CN106259081A - A kind of Apostichopus japonicus sex and the discrimination method of maturity of fish gonads - Google Patents
A kind of Apostichopus japonicus sex and the discrimination method of maturity of fish gonads Download PDFInfo
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- CN106259081A CN106259081A CN201610648409.3A CN201610648409A CN106259081A CN 106259081 A CN106259081 A CN 106259081A CN 201610648409 A CN201610648409 A CN 201610648409A CN 106259081 A CN106259081 A CN 106259081A
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- Prior art keywords
- collection tube
- apostichopus japonicus
- tissue
- radix morinae
- morinae bulleyanae
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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
- 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
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Abstract
A kind of Apostichopus japonicus sex and the discrimination method of maturity of fish gonads, belong to technical field of aquaculture, the present invention uses negative-pressure type tissue harvester to obtain the partial glandular tissue of Radix Morinae Bulleyanae after anesthesia, according to the color of Radix Morinae Bulleyanae male and female gonad and wherein sex and the maturity of fish gonads of Radix Morinae Bulleyanae are identified by the kind of sexual cell, size, quantity and architectural feature.Utilizing the method to identify sex and the maturity of fish gonads of Radix Morinae Bulleyanae, wound is little, it is not necessary to dissect Radix Morinae Bulleyanae parent, reduces parent loss and economic loss.
Description
Technical field
The invention belongs to technical field of aquaculture, be specifically related to a kind of Apostichopus japonicus sex and the discriminating of maturity of fish gonads
Method.
Background technology
Apostichopus japonicus, also known as Radix Morinae Bulleyanae, is top-quality one in Chinese 20 multiple eating Stichopus japonicuss, is listed in " marine products eight delicacies "
One of, cultivated the most widely in China.It is right to need during Radix Morinae Bulleyanae artificial increasing cultivation, stock breeding and scientific experiments
The male and female sex of Radix Morinae Bulleyanae and maturity of fish gonads differentiate.But Radix Morinae Bulleyanae male and female there is no significant difference in appearance, and gonad
Maturity is from apparent None-identified, thus the discriminating to Radix Morinae Bulleyanae sex and maturity of fish gonads is in Radix Morinae Bulleyanae artificial breeding work process
Difficult point.In the past, could identify, along with Radix Morinae Bulleyanae is artificial after Radix Morinae Bulleyanae sex can only be used dissection parent and parent row's essence ovulation
The development of raising technology, ultrasonic technology is also applied in Radix Morinae Bulleyanae sex identification (application number: 201410142943.8).By
In Radix Morinae Bulleyanae live body sex apparent qualification difficulty, male and female are difficult to differentiate, and when carrying out Radix Morinae Bulleyanae artificial breeding, need cultivation and accelerating big
The parent of amount is to ensure breeding effect, if hero is many, then germ cell is few, abnormal rate is high, and needs germ cell is carried out manual cleaning；
If female many, then rate of fertilization is low, reduces emergence rate, and these the most greatly increase the amount of labour during artificial ripening and produce into
This.Maturity of fish gonads is difficult to judge simultaneously, and utilizes the method dissecting parent to observe the maturity of fish gonads of Radix Morinae Bulleyanae, increases
Economic loss during Radix Morinae Bulleyanae artificial breeding.Thus seek the accurately practical discrimination method of a kind of Radix Morinae Bulleyanae sex to reducing thorn
Production cost during ginseng artificial breeding has important meaning.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of Apostichopus japonicus sex and the discrimination method of maturity of fish gonads, this
Invention uses negative-pressure type tissue harvester to obtain the partial glandular tissue of Radix Morinae Bulleyanae after anesthesia, according to the color of Radix Morinae Bulleyanae male and female gonad and
Wherein sex and the maturity of fish gonads of Radix Morinae Bulleyanae are identified by the kind of sexual cell, size, quantity and architectural feature.Utilizing should
Sex and the maturity of fish gonads of Radix Morinae Bulleyanae are identified by method, and wound is little, it is not necessary to dissect Radix Morinae Bulleyanae parent, reduce parent loss and warp
Ji loss.
The present invention completes according to following operating technology:
A kind of negative-pressure type tissue harvester, it includes push rod, piston, cylinder, fixed cover, collection tube, trocar sheath and collection
Protection of pipe set；Wherein push rod is connected with piston, is placed in cylinder, and collection tube is side-suction type, and collection tube one end is sharp-pointed, gathers tip
Sharp end side has aperture, and aperture lower limb is sharp blade structure；Collection tube is connected with cylinder body bottom by fixed cover, adopts
Collector sharp end is paperwrapped in outside collection tube away from cylinder, trocar sheath, and trocar sheath can slide up and down along collection tube, collection tube
Protection set is positioned at outside trocar sheath, and protection collection tube damages from external force.
