CN111543392B - Method and device for large-scale raising of fruit fly parasitic wasps - Google Patents
Method and device for large-scale raising of fruit fly parasitic wasps Download PDFInfo
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- 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
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/105—Aliphatic or alicyclic compounds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/111—Aromatic compounds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/24—Compounds of alkaline earth metals, e.g. magnesium
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/90—Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
Abstract
The invention relates to the field of agricultural pest biological control, and aims to provide a large-scale feeding method and device for fruit fly parasitic wasps. The method comprises the steps of host inoculation and propagation, fruit fly parasitic wasp inoculation and propagation, fruit fly culture medium removal, fruit fly removal, parasitic wasp pupa collection and wasp seed preservation; the method utilizes the characteristic that the emergence time of parasitic fruit fly parasitic wasps is later than that of normal fruit flies, and controls the fruit fly food to be thrown in, so that the newly-emerged fruit flies die before the emergence of the parasitic wasps, and dead fruit fly imagoes are separated. The feed can be separated from the fruit flies and the parasitic wasps by the feeding device; avoid the generation overlapping phenomenon of the fruit flies, and is beneficial to the collection of eclosion parasitic wasps in the later period. The operation is convenient, the breeding process can be standardized, and the large-scale propagation of parasitic wasps is facilitated. The invention can collect parasitic bee pupae in large scale, and can be directly used for large-scale release application in fields.
Description
Technical Field
The invention relates to the field of agricultural pest biological control, in particular to a method and a device for breeding fruit fly parasitic wasps on a large scale.
Background
Drosophila melanogaster (Drosophila suzukii Matsumura), also known as Linnaeus, was first discovered in strawberry orchards in sorb county, Japan. Different from common fruit flies, the spotted wing fruit flies can directly lay eggs in mature or to be mature fruits with soft peels such as cherries, peaches, European plums, grapes, strawberries, raspberries, blueberries, persimmons, tomatoes and the like besides eating fallen fruits or damaged fruits, and larvae eat in the fruits to cause serious loss to orchards. At present, the spotted wing fruit fly causes serious harm to fruit industry of more than 30 countries such as the United states and the like all over the world, and is listed as an important quarantine pest by a plurality of countries such as the United states, Japan, Australia, New Zealand, England, Germany and the like. In order to reduce the use of chemical pesticides, agricultural practitioners have expressed a great interest in sustainable control means, mainly biological control. The parasitic wasp is an important natural enemy of the spotted wing fruit fly and plays an important role in sustainable green prevention and control of the spotted wing fruit fly. Therefore, the research on the prevention and control capacity of the drosophila parasitism resource is receiving more and more attention.
There are a number of parasitic wasps known in the world that are capable of parasitizing fruit flies, about 4 families, more than 50 species of the 16 genera, including larval parasitic Braconidae and figitae species of the ceyloides; parasitic species of the family cuprinidae diaphoridae and the family ceramoedae Pteromalidae in the pupal stage. In nature, the fruit fly parasitic wasp can effectively control the fruit fly population, and the parasitic rate can reach 90%. Therefore, the parasitic wasps have great control potential for controlling fruit flies (especially, the spotted wing fruit flies).
Chinese invention patent CN102578052A discloses a method for artificially feeding black belly drosophila melanogaster in a waxberry garden and pupal parasitic wasps, namely chrysalis ceratosporum nudum, and Chinese invention patents CN105794728A, CN107439491A and CN109169526A respectively disclose a method for breeding trichogramma trichogrammatid in pupal parasitic wasps and trichogramma in larval parasitic wasps in drosophila melanogaster in open arm and reverse japonicas in indoor. The above-mentioned patent can implement indoor small-scale breeding of fruit fly parasitic wasp, and its operation is simple and easy. However, in the large scale propagation of parasitic wasps using these techniques, disadvantages have been found: 1. the Chinese invention patent CN102578052A utilizes an insect-raising cage to propagate fly pupae ceratophaga. In the breeding process of parasitic wasps, fruit fly pupae need to be collected separately and transferred into a parasitic cage to serve as a parasitic place, the step greatly increases the workload during large-scale propagation, and the insect cage is not beneficial to the collection of parasitic wasp adult wasps. 2. The Chinese invention patents CN105794728A, CN107439491A and CN109169526A utilize a fruit fly culture bottle to culture fruit flies and then propagate parasitic wasps, and the parasitic wasps and the non-parasitic fruit flies grow together, so that the generation overlapping phenomenon of the fruit flies is inevitable, and the later stage emergence parasitic wasp collection is not facilitated. Meanwhile, the fruit fly feed can mildew and grow bacteria after being placed for a long time, the humidity in the fruit fly culture bottle is greatly increased and even reaches the sticky degree, the growth and development of parasitic wasps are not facilitated, and the parasitic wasps are collected and lost.
