Method for artificially mass and rapid propagation of the fruit fly corethrobia (Pterocomma pilosum)

Abstract

The present application relates to the field of biological pest control, and particularly relates to a method for artificially mass and rapid propagation of Drosophila suzukii Coptera. The method comprises the following steps: 1) preparation of Drosophila biarmipes and Drosophila suzukii Coptera feeding tank; 2) collection of Drosophila biarmipes pupae; 3) parasitization of Drosophila suzukii Coptera on Drosophila biarmipes pupae; 4) eclosion and feeding of Drosophila biarmipes pupae parasitized by Drosophila suzukii Coptera; 5) feeding of Drosophila suzukii Coptera adults; and 6) repeating steps 1) to 5), so that a large amount of Drosophila suzukii Coptera can be obtained in a short period of time. The method provided by the present application is time-saving, labor-saving, space-saving, and requires simple devices, and can mass propagate Drosophila suzukii Coptera in a short period of time, so as to meet the application requirements of biological characteristic research test or field control of Drosophila suzukii.

Description

Technical Field

[0001] This invention belongs to the field of biological pest control, specifically relating to a method for the rapid and large-scale artificial propagation of fruit fly horned wasps. Background Technology

[0002] Spotted-winged fruit fly ( Drosophila suzukii Matsumura Also known as Suzuki's fruit fly, cherry fruit fly, blueberry fruit fly, etc., it belongs to the order Diptera, suborder Cyclorrhapha, family Drosophilidae, and genus Drosophila. Drosophila Fruit fly subgenus Sophophora Native to Southeast Asia, this fruit fly is one of the few pests in the genus that can damage fresh, healthy fruit. In recent years, it has spread globally, seriously damaging more than 240 kinds of economic crops, including blueberries, strawberries, cherries, grapes, and bayberries, causing huge losses to the global fruit industry.

[0003] Currently, chemical control remains the primary method for controlling the spotted-winged fruit fly, but its drawbacks are becoming increasingly apparent. Therefore, in recent years, both domestic and international efforts have focused on biological control research for this fly, aiming to develop a sustainable integrated control method that combines biological control with other environmentally friendly methods. (The text also mentions a fruit fly, *Hemiberlesia lataniae*, but this appears unrelated to the main topic and is likely a separate, incomplete sentence.) Trichopriadrosophilae Perkins The pupae of the parasitic fruit fly are widely distributed in Asia, Europe, and North America, and have broad application prospects. There is an urgent need for large-scale artificial propagation for production.

[0004] The spotted-winged fruit fly harms healthy fresh fruit, and large-scale artificial rearing is difficult. Using fresh fruit to rear the spotted-winged fruit flies and then artificially propagating the fruit fly *Hypterus xanthipes* is costly and inefficient, failing to meet production needs. This invention uses the double-spined fruit fly as an alternative host, providing a method for rapidly propagating the fruit fly *Hypterus xanthipes*. When using *Drosophila melanogaster* to breed *Hylocereus ferruginosa*, compared with existing patents that use *Drosophila melanogaster* instead of *Drosophila spp.* as the host, the advantages of using *Drosophila melanogaster* as the alternative host are: 1. *Drosophila melanogaster* has a strong xerotropism and pupates in cornmeal, and the pupae do not stick together, making them easy to collect; while the secretions of *Drosophila melanogaster* during pupation cause the pupae to stick together, making them difficult to separate and collect, and the quality of the pupae is affected after mechanical separation; 2. The weight and size of *Drosophila melanogaster* pupae are smaller than those of *Drosophila melanogaster* pupae, but this does not affect the quality of *Hylocereus ferruginosa*. Therefore, compared with *Drosophila melanogaster*, the same number of pupae of the same weight and quantity yield more parasitic wasps; 3. Studies have shown that the lifespan of both male and female *Hylocereus ferruginosa* bred using *Drosophila melanogaster* as the host is longer than that of male and female *Hylocereus ferruginosa* bred using *Drosophila melanogaster* as the host; 4. When selecting hosts, the results showed that the fruit fly larvae bred on the host *Drosophila melanogaster* (black-bellied fruit fly) preferred to parasitize the original host (i.e., *Drosophila melanogaster* pupae), while those bred on the host *Drosophila bisporus* (two-spined fruit fly) showed no obvious host preference. Therefore, in field applications, the fruit fly larvae bred on the host *Drosophila bisporus* are more effective in controlling *Drosophila melanogaster*. Summary of the Invention

[0005] The purpose of this invention is to provide a method for the rapid and large-scale artificial propagation of fruit fly horn bees.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] A method for the rapid and large-scale artificial propagation of the fruit fly horned wasp involves using the fruit fly *Drosophila melanogaster* as the host to propagate the fruit fly horned wasp.

