CN108077118B - Artificial regulation and control method for repeated spermiation and ovulation in propagation season of amphioxus - Google Patents
Artificial regulation and control method for repeated spermiation and ovulation in propagation season of amphioxus Download PDFInfo
- Publication number
- CN108077118B CN108077118B CN201711311481.8A CN201711311481A CN108077118B CN 108077118 B CN108077118 B CN 108077118B CN 201711311481 A CN201711311481 A CN 201711311481A CN 108077118 B CN108077118 B CN 108077118B
- Authority
- CN
- China
- Prior art keywords
- ovulation
- algae
- seawater
- spermiation
- rejuvenation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 230000016087 ovulation Effects 0.000 title claims abstract description 55
- 238000000034 method Methods 0.000 title claims abstract description 32
- 241000251522 Cephalochordata Species 0.000 title claims abstract description 27
- 241000251468 Actinopterygii Species 0.000 claims abstract description 58
- 230000003716 rejuvenation Effects 0.000 claims abstract description 54
- 238000011534 incubation Methods 0.000 claims abstract description 14
- 241000195493 Cryptophyta Species 0.000 claims description 82
- 239000013535 sea water Substances 0.000 claims description 50
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 32
- 210000002149 gonad Anatomy 0.000 claims description 14
- 239000004576 sand Substances 0.000 claims description 13
- 230000003203 everyday effect Effects 0.000 claims description 12
- 230000008569 process Effects 0.000 claims description 12
- 230000009471 action Effects 0.000 claims description 11
- 241000091751 Chaetoceros muellerii Species 0.000 claims description 10
- 241000196316 Tetraselmis subcordiformis Species 0.000 claims description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 240000009108 Chlorella vulgaris Species 0.000 claims description 7
- 235000007089 Chlorella vulgaris Nutrition 0.000 claims description 7
- 238000009835 boiling Methods 0.000 claims description 5
- 230000006872 improvement Effects 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 229930003231 vitamin Natural products 0.000 claims description 5
- 235000013343 vitamin Nutrition 0.000 claims description 5
- 239000011782 vitamin Substances 0.000 claims description 5
- 229940088594 vitamin Drugs 0.000 claims description 5
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 4
- 239000001963 growth medium Substances 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 230000035800 maturation Effects 0.000 claims description 3
- 238000004064 recycling Methods 0.000 claims description 3
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 2
- 229910052782 aluminium Inorganic materials 0.000 claims description 2
- 229960002685 biotin Drugs 0.000 claims description 2
- 235000020958 biotin Nutrition 0.000 claims description 2
- 239000011616 biotin Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 239000012528 membrane Substances 0.000 claims description 2
- 239000006228 supernatant Substances 0.000 claims description 2
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims 3
- 241000030251 Chelmon muelleri Species 0.000 claims 1
- 241000168525 Haematococcus Species 0.000 claims 1
- 241001466487 Phaeocystis Species 0.000 claims 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims 1
- 229910052925 anhydrite Inorganic materials 0.000 claims 1
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 claims 1
- YYRMJZQKEFZXMX-UHFFFAOYSA-N calcium;phosphoric acid Chemical compound [Ca+2].OP(O)(O)=O.OP(O)(O)=O YYRMJZQKEFZXMX-UHFFFAOYSA-N 0.000 claims 1
- 230000009027 insemination Effects 0.000 claims 1
- 239000011148 porous material Substances 0.000 claims 1
- 239000002426 superphosphate Substances 0.000 claims 1
- 230000012447 hatching Effects 0.000 abstract description 9
- 241000251534 Branchiostoma Species 0.000 abstract description 8
- 230000004720 fertilization Effects 0.000 abstract description 2
- 230000004083 survival effect Effects 0.000 abstract description 2
- 241000251537 Branchiostoma belcheri Species 0.000 description 9
- 238000009395 breeding Methods 0.000 description 6
- 230000001488 breeding effect Effects 0.000 description 6
- 241001078425 Isochrysis zhanjiangensis Species 0.000 description 5
- 238000007599 discharging Methods 0.000 description 4
- 238000012258 culturing Methods 0.000 description 3
- 235000013601 eggs Nutrition 0.000 description 3
- 210000000582 semen Anatomy 0.000 description 3
- 229930003270 Vitamin B Natural products 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 238000006213 oxygenation reaction Methods 0.000 description 2
- 235000019156 vitamin B Nutrition 0.000 description 2
- 239000011720 vitamin B Substances 0.000 description 2