Disclosure of Invention
The invention aims to solve the technical problem of overcoming the defects of the prior art and provides a method and a device for breeding fruit fly parasitic wasps on a large scale.
In order to solve the technical problem, the solution of the invention is as follows:
provides a large-scale feeding method of drosophila parasitifer, which comprises the following steps:
(1) host inoculation and propagation
Loading fruit fly food into the breeding and propagation device, and transferring the adult fruit flies which are eclosion for about one week and are fully mated into the breeding and propagation device; moving to a constant-temperature insect breeding room at 25 +/-1 ℃ for breeding, and taking out the fruit flies after 1 day of oviposition; moving the breeding and expanding device with the drosophila melanogaster eggs to a constant-temperature insect-culturing room with the temperature of 25 +/-1 ℃ and the RH of 50 percent for continuous culture, and observing and recording the quantity and the development condition of the drosophila melanogaster eggs;
(2) inoculating and breeding fruit fly parasitic wasp
When the drosophila melanogaster eggs grow to the inoculation age suitable in the step (1), selecting parasitic bee colonies to transfer into a breeding and propagation device according to the proportion of 1: 10 of the number of female bees and drosophila melanogaster eggs, and adding parasitic bee food; observing the parasitism condition of the parasitic wasps on the fruit fly larvae or pupae, and taking out the parasitic wasps after parasitizing for 1 day;
(3) drosophila culture medium removal
Moving the breeding and expanding device to a constant-temperature insect room with the temperature of 25 +/-1 ℃ and the RH of 50 percent for continuous culture, and removing fruit fly food after fruit fly larvae grow into pupae;
(4) drosophila removal
Under the conditions of 25 +/-1 ℃ and 50% RH, the emergence of parasitic fruit fly parasitic wasps is 6-8 days later than that of normal fruit flies, and the fruit flies which are not parasitic successfully emerge first. Because the fruit fly food is removed, the newly-feathered fruit flies can all die before the parasitic wasps eclosion, and dead fruit fly imagoes are removed;
(5) parasitic bee pupa collection
Collecting parasitized Drosophila pupae before eclosion of parasitic wasps, storing in a breeding and propagation device, and transferring to a constant temperature incubator with 16 + -1 deg.C and RH 50%;
(6) bee seed preservation
After the parasitic wasp emerges for about 18 days, using CO2Anaesthetizing and emerging parasitic wasps to form bees, quickly collecting the parasitic wasps, putting the parasitic wasps into a culture bottle filled with parasitic wasp food, and storing the parasitic wasps in a 16 +/-1 ℃ culture box with RH 50% for inoculation in the next breeding.
In the invention, the parasitic wasp is a drosophila larva parasitic wasp or a drosophila pupal-stage parasitic wasp; the drosophila larvae parasitoid is a leptospirillum Leptopilina or a Chondrograpus Asobara parasitoid; the fruit fly pupal stage parasitic wasps are Cerrio cottonii Spalangia, Cerrio trichogramma Pachybotrys or Cerrio trichogramma Trichopria parasitic wasps.
In the invention, in the step (2), the parasitic wasps are ensured to be in the eclosion period when the parasitic wasps are inoculated with the fruit flies; for parasitic wasps of drosophila larvae, the drosophila inoculation instar is 1-2 instars of larvae; for the fruit fly pupal parasitic wasps, the fruit fly inoculation age-adapted period is 1-2 days in pupal period.
