CN111838033A - Method for detackifying and hatching fertilized eggs of thamnaconus modestus - Google Patents
Method for detackifying and hatching fertilized eggs of thamnaconus modestus Download PDFInfo
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- CN111838033A CN111838033A CN202010668295.5A CN202010668295A CN111838033A CN 111838033 A CN111838033 A CN 111838033A CN 202010668295 A CN202010668295 A CN 202010668295A CN 111838033 A CN111838033 A CN 111838033A
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- 235000013601 eggs Nutrition 0.000 title claims abstract description 142
- 238000000034 method Methods 0.000 title claims abstract description 31
- 230000012447 hatching Effects 0.000 title claims abstract description 29
- 241001676702 Thamnaconus modestus Species 0.000 title claims abstract description 6
- 239000013535 sea water Substances 0.000 claims abstract description 26
- 210000004681 ovum Anatomy 0.000 claims abstract description 17
- 241000075387 Thamnaconus septentrionalis Species 0.000 claims abstract description 15
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 5
- 108091005658 Basic proteases Proteins 0.000 claims abstract description 5
- 238000003756 stirring Methods 0.000 claims abstract description 4
- 238000000926 separation method Methods 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 241000251468 Actinopterygii Species 0.000 claims description 12
- 238000011534 incubation Methods 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- 239000012452 mother liquor Substances 0.000 claims description 7
- 241000980706 Takifugu obscurus Species 0.000 claims description 4
- 241001441722 Takifugu rubripes Species 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 229910052760 oxygen Inorganic materials 0.000 claims description 4
- 239000001301 oxygen Substances 0.000 claims description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 3
- 230000000694 effects Effects 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 210000003205 muscle Anatomy 0.000 claims description 3
- 239000012266 salt solution Substances 0.000 claims description 3
- 235000002639 sodium chloride Nutrition 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 230000000384 rearing effect Effects 0.000 claims 1
- 210000003097 mucus Anatomy 0.000 abstract description 7
- 239000007788 liquid Substances 0.000 abstract description 3
- 238000009360 aquaculture Methods 0.000 abstract description 2
- 244000144974 aquaculture Species 0.000 abstract description 2
- 238000009395 breeding Methods 0.000 description 5
- 230000001488 breeding effect Effects 0.000 description 5
- 239000012535 impurity Substances 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 3
- 241001627955 Tetraodon lineatus Species 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000000645 desinfectant Substances 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- CPKVUHPKYQGHMW-UHFFFAOYSA-N 1-ethenylpyrrolidin-2-one;molecular iodine Chemical compound II.C=CN1CCCC1=O CPKVUHPKYQGHMW-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241001659573 Centriscus scutatus Species 0.000 description 1
- 229920000153 Povidone-iodine Polymers 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002149 gonad Anatomy 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 230000009027 insemination Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000006213 oxygenation reaction Methods 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 229960001621 povidone-iodine Drugs 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000002791 soaking 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
- A01K61/10—Culture of aquatic animals of fish
- A01K61/17—Hatching, e.g. incubators
-
- 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 provides a detackifying and hatching method for fertilized eggs of navodon septentrionalis, belonging to the field of aquaculture, and the method is characterized in that alkaline protease derived from bacillus licheniformis is dissolved in seawater to prepare 5g/L detackifying solution; collecting fertilized eggs of the thamnaconus modestus, putting the fertilized eggs into a mucus removing liquid, continuously stirring and uniformly mixing, stopping the removal of the mucus when the fertilized eggs are not adhered to each other or the walls of a barrel and are in a certain bright and transparent state, separating the fertilized eggs from unfertilized eggs by increasing the salinity of seawater after the fertilized eggs are removed, and hatching the fertilized eggs according to a normal procedure after the separated fertilized eggs are washed clean. The method can reduce the germ pollution in the hatching and seedling raising processes and improve the hatching rate and the seedling emergence rate.
Description
Technical Field
The invention relates to the field of aquaculture, in particular to a method for detackifying and hatching fertilized eggs of navodon septentrionalis.
Background
Takifugu obscurus (Thamnaconus septentrionalis) is commonly called as 'skinned fish', 'shrimpfish' and 'breaded fish', is distributed in Bohai sea, yellow sea and east sea, has delicious meat and rich nutrition, and is excellent economic fish in coastal areas of China. The cultivation property of the navodon septentrionalis is excellent, the survival temperature is 9-30 ℃, the omnivorous property is easy to domesticate and cultivate, the feed utilization rate is high, the growth speed is high, and various modes such as factory workshop cultivation, pond cultivation, net cage cultivation and the like can be developed. The product has various specifications, can be sold after 100g, can be cultivated and marketed in the north early seedling in the same year, can be cultivated all the year in the south sea area of the Fujian, and has very wide market prospect.
