CN101803581B - A method for cultivating large nucleated pearls - Google Patents
A method for cultivating large nucleated pearls Download PDFInfo
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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
The invention belongs to the technical field of cultivation, and relates to a method for cultivating large granular nuclear pearls. In order to solve the problem that in a traditional process for cultivating nuclear pearls, after pearl cores are inserted, pearl cultivation creature (e.g. freshwater mussel and the like) generally has high pearl production rate, the invention provides a method for cultivating large granular nuclear pearls. The method comprises the following steps that: a. outer mantle epithelial cells are obtained from mussel body; b. the pearl cores are incubated from the obtained outer mantle epithelial cells, and incubated pearl cores are obtained; c. the incubated pearl cores are implanted into pearl cultivating mussel to be cultivated, wherein, step c is divided into the following sub-steps that: (1) a sac for containing the incubated pearl cores is arranged in the pearl cultivating mussel; (2) the incubated pearl cores are stuffed into the sac, and the opening of the sac is sutured by a biological suture; and (3) the pearl cultivating mussel treated in step (2) is cultivated. The method of the invention can smoothly solve the technical problem, so that the diameter of the cultivated pearls is generally above 10mm.
Description
Technical field
The invention belongs to the cultural technique field, relate to a kind of method of cultivating nucleated pearl, the method of the invention is used for cultivating diameter and examines on 8-10mm the pearl that hatches, thereby can make the pearl that cultivates have larger diameter (generally can reach more than the 10mm), employed cultivation biology is generally freshwater shellfish, such as hydriopsis cumingii etc.
Background technology
China's freshwater resources are abundant, and freshwater shellfish is grown cultured pearls and had a extensive future.At present, more than China's fresh water pearl annual production kiloton, but hundred tons of high-qualitys, high-grade pearl less than, diameter is that the above pearl of 8mm is difficult to satisfy the requirement of international market in a short time.The size of pearl particles has determined the economic benefit that pearl produces in a lot of degree.Be still a difficult problem for freshwater shellfish bulky grain pearl cultivating in production.Over nearly 10 years, the theoretical foundation of artificial pearl culture technique has obtained new development: " cultivation of nucleus pearl by using free cell implantation method " reaches " visceral mass grow cultured pearls technology " and obtained breakthrough cultivating fresh water genuine pearl with pearl, for the cultivation of high quality pearl provides good Basic of Biology, also for the bulky grain pearl cultivating space is provided may.But bulky grain pearl nuclear is inserted and is told the high and wound of pearl rate behind the nuclear and be difficult to heal and troubling the cultivation of large granular nuclear pearls always, in the urgent need to carrying out technological innovation in production technology, reduces and inserts the risk that nuclear is performed the operation and brought to pearl sector.
Summary of the invention
Technical problem underlying to be solved by this invention is in the process of existing visceral mass cultivation nucleated pearl, after inserting pearl nuclear, pearl cultivating biological (such as hydriopsis cumingii etc.) generally has the problem that height is told the pearl rate, and provides a kind of method of cultivating nucleated pearl for this problem.
The technical solution adopted in the present invention is:
A kind of method of cultivating large granular nuclear pearls comprises: a, obtain the outer embrane exocuticle from the freshwater mussel body, b, hatch pearl nuclear with the outer embrane exocuticle that obtains, obtain hatching pearl nuclear, c, will hatch in the pearl renucleation pearl culturing clam and cultivate; The described cultivation of c step comprises:
1., in pearl culturing clam, open one and be used for placing the pouch of hatching pearl nuclear;
2., pearl nuclear be will hatch and pouch, the biological suture of pouch opening part filled in;
3., the pearl culturing clam after 2. step process is cultivated.
The pearl rate of telling of pearl cultivating biology, if on time course, two kinds of situations before and after can being divided into, the previous case refers to, fills in the process of pouch hatching pearl nuclear, tells pearl phenomenon (namely hatch pearl examine be difficult to fill in pouch); Latter event is, hatching after pearl nuclear fills in pouch, tells the pearl phenomenon.These two kinds of phenomenons all can cause the pearl rate of telling of pearl cultivating biology to increase, and work brings difficulty to pearl cultivating.
Prior art is when running into above-mentioned difficulties of the present invention, the problem that the pearl rate of telling of generally can only conscious solution latter event bringing improves, the scheme of taking normally reduces the pearl nuclear diameter of inserting, although this way can solve the problem that the pearl rate of telling that latter event brings improves to a certain extent, but this scheme has been brought new technical problem when dealing with problems, the pearl nuclear diameter of namely inserting reduces, the pearl particles that cultivates is also just smaller, does not satisfy the demand of international market.