Further, collection tube specification used by described negative-pressure type tissue harvester is: external diameter 0.81mm, internal diameter 0.5mm, length
60mm。
A kind of Apostichopus japonicus sex and the discrimination method of maturity of fish gonads, adopt including Apostichopus japonicus anesthesia, Apostichopus japonicus gonadal tissue
Collection and Apostichopus japonicus three steps of recovery；
Described Apostichopus japonicus is anaesthetized: with clean sea water configuration 20g/l MgCl2Solution, Apostichopus japonicus to be detected is put into containing
In the tank of sea water, make sea water not have Radix Morinae Bulleyanae, in tank, be then continuously added the 20g/l MgCl configured2Solution, makes
MgCl2The final concentration of 10g/l of solution, is constantly slowly stirred in adition process, makes MgCl2Solution is uniformly distributed, and anesthesia duration is
30min～1h, unfolds to Apostichopus japonicus tentacle naturally with health, stimulates reactionless to external world；
Described tissue obtaining method: the negative-pressure type tissue harvester described in utilization is acquired, takes off negative-pressure type tissue and adopts
The collection tube protection set of storage, at Apostichopus japonicus gonad gonopore rear 2-3cm, dorsal part is acquired to anus end, and collection tube is tight
Inside patch apostichopus japonicus body-wall, outwards twitch push rod 1-2cm, in making cylinder, produce negative pressure, treat that gonadal tissue sucks collection tube inner chamber
After, push down on trocar sheath, make in-vivo tissue separate with collection tube lumenal tissue；Collection tube and trocar sheath are extracted out, will gather
Gonadal tissue be placed on microscope slide and carry out microexamination；
Described Apostichopus japonicus is recovered: is put into by the Apostichopus japonicus after sampling in the sea water of cleaning and supports temporarily, until Apostichopus japonicus is recovered also
The most movable.
Further, during negative-pressure type tissue harvester collection tissue used, aperture is downward, and collection tube is close in Radix Morinae Bulleyanae body wall
Side.
Further, after gonadal tissue enters collection tube inner chamber under suction function, trocar sheath need to be pushed down on, make trocar sheath
Lower limb exceed the lower limb of collection tube aperture, described lower limb refers to the tip side near collection tube.
Apostichopus japonicus male and female sex characteristics be female gonads be faint yellow or Chinese red, with maturity of fish gonads increase and gradually add
Deeply, internal reproductive cell is oogonium and oocyte, mature egg seen from the period of maturation.The female gonads in period of maturation is Exocarpium Citri Rubrum
Color, also there is mature egg in internal full oocyte, more than oocyte diameter 90um, oocyte and germ-vesicle profile
Clearly, germ-vesicle is big, accounts for the 30%-50% of oocyte volume.Male gonad internal reproductive cell is that spermatogonium, essence are female thin
Born of the same parents and spermatid, the male gonad in period of maturation is milky, internal full substantial amounts of sperm.Study according to forefathers, Radix Morinae Bulleyanae
Gonad development degree, can be divided into four periods by Radix Morinae Bulleyanae maturity of fish gonads: resting stage, trophophase, period of maturation and draining period, according to
The maturity of fish gonads of Radix Morinae Bulleyanae can be made a determination by the feature in each period.
Present invention beneficial effect compared with prior art:
The inventive method is by conscientiously observing color and the wherein kind of sexual cell, size, the quantity of the gonad of Radix Morinae Bulleyanae
And sex and the maturity of fish gonads thereof of Radix Morinae Bulleyanae identified by architectural feature.Use the method to the sex of Radix Morinae Bulleyanae and gonad maturity
Degree carries out qualification and reaches Wicresoft, need not dissect Radix Morinae Bulleyanae parent, reduces parent loss, reaches to reduce the mesh of economic loss
's.Identifying Radix Morinae Bulleyanae sex and maturity of fish gonads by this method, reaching precise and high efficiency, technology is easily grasped, and has Radix Morinae Bulleyanae nursery
The skilled industrial worker of experience can identify, accuracy is up to 100%.Operability of the present invention is good, is equipped with negative-pressure type tissue
Harvester, volume is little, portable, easily buys and, low cost, can repeated multiple times use after cleaning and sterilizing.
