CN101664018B - Bee breeding artificial insemination culturing method - Google Patents
Bee breeding artificial insemination culturing method Download PDFInfo
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- CN101664018B CN101664018B CN2009101529064A CN200910152906A CN101664018B CN 101664018 B CN101664018 B CN 101664018B CN 2009101529064 A CN2009101529064 A CN 2009101529064A CN 200910152906 A CN200910152906 A CN 200910152906A CN 101664018 B CN101664018 B CN 101664018B
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Abstract
The invention discloses a bee breeding artificial insemination culturing method, comprising the following steps: (1) male bee culturing: culturing fine-quality bee breeding by using excellent bee breeding, and culturing a virgin queen before sexual maturity of male bees; (2) semen collection: preparing a semen diluent, placing in a bottle, sterilizing in high temperature, placing the semen diluent in a collection pipe of a semen collector, scraping the semen and mucus of the male bees to the diluent by a sterilized glass rod, and placing the collector into a centrifuge tube of a centrifuge for centrifugation so as to separate semen when enough semen is rinsed in the diluent; and (3) insemination: selecting the virgin queen of sexual maturity, placing in an anesthetic room, carrying out artificial insemination when the virgin queen is in a comatose state, and removing narcosis after insemination. The swarm cultured by the method in the invention is improved in the aspects of collection ability, disease resistance, stress resistance, oviposition rate, bee milk yield and the like, and the collected royal jelly and pollen yield are enhanced.
Description
Technical field
The present invention relates to the bee artificial insemination method.
Background technology
The Along with people's growth in the living standard, people's health care consciousness strengthens, and market, home and abroad is increasing the demand of royal jelly rapidly, and existing honeybee kind, the number of animals raised of royal jelly output and bee colony can not satisfy this demand far away.Simultaneously, honeybee is raised through long-term, often causes kind of a property to mix, degenerate, and the ability of keeping group gesture weakens, delivery and feed infant power (queen to lay eggs power and worker bee nurture power), premunition and resistance decline, and productivity descends.For a long time, the beekeeper just wants to increase the income of oneself with the output that improves royal jelly through raising royal jelly high yield honeybee kind; But all there is certain defective in the royal jelly high yield honeybee kind on the market: the royal jelly high yield, and collection power is low, or the robber honeybee becomes second nature; Or premunition is too poor, maybe can not keep strong crowd, and these all are the big factors that hinders the development of crowd's gesture and produce; Therefore need further carry out purification, the rejuvenation of honeybee kind, cultivate the honeybee kind of high-yield disease resisting, improve colony productivity comprehensively.
In the practice that premodern agriculture is produced, the hybridization of the yielding ability of different cultivars, strain can increase productivity within a short period of time greatly, and on bee-keeping was produced, making full use of hybrid vigour also was an effective well stimulation.According to relevant report, the first generation after two yuan of hybridization of external honeybee kind, second generation honey yield improve more than three one-tenth than the Apis mellifera of high yield, and the raising the output of three way cross is more.Through choosing seeds and breeding two steps, can improve its productivity, and material is provided for cultivating high-yield hybrid kind and new varieties.
Summary of the invention
Technical problem to be solved by this invention just provides a kind of bee breeding artificial insemination culturing method, to cultivate the honeybee kind of high-yield disease resisting, improves colony productivity comprehensively.Its bee colony of cultivating all is greatly improved at aspects such as collection power, premunition, resistance, spawning rate, paste producing amounts, and improves royal jelly and the pollen yield of gathering.
For solving the problems of the technologies described above, the present invention adopts following technical scheme: bee breeding artificial insemination culturing method is characterized in that comprising the steps:
1) drone is cultivated: utilize the good honeybee kind of introducing to cultivate the high-quality honeybee kind that makes new advances, before the drone sexual maturity, cultivate virgin king simultaneously;
2) semen collection: be mixed with spermatozoa diluent with sodium chloride, potassium chloride, glucose and dual distilled water; Bottling back high-temperature sterilization; Pack spermatozoa diluent in the collection tube of semen collection device into then; Scrape in the dilution with the seminal fluid and the mucus of the glass bar of sterilizing with drone, when rinsing in the dilution had enough seminal fluid, the centrifuge tube of collector being put into centrifuge carried out centrifugation to isolate seminal fluid;
3) insemination: the ripe virgin king of selectivity, the anaesthetic room of packing into is installed on the geometrical clamp of insemination appearance base, opens CO
2Flow when virgin king gets into comatose state, carries out artificial insemination, and insemination finishes and can remove anesthesia;
4) queen bee that will inseminate is placed on to have in the above nuclear colony of 3 frames foots honeybee and lays eggs.