In the invention, the preparation method of the fruit fly food comprises the following steps: dissolving 120g of dry yeast in 1000ml of warm water, weighing 318.75g of corn flour, 57g of agar, 60g of peptone, 90g of brown sugar, 180g of glucose and 60g of yeast extract, adding 5000ml of warm water, fully stirring, uniformly mixing, adding 1000ml of dissolved yeast solution, heating and stirring; when the mixture is cooked, 1.5g of MgSO (MgSO) are added in turn4·7H2O and 1.5g CaCl2·2H2Boiling, naturally cooling to 50-60 deg.C, adding methyl hydroxybenzoate solution 180ml and propionic acid 50ml, stirring,cooling for 6-7h, sealing with gauze, and refrigerating in a refrigerator at 4 deg.C; the methyl paraben solution is prepared by dissolving 100g of methyl paraben in 900ml of 95% ethanol.
In the invention, the preparation method of the parasitic wasp food comprises the following steps: adding 27g of agar into 1000ml of water, fully and uniformly stirring, sterilizing by high-temperature steam at 120 ℃, then adding 33g of brown sugar and 330ml of pure apple juice, fully and uniformly stirring, naturally cooling to 60 ℃, adding 20ml of methyl hydroxybenzoate solution, cooling to obtain a mixture, fully and uniformly stirring, and refrigerating at 4 ℃ for later use; the methyl paraben solution is prepared by dissolving 100g of methyl paraben in 900ml of 95% ethanol.
In the present invention, the drosophila is drosophila melanogaster, drosophila maculatus d.suzukii, or drosophila melanogaster d.virilias.
The invention further provides a raising and propagation expanding device for the large-scale raising method of the fruit fly parasitic wasps, which comprises a container body; the top of the container body is provided with an upper cover, and the bottom of the container body is provided with a detachable bottom box; the upper cover is provided with an air hole, and a sponge plug is filled in the air hole; a box body for containing parasitic wasp food is arranged inside the container body;
the breeding device also comprises a fly pupa collecting box which can be matched with the bottom of the container body, a pull cover is arranged at the bottom air hole of the fly pupa collecting box, and a gauze is arranged at the upper opening of the fly pupa collecting box. The aperture of the gauze is 20-40 meshes according to the size of the adult parasitic wasps.
In the invention, the container body is cylindrical or cubic, and the bottom box and the fly pupa collecting box are in shapes matched with the bottom box and the fly pupa collecting box.
In the invention, the box body for containing parasitic wasp food is hung below the upper cover through a string; or to the container wall by telescopic rails.
In the invention, the bottom box and the fly pupa collecting box are arranged at the bottom of the container body in a threaded connection or buckling sleeving manner.
Compared with the prior art, the invention has the beneficial effects that:
1. the feed can be separated from the fruit flies and the parasitic wasps by the feeding device; avoid the generation overlapping phenomenon of the fruit flies, and is beneficial to the collection of eclosion parasitic wasps in the later period.
2. The method is convenient and fast to operate, can standardize the breeding process, and is favorable for large-scale propagation of parasitic wasps.
3. The invention can collect parasitic bee pupae in large scale, and can be directly used for large-scale release application in fields.
Drawings
Fig. 1 is a schematic structural diagram of a fruit fly parasitic wasp propagation device in the invention.
Fig. 2 is a schematic view of the usage of the fruit fly parasitic wasp collecting device in the invention.
Reference numerals: 1a container body; 2, air holes are formed; 3, sponge plug; 4 a string; 5, a box body; 6 fixing a lantern ring; 7, a bottom box; 8, collecting fly pupae; 9, a gauze; 10 pulling the cover.
Detailed description of the preferred embodiments
The following detailed description of the embodiments of the present invention will be provided to assist those skilled in the art in more completely, accurately and deeply understanding the technical solutions of the present invention, by describing the embodiments with reference to the accompanying drawings.
The breeding and propagation device comprises a cylindrical or cubic container body 1 (cylindrical in the figure), an upper cover is arranged at the top of the container body 1, and a detachable bottom box 7 is arranged at the bottom of the container body; the upper cover is provided with an air vent 2, and a sponge plug 3 is filled in the air vent; be equipped with the box body 5 that is used for the parasitic bee food of splendid attire (box body 5 takes the aperture, and the aperture is convenient for parasitic bee to get food) in the inside of vessel 1, box body 5 is hung on the fixed lantern ring 6 of locating the upper cover below through string 4, perhaps through on the flexible track mounting container wall. The device also comprises a fly pupa collecting box 8 which can be matched and arranged with the bottom of the container body 1, a pull cover 10 is arranged at the air hole at the bottom of the fly pupa collecting box, and a gauze 9 is arranged at the opening at the upper part of the fly pupa collecting box. The bottom box 7 and the fly pupa collecting box 8 have a shape matched with the container body 1 and can be installed at the bottom of the container body 1 in a threaded connection or a buckling and sleeving manner. The pull cover 10 is used for picking and placing fruit fly pupae, and the gauze 9 is used for separating the fruit fly pupae and eclosion parasitic wasps.
