN104823882A CN104823882A CN201510165586.1A CN201510165586A CN104823882A CN 104823882 A CN104823882 A CN 104823882A CN 201510165586 A CN201510165586 A CN 201510165586A CN 104823882 A CN104823882 A CN 104823882A
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- China
- Prior art keywords
- egg
- pomacea canaliculata
- container
- ovum grain
- dispersion
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Links
- 241000570011 Pomacea canaliculata Species 0.000 title claims abstract description 93
- 238000000790 scattering method Methods 0.000 title abstract 3
- 239000007853 buffer solution Substances 0.000 claims abstract description 34
- 239000002270 dispersing agent Substances 0.000 claims abstract description 29
- 239000012459 cleaning agent Substances 0.000 claims abstract description 16
- 238000001035 drying Methods 0.000 claims abstract description 7
- 102000002322 Egg Proteins Human genes 0.000 claims description 87
- 108010000912 Egg Proteins Proteins 0.000 claims description 87
- 210000004681 ovum Anatomy 0.000 claims description 87
- 238000000034 method Methods 0.000 claims description 58
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 57
- 239000000243 solution Substances 0.000 claims description 43
- 239000006185 dispersion Substances 0.000 claims description 35
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 31
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 12
- 238000004140 cleaning Methods 0.000 claims description 12
- 239000011575 calcium Substances 0.000 claims description 11
- 210000001136 chorion Anatomy 0.000 claims description 9
- KWYUFKZDYYNOTN-UHFFFAOYSA-M potassium hydroxide Substances [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 9
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 6
- 230000007613 environmental effect Effects 0.000 claims description 6
- 239000011435 rock Substances 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 2
- -1 saccharide compound Chemical class 0.000 claims description 2
- 239000008187 granular material Substances 0.000 abstract 8
- 230000008827 biological function Effects 0.000 abstract 1
- 230000000877 morphologic effect Effects 0.000 abstract 1
- 238000002791 soaking Methods 0.000 abstract 1
- 235000013601 eggs Nutrition 0.000 description 60
- 239000011734 sodium Substances 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- 239000012452 mother liquor Substances 0.000 description 8
- 150000001875 compounds Chemical class 0.000 description 7
- 239000012153 distilled water Substances 0.000 description 7
- 238000011534 incubation Methods 0.000 description 5
- 230000007935 neutral effect Effects 0.000 description 5
- 238000002156 mixing Methods 0.000 description 4
- 210000003097 mucus Anatomy 0.000 description 4
- 230000008859 change Effects 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000001186 cumulative effect Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 210000004400 mucous membrane Anatomy 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 238000005411 Van der Waals force Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000008144 egg development Effects 0.000 description 2
- 230000012447 hatching Effects 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000237858 Gastropoda Species 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000011095 buffer preparation Methods 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 235000011116 calcium hydroxide Nutrition 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007614 genetic variation Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000005868 ontogenesis Effects 0.000 description 1
- 230000017448 oviposition Effects 0.000 description 1
- 230000000505 pernicious effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000012502 risk assessment Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
-
- 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Dental Preparations (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a golden apple snail egg mass scattering method, consisting of the following steps: (1) placing golden apple snail egg masses to be scattered in a container, and filling in dispersants and evenly shaking the container, wherein the size ratio of the dispersants to the golden apple snail egg masses is (2-8):1; (2) shaking or swinging the container until the golden apple snail egg masses are completely scattered into egg granules; (3) removing the dispersants and scraps in the container and leaving the egg granules obtained through scattering in the container; (4) filling cleaning agents into the container to clean the egg granules; (5) filling buffer solutions with the pH value as 6.0-8.0 into the container, and soaking the egg granules for at least 3min, wherein the size ratio of the filled buffer solutions to the dispersants filled in step (1) is (1-3):1; (6) filling in the cleaning agents to clean the egg granules; (7) drying or baking the egg granules. The golden apple snail egg mass scattering method can evenly scatter the golden apple snail egg masses and enables the scattered egg granules to keep the complete morphological structure and the biological function and characteristics of the egg granules.
Description
Technical field
The present invention relates to a kind of method of disperseing pieces of an egg, be specifically related to a kind of method of disperseing Pomacea canaliculata pieces of an egg.