Fertilized eggs of the thamnaconus modestus are sticky and sinking, and are adhered and fixed on other objects through mucous membranes and sticky wires on the surfaces of the eggs. Artificial propagation of takifugu obscurus mostly adopts a natural spawning mode, and in the prior art, fertilized eggs are collected by placing an attachment base at the bottom of a pond and then transferred into a hatching pond for hatching. Because the fertilized eggs are sticky, a large amount of impurities such as the feces of the parent fish and the like are adhered to the surfaces of the fertilized eggs, and germs are easy to breed in the hatching pond. In addition, the method can not well separate fertilized eggs from unfertilized eggs, and unfertilized eggs are easy to rot in the hatching pond, so that the germ reproduction is aggravated, and the hatching of the fertilized eggs and the subsequent seed cultivation are influenced.
Therefore, the research and development of the technical method for effectively removing the stickiness of the fertilized eggs and separating the fertilized eggs from unfertilized eggs is very important for perfecting the breeding technology of the takifugu rubripes seedlings.
Disclosure of Invention
The invention provides a method for debonding and incubating fertilized eggs of navodon septentrionalis, which can effectively remove mucus on the surfaces of the fertilized eggs, separate the fertilized eggs from unfertilized eggs, reduce germ pollution in the incubation and seedling raising processes, and improve the incubation rate and the emergence rate.
The invention is realized by the following technical scheme:
a method for detackifying fertilized eggs of Navodon septentrionalis comprises the following steps:
(1) Preparing a detackifying solution, dissolving alkaline protease (bacillus licheniformis) in seawater to prepare 5g/L of detackifying solution;
(2) collecting fertilized eggs of the thamnaconus modestus, putting the fertilized eggs into the mucus removing liquid, continuously stirring and uniformly mixing, and stopping the debonding when the fertilized eggs are not mutually bonded or adhered to the barrel wall and are in a certain bright and transparent state.
Further, the ratio of the fertilized eggs to the detackifying solution is 30-35 g/L
The invention also provides a hatching method after the debonding of the fertilized eggs of the navodon septentrionalis, the fertilized eggs which are cleaned after the debonding are transferred into a separation container, high-salt seawater mother liquor is gradually added into the separation container, the mixture is uniform, and the separation condition of the fertilized eggs is observed; stopping adding the high-salt seawater mother liquor when a large number of bright and transparent fertilized eggs float upwards, and standing; when all fertilized eggs float on the upper layer of the water body, unfertilized eggs sink to the bottom of the separation container; collecting upper fertilized eggs by using a screen, continuously washing the fertilized eggs by using clean seawater, removing high-salt seawater on the surfaces of the fertilized eggs, and incubating the collected fertilized eggs according to a normal procedure.
Furthermore, the preparation method of the high-salt solution is to add sea salt into seawater to prepare the high-salt solution.
Further, the normal procedure comprises the steps of placing an incubation net cage in an incubation pool, placing fertilized eggs in the net cage, wherein the water temperature is 18-20 ℃, the salinity is 27-30, the dissolved oxygen is 6.0-8.0 mg/L, and the PH is 7.8-8.2; after 40 hours, the fertilized eggs develop to the muscle effect period, and the fertilized eggs are collected by using a 80-mesh screen, wherein the fertilized eggs still contain undeveloped dead eggs and bad eggs, and the fertilized eggs are separated again by using the same method for separating the fertilized eggs and the unfertilized eggs to obtain the normally-developed fertilized eggs. And (4) putting the cleaned fertilized eggs into a fry breeding pond, and continuously breaking membranes of the newly hatched fries.
Compared with the prior art, the invention has the beneficial effects that:
1. compared with the prior art, the method for detackifying the fertilized eggs of the navodon septentrionalis is developed for the first time. The liver of the parent fish of the navodon septentrionalis covers the outer side of the gonad, so the artificial insemination operation easily loses the liver, and the parent fish dies, therefore, the artificial breeding of the navodon septentrionalis mostly adopts a natural spawning mode at present. Fertilized eggs are sticky and sinking and are often attached to the bottom and the wall of the parent fish pool, and a large amount of impurities such as feces of the parent fish and the like are adhered to the surface of the fertilized eggs. In the prior art, collected fertilized eggs are cleaned and disinfected firstly, and then are transferred to a hatching pond for hatching. Although the germ can be killed temporarily, a large amount of impurities are still adhered to the surface of the fertilized eggs for germ reproduction, and the germs can be bred again along with the time, so that the hatching of the fertilized eggs and the growth of seedlings are influenced. According to the invention, through alkaline protease (bacillus licheniformis), mucus on the surface of the fertilized egg can be efficiently removed, and the breeding of bacteria on the surface of the fertilized egg is reduced.