Accompanying drawing explanation
Fig. 1 is the structural representation of negative-pressure type tissue harvester: 1, push rod, 2, piston, 3, cylinder, 4, fixed cover, 5, adopt
Collector, 6, trocar sheath, 7, collection tube protection set, 8, aperture.
Detailed description of the invention
The technical method of the present invention is described in detail below by embodiment:
Embodiment 1:
Negative-pressure type tissue harvester, as it is shown in figure 1, it includes push rod 1, piston 2, cylinder 3, fixed cover 4, collection tube 5, outer
Sleeve pipe 6 and collection tube protection set 7；Wherein push rod 1 is connected with piston 2, is placed in cylinder 3, and collection tube 5 is side-suction type, collection tube
One end is sharp-pointed, and collection tube sharp end side has aperture 8, and aperture lower limb is sharp blade structure；Collection tube 5 is by fixing
Set 4 is connected bottom cylinder 3, and collection tube sharp end is paperwrapped in outside collection tube 5 away from cylinder, trocar sheath 6, and trocar sheath can
Sliding up and down along collection tube, collection tube protection set 7 is positioned at outside trocar sheath 6, and protection collection tube 5 damages from external force.Used negative
The impact on Radix Morinae Bulleyanae sex identification of the collection tube specification of pressure type tissue harvester.
Experiment use 25G4# (0.4mm*0.2mm*40mm, external diameter * internal diameter * length, lower with), 23G6# (0.64mm*
0.36mm*60mm), the adopting of 21G8# (0.81mm*0.5mm*60mm) and 19G10# (1.06mm*0.72mm*60mm) 4 kinds of specifications
The sex collecting the effective this method Radix Morinae Bulleyanae (207 ± 18g) to having carried out artificial ripening in 120 same culturing pools is identified,
Every kind of specification collection tube identifies 30, records qualification result, after qualification, dissects and has identified Radix Morinae Bulleyanae, and confirmatory experiment result is real
Test and the results are shown in Table 1.
The impact on Radix Morinae Bulleyanae sex identification of the table 1 different size collection tube
According to experimental result, through repeatedly testing certification, utilize the method for the present invention that Radix Morinae Bulleyanae male and female sex is identified
The suitableeest collection tube specification is external diameter 0.81mm, internal diameter 0.5mm, and accuracy rate is up to 100%.
The impact on Radix Morinae Bulleyanae sex identification of embodiment 2 different development stage
Utilize the inventive method (collection tube specification external diameter 0.81mm, internal diameter 0.5mm), respectively in November, 2015,2016
Radix Morinae Bulleyanae (256 ± 15g) sex in Weihai, Yantai cultivating pool is identified by March in year, May and August, and each time respectively takes
30 parent participations.Record qualification result, dissects Radix Morinae Bulleyanae after qualification, confirmatory experiment result, and experimental result is shown in Table 2.
The impact on Apostichopus japonicus sex identification of table 2 different development stage
According to experimental result, through certification repeatedly, the inventive method is accurate to the sex identification growing Apostichopus japonicus gonad
Rate is up to 100%.
The impact on Apostichopus japonicus male and female sex identification of the embodiment 3 different acquisition position
Utilize the inventive method (collection tube specification external diameter 0.81mm, internal diameter 0.5mm), respectively from Radix Morinae Bulleyanae gonad position
Dorsal part is anterior, the sex of Radix Morinae Bulleyanae is identified by middle part and position, three, rear portion, in the middle part of described dorsal part namely Apostichopus japonicus
At gland gonopore rear 2-3cm, dorsal part gathers gonad, each collection location detection Radix Morinae Bulleyanae 30 to anus end, and specification is 236 ±
27g, records experimental result, identifies and dissects Apostichopus japonicus after terminating, and being authenticated experimental result, experimental result is shown in Table 3.
According to experimental result, through certification repeatedly, from imitative thorn when utilizing the inventive method that Apostichopus japonicus sex is identified
Gathering in the middle part of the dorsal part of ginseng gonad position and be obtained in that preferable Detection results, accuracy rate is up to 100%.