When in use, the box body 5 is used for containing parasitic wasp food; the bottom box 7 is used for containing fruit fly food and can be disassembled to be replaced by a new culture-free substrate box so as to remove the fruit fly food. The bottom box 7 is used for the inoculation and breeding process of the fruit fly parasitic wasps, and the fly pupa collecting box 8 is used for collecting the parasitic wasps after the fruit fly parasitic wasps are inoculated and bred. By being combined with the container body 1, the collected fruit fly pupae are put into the fly pupae collecting box 8 through the pull cover 10. The eclosion parasitic wasps rapidly pass through the gauze 9 (20-40 meshes of gauze is selected according to the size of the imago bodies of the parasitic wasps), and are separated from the fruit fly pupae in the fly pupae collecting box 8, so that the aim of rapidly collecting the eclosion parasitic wasps is fulfilled.
Example 1 Large-Scale propagation of parasitic wasps of Drosophila larvae
(1) Host inoculation and propagation:
(A) preparing fruit fly food: dissolving 120g of dry yeast in 1000ml of warm water, weighing 318.75g of corn flour, 57g of agar, 60g of peptone, 90g of brown sugar, 180g of glucose and 60g of yeast extract, adding 5000ml of warm water, fully stirring, uniformly mixing, adding 1000ml of dissolved yeast solution, heating and stirring; when the mixture is cooked, 1.5g of MgSO (MgSO) are added in turn4·7H2O and 1.5g CaCl2·2H2And O, boiling, naturally cooling to 50-60 ℃, adding 180ml of methyl hydroxybenzoate solution (prepared by dissolving 100g of methyl hydroxybenzoate in 900ml of 95% ethanol) and 50ml of propionic acid, stirring uniformly, cooling for 6-7h, sealing with gauze, and refrigerating in a refrigerator at 4 ℃ for later use.
(B) Fruit fly food is filled in the bottom box 7; 1000 adult Drosophila melanogaster which has been eclosized for about one week and has been mated thoroughly is taken and treated with CO2After anesthesia, transferring the fruit fly into a breeding and expanding device, transferring the fruit fly into a constant-temperature insect breeding room at 25 +/-1 ℃ to lay eggs for 1 day, and taking out the fruit fly;
(C) and (C) placing the breeding and expanding device with the drosophila melanogaster eggs in the step (B) in a constant-temperature insect room with the temperature of 25 +/-1 ℃ and the RH of 50 percent for culturing, and observing and recording the quantity and the development condition of the drosophila melanogaster eggs.
(2) Parasitic wasp inoculation and reproduction
(A) Preparing parasitic bee food: adding 27g of agar into 1000ml of water, stirring uniformly, sterilizing with high-temperature steam at 120 ℃, adding 33g of brown sugar and 330ml of pure apple juice, stirring uniformly, naturally cooling to 60 ℃, adding 20ml of methyl hydroxybenzoate solution (dissolving 100g of methyl hydroxybenzoate in 900ml of 95% ethanol), cooling to obtain a mixture, stirring uniformly, and refrigerating at 4 ℃ for later use.
(B) Inoculating and breeding parasitic wasps: after the drosophila ova are incubated for 1-2 days in the step (1), selecting the drosophila larva parasitic wasp preserved in a laboratory according to the ratio of 1: 10 of female wasps and drosophila ova, and transferring the drosophila larva parasitic wasp into a drosophila parasitic wasp propagation device.
When parasitic wasp inoculation is carried out, the parasitic wasp is ensured to be in the eclosion stage, and for parasitic wasps, the fruit fly instar is ensured to be in the 1-2 instar larval stage.
(C) In order to improve the parasitic rate, the parasitic wasp food in the step (A) is added into the box body 5 of the fruit fly parasitic wasp propagation device to be used as nutrition supplement, the parasitic wasps are observed to parasitize fruit fly larvae, and the parasitic wasps are taken out after 1 day.