Solution thinking of the present invention is to tell two high aspects of pearl rate for the pearl cultivating biology simultaneously to be in full swing, on the one hand, for latter event, the technological means that the present invention sews up by biology can fundamentally be stopped pearl culturing clam after inserting pearl nuclear, tells the pearl phenomenon; On the other hand, for the previous case, the present invention's process research discovery, the opening part of hatching pearl nuclear pouch are not (take bivalves as example, cultivating organ is the visceral mass of freshwater mussel body) that is arranged on the cultivation organ of pearl cultivating biology, such organ does not often have good contraction of muscle performance, hatch after pearl nuclear fills in wherein, the pearl nuclear saturation of pouch opening part does not have much variations, in this case, hatch pearl and be easy to give prominence to from the pouch opening part, and finally break away from the freshwater mussel body.The present invention further finds, take bivalves as example, the intersection of freshwater mussel body visceral mass and abdominal foot has preferably contraction of muscle performance, therefore, the pouch opening part is arranged on this position, can increase greatly the pearl nuclear saturation of pouch opening part, hatch pearl nuclear and fill in the pouch process and become easy thereby make.
Use technical scheme of the present invention not only can solve the biological high technical problem of telling the pearl rate of pearl cultivating, but also obtained an additional technique effect, that is exactly the pouch opening part owing to can use biological suture, therefore the pouch opening part can be opened greatly (not affecting nucleated pearl cultivates under the prerequisite of effect) as much as possible, examine to hold the larger pearl that hatches of diameter, thereby create primary condition for cultivating large diameter pearl, in general, diameter of hatching pearl nuclear of the present invention is 8-10mm.
Certainly, after the biological suture, filling in mouthful (being the pouch opening part) also needs to carry out necessary processing, mouthful is unlikely to be infected and produces pathology so that fill in, thereby have a strong impact on the pearl cultivating effect, perhaps causes the death of pearl culturing clam.
For this reason, the c step in the inventive method is in 2. step with 3. also set up the cleaning and sterilizing step between the step:
Clean at the 2. pouch opening part of step with antibiotic first, then will urge muscle and produce on the pouch opening part that medicine is applied to 2. step so that the muscle Fast Restoration of pouch opening part.
Description of drawings
Fig. 1 is the described pearl nuclear that will adhere to the outer embrane cell of embodiment is filled in visceral mass pouch depths with tweezers operation chart.
Embodiment
In order to make those skilled in the art well understand concrete technical characterictic of the present invention, be beneficial to enforcement of the present invention, it is existing that the present invention will be further explained.
A kind of method of cultivating large granular nuclear pearls, comprise: the first, obtain the outer embrane exocuticle from the freshwater mussel body, the second, hatch pearl nuclear with the outer embrane exocuticle that obtains, obtain hatching pearl nuclear, the 3rd, will hatch in the pearl renucleation pearl culturing clam and cultivate.
Wherein comprise again some little steps in each step, for third step, it can be divided into again:
1, in pearl culturing clam, opens one and be used for placing the pouch of hatching pearl nuclear;
2, pearl nuclear be will hatch and pouch, the biological suture of pouch opening part filled in;
3, the pearl culturing clam after 2 step process is cultivated.
What the technical staff can understand learns from this specification summary of the invention, a, the b of the respectively corresponding summary of the invention part of first, second, third, 1,2 and 3 steps above-mentioned, c, 1., 2. and 3. step, the technical staff can be applied to this embodiment for the improved technical schemes that produce the relevant art effect and provide with related various of summary of the invention part.
The main purpose of this embodiment is exactly on the basis of above technical scheme, technical scheme is further expanded again or improves, and realizes better technique effect to help those skilled in the art.
One, this embodiment is carried out further refinement to first step, and the first step after the refinement comprises:
I, separate outer embrane exocuticle tissue from the freshwater mussel body;
II, outer embrane exocuticle tissue is put into trypsinization liquid digestion, collect the outer embrane exocuticle.
In order to reduce harmful pathogens such as bacterium to the infection of outer embrane exocuticle tissue, can further rinsing step be set between I step and II step:
Mixing washing lotion flushing outer embrane exocuticle tissue with PBS and antibiotic (such as the mixture of penicillin and streptomycin etc.).
And preferably the mass percent concentration with the trypsinization liquid in the II step is set between the 0.25%-0.32%, this embodiment is found, such concentration setting can be peeled off the outer embrane exocuticle fully from outer embrane exocuticle tissue, and don't damages for the outer embrane exocuticle.
Its two, this embodiment is carried out further refinement to second step, the second step after the refinement comprises:
(1), processes pearl nuclear with poly-D-lysine;
(2), with the outer embrane exocuticle that first step obtains, hatch the pearl nuclear that (1) step obtains, make the outer at least one deck outer embrane exocuticle that adheres to of pearl nuclear, form and hatch pearl nuclear.