Background technology
Pomacea canaliculata was just classified as one of 100 kinds of species of world's most invasive by World Conservation Union (IUCN) the Invasive Alien Species Committee of Experts as far back as 2000, was also one of first batch of 16 kinds of pernicious biological invasions of China listed by national environmental protection portion in 2003.Intrude into many countries and regions in the world at present, also extensively distributed in the Changjiang river areas to the south of China, while endangering the agricultural production of China, ecology of water safety, also threaten health.So become more and more important about the research of Pomacea canaliculata in recent years, this wherein the population dynamic of Pomacea canaliculata, genetic variation, the ontogeny even research of the problem such as risk assessment all be unable to do without the separation to spiral shell ovum, but traditional mechanical separation method waste time and energy and be difficult to complete, nondestructively the Pomacea canaliculata ovum grain of adhesion is separated.During Amazonian snail oviposition together with time secretion colloid mucus; namely mucus dry solidification forms glued membrane later, makes ovum grain close adhesion form pieces of an egg, thus plays fixing and protective effect to ovum grain; in addition the main component of spiral shell chorion is the calcium carbonate of stable in properties, thus the strong stress resistance of Pomacea canaliculata ovum.Complete, be nondestructively separated Pomacea canaliculata ovum grain, key to destroy the mucous membrane between ovum grain, and the main component of mucus is water and glycoprotein.
As everyone knows, the active force of the Protein requirement primary structure mainly covalent bond such as peptide bond and disulfide bond; And the active force of the higher structure of the Protein requirement mainly non-covalent bond such as hydrogen bond, hydrophobic bond, ionic bond and Van der Waals force.Wherein hydrogen bond be Protein requirement secondary structure structure as alpha-helix, the active force of the conformations such as beta sheet.Hydrophobic bond is the hole that amino acid whose non-polar sidechain that on polypeptide chain, hydrophobicity is stronger is avoided aqueous phase adhesion and aggregation and formed together, plays a part to be even more important in the tertiary structure of Protein requirement.Sat linkage is the active force formed by electrostatic attraction by the side-chain radical of positive and negative charge in protein.Van der Waals force is intermolecular attraction.The active force of Protein requirement structure can be destroyed and change to some physical factor or chemical factor, makes the structure of protein molecule that irreversible change occur, thus cause the physicochemical property of protein and physiological function to change.Usual physical factor has: heating, pressurization, stirring, vibration, Ultraviolet radiation, ultrasonic etc., and chemical factor has: strong acid, highly basic, heavy metallic salt, trichloroacetic acid, ethanol, acetone etc.
Thus find and utilize suitable physics, chemical factor to make protein denaturation, the mucous membrane especially solidified departs from from chorion surface, and the integrality that the mucus destroyed between ovum grain still retains ovum grain can reach the object being separated Pomacea canaliculata ovum grain.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art part and a kind of method of disperseing Pomacea canaliculata pieces of an egg is provided, simple and its stable components of the method agents useful for same, adopt the method Pomacea canaliculata pieces of an egg can be uniformly dispersed, and the ovum grain after dispersion can retain the complete and biology function characteristic of the morphosis of ovum grain itself.
For achieving the above object, the technical scheme that the present invention takes is: a kind of method of disperseing Pomacea canaliculata pieces of an egg, comprises the following steps:
(1) Pomacea canaliculata pieces of an egg to be disperseed are placed in container, add dispersant and shake up, add dispersant and Pomacea canaliculata pieces of an egg volume ratio be 2 ~ 8:1;
(2) vibrate or rock container and be separated into ovum grain completely to Pomacea canaliculata pieces of an egg;
(3) removing the dispersant in container and chip, staying in container by disperseing the ovum grain obtained;
(4) in container, cleaning agent is added, cleaning ovum grain (remove cleaning agent after cleaning ovum grain, stayed in container by ovum grain);
(5) in container, add the buffer solution that pH value is 6.0 ~ 8.0, infiltrate ovum grain at least 3 minutes, the buffer solution added and step (1) add dispersant volume ratio be 1 ~ 3:1;
(6) cleaning agent cleaning ovum grain is added;
(7) dry or dry ovum grain.
The present invention, by after pieces of an egg and dispersant, makes Pomacea canaliculata pieces of an egg be separated into ovum grain completely through vibrating or rocking; In disperseing to add buffer solution and cleaning agent in the ovum grain obtained, object is to regulate the pH value on ovum grain top layer to it for neutral, and in step (6), the cleaning agent of last cleaned ovum grain is that namely neutrality show that the pH value on ovum grain top layer is for neutral.
