CN113925005B - Efficient spawning induction method for holothuria leucospilota - Google Patents
Efficient spawning induction method for holothuria leucospilota Download PDFInfo
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- CN113925005B CN113925005B CN202111171330.3A CN202111171330A CN113925005B CN 113925005 B CN113925005 B CN 113925005B CN 202111171330 A CN202111171330 A CN 202111171330A CN 113925005 B CN113925005 B CN 113925005B
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- 241001213462 Holothuria leucospilota Species 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 33
- 230000006698 induction Effects 0.000 title claims abstract description 20
- 241000251511 Holothuroidea Species 0.000 claims abstract description 97
- 241000208340 Araliaceae Species 0.000 claims abstract description 76
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 claims abstract description 76
- 235000003140 Panax quinquefolius Nutrition 0.000 claims abstract description 76
- 235000008434 ginseng Nutrition 0.000 claims abstract description 76
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 45
- 230000002196 ecbolic effect Effects 0.000 claims abstract description 32
- 238000002347 injection Methods 0.000 claims abstract description 31
- 239000007924 injection Substances 0.000 claims abstract description 31
- 238000001035 drying Methods 0.000 claims abstract description 23
- 230000000638 stimulation Effects 0.000 claims abstract description 20
- 238000003860 storage Methods 0.000 claims abstract description 14
- 230000017448 oviposition Effects 0.000 claims abstract description 12
- 230000032696 parturition Effects 0.000 claims abstract description 11
- 239000004677 Nylon Substances 0.000 claims description 43
- 229920001778 nylon Polymers 0.000 claims description 43
- 239000013535 sea water Substances 0.000 claims description 31
- 238000009395 breeding Methods 0.000 claims description 11
- 230000001488 breeding effect Effects 0.000 claims description 11
- 230000004936 stimulating effect Effects 0.000 claims description 10
- 238000005273 aeration Methods 0.000 claims description 7
- CKOMXBHMKXXTNW-UHFFFAOYSA-N 6-methyladenine Chemical compound CNC1=NC=NC2=C1N=CN2 CKOMXBHMKXXTNW-UHFFFAOYSA-N 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 4
- 241000258195 Holothuria Species 0.000 claims description 3
- 229940090044 injection Drugs 0.000 description 23
- 230000008859 change Effects 0.000 description 5
- 230000000384 rearing effect Effects 0.000 description 5
- 241000208125 Nicotiana Species 0.000 description 4
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 4
- 235000013601 eggs Nutrition 0.000 description 4
- XNOPRXBHLZRZKH-MQYCRUOZSA-N oxytocin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSC[C@H](N)C(=O)N1)C(=O)N1C(CCC1)C(=O)N[C@@H](CC(C)C)C(=O)NCC(N)=O)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 XNOPRXBHLZRZKH-MQYCRUOZSA-N 0.000 description 4
- 229940082408 oxytocin injection Drugs 0.000 description 4
- XNOPRXBHLZRZKH-UHFFFAOYSA-N Oxytocin Natural products N1C(=O)C(N)CSSCC(C(=O)N2C(CCC2)C(=O)NC(CC(C)C)C(=O)NCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(CCC(N)=O)NC(=O)C(C(C)CC)NC(=O)C1CC1=CC=C(O)C=C1 XNOPRXBHLZRZKH-UHFFFAOYSA-N 0.000 description 3
- 101800000989 Oxytocin Proteins 0.000 description 3
- 102100031951 Oxytocin-neurophysin 1 Human genes 0.000 description 3
- XNOPRXBHLZRZKH-DSZYJQQASA-N oxytocin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSC[C@H](N)C(=O)N1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)NCC(N)=O)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 XNOPRXBHLZRZKH-DSZYJQQASA-N 0.000 description 3
- 229960001723 oxytocin Drugs 0.000 description 3
- 206010041497 Spermatorrhoea Diseases 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000021595 spermatogenesis Effects 0.000 description 2
- 208000035404 Autolysis Diseases 0.000 description 1
- 206010057248 Cell death Diseases 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- 241000218196 Persea Species 0.000 description 1
- 241000195474 Sargassum Species 0.000 description 1
- 206010047791 Vulvovaginal dryness Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000002149 gonad Anatomy 0.000 description 1
- 230000026109 gonad development Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000028043 self proteolysis Effects 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
Classifications
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- 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
- A01K61/30—Culture of aquatic animals of sponges, sea urchins or sea cucumbers
-
- 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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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention discloses a high-efficiency spawning induction method for holothuria leucospilota. The method comprises the steps of parent ginseng selection, parent ginseng storage and cultivation, drying stimulation in the shade, exposed running water stimulation, oxytocic injection, parent ginseng barrel for parturition, parent ginseng oviposition and the like. The spawning induction method can efficiently stimulate the holothuria leucospilota parent sea cucumber female to spawn, and obviously improve the proportion of the spawned female sea cucumber.