Second step generally need to carry out sterilization treatment to pearl nuclear before above-mentioned (1), (2) work.
(2) step is very huge in the effect aspect this embodiment of guarantee pearl cultivating effect, concrete:
A large amount of outer embrane exocuticles evenly adhere to adventitia epicuticle cell, make exocuticle can evenly wrap up pearl nuclear in the birth process of dividing a word with a hyphen at the end of a line, and nacreous secretion and deposition distribution are even, have avoided the defect pearl, and effectively shorten and grow cultured pearls the time.
Know how to judge the outer at least one deck outer embrane exocuticle that adhered to of pearl nuclear for the ease of those skilled in the art, and can guarantee that the outer embrane exocuticle is to be attached to uniformly on the pearl nuclear outer surface, this embodiment provides a kind of criterion:
Pearl after hatching nuclear is under inverted microscope, and the technical staff can observe: the outer embrane exocuticle has been paved with pearl nuclear surface, forms the cell monolayer layer, and at aqueous phase springing pearl nuclear gently, cell can be from the disengaging of pearl nuclear.
Can carry out smoothly in order to guarantee the work of hatching in (2) step, must be in advance process with adhesive (for example poly-D-lysine in (1) step) on pearl nuclear surface so that contact at first the outer embrane exocuticle on pearl nuclear surface and can stick to smoothly on the pearl nuclear surface, and then utilize these to be attached to the surperficial outer embrane exocuticles of pearl nuclear and breed cultivation.
In actual mechanical process, be conditional to the selection of adhesive viscosity size, in general, viscosity is too large, is not easy to the operation that pearl nuclear is hatched, and too the ability that is attached on the pearl nuclear of cellule is less for viscosity.
The means of the regulation and control adhesive viscosity that adopts, generally there are two kinds, for the poly-D-lysine that this embodiment is mentioned, can come adjusting viscosity from the molecular weight of poly-D-lysine and this two aspect of concentration of poly-D-lysine, any regulating measure can be used separately, when but two kinds of regulating measures were used simultaneously, those skilled in the art generally can obtain more accurately viscosity regulating effect.
Adhere to the needs of outer embrane exocuticle on pearl nuclear surface for satisfying this embodiment, the molecular weight of the poly-D-lysine of (1) step is between 10000-150000, and the concentration of poly-D-lysine can be set as 0.1-0.2mg/ml.
The growth and breeding that the temperature of (2) hatching the pearl nuclear that (1) step obtains in the step generally can adapt to the outer embrane exocuticle gets final product, and this embodiment is 25 ℃-27 ℃ with this Temperature Setting.
The below demonstrates down to those skilled in the art again, uses the embodiment of this embodiment.
Embodiment
That the present embodiment adopted about 1 age is healthy, energetic little freshwater mussel is the source that obtains of outer embrane exocuticle tissue.
The first, adopt strip masking process to separate the outer embrane exocuticle tissue of freshwater mussel.The strip masking process that adopts can make the mantle tissue composition of acquisition single, does not contain endepidermis tissue or cell, thereby helps the acquisition of high-purity outer embrane exocuticle.
The second, in desinfection chamber, get the outer embrane exocuticle and be organized in PBS solution and (contain 8g/LNaCl, 0.2g/LKCl, 1.44g Na
2HPO
4/ L and 0.24g/L KH
2PO
4, pH is 7.4) in washing, then each washes and organizes once with the order of two anti-, the two anti-washing lotions of PBS+200IU/mL of PBS+200IU/mL two anti-(penicillin and streptomycins), PBS+1000IU/mL, again washes with PBS at last.
Three, get the tissue of processing and be cut into 2mm
3The fritter of size washes small pieces 2 times with PBS, then adds trypsinization liquid digestion 30min under 37 ℃ of 0.3% in organizing small pieces, stops pancreatin with perfect medium and reacts and cell dispersion the phase of fetching water behind the static 10min, centrifugal rear collecting cell.With blood counting chamber cell suspension is counted the adjustment cell number 5 * 10
5-1 * 10
6Individual/mL, place 26 ℃ of CO
2The constant incubator cultured cell.
Four, the spheroidal that grinds of the shell of employing and pearl homogeneity, pancake pearl nuclear, diameter is 8mm, high pressure steam sterilization after the washing, dry after with poly-D-lysine (molecular weight is 15, the 000) immersion of 0.1mg/mL 8 hours.Take out pearl nuclear, use thermostatic drying chamber 60 ℃ lower dry, during guarantee that processing procedure is without fungi pollution.In cell culture chamber, the exocuticle that usefulness obtains and culture fluid are at CO
2Hatch the pearl nuclear of processing in the constant incubator, 26 ℃ of temperature, incubation time is 8h, judges with the criterion that embodiment provides, and makes pearl nuclear fill part outward and adheres to a large amount of outer embrane exocuticles.