As preferably, to remove group interior drone in the said step 1) drone cultivating process, and the drone obstructing instrument is installed; Extraneous drone can not be sneaked into, add the empty honeycomb of drone again, use single spleen button law of the country; Make this sky of queen bee only on the drone spleen, produce fertilized egg; Before the drone sexual maturity, remove the drone obstructing instrument, adopt the drone cage that circles in the air, the clarity of control drone.
As preferably, said step 2) in the spermatozoa diluent weight ratio of sodium chloride, potassium chloride, glucose be 8: 1: 3, the pH value of spermatozoa diluent transfers to 7.5, the back 115.6 ℃ of pressure sterilizings 15 minutes of bottling.
As preferably, said step 2) centrifuge separated 10 minutes with the speed that per minute 2500 changes in.
As preferably, said step 2) if in store at normal temperatures more than 2 days after the seminal fluid centrifugation, need the openend of collection tube is sealed on alcolhol burner.
As preferably, 8 μ l seminal fluid are injected in disposable insemination in the said step 3), if divide the secondary insemination, then respectively inject 4～5 μ l seminal fluid at every turn, and are 24～28 hours with the blanking time of inseminating for the second time for the first time.
As preferably, CO in the said step 3)
2Flow flows out 300 bubbles, anesthesia duration 30 seconds through control valve control per minute.
As preferably, every queen bee finishes must be controlled in 5 minutes from anesthesia to insemination in the said step 3), and the back of the body web of queen bee is once opening, inserting needle injection in time.
The present invention is owing to adopted technique scheme to carry out bee breeding artificial insemination culturing, and its bee colony of cultivating all is greatly improved at aspects such as collection power, premunition, resistance, spawning rate, paste producing amounts, and improves royal jelly and the pollen yield of gathering.
Embodiment
Below be the embodiment of bee breeding artificial insemination culturing method of the present invention, its process is following:
1, drone is cultivated: utilize the good honeybee kind of introducing to cultivate the high-quality honeybee kind that makes new advances.In this process, remove drone in the crowd, the drone obstructing instrument is installed, extraneous drone can not be sneaked into, add the empty honeycomb of drone again, use single spleen button law of the country, make this sky of queen bee only on the drone spleen, produce fertilized egg.The drone that goes out later on all is to go out room (also have on a small quantity regular hour poor) on the same day, and the quantity of controlled ripe drone is convenient in sexual maturity on the same day.Before the drone sexual maturity, remove the drone obstructing instrument, adopt the drone cage that circles in the air, the clarity (not containing extraneous honeybee kind) of control drone is cultivated virgin king with the time shift worm, makes drone and virgin king's sexual maturing period identical or close (differ and be no more than two days).We adopt low energy optically focused daylighting method on the artificial insemination appearance, and this light can reach artificial insemination daylighting requirement, queen bee is heated physiological variation takes place.
2, semen collection: after drone sexual maturity in 12 days, prepare spermatozoa diluent, spermatozoa diluent is prepared like following table.
That tighter, the belly of selecting abdominal segment to shrink seems is hard, wing is more bigger with the ratio of belly, take action relatively that the drone of sensitivity carries out semen collection.Left hand thumb, food, middle finger its chest is exerted pressure backward gently during semen collection, and make afterbody up, hand thumb is by saving at drone belly back side 1-3; Exert pressure with thumb, the forefinger while ventrad tail end of left hand, make the hemolymph in abdominal cavity overstock interior adeagus, force adeagus to turn up gradually; What at first rout up is the adeagus substrate, is that a diagonal angle capsule and pinniform are prominent then, is the adeagus ball at last; Ejaculatory duct is opened on the adeagus ball, when adeagus ball end is emerged khaki seminal fluid, stops extruding.Firmly safe during pressurization, be sure not to act with undue haste, too big as firmly, cause seminal fluid to splash easily or the adeagus ball fractured, make seminal fluid sneak into body fluid.In the drone adeagus turns up process, be sure not to make the adeagus ball and the seminal fluid that turn up to touch drone surface and operator's finger, in order to avoid pollute.During semen collection, collector is vertically fixed on the support, the spermatozoa diluent of having one's face covered with in seven minutes is housed in the collection tube, scrape in the dilution, utilize the semen collection device once can gather tens of drone seminal fluid with the seminal fluid and the mucus of the glass bar of sterilizing with drone.When rinsing in the dilution has enough seminal fluid, collector is put into the centrifuge tube of centrifuge, separated 10 minutes with the speed that per minute 2500 changes.After centrifugal, dilution is positioned at above the seminal fluid, and mid portion is a seminal fluid, and mucus deposits is in the bottom of collection tube.With blade the polyethylene collection tube is cut off, collection tube and funnel are separated.If ought in a few days inseminate to queen bee after the centrifugation,, suck or injection insemination syringe needle with the syringe holder seminal fluid with the blind end incision of collection tube with this seminal fluid.If store at normal temperatures after the centrifugation more than 2 days, need the openend of collection tube is sealed on alcolhol burner.Semen collection was at that time inseminated at that time, and opportunities for contamination is few, and sperm is short in the external holdup time, and vigor is stronger, generally need not check the sperm viability situation.
