CN115176767A - Artificial hatching method for bee frozen eggs - Google Patents
Artificial hatching method for bee frozen eggs Download PDFInfo
- Publication number
- CN115176767A CN115176767A CN202210969368.3A CN202210969368A CN115176767A CN 115176767 A CN115176767 A CN 115176767A CN 202210969368 A CN202210969368 A CN 202210969368A CN 115176767 A CN115176767 A CN 115176767A
- Authority
- CN
- China
- Prior art keywords
- eggs
- bee
- hatching
- frozen
- paraffin oil
- 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.)
- Pending
Links
- 235000013601 eggs Nutrition 0.000 title claims abstract description 153
- 230000012447 hatching Effects 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 26
- 239000005662 Paraffin oil Substances 0.000 claims abstract description 42
- 238000004113 cell culture Methods 0.000 claims abstract description 17
- 239000011248 coating agent Substances 0.000 claims abstract description 16
- 238000000576 coating method Methods 0.000 claims abstract description 16
- 238000007710 freezing Methods 0.000 claims abstract description 10
- 230000008014 freezing Effects 0.000 claims abstract description 10
- 241000257303 Hymenoptera Species 0.000 claims abstract description 9
- 241000256844 Apis mellifera Species 0.000 claims description 96
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 27
- 238000011282 treatment Methods 0.000 claims description 19
- 239000002250 absorbent Substances 0.000 claims description 5
- 230000002745 absorbent Effects 0.000 claims description 5
- 238000010257 thawing Methods 0.000 claims description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 241000256837 Apidae Species 0.000 claims 4
- 230000001681 protective effect Effects 0.000 abstract description 11
- 230000004083 survival effect Effects 0.000 abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 7
- 102000002322 Egg Proteins Human genes 0.000 abstract description 5
- 108010000912 Egg Proteins Proteins 0.000 abstract description 5
- 210000004681 ovum Anatomy 0.000 abstract 3
- 238000011534 incubation Methods 0.000 description 22
- 230000000052 comparative effect Effects 0.000 description 12
- 210000003278 egg shell Anatomy 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000005286 illumination Methods 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000238631 Hexapoda Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 238000005138 cryopreservation Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000010902 straw Substances 0.000 description 2
- 206010067482 No adverse event Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 235000013871 bee wax Nutrition 0.000 description 1
- 239000012166 beeswax Substances 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000002577 cryoprotective agent Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 210000002969 egg yolk Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 229940099259 vaseline Drugs 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 210000001534 vitelline membrane Anatomy 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
Abstract
The invention discloses an artificial hatching method for frozen bee eggs, and relates to the technical field of biology. Which comprises the following steps: coating the eggs of the bees to be hatched after freezing with paraffin oil, and then hatching the bee eggs coated with the paraffin oil in a cell culture plate. The outer surface of the bee ovum is completely covered by the paraffin oil by coating the bee frozen ovum to be hatched with the paraffin oil, so that a layer of protective film is formed outside the bee ovum again. The eggs form a layer of closed protective shell, the protective shell enables the eggs to be exposed in the air, the phenomena of shriveling and water loss can not occur, and the survival rate of the frozen eggs of the bees when the eggs are transferred to hatching equipment can be obviously improved.
Description
Technical Field
The invention relates to the technical field of biology, in particular to an artificial hatching method for frozen bee eggs.
Background
The bee is an important economic insect and can produce various bee products, meanwhile, the bee is also an ecological insect, and the ecological value of the bee for pollinating crops is far higher than the economic value of the bee products.
The bee germplasm resource protection generally adopts living body breed conservation technology on a colony level, but the living body breed conservation technology on the colony level is easily influenced by external adverse factors such as weather, natural enemies and the like, is greatly influenced by the external factors and has high living body preservation risk. In order to meet the need of breed conservation, the genetic material of the bee sperms and eggs is necessarily preserved. At present, the preservation of the genetic materials of the honeybee sperms and eggs mainly adopts an ultra-low temperature cryopreservation method.
The bee egg structure mainly comprises egg shell, yolk membrane, egg core, yolk and the like, and the egg shell is the outermost layer structure of the egg and mainly plays a role in protecting bee eggs. The egg shells of the bees have hydrophobicity, and when the bee eggs are frozen and preserved at ultralow temperature, firstly, the egg shells are subjected to infiltration treatment, and only the egg shells are removed, so that the cryoprotectant can enter the insides of the bee eggs, and then the bee eggs are protected correspondingly. And once the egg shells of the bee eggs are removed, the protective barriers are lost, and when the bee eggs are exposed to the air for a short time after being thawed, the bee eggs are easy to dehydrate and shrink, so that the bee eggs are easy to die.