[0008] The above-mentioned method for the rapid and large-scale artificial propagation of fruit fly horn beetles includes the following steps:

[0009] 1) Preparation of rearing tanks for *Drosophila melanogaster* and *Drosophila spp.*

[0010] 2) Collection of Drosophila melanogaster pupae;

[0011] 3) The fruit fly *Drosophila melitica* parasitizes the pupae of the double-spined fruit fly;

[0012] 4) The emergence and rearing of *Drosophila melanogaster* pupae parasitized by *Drosophila melanogaster*.

[0013] 5) Rearing of adult fruit fly gnats;

[0014] 6) Repeat steps 1) to 5) to obtain a large number of fruit fly horn bees in a short period of time.

[0015] In step 1) above, the same transparent plastic container is used to raise adult fruit flies and adult fruit fly hammerhorn wasps. Before raising the insects, the container needs to be cleaned and dried. When using it, the container should be placed horizontally on the table and sealed with 140-mesh nylon netting and rubber bands.

[0016] Step 2) above is as follows: Place the pupae of the fruit fly *Drosophila melanogaster* in a rearing tank and provide artificial feed and water. Five days after the fruit flies begin to emerge, place a plastic box containing artificial feed in the rearing tank for 24 hours to collect enough eggs. Place the plastic box containing the mature larvae and pupae of the fruit fly in a multi-functional fruit fly adult rearing cage and provide water and yeast powder to nourish the larvae. Utilize the desiccating nature of the mature larvae to collect and separate the pupae of the same age as the fruit fly using corn starch for parasitism.

[0017] Step 3) above is as follows: On the day after the emergence of the fruit fly larvae, provide double-spined fruit fly pupae for parasitism. The double-spined fruit fly pupae are laid flat in an empty plastic box. Each time, select 3 boxes of 1-day-old double-spined fruit fly pupae, each box containing 12,500 double-spined fruit fly pupae, and provide them to 7,500 fruit fly larvae for parasitism for 24 hours. Parasitize a batch every 24 hours. Each batch of fruit fly larvae can continuously parasitize 25-30 boxes of double-spined fruit fly pupae for large-scale propagation.

[0018] Step 4 above is as follows: Transfer each batch of *Drosophila melanogaster* pupae parasitized by the fruit fly horn wasp to an empty plastic box. Seal the plastic box with a 140-mesh nylon net and rubber bands. After 6 days, once the unparasitized *Drosophila melanogaster* have emerged and starved to death, remove the nylon net and place the plastic box containing the *Drosophila melanogaster* pupae parasitized by the fruit fly horn wasp into a rearing tank for development, waiting for the parasitic wasp to emerge.

[0019] Step 5 above is as follows: After the adult fruit fly horn bee emerges, provide 10wt% honey water in the rearing tank of the adult fruit fly horn bee, and replace it with fresh 10wt% honey water every 5 days.

[0020] The significant advantages of this invention are:

[0021] Existing technologies use *Drosophila melanogaster* as the host, while this invention uses *Drosophila bisporus* as the host to propagate *Drosophila melanogaster*, which has the following advantages:

[0022] 1) The Drosophila melanogaster has a strong tendency to burrow into corn flour to pupate, and the pupae do not stick together, making them easy to collect. In contrast, the secretions of the Drosophila melanogaster during pupation cause the pupae to stick together, making them difficult to separate and collect. The quality of the pupae will be affected after using instruments to separate them.

[0023] 2) The weight and size of the pupae of the double-spined fruit fly are smaller than those of the black-bellied fruit fly, but this does not affect the quality of the fruit fly horn wasp. Therefore, compared with the black-bellied fruit fly, the same number of pupae can produce more parasitic wasps. 3) The lifespan of the fruit fly horn wasps bred with the double-spined fruit fly as the host is longer for both male and female wasps than that of the fruit fly horn wasps bred with the black-bellied fruit fly as the host.