(3) Drosophila culture medium removal
Drosophila larvae have a Wandering period (Wandering stage) before pupation, i.e. the third instar larvae wander on the bottle wall and finally pupate on the bottle wall. According to the biological characteristics, the parasitic fruit fly larvae in the step (2) are placed in a constant-temperature insect chamber with the temperature of 25 +/-1 ℃ and the RH of 50 percent, and after the fruit fly larvae in the step (2) enter pupae, the bottom box 7 with fruit fly food is directly replaced by a clean bottom box 7 (the generation overlapping of fruit flies in the next step is avoided).
(4) Drosophila removal
Generally, the whole growth period of fruit fly at 25 deg.C via egg-larva-pupa-imago takes about 10 days. After parasitic wasps parasitize fruit flies, host development duration can be prolonged. The emergence of parasitic fruit fly parasitized bee is 6-8 days later than normal fruit fly under the conditions of 25 + -1 deg.C and RH of 50%. Thus, the fruit flies that did not succeed in parasitizing will emerge first before the parasitizing bees emerge. Because the fruit fly food is removed in step (3), the newly emerged fruit fly has no food source, so that the newly emerged fruit fly can die completely before the parasitic wasp emerges. Dead adult drosophila flies were removed about 14 days after parasitization.
(5) Parasitic bee pupa collection
And (3) scraping the parasitized fruit fly pupae in the step (4) away from the side wall of the breeding and propagation device by using a soft brush, and quickly collecting the parasitized fruit fly pupae in a fly pupae collecting box 8 (which can be stored in a constant temperature box with the temperature of 16 +/-1 ℃ and the RH of 50 percent for later use so as to be directly used for large-scale field release in the subsequent process).
(6) Bee seed preservation
(A) Fixing the fly pupa collecting box 8 with the collected parasitic bee pupae in the step (5) at the bottom of the container body, and after the parasitic bee eclosion, utilizing CO2Anaesthetizing the emergence of parasitic wasps and quickly collecting the emergence of the parasitic wasps.
(B) And (3) putting the collected parasitic wasps into a culture bottle filled with parasitic wasp food, and storing the culture bottle in a constant temperature box with the temperature of 16 +/-1 ℃ and the RH of 50 percent for next propagation and inoculation.
In this example, drosophila melanogaster was used, but also drosophila melanogaster d.suzukii, drosophila melanogaster d.virilias, or other drosophila species of drosophila. The Drosophila larvae parasitic wasp can be selected from Leptopila sp.
Example 2 Large-scale propagation of Drosophila parasitism in pupal stage
(1) Host inoculation and propagation:
(A) the bottom box 7 was filled with fruit fly food (same formulation as in example 1); 1000 adult Drosophila melanogaster which has been eclosized for about one week and has been mated thoroughly is taken and treated with CO2After anesthesia, the fruit flies are transferred into a breeding and propagation device and lay eggs in an incubator at the temperature of 25 +/-1 ℃ for 1 day and then are taken out;
(C) and (C) transferring the breeding and propagation device with the drosophila melanogaster eggs in the step (B) to a constant-temperature incubator with the temperature of 25 +/-1 ℃ and the RH of 50 percent for culture, and observing and recording the quantity and the development condition of the drosophila melanogaster eggs.
(2) Parasitic wasp inoculation and reproduction
(A) Inoculating and breeding the parasitic wasps in the pupal stage: and (3) when the fruit flies enter the pupal stage for 1-2 days in the step (1), selecting the fruit fly pupal stage parasitic wasps preserved in a laboratory according to the ratio of 1: 10 of female wasps and fruit flies, and transferring the fruit fly pupal stage parasitic wasps into a breeding and expanding device.
When the parasitic wasp is inoculated, the parasitic wasp is ensured to be in the eclosion stage, and the parasitic fruit fly is ensured to be 1-2 days fruit fly pupae for the parasitic wasp in the pupal stage.
(B) In order to increase the parasitic rate, parasitic wasp food (same formula as in example 1) is added into the box body 5 of the breeding and propagation device to supplement nutrition, the parasitic wasps are observed to parasitize the pupae of the fruit flies, and the parasitic wasps are taken out after 1 day.