In the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, the volume ratio adding dispersant and Pomacea canaliculata pieces of an egg in step (1) is 2 ~ 8:1, so that dispersant fully infiltrates pieces of an egg; Vibration can be preferable on shaking table in step (2) or manually to rock container, vibrating or rock container and can accelerate the dispersion of Pomacea canaliculata pieces of an egg, but vibration or shaking speed or movement range unsuitable excessive, in order to avoid destroy ovum grain; Other impurity etc. that the mucous membrane that chip source in step (3) solidifies between a small amount of damaged ovum grain, ovum grain and pieces of an egg adhere to.
The present invention's dispersant, buffer solution and cleaning agent used can be prepared under normal temperature and pressure; Wherein, in buffer preparation process, first can prepare mother liquor (mother liquor refers to the buffer component of high concentration) respectively, then mother liquor is got respectively according to certain volume ratio as required, mixing, if need the buffer solution of low concentration, dilutes mother liquor mixture by a certain percentage.The store method of reagent of the present invention is: dispersant can seal at ambient temperature and preserve in vial (rubber stopper) or plastic bottle; The mother liquor of buffer solution can seal preservation in brown bottle; Cleaning agent can store at ambient temperature.
As the preferred embodiment of the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, described dispersant is the strong base solution do not reacted with Pomacea canaliculata chorion, and in this solution, the mass percent concentration of highly basic is 3 ~ 10%.Dispersant should not react with chorion, and the main component of Pomacea canaliculata chorion is the calcium carbonate of stable in properties, pretend into dispersant strong base solution should not with calcium carbonate CaCO
3, calcium phosphate Ca
3(PO
4)
2with slaked lime Ca (OH)
2etc. reacting, in order to avoid destroy chorion.
As the preferred embodiment of the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, described dispersant is KOH, NaOH or Ca (OH)
2solution.
As the preferred embodiment of the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, described buffer solution not with saccharide compound and calcium carbonate reaction.In the present invention, buffer solution should not easily form double salt with compounds such as carbohydrates and with calcium carbonate reaction, chorion be decomposed, and buffer solution preferably can suppress the solution of the catalytic reaction of some enzyme; Best, select, to chorion, there is certain protective effect and under different temperature condition pH value comparatively stable, the simple buffer solution of composition.
As the preferred embodiment of the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, described buffer solution is Na
2hPO
4-NaH
2pO
4buffer solution.Na
2hPO
4-NaH
2pO
4the compound method of buffer solution is preferably: first prepare 0.2mol/L Na respectively
2hPO
4with 0.2mol/L NaH
2pO
4(0.2mol/L Na
2hPO
4with 0.2mol/L NaH
2pO
4be mother liquor), then get 0.2mol/LNa respectively according to certain volume ratio as required
2hPO
4with 0.2mol/L NaH
2pO
4, mixing, if need the buffer solution of low concentration, dilutes mother liquor mixture by a certain percentage.
As the preferred embodiment of the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, described cleaning agent is water.More preferably, described water is pure water or distilled water.
As the preferred embodiment of the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, in described step (1), container is cylindrical.More preferably, container is centrifuge tube or test tube.
As the preferred embodiment of the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, in described step (3), remove dispersant in container and chip by the method for drawing or filtering.
As the preferred embodiment of the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, in described step (4) and step (6), the volume ratio of the dispersant that cleaning agent and step (1) add is 3 ~ 8:1.
As the preferred embodiment of the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, in described step (7), dry ovum grain in 20 ~ 25 DEG C or dry ovum grain in 20 ~ 30 DEG C, and the difference of the temperature and environmental temperature of drying ovum grain is not more than 5 DEG C.
Beneficial effect of the present invention is: the invention provides a kind of new method of disperseing Pomacea canaliculata pieces of an egg; Pomacea canaliculata pieces of an egg can be uniformly dispersed by the method; and better to the protected effect of Pomacea canaliculata ovum grain during dispersion Pomacea canaliculata pieces of an egg, the ovum grain after dispersion can retain the complete and biology function characteristic of the morphosis of ovum grain itself.Method technique provided by the invention is simple and applicability is strong, successfully solves the difficult problem that Pomacea canaliculata pieces of an egg are difficult to efficient and complete dispersion.