2. Compared with the prior art, the method for separating the fertilized eggs and the unfertilized eggs of the Navodon septentrionalis, and the method for separating the normally developed fertilized eggs, the undeveloped dead eggs and the damaged eggs are developed for the first time on the basis of unsticking the fertilized eggs. In the prior art, the fertilized eggs are not detackified, impurities are adhered to the surfaces of the eggs, the fertilized eggs and the unfertilized eggs are not easy to distinguish, and the separation of the fertilized eggs is difficult to carry out by adjusting the density of seawater, so that a large amount of unfertilized eggs, dead eggs and bad eggs are always remained in the water body for hatching and seedling cultivation, and the water quality is deteriorated. After the fertilized eggs are detackified, the specific gravity of the fertilized eggs and unfertilized eggs is different, and the separation can be carried out by changing the density of seawater, so that the number of unfertilized eggs, dead eggs and bad eggs is reduced, the water quality of hatching and seedling growth is improved, and the hatching rate and the emergence rate are improved.
The specific implementation mode is as follows:
the following examples are provided to illustrate the present invention in detail, but the scope of the present invention should not be limited by the examples in any way.
Example 1:
the method for detackifying and hatching fertilized eggs of Navodon septentrionalis comprises the following steps:
(1) preparing a detackifying liquid and detackifying fertilized eggs: and in 2018, 5, 23 and 19 ℃, collecting fertilized eggs from the parent fish pond of the Navodon septentrionalis, wherein a large amount of fertilized eggs sink to the bottom of the pond due to viscosity and sinking of the fertilized eggs. And (4) reducing the water level of the parent fish pond to 20cm, and collecting fertilized eggs at the bottom of the pond through a dip net. Dissolving 25g of alkaline protease (Bacillus licheniformis) (Solarbio) in 5L of seawater, and mixing to obtain 5g/L of mucus.
Putting 163g of fertilized eggs (about 65.2 ten thousand fertilized eggs) into the debonding solution, continuously stirring and uniformly mixing for 20min, and stopping debonding when the fertilized eggs are not mutually bonded and are adhered to the barrel wall and present a certain bright and transparent state.
(2) Cleaning fertilized eggs: collecting the debonded fertilized eggs by using a 80-mesh screen, continuously washing the fertilized eggs by using clean seawater, and washing off mucus separated from the surfaces of the fertilized eggs until the fertilized eggs are in a bright and transparent state.
(3) Fertilized and unfertilized eggs are separated: 2kg of sea salt is put into 10L of sea water to prepare high-salt sea water mother liquor, the cleaned fertilized eggs are transferred into a separation barrel, the high-salt sea water mother liquor is gradually added into the separation barrel, the mixture is uniformly mixed, and the separation condition of the fertilized eggs is observed. When a large amount of bright and transparent fertilized eggs float upwards, the high-salt seawater mother liquor is stopped to be added, and the mixture is kept stand. When all fertilized eggs float on the upper layer of the water body, unfertilized eggs sink on the water bottom and are collected by a 80-mesh screen. And continuously washing the collected fertilized eggs with clean seawater, and removing high-salt seawater on the surfaces of the eggs to obtain 149g of high-quality fertilized eggs.
(4) Hatching fertilized eggs: placing an incubation net cage (58cm is multiplied by 48cm is multiplied by 45cm) in an incubation pool, placing fertilized eggs into the net cage, and arranging two air stones in the net cage for oxygenation, wherein the water temperature is 18-20 ℃, the salinity is 27-30, the dissolved oxygen is 6.0-8.0 mg/L, and the PH is 7.8-8.2. After 40 hours, the fertilized eggs develop to the muscle effect stage, and the fertilized eggs are collected by a 80-mesh screen. In this case, the fertilized eggs still contain undeveloped dead eggs and bad eggs, and need to be separated again. Since the proportion of normally developing fertilized eggs is lower than that of dead and bad eggs, the normally developing fertilized eggs can be isolated and obtained in the same manner as the separation of fertilized and unfertilized eggs. And (4) putting the cleaned fertilized eggs into a fry breeding pond, and continuously breaking membranes of the newly hatched fries.
(5) The fertilized eggs of the globefish are detackified and incubated by the method, the hatching rate of the fertilized eggs reaches 92.6 percent, the survival rate of the newly-incubated fry reaches 90.8 percent, 25.6 thousands of healthy fries with the length of 43-69 mm are obtained after fry incubation for 50 days, and the emergence rate is 39.2 percent.