Description
Technical Field
The invention belongs to the technical field of aquaculture, and particularly relates to a method for efficiently inducing spawning of holothuria leucospilota.
Background
Holothuria leucospilota (Holthus leucospilota) is a common holothurian in the south China sea area, has the advantages of strong ecological adaptability, transportation resistance, difficult spitting dirt, difficult autolysis and the like, and is particularly suitable for the restoration of marine ecological environment and the restoration of holothurian resources.
In the early stage, the large-scale artificial breeding technology of the holothuria leucospilota (Huang et al,2018) is broken through, but in the breeding process of the holothuria leucospilota, the holothuria leucospilota is not synchronous in gonad development, matured in batches and does not have a lunar phase spawning rule, and on the basis of mastering the rules, the method (ZL 20181179112.2) for improving the spawning efficiency of the holothuria leucospilota through continuous stimulation is invented, in the method, the spawning proportion of mature females reaches 10.7%, the female-male ratio capable of generating gametes is 3/7, and therefore, the number of the spermatogenesis males and the spermatogenesis males is far higher than the number of the spawning females. Therefore, the shortage of the number of the egg-laying females is still an important factor for limiting the egg-laying scale, so that a new efficient spawning induction method needs to be explored to further improve the proportion of the egg-laying females so as to improve the utilization rate of parents, save the cost of manpower and material resources and improve the efficiency of breeding work.
Disclosure of Invention
The invention aims to provide a method for inducing the high-efficiency spawning of holothuria leucospilota aiming at the defects. The efficient spawning induction method for holothuria leucospilota provided by the invention obviously improves the ratio of spawning female ginseng and spermiation male ginseng in the group, and greatly improves the individual ratio of effective spawning and spermiation female ginseng and spermiation male ginseng.
The technical scheme adopted by the invention is as follows:
a holothuria leucospilota efficient spawning induction method comprises the following steps:
(1) transporting wild caught holothurian leucospilota to a breeding base, and carrying out storage culture in a temporary culture pond, wherein baits are thrown during the storage culture period, and the storage culture time of the holothurian is 5-10 days;
(2) drying in the shade and stimulating: removing the sea cucumbers from the temporary rearing pond, flatly paving the sea cucumbers on nylon bolting silk, and drying the sea cucumbers in the shade for 0.5 to 1.5 hours;
(3) exposure to running water stimulation: after the shade drying of the sea cucumber is finished, drawing four sides of the nylon bolting silk into a wrapping shape, pricking a seawater outlet pipe into the wrapping of the nylon bolting silk, and tightening the seawater outlet pipe by using a rope to ensure that water flows out of meshes of the nylon bolting silk, and stimulating the flowing water for 0.2-1 hour;
(4) injection of oxytocic: after the running water stimulation is finished, injecting an oxytocic into the sea cucumber, wherein the oxytocic is 0.1-0.2 mM N6-methyladenine;
(5) parent ginseng barrel awaiting parturition: after the oxytocic is injected, immediately putting the sea cucumbers into parent sea cucumber buckets filled with clean seawater for spawning, putting 15-30 heads of the sea cucumbers into each bucket, and not aerating;
(6) parent sea cucumbers lay eggs: when the male ginseng with the sperm discharge is found, the male ginseng is immediately taken out and is independently placed in a male gamete barrel; the female ginseng with egg laying is found, immediately taken out and separately placed in a female gamete bucket.