The impact on Radix Morinae Bulleyanae sex identification of the table 3 different acquisition position
Embodiment 4
In April, 2016 carries out Apostichopus japonicus male and female sex identification in Rushan, Shandong Province nursery factory, step include Apostichopus japonicus anesthesia,
Apostichopus japonicus gonadal tissue gathers and Apostichopus japonicus three steps of recovery；
Described Apostichopus japonicus is anaesthetized: with clean sea water configuration 20g/l MgCl2Solution, Apostichopus japonicus to be detected is put into containing
In the tank of sea water, make sea water not have Radix Morinae Bulleyanae, in tank, be then continuously added the 20g/l MgCl configured2Solution, makes
MgCl2The final concentration of 10g/l of solution, is constantly slowly stirred in adition process, makes MgCl2Solution is uniformly distributed, and anesthesia duration is
30min, unfolds to Apostichopus japonicus tentacle naturally with health, stimulates reactionless to external world；
Described tissue obtaining method: take off the collection tube protection set of negative-pressure type tissue harvester, from Apostichopus japonicus gonad reproduction
At the 2cm of rear, hole, dorsal part is acquired to anus end, and collection tube is close to inside apostichopus japonicus body-wall, outwards twitches push rod 1cm, makes cylinder
Internal generation negative pressure, after gonadal tissue sucks collection tube inner chamber, pushes down on trocar sheath, makes in-vivo tissue and collection tube inner chamber
Separate tissue；Collection tube and trocar sheath are extracted out, the gonadal tissue of collection is placed on microscope slide and carries out microexamination；
Described Apostichopus japonicus is recovered: is put into by the Apostichopus japonicus after sampling in the sea water of cleaning and supports temporarily, until Apostichopus japonicus is recovered also
The most movable.
Evaluation apparatus is the negative-pressure type tissue harvester of sterilizing, outfit external diameter 0.81mm, and the collection tube of internal diameter 0.5mm is right
20 Radix Morinae Bulleyanaes of weight in wet base 150g to 300g carry out sex identification, and gathering position is in the middle part of the dorsal part of Radix Morinae Bulleyanae gonad position, also
At i.e. Apostichopus japonicus gonad gonopore rear 2cm, dorsal part also records Radix Morinae Bulleyanae sex and maturity of fish gonads to anus end, microexamination.Right
Radix Morinae Bulleyanae after qualification is dissected, confirmatory experiment result, identifies that accuracy is up to 100%.
Embodiment 5
In April, 2016 carries out Radix Morinae Bulleyanae male and female sex identification in Yantai, Shandong nursery factory, and authentication method is with embodiment 4, mirror
Locking equipment is the self-control negative-pressure type tissue harvester of sterilizing, is equipped with external diameter 0.81mm, and the collection tube of internal diameter 0.5mm, to carrying out
The weight in wet base of artificial ripening is that 10 Radix Morinae Bulleyanaes of 200g to 300g carry out sex identification, and gathering position is Radix Morinae Bulleyanae gonad position
In the middle part of dorsal part, namely at Apostichopus japonicus gonad gonopore rear 2cm, dorsal part gathers gonad to anus end, and microexamination also records Radix Morinae Bulleyanae
Sex and maturity of fish gonads.Radix Morinae Bulleyanae after identifying is dissected, verifies qualification result, identify that accuracy is up to 100%.
Sample result shows, when selecting suitable collection tube specification, suitably collection position and suitable gonad development
The sex of Radix Morinae Bulleyanae and maturity of fish gonads can be identified by the phase accurately, use the inventive method to Radix Morinae Bulleyanae sex and gonad maturity
Degree carries out identifying conveniently, low cost, and to Radix Morinae Bulleyanae Wicresoft, does not cause parent participation to lose, and precise and high efficiency, technology is easily grasped, permissible
It is widely used in the sex in Radix Morinae Bulleyanae artificial breeding and maturity of fish gonads is identified.
Claims (5)
1. a negative-pressure type tissue harvester, it is characterised in that it include push rod (1), piston (2), cylinder (3), fixed cover (4),
Collection tube (5), trocar sheath (6) and collection tube protection set (7)；Wherein push rod (1) is connected with piston (2), is placed in cylinder (3),
Collection tube (5) is side-suction type, and collection tube one end is sharp-pointed, and collection tube sharp end side has aperture (8), and aperture lower limb is sharp
Blade structure；Collection tube (5) is connected with cylinder (3) bottom by fixed cover (4), and collection tube sharp end is away from cylinder, overcoat
Pipe (6) is paperwrapped in collection tube (5) outside, and trocar sheath can slide up and down along collection tube, and collection tube protection set (7) is positioned at overcoat
Pipe (6) outside, protection collection tube (5) damages from external force.
A kind of negative-pressure type tissue harvester the most according to claim 1, it is characterised in that described negative-pressure type tissue harvester
Collection tube specification used is: external diameter 0.81mm, internal diameter 0.5mm, length 60mm.