Five, choose healthy great-hearted 3 age pearl culturing clam, it reaches 13cm, the wide 10cm of reaching, shell up to more than the 3.5cm, clean shell earth and algae, cracker is opened behind the shell opening with " U " type frame support shell, the cotton ball soaked in alcohol of use 75% carries out wiping to operative site, open approximately 15mm opening 4 with the opening pin at visceral mass 2 and abdominal foot 3 intersection horizontal hooks, and use logical sheet pin to open the packed pouch of a bite to visceral mass 2 directions, will adhere to the pearl nuclear 1 usefulness tweezers of outer embrane cell and fill in visceral mass pouch depths (insert the nuclear position and see Fig. 1).After slotting nuclear is finished, adopt singlehanded ropework with special biology line wound to be sewed up, anti-antiemetic pearl is attached to together the wound tissue, cleaned and smear to reduce wound infection at the wound suture place with antibiotic and short muscle growth medicine simultaneously, promoted anathrepsis.Through postoperative sew up pearl culturing clam vertically keep hanging on after in clear water reserviors, supporting 8 hours temporarily under water among the 20cm, carry out pearl cultivating.
In addition, the pearl nuclear diameter that the present embodiment is implanted is large (being 8mm), checks the mechanical damage of visceral mass in order to reduce pearl, reduces the lethality of freshwater mussel, and the pearl nuclear of implantation adopts the mode of a freshwater mussel one pearl.Its cultivation result can reach the bead yield rate about 75%, and more than 10mm, the cultivation time is about two-and-a-half years to acquisition pearl diameter at least.
Claims (1)
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| CN2010101628363A CN101803581B (en) | 2010-05-05 | 2010-05-05 | A method for cultivating large nucleated pearls |
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| CN2010101628363A CN101803581B (en) | 2010-05-05 | 2010-05-05 | A method for cultivating large nucleated pearls |
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| CN102461471B (en) * | 2010-11-11 | 2013-06-05 | 湖南文理学院 | Culture method of amorphous pearls |
| CN102578769B (en) * | 2012-03-27 | 2015-03-11 | 胡开吉 | Large-diameter bezoar pearl and preparation method |
| CN103098739B (en) * | 2013-03-13 | 2014-06-11 | 广东海洋大学 | Method for improving pearl forming rate and pearl quality of nucleated pearl |
| CL2015001349A1 (en) * | 2015-05-19 | 2017-06-02 | Univ Antofagasta | Abalone-free pearl production process |
| CN105284674B (en) * | 2015-06-07 | 2017-09-29 | 福建师范大学 | A kind of pearl containing noble metal and preparation method thereof |
| CN105309346B (en) * | 2015-06-07 | 2017-12-15 | 福建师范大学 | A kind of method that pearl is cultivated with tissue engineering bracket pearl core |
| CN105265353B (en) * | 2015-06-07 | 2018-07-10 | 肖义军 | There are pearl nucleus of the hydrogel layer containing external carbon dosage and preparation method thereof on a kind of surface |
| CN105284673B (en) * | 2015-06-07 | 2018-03-27 | 福建师范大学 | A kind of method of old pearl renovation |
| CN105309345B (en) * | 2015-06-07 | 2017-10-31 | 福建师范大学 | Outer embrane free cell plants the method that core cultivates pearl after being co-cultured in vitro with tissue engineering bracket pearl core |
| CN105075958B (en) * | 2015-10-05 | 2017-10-31 | 江毓锋 | A kind of method for shortening producing nucleated pearls phase and increase nucleated pearl diameter |
| CN111528148A (en) * | 2020-05-30 | 2020-08-14 | 抚州市水产科学研究所 | Method for rapidly cultivating high-quality nucleated pearls by using hyriopsis schlegeli |
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|---|---|---|---|---|
| CN1044203A (en) * | 1989-06-05 | 1990-08-01 | 欧阳育斌 | Free state pictographic pearl |
| CN1965644A (en) * | 2005-11-15 | 2007-05-23 | 施志仪 | Cultivation of nucleus pearl by using free cell implantation method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1163120C (en) * | 2000-06-20 | 2004-08-25 | 谢绍河 | Pearl seed inserting method for pearl culture in fresh water mussel |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1044203A (en) * | 1989-06-05 | 1990-08-01 | 欧阳育斌 | Free state pictographic pearl |
| CN1965644A (en) * | 2005-11-15 | 2007-05-23 | 施志仪 | Cultivation of nucleus pearl by using free cell implantation method |
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| 贺龙强.有核珍珠操作技术.《内陆水产》.1998,(第12期),第15页. * |
| 骆年华等.再生珍珠培育新技术.《科学养鱼》.2006,(第8期),第25页. * |
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