In the step (1), the adult quantity of the wild caught holothuria leucospilota parent sea cucumber is more than 250g, and the wild caught holothuria leucospilota parent sea cucumber is plump and has no damage.
In the step (1), the stocking density of the stocking is 10-20 heads/m 2 。
In the step (1), the bait is sargassum powder.
In the step (1), during the accumulation period, water is changed once every day, and the water change amount is 20% v/v.
In the step (2), the mesh diameter of the nylon bolting-silk is 50-70 meshes, and the area of the nylon bolting-silk is 2-3 m 2 。
In the step (4), the injection amount of the oxytocin is 0.5-1.0mL per sea cucumber according to the individual size.
In the step (4), the injection method comprises the following steps: after the running water stimulation is finished, the rope is loosened, the sea cucumber is held by the two hands, another person carries out oxytocic injection, the injection needle penetrates into the back of the sea cucumber at an angle of 45 degrees and is separated from one third of the head, and the length of the injection needle extending into the body cavity is 8-12 mm.
In the step (5), the capacity of the parent sea cucumber barrel is 200-300 liters, and 15-30 heads of sea cucumbers are put into each barrel.
In the step (6), 100-120 liters of clean seawater is added into both the male gamete barrel and the female gamete barrel.
The invention is different from the published sea cucumber spawning induction method in that:
(1) the parent ginseng is stored for a long time. In the past mode, the sea cucumber is temporarily raised for 1-2 days before transportation, the parent ginseng cannot obtain food, and in addition, the parent ginseng is in a tired state after long-distance transportation, such as immediate spawning induction, and the spawning rate of the parent ginseng is extremely low. After the invention is cultivated for 5-10 days, the parent ginseng obtains sufficient food, and the physical ability and the physical function are recovered. In addition, because the sea cucumbers are stocked in an indoor stable environment without spawning and spermiation conditions of a natural sea area, the gonads of the sea cucumbers are gradually developed uniformly, and the quantity of the sea cucumbers in producible and to-be-produced conditions is accumulated.
(2) Different drying in the shade. The invention does not carry out shade drying in the parent ginseng barrel, but is flatly paved on the nylon bolting silk for indoor shade drying, compared with the shade drying in the parent ginseng barrel, the seawater is more thoroughly discharged.
(3) Longer exposure to running water stimulation was used. In the common oxytocic process of holothuria leucospilota, water is directly added after the parent holothuria is dried in the shade, the time of running water stimulation is basically ignored, and the holothuria leucospilota is always in the water environment.
(4) And (4) injecting an oxytocic. In the prior art, the oxytocic is not adopted in the oxytocic of the holothuria leucospilota, but the oxytocic also plays an important role in the invention.
(5) Comprehensive measures are adopted to carry out efficient spawning induction on the holothuria leucospilota, and a mode of drying in the shade is mainly adopted in the past spawning induction on the holothuria leucospilota.
The invention has the beneficial effects that:
the efficient spawning induction method for holothuria leucospilota provided by the invention obviously improves the ratio of spawning female ginseng and spermatorrhea male ginseng in the group, and greatly improves the individual ratio of effective spawning and spermatorrhea female and male ginseng.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Example 1
In 2021, 8 months and 13 days, the following steps were adopted to induce spawning of parent ginseng, and the experimental group was set as 1:
(1) selecting parent parameters: the wild caught parent holothuria leucospilota reaches more than 250g of adults, and the individuals are plump without damage.
(2) And (3) storing and culturing parent ginseng: transporting wild harvested holothuria leucospilota to a breeding base, and carrying out storage culture in a temporary culture pond with the storage culture density of 15 heads/m 2 During the sea cucumber stocking period, gulfweed powder (Haifu feed Co., Ltd., Shandong tobacco platform, Laizhou) is fed, and water is changed once a day, wherein the water change amount is 20% v/v, and the sea cucumber stocking time is 7 days.
(3) Drying in the shade and stimulating: laying a nylon bolting silk with a mesh aperture of 60 meshes on the ground indoors, wherein the area of the nylon bolting silk is 2.5m 2 The sea cucumber is moved out of the temporary rearing pond, is laid on a nylon bolting silk and is dried in the shade for 1.0 hour.