The bee egg freezing preservation technology has been developed for decades, and although the bee egg freezing preservation technology has been successful to a certain extent, the hatching rate and the larva survival rate of the thawed bee eggs are not high, and the effect of breeding bees by using the frozen bee eggs is not ideal. Among them, hatching of eggs and larva culture become a technical problem of the neck clamp. Therefore, establishing the artificial hatching and breeding method of the bee frozen eggs and perfecting the bee egg freezing storage and utilization technology is very necessary for improving the bee resource protection and development and utilization efficiency.
In view of this, the invention is particularly proposed.
Disclosure of Invention
The invention aims to provide an artificial hatching method for bee frozen eggs, which is used for improving the hatching rate and the larva survival rate of the frozen bee eggs. Thereby better improving the efficiency of bee resource protection and development and utilization.
The invention is realized by the following steps:
the invention provides an artificial hatching method for frozen bee eggs, which comprises the following steps: coating the eggs of the bees to be hatched after freezing with paraffin oil, and then hatching the bee eggs coated with the paraffin oil in a cell culture plate.
The inventor finds that the paraffin oil is used for coating the frozen bee eggs to be hatched, so that the outer surfaces of the bee eggs are completely covered by the paraffin oil, and a protective film is formed outside the bee eggs again. The egg is enabled to form a layer of closed protective shell, the protective shell enables the egg to be exposed in the air, the phenomenon of shriveling and water loss can not occur, and the survival rate of the frozen egg when the frozen egg is transferred to hatching equipment can be remarkably improved.
Through verification, the bee eggs are coated after being frozen by the paraffin oil, so that the bee eggs can not be shriveled after being exposed in the air for more than 300 s.
And the paraffin oil coated before eggs are hatched by the inventor has no toxic effect on bee eggs, the cost of the paraffin oil is low, and the coating treatment step is simple and easy.
In a preferred embodiment of the present invention, the incubation temperature is controlled to 35 ℃ ± 0.5 ℃, and the incubation humidity is controlled to 90-95%.
In order to further improve the hatchability and larva survival rate of the frozen bee eggs, the inventor also conducts a large amount of grope for a long time on the microecological conditions of the frozen bee eggs. The discovery shows that the hatching temperature of the frozen eggs is accurately controlled to be 35 +/-0.5 ℃, and the hatching humidity is controlled to be 90-95%, so that the hatching rate of the eggs can be obviously improved.
In a preferred embodiment of the present invention, the incubation humidity is controlled by setting sulfuric acid with a volume concentration of 10% in the incubator.
In order to simulate the micro-ecological conditions in bee colonies more truly, the humidity is accurately controlled by using 10% sulfuric acid, and the humidity in the hatching process is accurately controlled by using the water absorption of the sulfuric acid.
In other embodiments, in order to realize temperature control, the incubator is used for controlling temperature, and the cell culture plate is sealed at the same time, so that the hatching rate of the eggs can be obviously improved by adopting the technical means.
For example, the incubation temperature of the frozen eggs is set to 35 ℃. + -. 0.5 ℃.
In a preferred embodiment of the present invention, the incubation is performed with sulfuric acid disposed in the lower chamber of the cell culture plate and bee eggs disposed above the lower chamber. The cell culture plate is a 24-well cell culture plate.
The inventor creatively uses a 24-hole cell culture plate to hatch the bee eggs. Has the following advantages: 1) Each hole of the 24-hole microbial culture plate is independently distributed at intervals, so that honeybee eggs in part of the holes die and cannot infect other eggs; 2) The hole size of the microbial culture plate with 24 holes is larger, so that the egg moving and taking are more convenient, and convenience in operation is brought to experimenters; 3) After the eggs are hatched into larvae, the eggs are not moved to other tools and can be directly raised on a culture plate.
In an alternative embodiment, 1-2 eggs are placed in each well.
In a preferred embodiment of the present invention, the incubation time is 16-24h. As the frozen eggs are generally 48-54 h day-old eggs, the sealed eggs can be incubated more fully within the incubation time. In an alternative embodiment, the eggs are observed to hatch larval after 18h incubation in the incubator every 1h, and the conversion to larvae is detected in time with the addition of the artificially prepared bee larva diet.