- 241000227752 Chaetoceros Species 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241001501885 Isochrysis Species 0.000 description 1
- 241001236817 Paecilomyces <Clavicipitaceae> Species 0.000 description 1
- 241001509341 Pyramimonas Species 0.000 description 1
- 241001509149 Pyramimonas sp. Species 0.000 description 1
- 210000004534 cecum Anatomy 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 241000894007 species Species 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
-
- 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
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Animal Husbandry (AREA)
- Wood Science & Technology (AREA)
- Marine Sciences & Fisheries (AREA)
- Food Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Environmental Sciences (AREA)
- Organic Chemistry (AREA)
- Botany (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Cell Biology (AREA)
- Biomedical Technology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Microbiology (AREA)
- Birds (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Health & Medical Sciences (AREA)
- Insects & Arthropods (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Physiology (AREA)
- Farming Of Fish And Shellfish (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to an artificial regulation and control method for repeated spermiation and ovulation in the propagation season of amphioxus. The method comprises the following steps: respectively selecting 4-5-year-old male and female Florida branchiostoma, placing the 4-5-year-old male and female Florida branchiostoma in an incubation cylinder for intensive cultivation according to the ratio of 10-18 male parent fishes to 10 female parent fishes until the parent fishes are observed to begin to perform first spermiation and ovulation; transferring the parent fish after the first spermiation and ovulation to a rejuvenation cultivation pond for postnatal rejuvenation cultivation until the parent fish is observed to have second spermiation and ovulation; the above method and the cultivation conditions are repeated until the parent fish is observed to begin to perform spermiation and ovulation for the fourth, fifth, sixth, seventh, eighth, ninth and tenth times. By adopting the method, the ovulation frequency of the amphioxus is high, the fertilization rate reaches 55-67%, the hatching rate of emergence of 10-12 h reaches 90-93%, the survival rate of 5 days reaches 70-75%, and sufficient fry are provided for the scale artificial propagation of the amphioxus.
Description
Technical Field
The invention relates to the technical field of artificial propagation of branchiostoma belcheri, in particular to an artificial regulation and control method for repeated spermiation and ovulation in a branchiostoma belcheri propagation season.
Background
In 1937, the indoor propagation technology of amphioxus was broken through in China first in the third week of childhood, etc., and in 1952, the normally-developed ovum was obtained by artificial fertilization. Later, many scholars at home and abroad develop research work on the laboratory feeding and breeding of the amphioxes successively, and the promotion achievement which has more influence on the artificial breeding of the amphioxes is as follows: yasui and the like complete the indoor feeding and reproduction of the Japanese branchiostoma by constructing a circulating water breeding system and breed offspring branchiostoma; wu et al have reported successful reproduction of Wenchang first filial generation in laboratory and raised in outdoor container to over 2 a; the Wangbei and Zhang, etc. obtain the second generation of the branchiostoma, and initially realize the full artificial culture of the branchiostoma in the laboratory.
The propagation seasons of the branchiostoma belcheri of different species or different geographical populations are different, the natural propagation season of the branchiostoma belcheri in northern hemisphere is 5-6 months, the natural propagation season of the branchiostoma belcheri in southern hemisphere is 11-12 months, and the general branchiostoma belcheri only has 1 semen excretion or ovulation. And the amphioxus in the tropical zone can naturally lay eggs for 2 times or more in one year. China is in temperate zones and subtropical zones, and the natural spawning of the amphioxus can be realized only 1 time in one year. Aiming at different growth periods of the amphioxes, the bait formula and different feeding management methods are changed, and comprehensive measures of artificial regulation are combined to regulate and control rejuvenation cultivation of the amphioxes after ovulation of sperms to mature gonads, so that repeated sperms and ovulation of the amphioxes are realized within the natural breeding season of the amphioxes.