(4) Exposure to running water stimulation: after the shade drying of the sea cucumber is finished, the four sides of the nylon bolting silk are folded into a wrapping shape, the seawater outlet pipe is tied into the wrapping of the nylon bolting silk and is tied tightly by a rope, the water is ensured to flow out of the meshes of the nylon bolting silk, and the flowing water is stimulated for 0.5 hour.
(5) Injection of oxytocic: a2 mL syringe is adopted, the oxytocic is 0.15mM N6-methyladenine, and each sea cucumber is injected with 0.5-1.0mL of the oxytocic according to the size of an individual. The injection method comprises the following steps: after the running water stimulation is finished, the rope is loosened, the sea cucumber is held by both hands, another person carries out oxytocic injection, the injection needle penetrates into the back of the sea cucumber at an angle of 45 degrees and is separated from one third of the head, and the length of the injection needle extending into the body cavity is 10 mm.
(6) Parent ginseng barrel awaiting parturition: after the oxytocic is injected, the sea cucumbers are immediately put into parent sea cucumber buckets filled with clean seawater for spawning, the volume of the parent sea cucumber buckets is 250 liters, and 20 heads of the sea cucumbers are put into each bucket without aeration.
(7) Parent ginseng spawning: when the male ginseng with the sperm discharge is found, the male ginseng is immediately taken out and is independently placed in a male gamete barrel with 110 liters of clean seawater; when the female ginseng is found to lay eggs, the female ginseng is immediately taken out and is separately placed in a female gamete bucket with 110 liters of clean seawater.
Example 2
In 2021, 8 months and 13 days, the parent ginseng was induced to spawn by the following procedure and set as experiment group 2:
(1) selecting parent parameters: the wild harvested holothuria leucospilota parent sea cucumber has the adult weight of more than 250g, and is full and free of damage.
(2) And (3) storing and culturing parent ginseng: transporting wild harvested holothuria leucospilota to a breeding base, and carrying out storage culture in a temporary culture pond with the storage culture density of 15 heads/m 2 During the sea cucumber stocking period, gulfweed powder (Haifu feed Co., Ltd., Shandong tobacco platform, Laizhou) is fed, and water is changed once a day, wherein the water change amount is 20% v/v, and the sea cucumber stocking time is 5 days.
(3) Drying in the shade and stimulating: laying a nylon bolting silk with a mesh aperture of 60 meshes on the ground indoors, wherein the area of the nylon bolting silk is 2.5m 2 The sea cucumber is moved out of the temporary rearing pond, is laid on a nylon bolting silk and is dried in the shade for 0.5 hour.
(4) Exposure to running water stimulation: after the shade drying of the sea cucumber is finished, the four sides of the nylon bolting silk are folded into a wrapping shape, the seawater outlet pipe is tied into the wrapping of the nylon bolting silk and is tied tightly by a rope, the water is ensured to flow out of the meshes of the nylon bolting silk, and the flowing water is stimulated for 0.2 hour.
(5) Injection of oxytocic: a2 mL syringe is adopted, the oxytocic is 0.15mM N6-methyladenine, and each sea cucumber is injected with 0.5-1.0mL of the oxytocic according to the size of an individual. The injection method comprises the following steps: after the running water stimulation is finished, the rope is loosened, the sea cucumber is held by the two hands, the other person carries out oxytocin injection, the injection needle penetrates into the back of the sea cucumber at an angle of 45 degrees and is separated from one third of the head, and the length of the injection needle extending into the body cavity is 10 mm.
(6) Parent ginseng barrel awaiting parturition: after the oxytocic is injected, the sea cucumbers are immediately put into parent sea cucumber buckets filled with clean seawater for spawning, the volume of the parent sea cucumber buckets is 250 liters, and 20 heads of the sea cucumbers are put into each bucket without aeration.
(7) Parent ginseng spawning: when the male ginseng with the essence discharge is found, the male ginseng is immediately taken out and is independently placed in a male gamete bucket with 110 liters of clean seawater; when the female ginseng is found to lay eggs, the female ginseng is immediately taken out and is separately placed in a female gamete bucket with 110 liters of clean seawater.