In a preferred embodiment of the present invention, the incubation time is 18-24h.
In a preferred embodiment of the present invention, the incubation is a dark sealed incubation.
In a preferred embodiment of the present invention, the incubation of the larvae further comprises culturing the larvae under the following conditions: the temperature is 34.5 +/-0.2 ℃, the humidity is 90-95 percent, and the air is intermittently ventilated.
The adoption of the cell culture plate can realize incubation and larva culture without changing the plate, and avoid the abnormal death of the larva caused by the transfer plate, thereby improving the survival rate of the larva to a certain extent. The larvae are supplied with the necessary oxygen for growing by intermittent ventilation. Under the above culture conditions, better larva cultivation can be achieved.
In a preferred embodiment of the present invention, the coating treatment of the frozen bee eggs with paraffin oil comprises: the bee frozen eggs are soaked in paraffin oil, and then the excessive paraffin oil is wiped off by absorbent paper.
In a preferred embodiment of the present invention, before coating bee eggs with paraffin oil, the method further comprises thawing frozen bee eggs to be hatched in a thawing solution (5% sucrose solution) at 37 ℃ for 1min.
The invention has the following beneficial effects:
the method for coating the frozen bee eggs to be hatched with the paraffin oil can ensure that the outer surfaces of the bee eggs are completely covered with the paraffin oil and a layer of protective film is formed outside the bee eggs again. The eggs form a layer of closed protective shell, and the protective shell enables the bee eggs to be exposed in the air without the phenomena of shriveling and water loss, so that the survival rate of the frozen bee eggs is remarkably improved when the bee eggs are transferred to hatching equipment. The inventor finds that the paraffin oil coating is beneficial to improving the hatchability of the frozen bee eggs.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
Figure 1 is a micrograph of a bee egg shriveled.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are conventional products which are not indicated by manufacturers and are commercially available.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The embodiment provides an artificial hatching method for frozen bee eggs, which comprises the following steps:
(1) And unfreezing the frozen bee eggs to be hatched.
(2) And then soaking the thawed bee eggs into paraffin oil for coating treatment.
(3) 1000ml of 10% sulfuric acid was added to the lower cavity of the incubator. And then the bee eggs coated with paraffin oil are transferred to a 24-hole cell culture plate (biological engineering (Shanghai) Co., ltd.), each hole is provided with an egg placing rack or an egg placing membrane which is arranged above a cavity below the hole, 1 egg is placed on each egg placing rack, and then the redundant paraffin oil is absorbed by absorbent paper.
(4) Controlling the incubation temperature to be 35.0 ℃ and the humidity to be 90 percent; illumination: and (5) under a light-tight condition, and placing the culture medium in a constant-temperature incubator for incubation culture.
(5) After the eggs are incubated in the incubator for 18 hours, the situation that the eggs are incubated into larvae is observed every 1 hour, the larvae are found to be converted into larvae, the artificially prepared bee feed is added in time, and the survival rate of the bee larvae is ensured.
(6) The culture conditions for the larvae were as follows: the temperature is 34.5 ℃, and the humidity is 90%; dark culture; intermittently ventilating to supply oxygen necessary for larva growth.
Example 2
The embodiment provides an artificial hatching method for frozen bee eggs, which comprises the following steps:
(1) And unfreezing the frozen bee eggs to be hatched.
(2) And then soaking the thawed bee eggs into paraffin oil for coating treatment.
(3) 1000ml of 10% sulfuric acid was added to the cavity below the incubator. And then transferring the bee eggs coated with the paraffin oil into a 24-hole cell culture plate, wherein each hole is provided with an egg placing frame or an egg placing film and is arranged above the cavity below, 1 egg is placed on each egg placing frame, and then the redundant paraffin oil is absorbed by absorbent paper.
(4) Controlling the incubation temperature to be 35.5 ℃ and the humidity to be 92 percent; illumination: and (5) under a light-tight condition, and placing the culture medium in a constant-temperature incubator for incubation culture.
(5) After the eggs are incubated in the incubator for 18 hours, the situation that the eggs are incubated into larvae is observed every 1 hour, the larvae are found to be converted into larvae, the artificially prepared bee feed is added in time, and the survival rate of the bee larvae is ensured.