(3) Drosophila culture medium removal
And (3) after the step (2) is finished, taking out the fruit fly food in the bottom box 7, or directly replacing the bottom box 7 with the fruit fly food with a clean bottom box 7 (avoiding the generation overlapping of fruit flies in the next step).
(4) Drosophila removal
Generally, the whole growth period of fruit fly at 25 deg.C via egg-larva-pupa-imago takes about 10 days. After parasitic wasps parasitize fruit flies, the host development period can be prolonged, so that the emergence time of the parasitic wasps is about 15 to 18 days after the parasitic wasps emerge. The emergence of parasitic fruit fly parasitized bee is 6-8 days later than normal fruit fly under the conditions of 25 + -1 deg.C and RH of 50%. Thus, the fruit flies that did not succeed in parasitizing will emerge first before the parasitizing bees emerge. Because the fruit fly food is removed in step (3), the newly emerged fruit fly has no food source, so that the newly emerged fruit fly can die completely before the parasitic wasp emerges.
Dead adult drosophila melanogaster were removed 5-6 days after parasitization.
(5) Parasitic bee pupa collection
And (3) scraping the parasitized fruit fly pupae in the step (4) away from the side wall of the culture device by using a soft brush, and quickly collecting the parasitized fruit fly pupae in a fly pupae collecting box 8 (which can be stored in a constant temperature box with the temperature of 16 +/-1 ℃ and the RH of 50 percent for later use and is directly used for large-scale field release).
(6) Bee seed preservation
(A) Fixing the fly pupa collecting box 8 with the collected parasitic bee pupae in the step (5) at the bottom of the container body, and after the parasitic bee eclosion, utilizing CO2And (4) intoxicating the adult wasps by the parasitic wasps with the feathering effect, and quickly collecting the feathered adult wasps.
(B) And (3) putting the collected parasitic wasps into a culture bottle filled with parasitic wasp food, and storing the culture bottle in a constant temperature box with the temperature of 16 +/-1 ℃ and the RH of 50 percent for next propagation and inoculation.
In this example, drosophila melanogaster was used, but also drosophila melanogaster d.suzukii, drosophila melanogaster d.virilias, or other drosophila species of drosophila. The fruit fly pupal parasitic wasp can be selected from Ceriporio cottonii Spalangia, Ceriporio pachyceidea or Ceriporio trichogramma parasitic wasp.
Claims (5)
1. A large-scale breeding method of fruit fly parasitic wasps is characterized by comprising the following steps:
(1) host inoculation and propagation
Loading fruit fly food into the breeding and propagation device, and transferring the adult fruit flies which are eclosion for about one week and are fully mated into the breeding and propagation device; move to 25 + -1oC, cultivating in a constant-temperature insect breeding room until 1 day of oviposition, and taking out fruit flies; moving the breeding and expanding device with the drosophila melanogaster eggs to 25 +/-1oC. Continuously culturing in a constant-temperature insect-raising room with the RH of 50 percent, and observing and recording the quantity and the development condition of the drosophila eggs;
the breeding and expanding device comprises a container body, wherein an upper cover is arranged at the top of the container body, and a detachable bottom box is arranged at the bottom of the container body; the upper cover is provided with an air hole, and a sponge plug is filled in the air hole; a box body for containing parasitic wasp food is arranged inside the container body; the breeding device also comprises a fly pupa collecting box which can be matched and installed with the bottom of the container body, wherein a pulling cover is arranged at an air hole at the bottom of the fly pupa collecting box, and a gauze is arranged at an opening at the upper part of the fly pupa collecting box; the aperture of the gauze is 20-40 meshes, and the gauze is selected according to the size of the adult parasitic wasps;
the container body is cylindrical or cubic, and the bottom box and the fly pupa collecting box are in shapes matched with the container body; the box body for containing parasitic wasp food is hung below the upper cover through a string; or the container wall is installed through a telescopic rail; the bottom box and the fly pupa collecting box are arranged at the bottom of the container body in a threaded connection or a buckling sleeving manner;