In addition, method agents useful for same provided by the invention is simple, its stable components, and is easy to produce and preparation, resistance to storage and transport.
Accompanying drawing explanation
Fig. 1 is in the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, and the required minimum jitter time of step (2) and Pomacea canaliculata egg development hatch the corresponding relation figure of degree;
When Fig. 2 is for employing the method for the invention dispersion Pomacea canaliculata pieces of an egg, the state diagram of Pomacea canaliculata ovum before and after dispersion.
Embodiment
For better the object, technical solutions and advantages of the present invention being described, below in conjunction with the drawings and specific embodiments, the invention will be further described.
In embodiment, agents useful for same and compound method as follows:
Dispersant: embodiment is preferably with KOH, NaOH or Ca (OH)
2solution is dispersant, and its concrete compound method is: according to mass percent concentration (3 ~ 10%) and the amount of solution to be prepared of highly basic in solution, calculates the quality of required highly basic, takes strong basic solid, add water, be stirred to it and dissolve completely, constant volume.Such as: to be the compound method of the NaOH solution of 5% be mass percent concentration: weighing sodium hydroxide 50g at ambient temperature, be placed in volumetric flask, add water and be stirred to NaOH and dissolve completely, be settled to 1000ml, namely obtain the NaOH solution that mass concentration is 5%.
Buffer solution: the preferred Na of embodiment
2hPO
4-NaH
2pO
4buffer solution, its compound method is: first compound concentration is the mother liquor of 0.2mol/L, namely prepares 0.2mol/LNa
2hPO
4solution and 0.2mol/LNaH
2pO
4solution, according to the pH value of required buffer solution, measures the Na of 0.2mol/L respectively
2hPO
4the NaH of solution and 0.2mol/L
2pO
4solution, mixing (if need the buffer solution of low concentration, also needing to dilute).Such as: pH value is the Na of 7
2hPO
4-NaH
2pO
4buffer solution is by mixing 61.0mL 0.2mol/LNa
2hPO
4with 39.0mL 0.2mol/LNaH
2pO
4and obtain.
Cleaning agent: pure water or distilled water.
Embodiment 1
The present invention disperses a kind of embodiment of the method for Pomacea canaliculata pieces of an egg, disperses the method for Pomacea canaliculata pieces of an egg to comprise the following steps described in the present embodiment:
(1) Pomacea canaliculata pieces of an egg to be disperseed are placed in the centrifuge tube that volume is 50mL, add the NaOH solution 6.0mL that mass percent concentration is 5%, shake up; Wherein, Pomacea canaliculata pieces of an egg are the pieces of an egg of laying eggs 7 hours, and its volume is 2.82cm
3, heavy 2.12g, add NaOH solution and Pomacea canaliculata pieces of an egg volume ratio be 2.1:1;
(2), after covering centrifuge tube lid, on constant-temperature table, at the uniform velocity shake (temperature is 28 DEG C, and shake speed is 80rpm) centrifuge tube and timing, after 79 seconds, Pomacea canaliculata pieces of an egg are separated into ovum grain completely;
(3) drawing out NaOH solution in centrifuge tube and chip with liquid-transfering gun, staying in container by disperseing the ovum grain obtained;
(4) in centrifuge tube, add 24mL pure water (volume ratio of the NaOH solution that pure water and step (1) add is 4:1), cleaning ovum grain once;
(5) in container, the Na that pH value is 7 is added
2hPO
4-NaH
2pO
4buffer solution 18mL, infiltrates ovum grain 4 minutes, the Na added
2hPO
4-NaH
2pO
4buffer solution and step (1) add NaOH solution volume ratio be 3:1;
(6) add 24mL pure water (volume ratio of the NaOH solution that pure water and step (1) add is 4:1) and clean ovum grain 2 times, the pure water of last cleaned ovum grain is that namely neutrality show that the pH value on spiral shell ovum surface is for neutral;
(7) dry ovum grain in 30 DEG C (difference of the temperature and environmental temperature of drying ovum grain is not more than 5 DEG C), drying the time used is 30 minutes.