(6) The culture conditions for the larvae were as follows: the temperature is 34.5 +/-0.5 ℃, and the humidity is 90 percent; dark culture; intermittently ventilating to supply oxygen necessary for larva growth. It should be noted that: the coexistence time of the larvae and the eggs does not differ by 2 hours, and the short-time temperature difference has no influence on the incubation of the eggs and the larvae.
Example 3
The embodiment provides an artificial hatching method for frozen bee eggs, which comprises the following steps:
(1) And unfreezing the frozen bee eggs to be hatched.
(2) And then soaking the thawed bee eggs into paraffin oil for coating treatment.
(3) 1000ml of 10% sulfuric acid was added to the incubator. And then transferring the bee eggs coated with the paraffin oil into a 24-hole cell culture plate, wherein each hole is provided with an egg placing frame or an egg placing film and is arranged above the cavity below, 1 egg is placed on each egg placing frame, and then the redundant paraffin oil is absorbed by absorbent paper.
(4) Controlling the incubation temperature to be 35.5 ℃ and the humidity to be 94 percent; illumination: and (5) under a light-tight condition, and placing the culture medium in a constant-temperature incubator for incubation culture.
(5) After the eggs are incubated in the incubator for 16 hours, the situation that the eggs are incubated into larvae is observed every 1 hour, the larvae are found to be converted into larvae, the artificially prepared bee feed is added in time, and the survival rate of the bee larvae is ensured.
(6) The culture conditions for the larvae were as follows: the temperature is 34.5 +/-0.2 ℃, and the humidity is 90 percent; dark culture; ventilating intermittently, and supplying oxygen necessary for larva cultivation.
Example 4
The only difference compared to example 1 is that step (3) is different and the remaining steps are the same. In this embodiment, sulfuric acid is not provided in step (3), but the incubator is used to directly control the humidity to 90% and the incubation temperature to 35.0 ℃.
Comparative example 1
Compared with example 1, the difference is that the paraffin oil treatment of step (2) is not performed, the thawed bee eggs are stored in a frozen straw, and then transferred to a 24-well cell culture plate for hatching culture according to step (3) of example 1, and 10% sulfuric acid is provided in a culture device.
Comparative example 2
The difference from example 1 is that the paraffin oil treatment of step (2) was not performed, the thawed bee eggs were stored in a frozen straw, and then transferred to a 24-well cell culture plate for hatching culture according to step (3) of example 1, and 10% sulfuric acid was not provided in the incubator.
Comparative example 3
The only difference compared to example 1 is that the paraffin oil treatment of step (2) was replaced by a treatment with the substance vaseline. The other steps are the same as in example 1.
Comparative example 4
The only difference compared to example 1 is that the paraffin oil treatment of step (2) was replaced by a treatment with the substance liquid beeswax. The other steps were the same as in example 1.
Experimental example 1
This example tests the effect of different treatments on egg deflation. The method specifically comprises the following steps:
control group: exposing the thawed 30 bee eggs to the air without any treatment; test groups: the thawed bee eggs 30 are exposed to air by adding paraffin oil "protective shell". The egg collapse was recorded for each group at different times of exposure to air, and 3 replicates for each group were made.
The bee eggs are plump without water loss (figure 1, left), the egg surface folds are shriveled with water loss (figure 1, right), and figure 1 is observed under a microscope.
The statistical results are shown in Table 1.
TABLE 1 number of eggs shriveled by different treatments exposed to air
| 15s | 30s | 45s | 60s | 300s | |
| Control group | 5.66±0.47 | 14.33±1.70 | 9.00±1.63 | 1.00±0.82 | 0 |
| Test group | 0 | 0 | 0 | 0 | 0 |
The results show that the eggs of the bees thawed in the control group are shriveled and dehydrated after being exposed in the indoor environment for 15s, all 30 eggs in 60s are shriveled and dehydrated, and the test group adopting the pretreatment is exposed in the indoor environment for 300s without the phenomena of shriveling and dehydration.
The method for wrapping the frozen eggs by the paraffin oil can ensure that the eggs are exposed in the indoor environment for a short time and do not lose water, and can remarkably improve the survival rate of the frozen eggs when the frozen eggs are transferred to hatching equipment.
Experimental example 2
This experimental example statistical experiments of hatchability of eggs were performed for the above-described examples 1 and 4 and comparative examples 1 to 4. Each example or comparative example was run on 30 eggs and repeated 3 times.
The results of the experiments are shown in Table 2.