[0024] 4) The fruit fly horn wasp bred with Drosophila melanogaster as the host prefers to parasitize the original host (i.e., Drosophila melanogaster pupae), while the fruit fly horn wasp bred with Drosophila bisporus does not have a clear host preference. Therefore, when applied in the field, the fruit fly horn wasp bred with Drosophila bisporus as the host has a better control effect on Drosophila melanogaster. Attached Figure Description

[0025] Figure 1 : Rearing tank for double-spined fruit flies and fruit fly hammer-horned bees.

[0026] Figure 2 A schematic diagram showing how the same-day-old pupae of the Drosophila melanogaster were obtained by collecting and separating them using corn starch, taking advantage of the stem-driing properties of mature larvae. Detailed Implementation

[0027] The invention will now be described in further detail with reference to specific examples.

[0028] In this invention, the formula of the artificial feed is: 50g / L corn starch, 50g / L white sugar, 25g / L yeast, and 5g / L agar powder.

[0029] Example 1

[0030] Step 1: Preparation of rearing tanks for *Drosophila melanogaster* and *Drosophila hammer-horn* wasps

[0031] Adult Drosophila melanogaster and adult Drosophila spp. were reared in the same transparent plastic container. Figure 1 The container has a capacity of 20 L, a height of 41 cm, a base diameter of 28 cm, and a mouth diameter of 13 cm. Before raising insects, the container must be cleaned and dried. When using, place the container horizontally on a table with the mouth perpendicular to the surface. Secure the container to the table with two rubber tubes to prevent it from rolling during use. Cover the mouth of the container with a 140-mesh nylon mesh and secure it with a rubber band.

[0032] Step 2: Collection of Drosophila pupae

[0033] 25 ml of *Drosophila melanogaster* pupae (approximately 500 pupae per ml) was placed in a prepared rearing container for development. The rearing temperature was 25±1℃, the relative humidity was 60±10%, and the photoperiod was 14L:10D. Inside the rearing container, a plastic box containing artificial feed (10cm long, 10cm wide, and 4cm high) with an unsealed lid was placed inside. A long strip of absorbent cotton was placed inside the conical flask, extending from the mouth to the bottom. Five days after the *Drosophila melanogaster* began to emerge, a plastic box containing artificial feed (10cm long, 10cm wide, and 4cm high, with an unsealed lid) was placed in the rearing container for 24 hours to collect sufficient eggs. Plastic boxes containing mature larvae and pupae of *Drosophila melanogaster* and artificial feed were transferred to multifunctional adult fruit fly rearing cages (CN201220177838.4) for development. The rearing temperature was 25±1℃, the relative humidity was 60±10%, and the photoperiod was 14L:10D. One conical flask containing water and one petri dish containing yeast powder were placed in the multifunctional adult fruit fly rearing cage to provide nutrition for the larvae. Qualitative filter paper with a diameter of 18cm was folded into a triangular funnel shape and placed in an empty plastic box (10cm long, 10cm wide, and 4cm high, with an unsealed lid). A layer of dry corn starch was spread evenly inside the filter paper funnel. The plastic boxes containing mature larvae and pupae of *Drosophila melanogaster* and artificial feed were then stacked at a 45-degree angle, alternating with the empty plastic boxes containing the filter paper funnels. Figure 2 The process involves placing a plastic box containing mature larvae and pupae of the fruit fly *Drosophila melanogaster* (also known as the Drosophila spp.) and artificial feed on top, and an empty plastic box containing a filter paper funnel on the bottom. The larvae mature 2-3 days after hatching. The mature larvae of *Drosophila melanogaster* are strongly attracted to dryness and will crawl out of the feed box along the inner wall of the artificial feed box and burrow into the corn starch spread in the filter paper funnel to wait for pupation. After pupation, the corn starch in the filter paper funnel is poured onto a 40-mesh sieve with a diameter of 0.45 mm, and the sieve is gently shaken to remove excess corn starch. The sieved *Drosophila melanogaster* pupae are then poured into an empty plastic box (17cm x 10cm x 1cm) and spread out. The pupae are sieved every 24 hours for subsequent parasitism.

[0034] Step 3: Fruit fly horn wasp parasitizes double-spined fruit fly pupae.