Embodiment 2
The present invention disperses a kind of embodiment of the method for Pomacea canaliculata pieces of an egg, disperses the method for Pomacea canaliculata pieces of an egg to comprise the following steps described in the present embodiment:
(1) Pomacea canaliculata pieces of an egg to be disperseed are placed in test tube, add mass percent concentration be 3% KOH solution shake up, add KOH solution and Pomacea canaliculata pieces of an egg volume ratio be 8:1;
(2) manual shaking test tube is separated into ovum grain completely to Pomacea canaliculata pieces of an egg;
(3) adopting the KOH solution in the method removal test tube filtered and chip, staying in test tube by disperseing the ovum grain obtained;
(4) in test tube, add distilled water, the volume ratio of the KOH solution that distilled water and step (1) add is 3:1, and cleaning ovum grain once;
(5) in container, the Na that pH value is 6.0 is added
2hPO
4-NaH
2pO
4buffer solution, infiltrates ovum grain 3 minutes, the buffer solution added and step (1) add KOH solution volume ratio be 2:1;
(6) add distilled water cleaning ovum grain 2 times, the volume ratio of the KOH solution that the cumulative volume adding distilled water adds with step (1) is 3:1, and the distilled water of last cleaned ovum grain is neutrally namely show that the pH value on spiral shell ovum surface is for neutrality;
(7) ovum grain is dried in 20 ~ 25 DEG C.
Embodiment 3
The present invention disperses a kind of embodiment of the method for Pomacea canaliculata pieces of an egg, disperses the method for Pomacea canaliculata pieces of an egg to comprise the following steps described in the present embodiment:
(1) Pomacea canaliculata pieces of an egg to be disperseed are placed in centrifuge tube, add the Ca (OH) that mass percent concentration is 10%
2solution shakes up, and institute adds Ca (OH)
2the volume ratio of solution and Pomacea canaliculata pieces of an egg is 2:1;
(2) centrifuge tube that vibrates is separated into ovum grain completely to Pomacea canaliculata pieces of an egg;
(3) method by drawing removes the Ca (OH) in centrifuge tube
2solution and chip, stay in centrifuge tube by disperseing the ovum grain obtained;
(4) in centrifuge tube, pure water is added, the Ca (OH) that pure water and step (1) add
2the volume ratio of solution is 8:1, and cleaning ovum grain once;
(5) in container, the Na that pH value is 8.0 is added
2hPO
4-NaH
2pO
4buffer solution, infiltrate ovum grain 6 minutes, the buffer solution added and step (1) add Ca (OH)
2the volume ratio of solution is 1:1;
(6) add pure water cleaning ovum grain 2 times, add the Ca (OH) that the cumulative volume of pure water and step (1) add
2the volume ratio of solution is 8:1, and the pure water of last cleaned ovum grain is that namely neutrality show that the pH value on spiral shell ovum surface is for neutral;
(7) dry ovum grain in 20 ~ 30 DEG C, and the difference of the temperature and environmental temperature of drying ovum grain is not more than 5 DEG C.
Embodiment 4
The present invention disperses a kind of embodiment of the method for Pomacea canaliculata pieces of an egg, disperses the method for Pomacea canaliculata pieces of an egg to comprise the following steps described in the present embodiment:
(1) Pomacea canaliculata pieces of an egg to be disperseed are placed in centrifuge tube, add mass percent concentration be 6% NaOH solution shake up, add NaOH solution and Pomacea canaliculata pieces of an egg volume ratio be 5:1;
(2) centrifuge tube that vibrates is separated into ovum grain completely to Pomacea canaliculata pieces of an egg;
(3) by the NaOH solution in the method removal centrifuge tube of absorption and chip, stay in centrifuge tube by disperseing the ovum grain obtained;
(4) in centrifuge tube, add pure water, the volume ratio of the NaOH solution that pure water and step (1) add is 5:1, and cleaning ovum grain once;
(5) in container, the Na that pH value is 7.0 is added
2hPO
4-NaH
2pO
4buffer solution, infiltrates ovum grain 4 minutes, the buffer solution added and step (1) add NaOH solution volume ratio be 2:1;
(6) pure water cleaning ovum grain is added 2 times, the volume ratio adding the NaOH solution that the cumulative volume of pure water and step (1) add is 5:1, and the pure water of last cleaned ovum grain is neutrally namely show that the pH value on the ovum grain top layer disperseing to obtain is for neutral;
(7) dry ovum grain in 20 ~ 30 DEG C, and the difference of the temperature and environmental temperature of drying ovum grain is not more than 5 DEG C.Embodiment 5
In the method for dispersion Pomacea canaliculata pieces of an egg of the present invention, it is relevant with incubation period residing for Pomacea canaliculata ovum that Pomacea canaliculata pieces of an egg are separated into ovum grain required time by step (2) completely.When incubation period residing for Pomacea canaliculata ovum is different, the shortest time of the required dispersion treatment of the method for the invention step (2) differs greatly, and minimum jitter time needed for it and Pomacea canaliculata egg development hatch the corresponding relation of degree as shown in Figure 1.Wherein, the incubation cycle that Pomacea canaliculata is complete is 12 days.