Disclosure of Invention
The artificial regulation and control method for repeated spermiation and ovulation in one year of a single individual amphioxus is realized by taking the amphioxus florida as a material and by adopting comprehensive technical schemes of bait improvement culture, seawater quality control, illumination and temperature regulation, after-natal rejuvenation and the like. The technology lays a good foundation for the scale artificial propagation of the amphioxus.
The technical scheme adopted for realizing the purpose of the invention is as follows:
1. parent fish selection
Selecting part of male and female florida Amphioxus (Branc jiosttoma. floridae) with the ages of 4-5 as parent fishes for later use.
2. First ovulation of sperm
Selecting parent fishes of the amphioxus florida, which are healthy, good in liver cecum development and good in gonad plump development level, placing the parent fishes in an incubation cylinder filled with seawater and fine sand with the thickness of 5-10 cm paved at the bottom according to the proportion of 10-18 male parent fishes to 10 female parent fishes, carrying out prenatal fish body intensified culture by adopting micro-water flow circulation and artificial light control, feeding intensified unicellular algae mixed algae 2 times every day in an equal amount in the intensified culture process, feeding the intensified unicellular algae mixed algae 0.5-0.6 million intensified unicellular algae per cubic seawater each time, and after the gonads of the parent fishes are fully developed in stage V, 16 pm every day: 00 observing whether the parent fish has the action of excreting sperm and ovulation until the parent fish is observed to begin to carry out the first action of excreting sperm and ovulation.
The temperature of seawater in the hatching tank is between 24 and 27 ℃, the salinity of the seawater is 28 to 33 per thousand, and the pH of the seawater is controlled to be between 8 and 9.
The feeding is carried out 2 times a day in an equal amount, the reinforced single-cell mixed algae is fed when the fixed time is 8:00-9:00 o 'clock every day for the first time, and the reinforced single-cell mixed algae is fed when the fixed time is 16:00-17:00 o' clock every day for the second time.
The Light quantity of the manual Light control is 10Dark between 08:30 and 18:30, and 14Light is 18:30 to 08: 30.
The micro water flow circulation is to strengthen the micro water flow circulation flow, and the micro water flow circulation flow rate is 1L/h. The process is as follows: the seawater in the high-position reservoir flows into the water inlet of the middle-position incubation cylinder or the rejuvenation culture pond through the rubber pipe by means of water level difference, the seawater flows into the low-position reservoir through the water outlet of the incubation cylinder or the rejuvenation culture pond, and the seawater in the low-position reservoir is pumped back to the high-position reservoir by the micro water pump for recycling.
The reinforced single-cell algae mixed algae is obtained by mixing the following algae species in parts by weight:
the intensified single-cell algae mixed algae is composed of Platymonas subcordiformis (Platymonas subcordiformis), Isochrysis zhanjiangensis (Isochrysis zhanjiangensis), Chlorella vulgaris (Chlorella vulgaris), Chaetoceros muelleri (Chaetoceros mueller) and Tachysolen algae (Pyramimonas sp), and is obtained by performing quality improvement by artificially culturing single-cell algae from algae research laboratory of institute of bioscience, university of Fujian.
The artificially cultured unicellular algae is obtained by adopting a self-prepared improved culture medium for culture.
The formula of the improved culture medium is as follows:
the sea mud extract is prepared by taking upper layer soft mud which has less sand on a beach and more organic matters but is not excessively silted black, removing sundries such as small branches and small stones in the upper layer soft mud, adding 2 parts of water into 1 part of mud by volume, fully and uniformly stirring, standing for l-2 min, pouring the upper layer mud into an aluminum pot after coarse sand and small stones sink, removing sundries such as bottom coarse sand and small stones, adding 1g of NaOH into every 1000ml of mud, boiling for 20-30 min, continuously stirring during boiling, standing for 24h after boiling, and absorbing supernatant for use.