[0035] The day after the emergence of the fruit fly *Drosophila melanogaster*, provide *Drosophila bisporus* pupae for parasitism. Place 25 ml of 1-day-old *Drosophila bisporus* pupae (approximately 500 pupae per ml) flat in an empty plastic box (17cm x 10cm x 1cm, unsealed). Place 3 boxes of *Drosophila bisporus* pupae in each *Drosophila melanogaster* rearing container, providing parasitism for 24 hours. Cover the opening of the rearing container with 140-mesh nylon mesh and secure with rubber bands. Each *Drosophila melanogaster* rearing container houses 7500 *Drosophila melanogaster* wasps. A batch can be parasitized every 24 hours, and each batch of *Drosophila melanogaster* can continuously parasitize 25-30 boxes of *Drosophila bisporus* pupae for mass propagation.

[0036] Step 4: Emergence and rearing of *Drosophila melanogaster* pupae parasitized by *Drosophila melanogaster*

[0037] Each batch of *Drosophila melanogaster* pupae parasitized by the fruit fly wasp *Schefflera spp.* were transferred to empty transparent plastic boxes (16cm x 16cm x 20cm), sealed with 140-mesh nylon mesh and secured with rubber bands. After 6 days, once the unparasitized *Schefflera spp.* emerged and starved to death, the nylon mesh was removed, and the pupae boxes were placed in a *Schefflera spp.* rearing tank for development. The rearing temperature was 25±1℃, the relative humidity was 60±10%, and the photoperiod was 14L:10D, while waiting for the parasitic wasps to emerge. After 14-15 days of rearing, the male parasitic wasps began to emerge, and the female wasps began to emerge the day after the males began to emerge. The peak emergence period was reached on the 3rd day after the males began to emerge.

[0038] Step 5: Rearing of adult fruit fly gnats

[0039] After the adult fruit fly horn bees emerge, place a 50ml conical flask in the rearing tank and add a 10wt% honey solution to the flask. Insert cotton wool from the bottom of the flask to the mouth of the flask for the fruit fly horn bees to feed on. Change the honey solution with fresh honey every 5 days.

[0040] Step 6:

[0041] Repeating steps 1 to 5 above can yield a large quantity of fruit fly horn bees for field application in a short period of time.

[0042] The relevant parasitic indicators are shown in the table below:

[0043]

[0044] Note: Data in the table are mean ± standard error.

[0045] As shown in the table above, using *Drosophila melanogaster* pupae as a host for breeding *F. melanogaster* can achieve a parasitism rate of 82.64% ± 0.88 and an emergence rate of 76.21% ± 1.06. Each rearing tank can hold 37,875-39,375 *Drosophila melanogaster* pupae, and each batch can yield 23,282.76-25,724.39 adult *F. melanogaster*, including 13,038.37-15,949.12 females.

[0046] This parasitic wasp breeding technology can quickly and easily propagate large quantities of fruit fly gnatella. The process is easy to master and implement, and the double-spined fruit fly is easy to raise, lays a large number of eggs, and the pupae have a strong tendency to dry out, making them easy to collect and store. It is suitable for rapid and large-scale breeding of wasps to meet the needs of biological characteristic research experiments and field control of spotted-winged fruit flies.

[0047] Finally, it should be stated that the above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made within the scope of the claims of the present invention should be included in the scope of the present invention.