As shown in Figure 1, the Pomacea canaliculata ovum in early development and latter stage is all easy to dispersion, and the Pomacea canaliculata ovum being in the hatching interstage needs the longer dispersion treatment time.
Embodiment 6
We have also investigated the effect adopting the method for the invention dispersion Pomacea canaliculata pieces of an egg, before and after dispersion, the state of Pomacea canaliculata ovum as shown in Figure 2.
As shown in Figure 2, the method for the invention is adopted Pomacea canaliculata pieces of an egg can be uniformly dispersed.Further, we also find in research, and before being in hatching, the spiral shell pieces of an egg in each stage can keep ovum grain complete after dispersion treatment, and the Pomacea canaliculata ovum grain after dispersion can go out young spiral shell and incubation rate still can reach more than 80% by normal incubation.
Finally to should be noted that; above embodiment is only in order to illustrate technical scheme of the present invention but not limiting the scope of the invention; although be explained in detail the present invention with reference to preferred embodiment; those of ordinary skill in the art is to be understood that; can modify to technical scheme of the present invention or equivalent replacement, and not depart from essence and the scope of technical solution of the present invention.
Claims (10)
1. disperse a method for Pomacea canaliculata pieces of an egg, it is characterized in that: comprise the following steps:
(1) Pomacea canaliculata pieces of an egg to be disperseed are placed in container, add dispersant and shake up, add dispersant and Pomacea canaliculata pieces of an egg volume ratio be 2 ~ 8:1;
(2) vibrate or rock container and be separated into ovum grain completely to Pomacea canaliculata pieces of an egg;
(3) removing the dispersant in container and chip, staying in container by disperseing the ovum grain obtained;
(4) in container, cleaning agent is added, cleaning ovum grain;
(5) in container, add the buffer solution that pH value is 6.0 ~ 8.0, infiltrate ovum grain at least 3 minutes, the buffer solution added and step (1) add dispersant volume ratio be 1 ~ 3:1;
(6) cleaning agent cleaning ovum grain is added;
(7) dry or dry ovum grain.
2. the method for dispersion Pomacea canaliculata pieces of an egg as claimed in claim 1, it is characterized in that: described dispersant is the strong base solution do not reacted with Pomacea canaliculata chorion, in this solution, the mass percent concentration of highly basic is 3 ~ 10%.
3. the method for dispersion Pomacea canaliculata pieces of an egg as claimed in claim 2, is characterized in that: described dispersant is KOH, NaOH or Ca (OH)
2solution.
4. the method for dispersion Pomacea canaliculata pieces of an egg as claimed in claim 1, is characterized in that: described buffer solution not with saccharide compound and calcium carbonate reaction.
5. the method for the dispersion Pomacea canaliculata pieces of an egg as described in claim 1 or 4, is characterized in that: described buffer solution is Na
2hPO
4-NaH
2pO
4buffer solution.
6. the method for dispersion Pomacea canaliculata pieces of an egg as claimed in claim 1, is characterized in that: described cleaning agent is water.
7. the method for dispersion Pomacea canaliculata pieces of an egg as claimed in claim 1, it is characterized in that: in described step (1), container is cylindrical.
8. the method for dispersion Pomacea canaliculata pieces of an egg as claimed in claim 1, is characterized in that: in described step (3), removes dispersant in container and chip by the method for drawing or filtering.
9. the method for dispersion Pomacea canaliculata pieces of an egg as claimed in claim 1, is characterized in that: in described step (4) and step (6), the volume ratio of the dispersant that cleaning agent and step (1) add is 3 ~ 8:1.