TABLE 2 comparison of hatchability of eggs after incubation and freezing by different methods
| Group of | Hatching rate (%) |
| Comparative example 1 | 21.36±3.16 |
| Comparative example 2 | 15.65±1.35 |
| Comparative example 3 | 7.31±2.63 |
| Comparative example 4 | 2.64±1.35 |
| Example 1 | 52.13±2.63 |
| Example 4 | 36.29±3.64 |
The experimental result shows that the hatching rate of the bee eggs after freezing can reach (52.13 +/-2.63)%, while the control example 1 does not carry out paraffin oil coating treatment, the hatching rate of the bee eggs after freezing is only (21.36 +/-3.16)%, and the hatching rate of the bee eggs in the example 1 is obviously higher than that of the control example 1, namely, the bee eggs are coated by paraffin oil to help to improve the hatching rate of the bee eggs. Comparative example 2 compared to example 1, the absence of paraffin treatment and the provision of 10% sulphuric acid resulted in a further decrease in bee egg hatchability after cryopreservation. If the substances for treating the bee eggs were changed, as shown in comparative examples 3-4, the hatchability of the eggs after hatching and freezing could not be improved. Therefore, the paraffin oil treatment and the sulfuric acid humidity control achieve remarkable technical effect of high hatching rate.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. An artificial hatching method of bee frozen eggs is characterized by comprising the following steps: coating the eggs of the bees to be hatched after freezing with paraffin oil, and then hatching the bee eggs coated with the paraffin oil in a cell culture plate.
2. The artificial hatching method for frozen eggs of honeybees according to claim 1, wherein the hatching temperature is controlled to 35 ℃ ± 0.5 ℃ and the hatching humidity is controlled to 90-95%.
3. The artificial hatching method for frozen eggs of bees of claim 2, wherein the humidity of hatching is controlled by setting sulfuric acid with a volume concentration of 10% in an incubator.
4. The artificial hatching method for frozen honeybee eggs according to claim 3, wherein during hatching, sulfuric acid is disposed in the lower chamber of the cell culture plate, and honeybee eggs are disposed above the lower chamber;
preferably, the cell culture plate is a 24-well cell culture plate;
preferably, 1-2 eggs are placed in each well.
5. The artificial hatching method for frozen eggs of honeybees according to claim 1, wherein the hatching time is 16-24h.
6. The artificial hatching method for frozen eggs of honeybees according to claim 5, wherein the hatching time is 18 to 24 hours.
7. The artificial hatching method for frozen eggs of honeybees according to claim 5, wherein the hatching is hatching protected from light.
8. Method for the artificial hatching of frozen eggs of bees according to any one of claims 1 to 7, characterized in that said hatching after obtaining larvae further comprises the cultivation of larvae under the conditions: the temperature is 34.5 +/-0.5 ℃, the humidity is 90-95%, and the air exchange is intermittent.
9. The artificial hatching method for frozen honeybee eggs according to claim 1, wherein the coating treatment of the frozen honeybee eggs with paraffin oil comprises: the bee frozen eggs are soaked in paraffin oil, and then the excessive paraffin oil is wiped off by absorbent paper.
10. The artificial hatching method for the bee frozen eggs according to claim 9, characterized by further comprising thawing the bee frozen eggs to be hatched in a thawing solution at 37 ℃ for 1min before coating the bee eggs with paraffin oil;
preferably, the thawing solution is selected from a 5% sucrose solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210969368.3A CN115176767A (en) | 2022-08-12 | 2022-08-12 | Artificial hatching method for bee frozen eggs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210969368.3A CN115176767A (en) | 2022-08-12 | 2022-08-12 | Artificial hatching method for bee frozen eggs |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115176767A true CN115176767A (en) | 2022-10-14 |
Family
ID=83522665