Example 3
In 2021, 8 months and 13 days, the following steps were used for induced spawning of parent ginseng, and the experimental group was set as 3:
(1) parent ginseng selection: the wild harvested holothuria leucospilota parent sea cucumber has the adult weight of more than 250g, and is full and free of damage.
(2) Parent ginseng is cultivated: transporting wild harvested holothuria leucospilota to a breeding base, and carrying out storage culture in a temporary culture pond with the storage culture density of 15 heads/m 2 During the sea cucumber stocking period, gulfweed powder (Haifu feed Co., Ltd., Shandong tobacco platform, Laizhou) is fed, and water is changed once a day, wherein the water change amount is 20% v/v, and the sea cucumber stocking time is 10 days.
(3) Drying in the shade and stimulating: indoors on the groundA nylon bolting silk is laid, the aperture of the net is 60 meshes, and the area of the nylon bolting silk is 2.5m 2 The sea cucumber is moved out of the temporary rearing pond, is laid on a nylon bolting silk and is dried in the shade for 1.5 hours.
(4) Exposure to running water stimulation: after the shade drying of the sea cucumber is finished, the four sides of the nylon bolting silk are folded into a wrapping shape, the seawater outlet pipe is tied into the wrapping of the nylon bolting silk and is tied tightly by a rope, the water is ensured to flow out of the meshes of the nylon bolting silk, and the flowing water is stimulated for 1.0 hour.
(5) Injection of oxytocic: a2 mL syringe is adopted, the oxytocin is 0.15mM N6-methyladenine, and each sea cucumber is injected with 0.5-1.0mL of the oxytocin according to the size of an individual. The injection method comprises the following steps: after the running water stimulation is finished, the rope is loosened, the sea cucumber is held by the two hands, the other person carries out oxytocin injection, the injection needle penetrates into the back of the sea cucumber at an angle of 45 degrees and is separated from one third of the head, and the length of the injection needle extending into the body cavity is 10 mm.
(6) Parent ginseng barrel awaiting parturition: after the oxytocic is injected, the sea cucumbers are immediately put into parent sea cucumber buckets filled with clean seawater for spawning, the volume of the parent sea cucumber buckets is 250 liters, and 20 heads of the sea cucumbers are put into each bucket without aeration.
(7) Parent ginseng spawning: when the male ginseng with the essence discharge is found, the male ginseng is immediately taken out and is independently placed in a male gamete bucket with 110 liters of clean seawater; the female ginseng with egg laying was found, immediately taken out, and placed alone in a female gamete bucket with 110 liters of clean seawater.
Example 4
On 8/13/2021, the following procedure was used to induce the labor of parent sea cucumber, and the control group 1 was set, and the control group 1 was distinguished from the experimental group 1 in that the oxytocin injection was not performed.
(1) Selecting parent parameters: the wild harvested holothuria leucospilota parent sea cucumber has the adult weight of more than 250g, and is full and free of damage.
(2) And (3) storing and culturing parent ginseng: transporting wild harvested holothuria leucospilota to a breeding base, and carrying out stocking with a stocking density of 15 heads/m 2 During the sea cucumber stocking period, gulfweed powder (Haifu feed Co., Ltd., Shandong tobacco platform, Laizhou) is fed, and water is changed once a day, wherein the water change amount is 20% v/v, and the sea cucumber stocking time is 7 days.
(3) Drying in the shade and stimulating:laying a nylon bolting silk with a mesh aperture of 60 meshes on the ground indoors, wherein the area of the nylon bolting silk is 2.5m 2 The sea cucumber is moved out of the temporary rearing pond, is laid on a nylon bolting silk and is dried in the shade for 1.0 hour.
(4) Exposure to running water stimulation: after the shade drying of the sea cucumber is finished, the four sides of the nylon bolting silk are folded into a wrapping shape, the seawater outlet pipe is tied into the wrapping of the nylon bolting silk and is tied tightly by a rope, the water is ensured to flow out of the meshes of the nylon bolting silk, and the flowing water is stimulated for 0.5 hour.