3. an Apostichopus japonicus sex and the discrimination method of maturity of fish gonads, it is characterised in that it includes Apostichopus japonicus anesthesia, Apostichopus japonicus
Gonadal tissue gathers and Apostichopus japonicus three steps of recovery；
Described Apostichopus japonicus is anaesthetized: with clean sea water configuration 20g/l MgCl2Solution, puts into Apostichopus japonicus to be detected containing sea water
Tank in, make sea water not have Radix Morinae Bulleyanae, in tank, be then continuously added the 20g/lMgCl configured2Solution, makes MgCl2Solution
Final concentration of 10g/l, is constantly slowly stirred in adition process, makes MgCl2Solution is uniformly distributed, and anesthesia duration is 30min～1h,
Naturally unfold with health to Apostichopus japonicus tentacle, stimulate reactionless to external world；
Described Apostichopus japonicus gonadal tissue acquisition method: utilize the negative-pressure type tissue harvester described in claim 1 or 2 to adopt
Collection, takes off the collection tube protection set of negative-pressure type tissue harvester, and at Apostichopus japonicus gonad gonopore rear 2-3cm, dorsal part is to anus
End is acquired, and collection tube is close to inside apostichopus japonicus body-wall, outwards twitches push rod 1-2cm, produces negative pressure, treat gonad in making cylinder
After tissue sucks collection tube inner chamber, push down on trocar sheath, make in-vivo tissue separate with collection tube lumenal tissue；By collection tube and
Trocar sheath is extracted out, the gonadal tissue of collection is placed on microscope slide and carries out microexamination；
Described Apostichopus japonicus is recovered: the Apostichopus japonicus after sampling is put in clean sea water and supported temporarily, until Apostichopus japonicus recovery is the most certainly
By activity.
Discrimination method the most according to claim 3, it is characterised in that during negative-pressure type tissue harvester collection tissue used, little
Hole is downward, and collection tube is close to inside Radix Morinae Bulleyanae body wall.
Discrimination method the most according to claim 3, it is characterised in that gonadal tissue enters in collection tube under suction function
Behind chamber, need to push down on trocar sheath, make the lower limb of trocar sheath exceed the lower limb of collection tube aperture, described lower limb refers to
Tip side near collection tube.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610648409.3A CN106259081B (en) | 2016-08-09 | 2016-08-09 | A kind of discrimination method of imitative stichopus japonicus gender and maturity of fish gonads |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610648409.3A CN106259081B (en) | 2016-08-09 | 2016-08-09 | A kind of discrimination method of imitative stichopus japonicus gender and maturity of fish gonads |
Publications (2)
| Publication Number | Publication Date |
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| CN106259081A true CN106259081A (en) | 2017-01-04 |
| CN106259081B CN106259081B (en) | 2019-03-29 |
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ID=57667353
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| CN201610648409.3A Expired - Fee Related CN106259081B (en) | 2016-08-09 | 2016-08-09 | A kind of discrimination method of imitative stichopus japonicus gender and maturity of fish gonads |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4919146A (en) * | 1988-10-25 | 1990-04-24 | Medrad, Inc. | Biopsy device |
| WO1993004631A1 (en) * | 1991-09-09 | 1993-03-18 | Sherwood Medical Company | Actuated biopsy cutting needle with removable stylet |
| CN202005769U (en) * | 2011-01-08 | 2011-10-12 | 吴新奎 | Tumor biopsy sampler |
| CN203591278U (en) * | 2013-11-09 | 2014-05-14 | 邢红梅 | Needle-tube-type living body sampler for tumour |
| CN204562230U (en) * | 2015-03-27 | 2015-08-19 | 吕静 | A kind of tumor needle tube type sampler |
-
2016
- 2016-08-09 CN CN201610648409.3A patent/CN106259081B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4919146A (en) * | 1988-10-25 | 1990-04-24 | Medrad, Inc. | Biopsy device |
| WO1993004631A1 (en) * | 1991-09-09 | 1993-03-18 | Sherwood Medical Company | Actuated biopsy cutting needle with removable stylet |
| CN202005769U (en) * | 2011-01-08 | 2011-10-12 | 吴新奎 | Tumor biopsy sampler |
| CN203591278U (en) * | 2013-11-09 | 2014-05-14 | 邢红梅 | Needle-tube-type living body sampler for tumour |
| CN204562230U (en) * | 2015-03-27 | 2015-08-19 | 吕静 | A kind of tumor needle tube type sampler |
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| CN106259081B (en) | 2019-03-29 |
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Granted publication date: 20190329 Termination date: 20190809 |