10. the method for dispersion Pomacea canaliculata pieces of an egg as claimed in claim 1, is characterized in that: in described step (7), dry ovum grain or dry ovum grain in 20 ~ 30 DEG C, and the difference of the temperature and environmental temperature of drying ovum grain is not more than 5 DEG C in 20 ~ 25 DEG C.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108834959A (en) * | 2018-07-06 | 2018-11-20 | 浙江海洋大学 | One kind drags ovum type Copepods ovum grain to be detached from method |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5133289A (en) * | 1991-09-05 | 1992-07-28 | Cornell Research Foundation, Inc. | Artificial system and method for breeding fleas |
| CN1675993A (en) * | 2004-04-01 | 2005-10-05 | 浙江省淡水水产研究所 | Nanjing fairy shrimp egg proliferating method |
| CN101831400A (en) * | 2009-03-09 | 2010-09-15 | 中国水产科学研究院东海水产研究所 | Method for collecting and separating medaka zygotes |
| CN102077802A (en) * | 2010-12-31 | 2011-06-01 | 刘立明 | Parent fish breeding and hatching method for greenlings |
| CN102106316A (en) * | 2010-12-29 | 2011-06-29 | 大连医科大学附属第一医院 | Cultivation method of lucilia sericata larva and disinfection method thereof |
| JP2013141431A (en) * | 2012-01-10 | 2013-07-22 | Ono Doboku Kogyo Ltd | Method for preventing oviposition of apple snail |
| CN103750035A (en) * | 2014-01-03 | 2014-04-30 | 中国水产科学研究院珠江水产研究所 | Method for causing bodies of ornamental fishes to go red by utilizing eggs of ampullaria gigas |
| KR20140084484A (en) * | 2012-12-26 | 2014-07-07 | 강창석 | Reef block |
| CN203735291U (en) * | 2014-01-03 | 2014-07-30 | 中国水产科学研究院珠江水产研究所 | Apple snail egg collection device |
| CN203985551U (en) * | 2014-05-27 | 2014-12-10 | 张俊彬 | Fish-egg separator |
-
2015
- 2015-04-09 CN CN201510165586.1A patent/CN104823882B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5133289A (en) * | 1991-09-05 | 1992-07-28 | Cornell Research Foundation, Inc. | Artificial system and method for breeding fleas |
| CN1675993A (en) * | 2004-04-01 | 2005-10-05 | 浙江省淡水水产研究所 | Nanjing fairy shrimp egg proliferating method |
| CN101831400A (en) * | 2009-03-09 | 2010-09-15 | 中国水产科学研究院东海水产研究所 | Method for collecting and separating medaka zygotes |
| CN102106316A (en) * | 2010-12-29 | 2011-06-29 | 大连医科大学附属第一医院 | Cultivation method of lucilia sericata larva and disinfection method thereof |
| CN102077802A (en) * | 2010-12-31 | 2011-06-01 | 刘立明 | Parent fish breeding and hatching method for greenlings |
| JP2013141431A (en) * | 2012-01-10 | 2013-07-22 | Ono Doboku Kogyo Ltd | Method for preventing oviposition of apple snail |
| KR20140084484A (en) * | 2012-12-26 | 2014-07-07 | 강창석 | Reef block |
| CN103750035A (en) * | 2014-01-03 | 2014-04-30 | 中国水产科学研究院珠江水产研究所 | Method for causing bodies of ornamental fishes to go red by utilizing eggs of ampullaria gigas |
| CN203735291U (en) * | 2014-01-03 | 2014-07-30 | 中国水产科学研究院珠江水产研究所 | Apple snail egg collection device |
| CN203985551U (en) * | 2014-05-27 | 2014-12-10 | 张俊彬 | Fish-egg separator |
Non-Patent Citations (3)
| Title |
|---|
| 吴帆 等: "入侵生物福寿螺抗逆性的研究进展", 《生物安全学报》 * |
| 罗渡 等: "不同生境福寿螺(Pomaceaca naliculata)的营养成分及其利用价值", 《生物学杂志》 * |
| 陈泽柠 等: "辣椒碱对外来入侵种福寿螺的急性毒性作用", 《湖北农业科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108834959A (en) * | 2018-07-06 | 2018-11-20 | 浙江海洋大学 | One kind drags ovum type Copepods ovum grain to be detached from method |
| CN108834959B (en) * | 2018-07-06 | 2021-01-05 | 浙江海洋大学 | Method for separating eggs of dragging-egg type copepods |
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|---|---|
| CN104823882B (en) | 2017-08-01 |
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