Claims

1. A method for the rapid and large-scale artificial propagation of fruit fly hornflies, characterized in that, The steps are as follows: 1) Preparation of rearing tanks for *Drosophila melanogaster* and *Drosophila spp.* The same transparent plastic container was used to raise adult Drosophila melanogaster and adult Drosophila spp. The container has a capacity of 20 L, a height of 41 cm, a bottom diameter of 28 cm, and a mouth diameter of 13 cm. Before raising the insects, the container should be cleaned and dried. When using it, the container should be placed horizontally on the table with the mouth perpendicular to the table. The container should be fixed to the table with two rubber tubes to prevent it from rolling during use. The mouth of the container should be covered with 140-mesh nylon netting and secured with rubber bands. 2) Collection of Drosophila melanogaster pupae; 25 ml of *Drosophila melanogaster* pupae were placed in a prepared rearing container for development. Each ml of *Drosophila melanogaster* pupae contained 500 pupae. The rearing temperature was 25±1℃, the relative humidity was 60±10%, and the photoperiod was 14L:10D. A plastic box containing artificial feed and a conical flask containing clean water were placed inside the rearing container for feeding. The plastic box was 10 cm long, 10 cm wide, and 4 cm high, with an unsealed lid. Long strips of absorbent cotton were placed inside the conical flask, extending from the mouth to the bottom. Five days after the *Drosophila melanogaster* began to emerge, a plastic box containing artificial feed was placed in the rearing container for 24 hours to collect sufficient eggs. The plastic box containing artificial feed was 10 cm long, 10 cm wide, and 4 cm high. The plastic box containing mature larvae and pupae of the fruit fly (Drosophila melanogaster) and artificial feed was left unsealed and transferred to a multifunctional fruit fly adult rearing cage for development. The rearing temperature was 25±1℃, the relative humidity was 60±10%, and the photoperiod was 14L:10D. A conical flask containing water and a petri dish containing yeast powder were placed in the multifunctional fruit fly adult rearing cage to provide nutrition for the larvae. A qualitative filter paper with a diameter of 18 cm was folded into a triangular funnel shape and placed in a 10 cm long, 10 cm wide, and 4 cm high container. In an empty plastic box (17 cm x 10 cm x 1 cm, unsealed), spread a layer of dry cornstarch inside a filter paper funnel. Then, place the plastic box containing mature Drosophila larvae and pupae, along with the box containing the filter paper funnel, at a 45-degree angle, alternating them. The box containing the mature Drosophila larvae and pupae and the box containing the artificial feed should be on top, with the box containing the filter paper funnel on the bottom. The hatched larvae will mature in 2-3 days. The mature Drosophila larvae are strongly attracted to dryness and will crawl out of the plastic box containing the artificial feed along the inner wall and burrow into the cornstarch layer in the filter paper funnel to pupate. After pupation, pour the cornstarch from the filter paper funnel onto a 40-mesh sieve with a 0.45 mm aperture, gently shake it left and right to remove excess cornstarch, and then pour the sieved Drosophila pupae into a 17 cm x 10 cm x 1 cm empty plastic box and spread them out. Sift the pupae every 24 hours for subsequent parasitism. 3) The fruit fly *Drosophila melitica* parasitizes the pupae of the double-spined fruit fly; The day after the emergence of the fruit fly hyacinth wasp, provide *Drosophila melanogaster* pupae for parasitism. Place 25 ml of 1-day-old *Drosophila melanogaster* pupae flat in an empty plastic box (17 cm × 10 cm × 1 cm, with the lid unsealed). Each ml of 1-day-old *Drosophila melanogaster* pupae contains 500 pupae. Place 3 boxes of *Drosophila melanogaster* pupae in each *Drosophila melanogaster* rearing tank and provide parasitism for 24 hours. Cover the opening of the rearing tank with 140-mesh nylon netting and secure it with rubber bands. Each *Drosophila melanogaster* rearing tank contains 7500 *Drosophila melanogaster* wasps. A batch can be parasitized every 24 hours. Each batch of *Drosophila melanogaster* wasps can continuously parasitize 25-30 boxes of *Drosophila melanogaster* pupae for mass propagation. 4) The emergence and rearing of *Drosophila melanogaster* pupae parasitized by *Drosophila melanogaster*. Each batch of *Drosophila melanogaster* pupae parasitized by *S. melanogaster* were transferred to empty, transparent plastic boxes (16cm in diameter). The boxes were sealed with 140-mesh nylon mesh and secured with rubber bands. After 6 days, once the unparasitized *Drosophila melanogaster* pupae had emerged and starved to death, the nylon mesh was removed, and the pupae boxes were placed in a *S. melanogaster* rearing tank for development. The rearing temperature was 25±1℃, the relative humidity was 60±10%, and the photoperiod was 14L:10D. The *S. melanogaster* pupae were kept until they emerged. After 14-15 days of rearing, the male *S. melanogaster* pupae began to emerge. The female pupae began to emerge the day after the males began to emerge, and the peak emergence period was reached on the 3rd day after the males began to emerge. 5) Rearing of adult fruit fly gnats; After the adult fruit fly horn bee emerges, place a 50 ml conical flask in the rearing tank of the adult fruit fly horn bee and add 10 wt% honey water to the conical flask. Insert absorbent cotton at the bottom and extend it to the mouth of the flask for the fruit fly horn bee to feed on. Change the honey water with fresh honey every 5 days. 6) Repeat steps 1) to 5) above to obtain a large number of fruit fly horn bees for field application in a short period of time.

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