The vitamin solution is prepared from 10.1g of vitamin B, 120.5 g of vitamin B, 0.5g of biotin and 1L of seawater.
The seawater refers to seawater obtained by filtering ordinary seawater from offshore through a filter membrane with the aperture of 0.4 m.
3. Rejuvenation cultivation and secondary ovulation
Transferring the parent fish after the first spermiation and ovulation to a rejuvenation cultivation pond containing seawater, and performing post-partum fish rejuvenation and gonad maturation by adopting micro-water flow circulation and artificial light control to perform rejuvenation cultivation, wherein the rejuvenation cultivation process regularly feeds equal amount of rejuvenation single-cell algae mixed algae 2 times a day, and the amount of the rejuvenated single-cell algae mixed algae fed each time is 0.5-0.6 ten thousand of rejuvenated single-cell algae mixed algae per cubic seawater. The regular feeding, the artificial light control, the seawater temperature, the seawater salinity and the seawater pH value in the rejuvenation cultivation process are the same as those in the step 2. When the gonad of the parent fish is in stage V, 16:00 observing whether the parent fish has the action of ovulation and spermiation for the second time until observing that the parent fish has the action of ovulation and spermiation for the second time. And after the second ovulation and spermiation, transferring all the spermiated parent fishes to the next rejuvenation cultivation pool for repeated rejuvenation cultivation.
The rejuvenation single-cell algae mixed algae is obtained by mixing the following algae species in parts by weight:
the above-mentioned rejuvenated unicellular algae mixed algae are Platymonas subcordiformis (Platymonas subcordiformis), Isochrysis zhanjiangensis (Isochrysis zhanjiangensis), Chlorella vulgaris (Chlorella vulgaris) and Chaetoceros muelleri (Chaetoceros muelleri), and are obtained by performing quality improvement by artificial culture of unicellular algae from algae research laboratory of institute of bioscience, university of Fujian province.
4. Repeated rejuvenation cultivation and third ovulation of semen
And (3) placing the parent fish after the second spermiation and ovulation in a rejuvenation culture pond, and repeating the method and the culture condition in the step (3) until the parent fish is observed to begin the fourth spermiation and ovulation.
And 3, continuously repeating the method and the cultivation condition in the step 3, and realizing repeated ejaculation and ovulation for multiple times in the breeding season. In order to keep the hatching tank or the rejuvenation cultivation pond clean, the fine sand in the hatching tank or the rejuvenation cultivation pond is washed every two weeks. Providing sufficient bait and timely micro-water flow for circular oxygenation.
By adopting the artificial regulation and control method, the sperms discharging and ovulation frequency of the amphioxus is high in 4-11 months, the sperms discharging and ovulation frequency of most of the amphioxus is 8 or 9 times, the highest sperms discharging and ovulation frequency can reach 10 times, the fertility rate reaches 55-67%, the hatching rate of fertilized eggs for 10-12 hours reaches 90-93%, the survival rate in 5 days reaches 70-75%, and sufficient fish fries are provided for large-scale artificial propagation of the amphioxus.
Detailed Description
Example 1
1. Selecting parent fish: 2016 (3/21/3), when spawning season comes, 5-year old (2011 incubation) Branchiostoma florida fish is screened out from sand, and at this time, healthy individuals with fully developed male and female gonads are carefully selected, and then 16 randomly selected (10 female and 6 male) fishes are used as parent fishes for standby. According to the sex, the body length, the number of the left and right gonads and the morphological characteristics of the fin chamber of the anterior fin of the hip of the selected branchiostoma, different parent fish individuals are marked and numbered, as shown in the following table:
table 1 initial data for experimental amphioxus (2016 year 3 month 21 day)
Placing fine sand which is cleaned and dried in advance and is rinsed by seawater into an incubation cylinder of the branchiostoma belcheri for paving the branchiostoma belcheri for 10cm thick, then placing fresh and clean seawater which is 5-10 cm higher than the sand surface, and placing the selected branchiostoma belcheri with 16 full gonads into the incubation cylinder.