(5) Parent ginseng barrel awaiting parturition: after the running water stimulation is finished, the sea cucumbers are immediately put into parent sea cucumber buckets filled with clean sea water for production, the volume of the parent sea cucumber buckets is 250 liters, and 20 heads of the sea cucumbers are put into each bucket without aeration.
(6) Parent ginseng spawning: when the male ginseng with the essence discharge is found, the male ginseng is immediately taken out and is independently placed in a male gamete bucket with 110 liters of clean seawater; the female ginseng with egg laying was found, immediately taken out, and placed alone in a female gamete bucket with 110 liters of clean seawater.
Example 5
On 12/8/2021, the following procedure was used to induce spawning of the parent ginseng, and the control group 2 was set as the experimental group 1, and the control group 2 was distinguished from the experimental group 1 in that the parent ginseng was not kept in stock and exposed to running water.
(1) Selecting parent parameters: the wild harvested holothuria leucospilota parent sea cucumber has the adult weight of more than 250g, and is full and free of damage. And (5) after the seeds are transported to a breeding base, hastening parturition on the same day.
(2) Drying in the shade and stimulating: laying a nylon bolting silk with a mesh aperture of 60 meshes on the ground indoors, wherein the area of the nylon bolting silk is 2.5m 2 The sea cucumber is spread on the nylon bolting silk and dried in the shade for 1.0 hour.
(3) Injection of oxytocic: a2 mL syringe is adopted, the oxytocic is 0.15mM N6-methyladenine, and each sea cucumber is injected with 0.5-1.0mL of the oxytocic according to the size of an individual. The injection method comprises the following steps: after the vaginal dryness is finished, the sea cucumber is held by two hands, another person carries out oxytocic injection, the injection needle is inserted into the back of the sea cucumber at an angle of 45 degrees and is separated from one third of the head, and the length of the injection needle extending into the body cavity is 10 mm.
(4) Parent ginseng barrel awaiting parturition: after the oxytocic is injected, the sea cucumbers are immediately put into parent sea cucumber buckets filled with clean seawater for spawning, the volume of the parent sea cucumber buckets is 250 liters, and 20 heads of the sea cucumbers are put into each bucket without aeration.
(5) Parent ginseng spawning: when the male ginseng with the essence discharge is found, the male ginseng is immediately taken out and is independently placed in a male gamete bucket with 110 liters of clean seawater; the female ginseng with egg laying was found, immediately taken out, and placed alone in a female gamete bucket with 110 liters of clean seawater.
Example 6
On 8/12/2021, the following procedure was used to induce the spawning of the parent ginseng, and the control group 3 was set as the control group 3, and the control group 3 was distinguished from the experimental group 1 in that the parent ginseng was not kept in stock, exposed running water stimulation was not performed, and the oxytocin injection was not performed.
(1) Selecting parent parameters: the wild caught parent holothuria leucospilota reaches more than 250g of adults, and the individuals are plump without damage. And (5) after the seeds are transported to a breeding base, hastening parturition on the same day.
(2) Drying in the shade and stimulating: laying a nylon bolting silk with a mesh aperture of 60 meshes on the ground indoors, wherein the area of the nylon bolting silk is 2.5m 2 The sea cucumber is spread on the nylon bolting silk and dried in the shade for 1.0 hour.
(3) Parent ginseng barrel awaiting parturition: after the cloudy drying is finished, the sea cucumbers are immediately put into parent sea cucumber buckets filled with clean seawater for production, the volume of the parent sea cucumber buckets is 250 liters, and 20 heads of the sea cucumbers are put into each bucket without aeration.
(4) Parent sea cucumbers lay eggs: when the male ginseng with the essence discharge is found, the male ginseng is immediately taken out and is independently placed in a male gamete bucket with 110 liters of clean seawater; the female ginseng with egg laying was found, immediately taken out, and placed alone in a female gamete bucket with 110 liters of clean seawater.
Example 7
The sperm discharging and egg laying proportion of the parent ginseng and the effective male and female individual ratio for egg laying and sperm discharging in the six embodiments are counted, and the results are shown in table 1.