3, insemination: install device for artificial insemination, adopt the daylighting of low energy optically focused, the ripe virgin king of selectivity, the anaesthetic room of packing into (stationary pipes) is installed on the geometrical clamp on the insemination appearance base, opens CO
2Flow flows out 300 bubbles through control valve with per minute and is advisable, and anesthesia duration is about 30 seconds, and virgin king gets into comatose state, finishes up to insemination, can remove anesthesia.In the time of queen bee anesthesia, regulate microscopical position and focal length, make virgin king's belly be presented on visual field central authorities clearly.Move back of the body abdomen hook control stick then; When the queen bee belly stops to twitch, move abdomen hook control stick, make the abdomen hook hook web; Then under the cooperation of exploration pin, make back of the body hook hook backboard; Move the back of the body, abdomen hook control stick simultaneously, back of the body chimoto " is expanded " partly reach queen bee stylet sheath base portion, and make in its three angle seams that embed base portion.After back of the body hook embeds, move back of the body abdomen hook control stick, be pulled to 5～6mm to back of the body web distance at this.After queen bee back of the body web drew back, the angle of adjustment queen bee made the longitudinal axis of queen bee parallel with the syringe longitudinal axis, and the queen bee longitudinal axis is 32 degree with the angle of insemination appearance vertical line.Regulate longitudinal rail and make pin near introitus, vaginal probe is inserted vagina, and draw close to the queen bee outside of belly, it is prominent to make probe push down the vagina lobe; And make the introitus expansion, and regulate longitudinal rail and make needle point get into vagina slowly along the north of probe, until median oviduct, take out probe; When depth of needle reaches 1.8mm, when inserting needle slightly feels to receive resistance, introitus that sagging sign is arranged; Stop inserting needle, then syringe needle is withdrawed from a bit slightly, the start injection seminal fluid; Disposable insemination is injection 8 μ l seminal fluid (if divide the secondary insemination, then respectively inject 4～5 μ l seminal fluid, be 24～28 hours with the blanking time of inseminating for the second time at every turn for the first time) usually.The notes essence finishes, and treats slightly to withdraw from syringe needle after 10～15 seconds, unloads anaesthetic room, carefully withdraws from queen bee, marks to queen bee, sends former nuclear colony back to after waiting to revive.And at separated king's grid of the nest doorway of little nuclear colony installation or shedding sheet, the king that prevents to inseminate flies out outside the nest.Inferior daily CO
2Anaesthetized once more 5～10 minutes, and promoted laying eggs early of insemination queen bee.
4, the insemination queen bee must be placed on and have in the above nuclear colony of 3 frames foot honeybee, the good heat insulating of this nuclear colony, and new queen to lay eggs was generally laid eggs in 2～5 days than very fast., the insemination king just becomes a new queen bee when beginning to lay eggs, the egg laying amount of record in one week.The ovum that new queen bee produces is observed the situation of capping through capping in 11 days, selects capping smooth, and the capping area is big, and the new queen bee that does not have the beggar is the commodity king, can sell.Must follow-up service after the commodity king sells, write down the economic performance of new Wang Qun.Premunition, anti-adversity, robber's property, docility property, and write down the color of drone, the morphological feature of worker bee etc.
The several Control point of artificial insemination operation; Technology of artificial insemination is a breadboard fine work; The honeybee kind of good experimental situation, superior experimental facilities, superior performance, profound experimental technique are the successful necessary conditions of artificial insemination, and the operation of profound technology of artificial insemination is successful key.Through the practice of artificial insemination operation, we have summed up following control point:
1, in the insemination operating process, must operate gently surely, accurately do not tremble, in order to avoid back of the body abdomen hook and syringe needle damage queen bee.
2, on top of stereomicroscope, the insemination appearance each component capabilities, must not leave under the situation of eyepiece at eyes, both hands can be operated freely.
3, every queen bee finishes must be controlled in 5 minutes from anesthesia to insemination, and the practician is greatly about about 2 minutes.The back of the body web of queen bee is once opening, and inserting needle injection in time is in order to avoid the introitus drying causes the inserting needle difficulty.It carries on the back the chance that web is opened for a long time can increase pollution again.