2. First ovulation of sperm
Carrying out prenatal fish body intensive cultivation by micro-water flow circulation and artificial light control, feeding the reinforced single-cell algae mixed algae in an equal amount 2 times a day in the reinforced cultivation process, wherein the amount of the reinforced single-cell algae mixed algae fed every time is 0.5-0.6 ten thousand reinforced single-cell algae mixed algae per cubic seawater, and when the gonad of the parent fish is fully developed in stage V, 16 pm every day: 00 observing whether the parent fish has the action of excreting sperm and ovulation until the parent fish is observed to begin to carry out the first action of excreting sperm and ovulation.
The temperature of seawater in the hatching tank is controlled to be 27 ℃, the salinity of the seawater is controlled to be 28 per mill, and the pH value of the seawater is controlled to be 8-9.
The feeding is carried out 2 times a day in an equal amount, the reinforced single-cell algae mixed algae is fed for the first time at 8:00-9:00 am every day at a fixed time, and the reinforced single-cell algae mixed algae is fed for the second time at 16:00-17:00 pm every day at a fixed time.
The Light quantity of the manual Light control is 10Dark between 08:30 and 18:30, and 14Light is 18:30 to 08: 30.
The reinforced single-cell algae mixed algae is obtained by the following method:
respectively selecting the intensified single-cell algae mixed algae artificially cultured in a laboratory as Platymonas subcordiformis (Platymonas subcordiformis), Isochrysis glabrata (Isochrysis zhanjiangensis), Chlorella vulgaris (Chlorella vulgaris), Chaetoceros muelleri (Chaetoceros muelleri) and Paecilomyces takawakii (Pyramimonas sp.) according to the following weight ratio:
mixing to obtain the strengthened unicellular algae mixture.
The micro water flow circulation is to strengthen the micro water flow circulation flow, and the micro water flow circulation flow rate is 1L/h. The process is as follows: the seawater in the high-position reservoir flows into the water inlet of the middle-position incubation cylinder or the rejuvenation culture pond through the rubber pipe by means of water level difference, the seawater flows into the low-position reservoir through the water outlet of the incubation cylinder or the rejuvenation culture pond, and the seawater in the low-position reservoir is pumped back to the high-position reservoir by the micro water pump for recycling.
3. Rejuvenation cultivation and secondary ovulation
Transferring the parent fish after the first spermiation and ovulation to a rejuvenation cultivation pool filled with seawater for rejuvenation cultivation, regularly feeding equal rejuvenated single-cell algae mixed algae for 2 times every day, wherein the quantity of the rejuvenated single-cell algae mixed algae fed every time is 0.5-0.6 ten thousand rejuvenated single-cell algae mixed algae per cubic seawater, and the feeding time, the illumination condition, the seawater temperature, the seawater salinity and the seawater pH value in the rejuvenation cultivation process are the same as those in the step 2. And carrying out post-partum fish rejuvenation and gonad maturation cultivation by micro-water flow circulation and artificial light control, when the gonad of the parent fish is fully developed in stage V, 16 in the afternoon each day: 00 observing whether the parent fish has the action of ovulation and spermiation for the second time until observing that the parent fish has the action of ovulation and spermiation for the second time. And after the second ovulation and spermiation, transferring all the spermiated parent fishes to the next rejuvenation cultivation pond for repeated rejuvenation cultivation.
Artificially culturing and rejuvenating the mixed algae of the single-cell algae in a laboratory, and matching and composing according to the following different algae species:
4. repeated rejuvenation cultivation and third ovulation of semen
And (3) placing the parent fish after the second spermiation and ovulation in a rejuvenation culture pond, and repeating the method and the culture condition in the step (3) until the parent fish is observed to begin the fourth spermiation and ovulation.