TABLE 1 comparison of induction effect of holothuria leucospilota in different modes
As can be seen from the table 1, the efficient spawning induction method for holothuria leucospilota provided by the invention obviously improves the ratio of spawning female ginseng and spermiation male ginseng in the group, and greatly improves the individual ratio of effective spawning and spermiation female ginseng and spermiation male ginseng.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and should be considered to be within the scope of the invention.
Claims (6)
1. The efficient spawning induction method for holothuria leucospilota is characterized by comprising the following steps:
(1) and (3) storing and culturing parent ginseng: transporting wild caught holothuria leucospilota to a breeding base, and carrying out storage culture in a temporary culture pond with the storage culture density of 10-20 heads/m 2 Feeding baits during the storage period, and changing water once a day, wherein the water changing amount is 20% v/v, and the sea cucumber storage time is 5-10 days;
(2) drying in the shade and stimulating: removing the sea cucumbers from the temporary culture pond, and flatly paving the sea cucumbers in a nylon bolting-silk area, wherein the mesh diameter of the sea cucumbers is 50-70 meshes and the nylon bolting-silk area is 2-3 m 2 Drying the nylon bolting silk in the shade for 0.5 to 1.5 hours;
(3) exposure to running water stimulation: after the shade drying of the sea cucumber is finished, drawing four sides of the nylon bolting silk into a wrapping shape, pricking a seawater outlet pipe into the wrapping of the nylon bolting silk, and tightening the seawater outlet pipe by using a rope to ensure that water flows out of meshes of the nylon bolting silk, and stimulating the flowing water for 0.2-1 hour;
(4) injection of oxytocic: after the running water stimulation is finished, injecting an oxytocic into the sea cucumbers, wherein the oxytocic is 0.1-0.2 mM N6-methyladenine, and the injection amount is that each sea cucumber is injected with 0.5-1.0mL according to the size of an individual;
(5) parent ginseng barrel awaiting parturition: after the oxytocic is injected, the sea cucumber is immediately put into a parent sea cucumber barrel filled with clean seawater for spawning without aeration;
(6) parent ginseng spawning: when the male ginseng with the sperm discharge is found, the male ginseng is immediately taken out and is independently placed in a male gamete barrel; the female ginseng with egg laying is found, immediately taken out and separately placed in a female gamete bucket.
2. The efficient spawning induction method for holothuria leucospilota according to claim 1, wherein in the step (1), the adult of the wild-caught holothuria leucospilota reaches more than 250g, and the individual is plump without damage.
3. The efficient spawning induction method for holothuria leucospilota according to claim 1, wherein in the step (1), the bait is gulfweed powder.
4. The holothuria leucospilota high-efficiency spawning induction method according to claim 1, characterized in that in the step (4), the injection method comprises the following steps: after the running water stimulation is finished, the rope is loosened, the sea cucumber is held by the two hands, another person carries out oxytocic injection, the injection needle penetrates into the back of the sea cucumber at an angle of 45 degrees and is separated from one third of the head, and the length of the injection needle extending into the body cavity is 8-12 mm.
5. The efficient spawning induction method for holothuria leucospilota according to claim 1, wherein in the step (5), the capacity of the parent holothuria tub is 200-300 liters, and 15-30 heads of holothuria are placed in each tub.
6. The efficient spawning induction method for holothuria leucospilota according to claim 1, wherein in the step (6), 100-120 liters of clean seawater is added into both the male gamete barrel and the female gamete barrel.
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| CN202111171330.3A CN113925005B (en) | 2021-10-08 | 2021-10-08 | Efficient spawning induction method for holothuria leucospilota |
| PCT/CN2021/140666 WO2022148246A1 (en) | 2021-10-08 | 2021-12-23 | Efficient spawning induction method for holothuria leucospilota |
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| KR20180047631A (en) * | 2016-11-01 | 2018-05-10 | 이재두 | Controlling method of the time of sea cucumber artificial seed production through management of parents sea cucumbers in land farm |
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| CN101595847A (en) * | 2009-07-01 | 2009-12-09 | 大连水产学院 | The method of inducing Stichopus Japonicus Selenka to lay eggs in vivo |
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