4, in the process that the queen bee that inseminates is observed, find that temperature is the key factor that influences the sperm transfer velocity.The insemination queen bee must be placed on to have in the above nuclear colony of 3 frames foot honeybee, and insulation, the temperature adjusting performance of this nuclear colony are relatively good, and the egg-laying period in advance.
Claims (7)
1. bee breeding artificial insemination culturing method is characterized in that comprising the steps:
1) drone is cultivated: utilize the good honeybee kind of introducing to cultivate the high-quality honeybee kind that makes new advances, before the drone sexual maturity, cultivate virgin king simultaneously;
2) semen collection: be mixed with spermatozoa diluent with sodium chloride, potassium chloride, glucose and dual distilled water; Bottling back high-temperature sterilization is packed spermatozoa diluent in the collection tube of semen collection device into then, scrapes in the dilution with the glass bar of sterilization seminal fluid and the mucus with drone; When rinsing in the dilution has enough seminal fluid; The centrifuge tube of collector being put into centrifuge carries out centrifugation to isolate seminal fluid, and the weight ratio of sodium chloride, potassium chloride, glucose is 8: 1: 3 in the wherein said spermatozoa diluent, and the pH value of spermatozoa diluent transfers to 7.5; Bottle back 115.6 ℃ of pressure sterilizings 15 minutes, centrifuge separated 10 minutes with the speed that per minute 2500 changes;
3) insemination: the ripe virgin king of selectivity, the anaesthetic room of packing into is installed on the geometrical clamp of insemination appearance base, opens CO
2Flow when virgin king gets into comatose state, carries out artificial insemination, and insemination finishes and can remove anesthesia; 4) queen bee that will inseminate is placed on to have in the above nuclear colony of 3 frames foots honeybee and lays eggs.
2. bee breeding artificial insemination culturing method according to claim 1; It is characterized in that: will remove group interior drone in the said step 1) drone cultivating process; And the drone obstructing instrument is installed, and extraneous drone can not be sneaked into, add the empty honeycomb of drone again; Use single spleen button law of the country, make this sky of queen bee only on the drone spleen, produce fertilized egg.
3. bee breeding artificial insemination culturing method according to claim 2 is characterized in that: before the drone sexual maturity, remove the drone obstructing instrument, adopt the drone cage that circles in the air, the clarity of control drone.
4. bee breeding artificial insemination culturing method according to claim 3 is characterized in that: said step 2) if store at normal temperatures more than 2 days after the middle seminal fluid centrifugation, need the openend of collection tube is sealed on alcolhol burner.
5. according to each described bee breeding artificial insemination culturing method of claim 1 to 4; It is characterized in that: 8 μ l seminal fluid are injected in disposable insemination in the said step 3); If divide the secondary insemination; Then respectively inject 4～5 μ l seminal fluid at every turn, and be 24～28 hours with the blanking time of inseminating for the second time for the first time.
6. bee breeding artificial insemination culturing method according to claim 5 is characterized in that: CO in the said step 3)
2Flow flows out 300 bubbles, anesthesia duration 30 seconds through control valve control per minute.
7. bee breeding artificial insemination culturing method according to claim 5 is characterized in that: every queen bee finishes must be controlled in 5 minutes from anesthesia to insemination in the said step 3), and the back of the body web of queen bee is once opening the injection of necessary inserting needle in time.
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Families Citing this family (7)
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| CN101960998B (en) * | 2010-08-18 | 2012-07-25 | 常仲乐 | Multifunctional microscope operating system |
| CN104604812B (en) * | 2015-01-26 | 2017-02-01 | 郭云胶 | Wasp artificial insemination operation method and insemination device of wasp artificial insemination operation method |
| CN105145546B (en) * | 2015-09-23 | 2017-05-24 | 中国农业科学院蜜蜂研究所 | Preserving fluid of drone semen and preparation method of preserving fluid of drone semen |
| CN105230607A (en) * | 2015-09-23 | 2016-01-13 | 中国农业科学院蜜蜂研究所 | Honeybee artificial insemination buffer solution, and preparation method thereof |
| CN106614388B (en) * | 2016-09-26 | 2020-11-03 | 广西田林县林子蚂蜂养殖科技有限公司 | Method for mating wasps |
| CN106900658B (en) * | 2017-02-24 | 2020-12-11 | 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） | Chinese bee breeding method |
| CN109984093A (en) * | 2019-03-29 | 2019-07-09 | 西南大学 | A kind of Groton bug rearging cage and Groton bug method for breeding |
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| CN1820592A (en) * | 2006-03-28 | 2006-08-23 | 衡水田益生防有限责任公司 | Standard artificial breeding technology for short tongue bumblebee |
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