And 3, continuously repeating the method and the cultivation condition in the step 3, and realizing repeated ejaculation and ovulation for multiple times in the breeding season. In order to keep the hatching tank or the rejuvenation cultivation pond clean, the fine sand in the hatching tank or the rejuvenation cultivation pond is washed every two weeks. Providing sufficient bait and timely micro-water flow for circular oxygenation.
Artificially culturing and rejuvenating the mixed algae of the single-cell algae in a laboratory, and matching and composing according to the following different algae species:
the experimental results are as follows: in 2016, in 16 amphioxus, the 9 th amphioxus died halfway after discharging sperms for 5 times, and other 15 (10 female parent and 5 male parent) amphioxus have ovulation and sperm discharge frequencies concentrated in 7-10 times in 4-11 months, wherein the spawning times of most of the amphioxus are 8 or 9 times, as shown in the following table:
ovulation and sperm discharge frequency of Table 2-year Florida amphioxus
Hatching condition: 45000 fertilized eggs are obtained in the test, and are incubated in an incubator for about 10-12 hours in the embryo development process under the conditions that the seawater temperature is 24.5-26.0 ℃ and the salinity is 28-33 per mill, and the incubation emergence rate reaches 91.8%.
Claims (5)
1. An artificial control method for repeated spermiation and ovulation in the propagation season of amphioxus, which is characterized in that:
1) parent fish selection
Selecting male and female Florida amphioxus of 4-5 ages respectively (Branc jiosttoma. floridae) The fish is used as parent fish for standby;
2) first ovulation of sperm
Placing 10-18 male parent fishes in proportion to 10 female parent fishes in an incubation cylinder filled with seawater and paved with fine sand with the thickness of 5-10 cm at the bottom, performing prenatal fish body intensive cultivation by adopting micro water flow circulation and artificial light control, feeding reinforced single-cell algae mixed algae in an equal amount 2 times every day in the reinforced cultivation process, wherein the amount of the reinforced single-cell algae mixed algae fed every time is 0.5-0.6 ten thousand reinforced single-cell algae mixed algae per cubic seawater, and when the gonads of the parent fishes are fully developed in a V stage, 16 pm every day: 00 observing whether the parent fish has the action of spermiation and ovulation until the parent fish is observed to start to perform the first spermiation and ovulation;
3) rejuvenation cultivation and secondary ovulation
Transferring the parent fish after the first spermiation and ovulation to a rejuvenation cultivation pool filled with seawater to perform post-partum fish rejuvenation and gonad maturation cultivation by adopting micro-water flow circulation and artificial light control, regularly feeding equal amount of rejuvenation single-cell algae mixed algae 2 times a day in the rejuvenation cultivation process, wherein the amount of the rejuvenation single-cell algae mixed algae fed every time is 0.5-0.6 ten thousand rejuvenation single-cell algae mixed algae per cubic seawater; the regular feeding, the artificial light control, the seawater temperature, the seawater salinity and the seawater pH value in the rejuvenation cultivation process are the same as those in the step 2), when the gonad of the parent fish is fully developed in the stage V, 16:00 observing whether the parent fish has the action of ovulation and spermiation for the second time until the parent fish is observed to have the action of ovulation and spermiation for the second time;
after finishing the second spermiation and ovulation, transferring the parent fish to the next rejuvenation cultivation pool for repeated rejuvenation cultivation; continuously repeating the method and the culture conditions of the step 3) until the fourth, fifth, sixth, seventh, eighth, ninth and tenth ovulation and spermiation of the parent fish are observed;