Example 2
Hatching fertilized eggs of the navodon septentrionalis by using a conventional method:
24 days in 5 months in 2018, the water temperature is 19 ℃, the water level of the parent fish pond of the takifugu obscurus is reduced to 20cm, and 152g (about 60.8 ten thousand fertilized eggs) of fertilized eggs are collected at the bottom of the pond through a plastic plate (an attaching base). Continuously washing the fertilized eggs with clean seawater for 3min, and washing off impurities on the surfaces of the fertilized eggs as far as possible. Soaking fertilized eggs in 200mg/L povidone iodine disinfectant for 5min, and continuously turning and mixing uniformly. Then cleaning fertilized eggs with clean seawater for 3min, and washing away the disinfectant. And uniformly spreading the sterilized fertilized eggs into an incubation pool, wherein the water temperature is 18-20 ℃, the salinity is 27-30, the dissolved oxygen is 6.0-8.0 mg/L, and the pH is 7.8-8.2.
And (2) hatching fertilized eggs of the globefish by using a conventional method, wherein the hatching rate of the fertilized eggs is 79.3%, the survival rate of the newly hatched fries is 75.6%, 11.9 thousands of healthy fries with the length of 38-65 mm are obtained after 50-day fry cultivation, and the emergence rate is 19.6%.
Claims (5)
1. A method for detackifying fertilized eggs of Navodon septentrionalis is characterized by comprising the following steps:
(1) preparing a detackifying solution, namely dissolving alkaline protease derived from bacillus licheniformis in seawater to prepare the detackifying solution with the concentration of 5 g/L;
(2) and (3) detackifying the fertilized eggs, collecting the fertilized eggs of the thamnaconus modestus, putting the fertilized eggs into a detackifying solution, continuously stirring and uniformly mixing, and stopping detackifying when the fertilized eggs are not adhered to each other, adhere to the barrel wall and present a certain bright and transparent state.
2. The method for detackifying fertilized eggs of takifugu obscurus according to claim 1, wherein the ratio of the fertilized eggs to the detackifying solution is 30-35 g/L.
3. A hatching method after the debonding of fertilized eggs of takifugu rubripes, which is characterized in that the fertilized eggs after the debonding of claim 1 or 2 are washed clean and transferred into a separation container, high-salt seawater mother liquor is gradually added into the separation container, the mixture is uniformly mixed, and the separation condition of the fertilized eggs is observed; stopping adding the high-salt seawater mother liquor when a large number of bright and transparent fertilized eggs float upwards, and standing; when all fertilized eggs float on the upper layer of the water body, unfertilized eggs sink to the bottom of the separation container; collecting upper fertilized eggs by using a screen, continuously washing the fertilized eggs by using clean seawater, removing high-salt seawater on the surfaces of the fertilized eggs, and incubating the collected fertilized eggs according to a normal procedure.
4. The method for hatching the debonded fertilized eggs of takifugu rubripes according to claim 3, wherein the high-salt solution is prepared by adding sea salt into seawater.
5. The method for hatching Fugu rubripes fertilized egg after detachment according to claim 3, wherein the normal hatching procedure comprises: the normal procedure comprises the steps of placing an incubation net cage in an incubation pool, placing fertilized eggs into the net cage, wherein the water temperature is 18-20 ℃, the salinity is 27-30, the dissolved oxygen is 6.0-8.0 mg/L, and the pH is 7.8-8.2; after 40 hours, the fertilized eggs develop to a muscle effect period, the fertilized eggs are collected by a 80-mesh screen, at this time, the fertilized eggs still contain undeveloped dead eggs and damaged eggs, the fertilized eggs are separated again by the same method as the separation of the fertilized eggs and the unfertilized eggs to obtain the fertilized eggs which normally develop, the cleaned fertilized eggs are placed into a seed rearing pond, and the newly hatched larva fish continuously breaks membranes.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116171899A (en) * | 2023-03-24 | 2023-05-30 | 广西壮族自治区水产引育种中心 | Efficient de-gluing and hatching method for pelteobagrus fulvidraco fertilized eggs |
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| CN101268764A (en) * | 2008-05-07 | 2008-09-24 | 中国水产科学研究院黄海水产研究所 | Method for separating high quality oosperm of asropyle platichthys stellatus pallas |
| CN101288384A (en) * | 2007-04-20 | 2008-10-22 | 中国水产科学研究院淡水渔业研究中心 | Black shrimp intraspecific hybridization method |
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- 2020-07-13 CN CN202010668295.5A patent/CN111838033A/en active Pending
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|---|---|---|---|---|
| CN101288384A (en) * | 2007-04-20 | 2008-10-22 | 中国水产科学研究院淡水渔业研究中心 | Black shrimp intraspecific hybridization method |
| CN101268764A (en) * | 2008-05-07 | 2008-09-24 | 中国水产科学研究院黄海水产研究所 | Method for separating high quality oosperm of asropyle platichthys stellatus pallas |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116171899A (en) * | 2023-03-24 | 2023-05-30 | 广西壮族自治区水产引育种中心 | Efficient de-gluing and hatching method for pelteobagrus fulvidraco fertilized eggs |
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