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210969368.3A Pending CN115176767A (en) | 2022-08-12 | 2022-08-12 | Artificial hatching method for bee frozen eggs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115176767A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1181180A (en) * | 1997-10-20 | 1998-05-13 | 敖济康 | Fresh bee chrysalis and prodn. method of its various kinds of preparation |
| CA2643220A1 (en) * | 2006-02-23 | 2007-08-30 | Leonard Moran | Method for incubation and hatching of eggs |
| CN107372361A (en) * | 2017-07-14 | 2017-11-24 | 吉林省养蜂科学研究所（吉林省蜂产品质量管理监督站、吉林省蜜蜂遗传资源基因保护中心） | A kind of honeybee ovum superfreeze method and its kit |
| CN212212368U (en) * | 2020-03-16 | 2020-12-25 | 云南蜜月追踪蜂业有限公司 | Bee pupa hatching cabinet |
| CN212971243U (en) * | 2020-04-13 | 2021-04-16 | 吉林省养蜂科学研究所（吉林省蜂产品质量管理监督站、吉林省蜜蜂遗传资源基因保护中心） | Bee incubator |
| CN216219540U (en) * | 2021-11-16 | 2022-04-08 | 华蜜智能装备(常州)有限公司 | Production system that batch production honeybee was bred |
-
2022
- 2022-08-12 CN CN202210969368.3A patent/CN115176767A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1181180A (en) * | 1997-10-20 | 1998-05-13 | 敖济康 | Fresh bee chrysalis and prodn. method of its various kinds of preparation |
| CA2643220A1 (en) * | 2006-02-23 | 2007-08-30 | Leonard Moran | Method for incubation and hatching of eggs |
| CN107372361A (en) * | 2017-07-14 | 2017-11-24 | 吉林省养蜂科学研究所（吉林省蜂产品质量管理监督站、吉林省蜜蜂遗传资源基因保护中心） | A kind of honeybee ovum superfreeze method and its kit |
| CN212212368U (en) * | 2020-03-16 | 2020-12-25 | 云南蜜月追踪蜂业有限公司 | Bee pupa hatching cabinet |
| CN212971243U (en) * | 2020-04-13 | 2021-04-16 | 吉林省养蜂科学研究所（吉林省蜂产品质量管理监督站、吉林省蜜蜂遗传资源基因保护中心） | Bee incubator |
| CN216219540U (en) * | 2021-11-16 | 2022-04-08 | 华蜜智能装备(常州)有限公司 | Production system that batch production honeybee was bred |
Non-Patent Citations (1)
| Title |
|---|
| 王丽华, 罗建能, 王涛, 赵寿江: "中华蜜蜂卵冷冻贮存技术", 福建农业大学学报, no. 04, pages 470 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR970000596B1 (en) | In vitro embryo culture technique | |
| Janik et al. | Overcoming a permeability barrier by microinjecting cryoprotectants into zebrafish embryos (Brachydanio rerio) | |
| CN105075992A (en) | Method for raising natural enemy insect southern minute pirate bug indoor in great deal and raising box | |
| Lardy et al. | A metabolic regulator in mammalian spermatozoa | |
| Simkiss | Eggshell porosity and the water metabolism of the chick embryo | |
| Pence | Cryopreservation of in vitro grown fern gametophytes | |
| Niino et al. | Development of V and D cryo-plate methods as effective protocols for cryobanking | |
| CN112753656B (en) | Breeding device and indoor seed preservation method for Hemerocallis ventricosus such as natural enemy of sugarcane striped rice borer | |
| CN115176767A (en) | Artificial hatching method for bee frozen eggs | |
| CN109122663A (en) | A kind of freeze-drying refrigeration for the very wide water flea resting egg in the long-term preservation Pacific Ocean | |
| CN104663439A (en) | Tissue culture and rapid propagation method of waterweed | |
| CN103960182A (en) | Method for establishing exopalaemon carinicauda family by applying in-vitro egg culture | |
| US2704531A (en) | Method of incubating eggs | |
| CN1080994C (en) | Small egg intermediate host industrial production process of oophagous trichogrammae | |
| Poole et al. | In vitro preservation of honey bee semen enhanced by storage at 13—15 C | |
| Radder et al. | Influence of incubation temperature and substrate on eggs and embryos of the garden lizard, Calotes versicolor (Daud.) | |
| CN113243358B (en) | Method for storing tussah eggs | |
| Salama et al. | Biological effects of Bacillus thuringiensis on the egg parasitoid, Trichogramma evanescens | |
| CN109588377B (en) | Silkworm egg hatching method | |
| CN207744591U (en) | A kind of tortoise egg is without medium incubator | |
| JP2020065508A (en) | Method for producing dormant Paramecium-containing dried plant | |
| Pence | Cryopreservation of shoot tips of Selaginella uncinata | |
| ONO et al. | Development of cultured quail embryos | |
| CN109592862A (en) | A kind of modifying agent for bottom material of pond for breeding aquatic products | |
| CN112913785B (en) | Method for improving parasitic rate of parasitic tenebrio molitor of dastarcus helophoroides by using longicorn soaking solution |
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 |