the reinforced single-cell algae mixed algae is obtained by mixing the following algae species in parts by weight:
platymonas subcordiformis 40-60
15-20 of Zhanjiang isoflagellate
5-10 parts of chlorella
Chaetoceros muelleri 10-15
10-15 parts of phaeocystis;
the intensified single-cell algae mixed algae is Platymonas subcordiformis (A. subcordiformis)Platymonas subcordiformis) Haematococcus, Zhanjiang and the likeIsochrysis zhanjiangensis) Chlorella (A) and (B)Chlorella vulgaris) Chaetoceros muelleri (C. mulleri) (C. muelleri)Chaetoceros muelleri) And Taocystis (a), (b), (c), (dPyramimonas sp.) The compositions are all obtained by carrying out quality improvement in an artificial culture mode of unicellular algae from an algae research laboratory of the institute of life science of the university of Fujian faculty;
the rejuvenation single-cell algae mixed algae is obtained by mixing the following algae species in parts by weight:
platymonas subcordiformis 20-25
25-30 of Zhanjiang isoflagellate
Chaetoceros muelleri 10-15
30-35 parts of chlorella;
the temperature of seawater in the incubation cylinder in the step 2) is between 24 and 27 ℃, the salinity of the seawater is 28 to 33 per thousand, and the pH of the seawater is controlled to be between 8 and 9;
feeding the mixture of the reinforced single-cell algae 2 times a day in the step 2), feeding the reinforced single-cell algae mixture at 8:00-9:00 am every day for the first time, and feeding the reinforced single-cell algae mixture at 16:00-17:00 pm every day for the second time;
the artificial Light control is performed, wherein the Light quantity is 10Dark between 08:30 and 18:30, and 14Light is 18:30 to 08: 30;
the micro water flow circulation is to strengthen the micro water flow circulation flow, the micro water flow circulation flow rate is 1L/h, and the process is as follows: the seawater in the high-position reservoir flows into the water inlet of the middle-position incubation cylinder or the rejuvenation culture pond through the rubber pipe by means of water level difference, the seawater flows into the low-position reservoir through the water outlet of the incubation cylinder or the rejuvenation culture pond, and the seawater in the low-position reservoir is pumped back to the high-position reservoir by the micro water pump for recycling.
2. The artificial regulation and control method for repeated ovulation and spermiation in the propagation season of amphioxus according to claim 1, characterized in that the artificially cultured unicellular algae are subjected to quality improvement and are obtained by adopting a self-prepared improved culture medium for culture;
the formula of the improved culture medium is as follows:
（NH4）2SO4 200mg
sea mud extract 20ml
Superphosphate [ Ca (H) ]2PO4)2·H2O+2(CaSO4·H2O)] 30mg
FeC6H5O7(1% solution) 0.5ml
Vitamin solution 20ml
1000ml of seawater.
3. The artificial control method for repeated insemination and ovulation in the propagation season of the amphioxus according to claim 2, characterized in that the sea mud extract is prepared by taking the upper soft mud which has less sand and more organic matters and is not excessively silted black on the beach, removing the sundries of small branches and small stones, adding 2 parts of water into 1 part of mud by volume, fully and uniformly stirring, standing for l-2 min, pouring the upper mud into an aluminum pot after the coarse sand and the small stones subside, removing the sundries of coarse sand and the small stones at the bottom, adding NaOH in an amount of 1g of NaOH into each 1000ml of mud, boiling for 20-30 min, continuously stirring during boiling, standing for 24h, and absorbing the supernatant for use.
4. The artificial regulation and control method for repeated ejaculation and ovulation in the propagation season of Amphioxus as claimed in claim 2, wherein the vitamin solution comprises vitamin B10.1g, vitamin B120.5g, biotin 0.5g, and 1L seawater.