when in use, the box body is used for containing parasitic wasp food; the bottom box is used for containing fruit fly food and can be detached and replaced by a new culture-free substrate box so as to remove the fruit fly food; the bottom box is used in the inoculation and propagation process of the fruit fly parasitic wasps, and the fly pupa collecting box is used for collecting the parasitic wasps after the fruit fly parasitic wasps are inoculated and propagated; the collected fruit fly pupae are put into a fly pupae collecting box through a pull cover by combining with the container body; the eclosion parasitic wasps rapidly pass through the gauze; the fly pupae are separated from the fruit fly pupae in the fly pupae collecting box, so that the aim of quickly collecting eclosion parasitic wasps is fulfilled;
(2) inoculating and breeding fruit fly parasitic wasp
When the drosophila melanogaster eggs grow to the inoculation age suitable in the step (1), selecting parasitic bee colonies to transfer into a breeding and propagation device according to the proportion of 1: 10 of the number of female bees and drosophila melanogaster eggs, and adding parasitic bee food; observing the parasitism condition of the parasitic wasps on the fruit fly larvae or pupae, and taking out the parasitic wasps after parasitizing for 1 day;
when inoculating fruit flies, parasitic wasps are ensured to be in the eclosion period; for parasitic wasps of drosophila larvae, the drosophila inoculation instar is 1-2 instars of larvae; for the fruit fly pupal-stage parasitic wasps, the fruit fly inoculation suitable age is 1-2 days in pupal stage;
(3) drosophila culture medium removal
Moving the breeding and expanding device to 25 +/-1 oCContinuously culturing in a constant-temperature insect culturing room with the RH of 50 percent, and removing fruit fly food after fruit fly larvae grow into pupae;
(4) drosophila removal
At 25 +/-1oC, under the condition that RH is 50%, the eclosion of parasitic drosophila parasitizing bees is 6-8 days later than that of normal drosophila; the fruit flies which are not parasitic successfully emerge first; because the fruit fly food is removed, the newly-feathered fruit flies can all die before the parasitic wasps eclosion, and dead fruit fly imagoes are removed;
(5) parasitic bee pupa collection
Collecting parasitized Drosophila pupae before emergence of parasitic wasp, storing in breeding device, and transferring to 16 + -1oC. RH 50% constant temperature incubator;
(6) bee seed preservation
After the parasitic wasp emerges for about 18 days, using CO2Anaesthetizing and emerging parasitic wasp, collecting rapidly, placing into culture bottle containing parasitic wasp food, and placing into 16 + -1oC. The culture medium is stored in an RH 50% incubator and is used for inoculation in the next breeding.
2. The method of claim 1, wherein the parasitic wasp is a drosophila larvae parasitic wasp or a drosophila pupaeParasitoid wasps; wherein said Drosophila larvae parasitoid is a Pediobolus cecidulaLeptopilinaOr open arm CoccomysAsobaraParasitic wasps; the fruit fly pupal stage parasitic wasp belongs to the genus Cerrio cottoniiSpalangiaGenus CerasiferaPachycrepoideusOr Leptospermum speciesTrichopriaParasitic wasps.
3. The method of claim 1, wherein the drosophila diet is formulated by: dissolving 120g of dry yeast in 1000ml of warm water, weighing 318.75g of corn flour, 57g of agar, 60g of peptone, 90g of brown sugar, 180g of glucose and 60g of yeast extract, adding 5000ml of warm water, fully stirring, uniformly mixing, adding 1000ml of dissolved yeast solution, heating and stirring; when the mixture is cooked, 1.5g of MgSO (MgSO) are added in turn4·7H2O and 1.5g CaCl2·2H2Boiling, naturally cooling to 50-60%oAdding 180ml of methyl hydroxybenzoate solution and 50ml of propionic acid, uniformly stirring, cooling for 6-7h, sealing with gauze, and refrigerating in a refrigerator at 4 ℃ for later use; the methyl paraben solution is prepared by dissolving 100g of methyl paraben in 900ml of 95% ethanol.
4. The method of claim 1, wherein the parasitic wasp food is formulated by: adding 27g agar into 1000ml water, stirring thoroughly, mixing well, and 120%oSterilizing with high temperature steam, adding 33g brown sugar and 330ml pure apple juice, stirring, naturally cooling to 60%oC, adding 20ml of methyl hydroxybenzoate solution into the cooled mixture, fully stirring and uniformly mixing, and 4oC, refrigerating for later use; the methyl paraben solution is prepared by dissolving 100g of methyl paraben in 900ml of 95% ethanol.
5. The method of claim 1, wherein the fruit fly is a drosophila melanogaster, a samara flyD. suzukiiOr black fruit flyD. virilis。
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