CN114128652A - Black spot medaka embryo hatching device and method - Google Patents
Black spot medaka embryo hatching device and method Download PDFInfo
- Publication number
- CN114128652A CN114128652A CN202111462740.3A CN202111462740A CN114128652A CN 114128652 A CN114128652 A CN 114128652A CN 202111462740 A CN202111462740 A CN 202111462740A CN 114128652 A CN114128652 A CN 114128652A
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- medaka
- embryo
- separation
- hatching
- culture
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- 241000276569 Oryzias latipes Species 0.000 title claims abstract description 71
- 230000012447 hatching Effects 0.000 title claims abstract description 60
- 210000001161 mammalian embryo Anatomy 0.000 title claims abstract description 55
- 238000000034 method Methods 0.000 title claims abstract description 18
- 206010027146 Melanoderma Diseases 0.000 title description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 44
- 241000196324 Embryophyta Species 0.000 claims abstract description 38
- 210000002257 embryonic structure Anatomy 0.000 claims abstract description 32
- 239000013535 sea water Substances 0.000 claims abstract description 27
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims abstract description 18
- 229910052760 oxygen Inorganic materials 0.000 claims abstract description 18
- 239000001301 oxygen Substances 0.000 claims abstract description 18
- 239000000725 suspension Substances 0.000 claims abstract description 14
- 239000013013 elastic material Substances 0.000 claims abstract description 8
- 238000000926 separation method Methods 0.000 claims description 41
- 241000251468 Actinopterygii Species 0.000 claims description 7
- 230000001174 ascending effect Effects 0.000 claims description 7
- 230000001418 larval effect Effects 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 241000238426 Anostraca Species 0.000 claims description 3
- 230000005484 gravity Effects 0.000 claims description 3
- 125000006850 spacer group Chemical group 0.000 description 3
- 238000005265 energy consumption Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 208000002874 Acne Vulgaris Diseases 0.000 description 1
- 206010000496 acne Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 231100000463 ecotoxicology Toxicity 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000001632 homeopathic effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
Images
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
- A01K61/10—Culture of aquatic animals of fish
- A01K61/17—Hatching, e.g. incubators
-
- 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
- A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
- A01K63/04—Arrangements for treating water specially adapted to receptacles for live fish
- A01K63/042—Introducing gases into the water, e.g. aerators, air pumps
-
- 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)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Zoology (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention discloses a medaka embryo hatching device, which comprises a culture tank, wherein a plurality of simulated aquatic weeds are arranged in the culture tank, the simulated aquatic weeds are made of light elastic materials, a heavy object is fixed at the bottom ends of the simulated aquatic weeds, the heavy object is placed at the bottom of the culture tank, the culture tank is filled with water, and stem leaves of the simulated aquatic weeds are in a suspended state in the water. The stem and leaf of the simulated aquatic weed are connected with a culture chamber, the culture chamber can be a sphere or cube and other regular three-dimensional structures, a cavity is formed in the culture chamber, a plurality of gaps are formed in the outer wall of the culture chamber, seawater enters the cavity from the gaps, the culture chamber is made of a light elastic material, a culture tank is filled with water, and the culture chamber is in a suspension state in the seawater. According to the method, the simulated aquatic weed and the oxygen pump are used for simulating the marine ecological environment, the hatching environment in the suspension water is built for the hatching of the medaka embryos, and the oxygen pump is used for supplying oxygen and assisting in simulating the suspension of the aquatic weed to provide power.
Description
Technical Field
The invention belongs to the technical field of fish culture, and relates to a method for hatching a medaka embryo and a hatching device used by the hatching method.
Background
The medaka is an ossiphila model organism which is increasingly widely used internationally in marine ecotoxicology research in recent years. The novel sea water culture device is characterized by small size, life in sea water for the whole life and easy feeding in a laboratory; the generation cycle is short; the spawning rule is regular, and the embryo is transparent; the males and females are easily distinguished by morphology; the growth rate of individuals in the same batch is consistent, and the like.
The method for hatching the medaka embryos in the black spot used in laboratories at present is set according to the life habits of the medaka embryos in the black spot, and has the defects that the hatching rate is not fixed due to a plurality of influence factors, the hatching rate is random, the number of the medaka fries in the black spot cannot be guaranteed, and the subsequent test progress is seriously influenced. So the hatching rate is stabilized and improved, which is a problem to be solved in the current laboratory.
Disclosure of Invention
The first purpose of the invention is to provide a medaka embryo hatching device which can stabilize and improve the medaka embryo hatching rate.
Another object of the invention is to provide a method for hatching a medaka embryo, which also stabilizes and improves the rate of hatching the medaka embryo.
The first technical scheme adopted by the invention is that a medaka embryo hatching device comprises:
the culture tank is provided with a water outlet pipe connected with a water pump and used for replacing seawater;
the simulated aquatic weeds are arranged in the culture tank, the simulated aquatic weeds are made of light elastic materials, weights are fixed at the bottom ends of the simulated aquatic weeds, the weights are placed at the bottom of the culture tank, and the stems and leaves of the simulated aquatic weeds are in a suspension state in water;
the culture chamber is internally provided with a cavity, the outer wall of the culture chamber is provided with a plurality of gaps for communicating the culture cylinder with the culture chamber, the culture chamber is connected with stem leaves of simulated aquatic weeds, a light elastic material is selected for manufacturing the culture chamber, and the culture chamber is in a suspension state in seawater.
According to another technical scheme adopted by the invention, the method for hatching the embryo of the medaka in black spots comprises the following steps:
and 4, transferring newly hatched larval fish after membrane emergence, and feeding the larval fish in seawater with the salinity of 30 per mill and the temperature of 28 +/-2 ℃, wherein the light-dark period ratio is 14: feeding the newly hatched brine shrimp twice a day for 10 h.
The invention is also characterized in that:
the device also comprises an aerobic air pump, the oxygen pump is connected with a conduit, the conduit extends into the culture tank, the conduit is placed beside a heavy object, the conduit is provided with air holes, the air holes discharge air bubbles, the air bubbles rise upwards to generate ascending water flow, and the ascending water flow surrounds the simulated aquatic weeds, so that the simulated aquatic weeds do not sink due to the gravity of the culture chamber and keep a suspension state.
Also comprises an embryo separating mechanism which is arranged above the culture tank.
Embryo separating mechanism is including the separating chamber, and the suction pump is connected to the separating chamber bottom, and the delivery pipe is connected to the suction pump exit end, and the conveyer pipe will carry the culture room through single embryo that the separation handled, and the separating chamber vertically is provided with the spacer plate, and the spacer plate is two-layer about being divided into the separating chamber, has seted up the separation hole on the spacer plate.
The diameter of the separation hole is smaller than that of the medaka embryo.
The lower layer of the separation chamber is longitudinally provided with a separation block, the side wall of the separation block is fixedly connected with the inside of the separation chamber, two ends of the separation block leave spaces with the separation plate and the separation chamber, and the separation block is transversely provided with a separation pipe.
The separating tube is a tube body formed by connecting a plurality of round balls in series, the diameter of each round ball is larger than that of the medaka embryo, and the diameter of the interface of every two adjacent round balls is smaller than that of the medaka embryo.
The inner wall of the separation tube is provided with a barb, the size of the barb is the same as the size of villus on the surface of the medaka embryo, and the direction of the barb is opposite to the water flow direction.
The invention has the beneficial effects that:
1. according to the invention, the simulated aquatic weed and the oxygen pump are utilized to simulate the marine ecological environment, the hatching environment in the suspension water is built for the hatching of the medaka embryos, the oxygen pump has the function of supplying oxygen on one hand, and on the other hand, the oxygen pump assists in simulating the suspension of the aquatic weed to provide power, the whole hatching device is simple in structure, and the energy consumption is reduced.
2. According to the device, negative pressure water flow is used as a power source, and the sticky black oryzias latipes embryos are separated one by one according to the principle that the oryzias latipes embryos are separated due to different resistance in the advancing process, so that the device is time-saving and labor-saving, and replaces the complicated operation of manually screening the oryzias latipes one by one under a microscope by laboratory manpower.
Drawings
FIG. 1 is a schematic structural view of the present invention;
FIG. 2 is a schematic diagram of the embryo separation mechanism of the present invention;
FIG. 3 is an enlarged view of portion A of the present invention;
fig. 4 is an enlarged view of part B of the present invention.
In the figure, 1 is an culture tank, 2 is a simulated aquatic weed, 3 is a culture chamber, 4 is an oxygen pump, 5 is a weight, 6 is an embryo separating mechanism, 7 is a guide pipe, 6-1 is a separating chamber, 6-2 is a separating plate, 6-3 is a conveying pipe, 6-4 is a separating block, and 6-5 is a separating pipe.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
The device for hatching the medaka embryos in black spots comprises a culture cylinder 1, wherein the culture cylinder 1 is provided with a water outlet pipe, the water outlet pipe is connected with a water pump and used for replacing seawater, and the culture cylinder 1 can be further provided with a constant-temperature heating rod for controlling the hatching temperature, as shown in figures 1, 2, 3 and 4. A plurality of simulated aquatic weeds 2 are arranged in the culture tank 1, the simulated aquatic weeds 2 are made of light elastic materials, a weight 5 is fixed at the bottom end of each simulated aquatic weed 2, the weight 5 is placed at the bottom of the culture tank 1, the culture tank 1 is filled with water, and stem leaves of the simulated aquatic weeds 2 are in a suspension state in the water.
Connect culture room 3 on the stem leaf of simulation pasture and water 2, adopt pull ring or couple class detachable connection structure, culture room 3 can be the regular spatial structure such as spheroid or square, and its inside is the cavity, and 3 outer walls in culture room open and have a plurality of spaces, and the sea water gets into the cavity from the space, and culture room 3 chooses for use light elastic material to make, and culture tank 1 fills with water, and culture room 3 is out of suspended state in the sea water.
The black spot medaka embryo hatching device further comprises an oxygen pump 4, the oxygen pump 4 is connected with a guide pipe 7, the guide pipe 7 extends into the culture tank 1, the guide pipe 7 is placed beside the heavy object 5, air holes are formed in the guide pipe 7, air bubbles are discharged from the air holes, the air bubbles rise upwards to generate ascending water flow, the ascending water flow surrounds the simulated aquatic weeds 2, the simulated aquatic weeds 2 hang on the culture chambers 3, the simulated aquatic weeds 2 can sink under the gravity borne by the simulated aquatic weeds 2, the ascending water flow pushes the simulated aquatic weeds 2 upwards, and the simulated aquatic weeds 2 are forced to keep a suspension state.
The simulated aquatic weed 2 also comprises an embryo separating mechanism 6, the embryo separating mechanism 6 is arranged above the culture cylinder 1, the main function of the mechanism is to separate the adhered medaka embryos into one single medaka, and a proper number of medaka embryos are placed in the same culture chamber 3 for hatching.
The embryo separating mechanism 6 comprises a separating chamber 6-1, the bottom of the separating chamber 6-1 is connected with a water pump, the outlet end of the water pump is connected with a conveying pipe 6-3, the conveying pipe 6-3 extends into the culture chamber 3, the conveying pipe 6-3 conveys a single embryo subjected to separation treatment to the culture chamber 3, a separating plate 6-2 is longitudinally arranged in the separating chamber 6-1, the separating chamber 6-1 is divided into an upper layer and a lower layer by the separating plate 6-2, a separating hole is formed in the separating plate 6-2, the diameter of the separating hole is smaller than that of a black point medaka embryo, a separating block 6-4 is longitudinally arranged in the lower layer of the separating chamber 6-1, the side wall of the separating block 6-4 is fixedly connected with the inside of the separating chamber 6-1, spaces are reserved between the two ends of the separating block 6-4 and the separating plate 6-2 and the separating chamber 6-1, a separating pipe 6-5 is transversely arranged on the separating block 6-4, the separating tube 6-5 is a tube body formed by connecting a plurality of round balls in series, the diameter of each round ball is larger than that of the medaka embryo at the black spot, and the diameter of the interface of every two adjacent round balls is smaller than that of the medaka embryo at the black spot.
The separation hole has a function of blocking the separation tube 6-4, the medaka embryos have elasticity, and in two bonded medaka embryos, one is extruded through the separation hole along with water flow generated by the water pump to enter the medaka embryo in a homeopathic manner, and the other is blocked by the separation hole; or the plurality of adhered medaka embryos are separated through the separating tubes 6-4 and the interfaces among the round balls, and the medaka embryos are separated due to different resistance in the advancing process. The inner wall of the separation tube 6-5 is provided with a barb, the size of the barb is the same as the size of villus on the surface of the medaka embryo at the black spot, and the tip of the barb is passivated to ensure that the embryo is not punctured. The barb increases the friction force of the inner wall of the separation tube 6-5, the barb faces to the direction opposite to the water flow direction, and the medaka embryo in black spot moves along the water flow direction, namely the barb faces to the direction opposite to the progress of the medaka embryo in black spot.
According to another technical scheme adopted by the invention, the method for hatching the embryo of the medaka in black spots comprises the following steps:
and 4, transferring newly hatched larval fish after membrane emergence, and feeding the larval fish in seawater with the salinity of 30 per mill and the temperature of 28 +/-2 ℃, wherein the light-dark period ratio is 14: feeding the newly hatched brine shrimp twice a day for 10 h.
The invention relates to a medaka embryo hatching device, which comprises the following working processes:
and (3) starting a water pump by turning on a power supply, generating negative pressure water flow by the water pump, driving the medaka in black spots to flow along with the water flow, sequentially separating the medaka in two acne measures of the separation hole and the separation pipe, and separating 30-40 medaka embryos and then entering a culture chamber. And turning on an oxygen pump, supplying oxygen and keeping the simulated aquatic weeds and the culture chamber suspended.
Compared with the conventional standing hatching method, the embryo hatching rate is more stable and obviously improved, and the specific results are shown in the table below.
TABLE 1 comparison of hatchability of medaka embryos in black spots within 15 days under different methods
Note: p < 0.05.
The medaka embryo hatching device has the advantages that:
according to the invention, the simulated aquatic weed and the oxygen pump are utilized to simulate the marine ecological environment, the hatching environment in the suspension water is built for the hatching of the medaka embryos, the oxygen pump has the function of supplying oxygen on one hand, and on the other hand, the oxygen pump assists in simulating the suspension of the aquatic weed to provide power, the whole hatching device is simple in structure, and the energy consumption is reduced. The device is time-saving and labor-saving, and replaces the complex operation of manually screening the black medaka embryos under a microscope one by manpower in a laboratory.
Claims (9)
1. A medaka embryo hatching device is characterized by comprising:
the culture tank (1) is provided with a water outlet pipe which is connected with a water pump and used for replacing seawater;
the simulated aquatic weeds (2) are arranged in the culture tank (1), the simulated aquatic weeds (2) are made of light elastic materials, weights (5) are fixed at the bottom ends of the simulated aquatic weeds (2), the weights (5) are placed at the bottom of the culture tank (1), and stem leaves of the simulated aquatic weeds (2) are in a suspension state in water;
the culture chamber (3) is internally provided with a cavity, the outer wall of the culture chamber (3) is provided with a plurality of gaps, the gaps are used for communicating the culture cylinder (1) with the culture chamber (3), the culture chamber (3) is connected with stem leaves simulating aquatic weeds (2) and made of light elastic materials, and the culture chamber (3) is in a suspension state in seawater.
2. A medaka embryo hatching device according to claim 1, characterized in that the device further comprises an aerobic air pump (4), the oxygen pump (4) is connected with a conduit (7), the conduit (7) extends into the culture tank (1), the conduit (7) is placed beside the weight (5), the conduit (7) is provided with air holes, air bubbles are discharged from the air holes, the air bubbles rise upwards to generate ascending water flow, and the ascending water flow surrounds the simulated aquatic weeds (2), so that the simulated aquatic weeds (2) do not sink due to the gravity of the culture chamber (3) and keep the suspended state.
3. A medaka embryo hatching device according to claim 1, further comprising an embryo separating mechanism (6), wherein the embryo separating mechanism (6) is arranged above the culture tank (1).
4. A medaka embryo hatching device according to claim 3, characterized in that the embryo separating mechanism (6) comprises a separating chamber (6-1), the bottom of the separating chamber (6-1) is connected with a water pump, the outlet end of the water pump is connected with a delivery pipe (6-3), the delivery pipe (6-3) delivers a single embryo which is subjected to separation treatment to the culture chamber (3), a separating plate (6-2) is longitudinally arranged in the separating chamber (6-1), the separating chamber (6-1) is divided into an upper layer and a lower layer by the separating plate (6-2), and a separating hole is formed in the separating plate (6-2).
5. A medaka embryo hatching device according to claim 4, wherein the diameter of the separation hole is less than the diameter of the medaka embryo.
6. A medaka embryo hatching device according to claim 3, characterized in that a separation block (6-4) is longitudinally arranged at the lower layer of the separation chamber (6-1), the side wall of the separation block (6-4) is fixedly connected with the inside of the separation chamber (6-1), spaces are reserved between the two ends of the separation block (6-4) and the separation plate (6-2) and the separation chamber (6-1), and a separation pipe (6-5) is transversely arranged on the separation block (6-4).
7. A medaka embryo hatching device according to claim 6, characterized in that the separation tube (6-5) is a tube body formed by connecting a plurality of round balls in series, the diameter of each round ball is larger than that of the medaka embryo, and the interface diameter of two adjacent round balls is smaller than that of the medaka embryo.
8. A medaka embryo hatching device according to claim 6, characterized in that the inner wall of the separating tube (6-5) is provided with barbs, the size of the barbs is the same as the size of villi on the surface of the medaka embryo, and the barbs face in the direction opposite to the water flow direction.
9. A method for hatching a medaka embryo, which uses the medaka embryo hatching device according to claims 1-8, and comprises the following steps:
step 1, carrying out separation operation on medaka embryos in black spots, and putting the medaka embryos after separation into a hatching device for hatching;
step 2, introducing clean seawater into the hatching device, replacing the clean seawater once a day, and hatching until membranes are produced, wherein the salinity of the clean seawater is 30 per mill, the water temperature is 28 +/-2 ℃, and the light-dark period ratio in the hatching process is 14: 10 h;
step 3, when clean seawater is replaced, taking out medaka embryos from the hatching device, and removing unfertilized embryos or dead embryos under a microscope;
and 4, transferring newly hatched larval fish after membrane emergence, and feeding the larval fish in seawater with the salinity of 30 per mill and the temperature of 28 +/-2 ℃, wherein the light-dark period ratio is 14: feeding the newly hatched brine shrimp twice a day for 10 h.
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