5. The method as claimed in claim 2, wherein the seawater is obtained by filtering normal seawater from offshore region with 0.4 m-pore size filter membrane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711311481.8A CN108077118B (en) | 2017-12-11 | 2017-12-11 | Artificial regulation and control method for repeated spermiation and ovulation in propagation season of amphioxus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711311481.8A CN108077118B (en) | 2017-12-11 | 2017-12-11 | Artificial regulation and control method for repeated spermiation and ovulation in propagation season of amphioxus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108077118A CN108077118A (en) | 2018-05-29 |
| CN108077118B true CN108077118B (en) | 2021-06-29 |
Family
ID=62174659
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711311481.8A Active CN108077118B (en) | 2017-12-11 | 2017-12-11 | Artificial regulation and control method for repeated spermiation and ovulation in propagation season of amphioxus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108077118B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114128663B (en) * | 2021-11-24 | 2023-02-10 | 大连海洋大学 | Thermo-optical regulation and control method for anti-season propagation of parent fish of takifugu rubripes and application of thermo-optical regulation and control method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106172101A (en) * | 2016-07-10 | 2016-12-07 | 龙南源头活水生态科技有限责任公司 | A kind of moon murrel Ecology breeding and efficient and healthful cultural method |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6584935B2 (en) * | 2001-03-16 | 2003-07-01 | University Of Maryland Biotechnology Institute | Process for culturing crabs in recirculating marine aquaculture systems |
| US9205284B2 (en) * | 2010-08-09 | 2015-12-08 | Allele Biotechnology & Pharmaceuticals, Inc. | Light-absorbing compositions and methods of use |
| CN104782577B (en) * | 2015-04-13 | 2018-07-27 | 临沂大学 | A method of with bottle,suction culture beak tip Magna |
| CN206061856U (en) * | 2016-06-06 | 2017-04-05 | 蔡东林 | Multi-functional batch production Aquatic product sapling multiplication device |
| CN107041330B (en) * | 2017-06-08 | 2019-07-23 | 福建师范大学 | A kind of method of lancelet artificial induced spawning fish-egg |
-
2017
- 2017-12-11 CN CN201711311481.8A patent/CN108077118B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106172101A (en) * | 2016-07-10 | 2016-12-07 | 龙南源头活水生态科技有限责任公司 | A kind of moon murrel Ecology breeding and efficient and healthful cultural method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108077118A (en) | 2018-05-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102090361A (en) | Artificial breeding method for Acrossocheilus fasciatus | |
| KR20180002549A (en) | Solid insulation fish tank system for fish preserve and aquaculture of high density in inner-city on land based | |
| CN106719184A (en) | A kind of artificial breeding method of Mactra chinensis | |
| Gao et al. | Transitions, challenges and trends in China's abalone culture industry | |
| CN101699998B (en) | Selection and culture method of excellent breed large-tooth flounder featuring fast growth and high survival rate | |
| CN102511424A (en) | Artificial eel propagation method | |
| JP2011244795A (en) | Method for producing on land of sand-submerged bivalves, such as littleneck clam | |
| CN108077118B (en) | Artificial regulation and control method for repeated spermiation and ovulation in propagation season of amphioxus | |
| CN103461224B (en) | The system of selection of seed ginseng in stichopus japonicus crossbreeding | |
| CN109287533B (en) | Large-scale breeding method and equipment for hybrid scallop offspring seeds | |
| CN107087560A (en) | Penaeus Vannmei and the foundation and its application of golden pomfret ecological polyculturing pattern | |
| CN204949087U (en) | Giant salamander hatching apparatus | |
| CN111316943A (en) | Pond seedling raising method for ostrea rivularis | |
| Yang et al. | Induced ovulation using LHRHa in anadromous obscure puffer Takifugu obscurus cultured entirely in freshwater | |
| CN103081828B (en) | Method for artificially propagating parabramis pekinensis | |
| CN105210937A (en) | A kind of loach ecological cultivation method | |
| CN210580538U (en) | Australia freshwater lobster circulating water breeding system | |
| JP2010041983A (en) | Method of producing breeder of pinctada fucata martensii and noble scallop, and its breeder | |
| CN104351085A (en) | Breeding method of river crabs | |
| CN111387095B (en) | Breeding method for improved variety parent of mullet | |
| CN114532264A (en) | Staged pond culture method for improving pearl quality of pinctada maxima | |
| CN107867757B (en) | Water quality improvement method for cultivating grouper fries | |
| CN108124802B (en) | Method for ecologically cultivating juvenile crabs into 1-year-old crab seeds in stage III of salt marsh wetland | |
| CN107125174B (en) | Method for promoting gonad development of blood clam | |
| CN112673998A (en) | Cyclic cultivation method for stichopus japonicus and urechis unicinctus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |