Artificial feed for larvae of monochamus alternatus and method for feeding and breeding generations

Abstract

The application discloses artificial feed for Anoplophora glabripennis larvae and a method for feeding and breeding in generations. The feed components include potato starch, sucrose, soybean powder, casein, cholesterol, Wies salt, beer yeast, microcrystalline cellulose, agar powder, citric acid, sorbic acid, 4-hydroxy benzoic acid methyl ester, soybean oil, distilled water, ascorbic acid and propionic acid. The method for feeding and breeding in generations is as follows: low-age larvae of Anoplophora glabripennis are collected and used as the initial larvae source, and the larvae are fed with the artificial feed until pupation and then eclosion; the eclosion adult insects are put into a container with Acer pseudosieboldianus branches in pairs according to the female and male; a wood block with gauze on the surface is arranged in the container as the mating and oviposition matrix of Anoplophora glabripennis; the eggs on the gauze are taken out and hatched, and the newly hatched larvae can be continuously fed with the artificial feed. The components and quality of the artificial feed for Anoplophora glabripennis larvae are uniform, the feeding cycle of the larvae is shortened to less than 200 days, fresh host wood segments do not need to be provided for the adult insects to lay eggs, and the workload and cost of feeding and breeding in generations are reduced.

Description

Technical Field

[0001] This invention belongs to the field of artificial insect rearing technology, specifically relating to an artificial feed for the larvae of the longhorn beetle and a method for its propagation. Background Technology

[0002] The bald-shouldered star beetle ( Anoplophora glabripennis Beetle (Ceratopteris) belongs to the order Coleoptera, family Cerambycidae, and genus Cerambycidus. Anoplophora The Asian longhorn beetle (Populus spp.), native to China, is a major forest pest. Its larvae bore into the trunk and main branches of the host plant, disrupting nutrient and water transport, leading to weakening and death of the entire tree. It has a wide host range; in China, it primarily affects plants in the genus Populus (…). Populus ), Salix ( Salix ), Acer ( Acer ) and elm ( Ulmus Trees.

[0003] The Asian longhorn beetle (Anoplophora glabripennis) is particularly prevalent in the Three-North Shelterbelt Project. Since the first phase of the Three-North Shelterbelt Project began in the 1980s, this pest has almost destroyed the first phase of the shelterbelts, which were mainly composed of poplar trees. Subsequent phases of poplar shelterbelt projects have also suffered severe damage. Even today, the Asian longhorn beetle continues to cause significant damage in the Three-North Shelterbelt area. Furthermore, due to the extensive planting of willows and maples in urban greening projects, the Asian longhorn beetle has also become a serious urban landscaping pest in recent years.

[0004] The entire larval and pupal stages of the Asian longhorn beetle are concealed within the trunk of the host tree, making control extremely difficult. Biological control is an important technique for controlling juvenile Asian longhorn beetles, such as using various parasitic wasps (…). Sclerodermus To control its young larvae, use the flower beetle (spp.) Dastarcushelophoroides Controlling mature larvae and pupae is crucial. Additionally, spraying microencapsulated contact and stomach poisons during the adult stage is currently the main method for controlling adult *Anoplophora glabripennis*. However, due to the poor host specialization of parasitic wasps such as *Lycoperdon perlatum* and *Pterygodium japonicum*, their overall effectiveness in controlling juvenile *Anoplophora glabripennis* is insufficient. This necessitates continuous and widespread application of chemical pesticides during the adult stage, causing damage to the ecological environment. Even after years of chemical control, the damage caused by this pest has not been alleviated in various regions. Therefore, utilizing highly specialized natural enemies may be an important way to effectively control this pest.

[0005] After years of research, several specialized parasitic wasps that parasitize the larvae of the longhorn beetle have been discovered, such as the broad-shouldered wasp ( ). Eurytoma sp.), stalked braconid wasp ( Spathius sp.), Aukin bee ( Oxysychus(sp.), etc. Utilizing these natural enemies to control the *Anoplophora glabripennis* in production requires high-quality artificial breeding techniques, and a continuous supply of high-quality *Anoplophora glabripennis* larvae as hosts is the foundation for large-scale artificial breeding of natural enemies. In most parts of China, the *Anoplophora glabripennis* has one generation every two years, with an extremely long larval stage. During the larval stage, it bores and damages trees under the bark or inside the xylem, making it extremely difficult to breed natural enemies in large quantities by collecting larvae from the wild. Therefore, it is necessary to artificially rear the larvae, and to conduct indoor artificial propagation, to continuously provide a consistent quality host for the breeding of natural enemies.

[0006] The April 1992 issue of the *Journal of Beijing Forestry University* published "Artificial Rearing of the Asian Longhorn Beetle" by He Ping and Huang Jingfang, which disclosed a semi-artificial feed for Asian Longhorn Beetle larvae made from poplar powder mixed with various chemical reagents. However, it did not propose a method for the generation of the beetle. The July 1999 issue of the same journal published "Artificial Rearing of the Asian Longhorn Beetle (I)" and "Artificial Rearing of the Asian Longhorn Beetle (II)" by Zhao Jun, Ogura Nobuo, and Isono Masahiro. These articles similarly disclosed a semi-artificial feed for Asian Longhorn Beetle larvae made from poplar powder mixed with various chemical reagents. Their proposed method for generation was to provide host wood segments for the adults to lay eggs after they emerged from the artificially raised beetles. Several days later, the wood segments were dissected to extract the eggs or larvae for continued feeding. The above methods have the following shortcomings: First, the published feeds are all semi-artificial, with poplar powder as the base material, the supply of which is easily limited by region and season. Furthermore, the physiological conditions of poplar trees vary across different regions, years, and seasons, making it impossible to standardize the quality of artificial feed made from them. The developmental progress of longhorn beetles after consuming this feed is also highly uneven, increasing the difficulty of large-scale breeding. Secondly, the published method for breeding longhorn beetles requires providing host wood segments for adults to lay eggs, and then dissecting the eggs or larvae from the wood segments to continue reproduction. This method is cumbersome, requiring the collection of large quantities of wood segments from the wild, increasing the cost of breeding and making it unsuitable for large-scale propagation.

[0007] Therefore, there is an urgent need to develop a standardized and high-quality artificial feed for the larvae of the longhorn beetle, and to develop more economical and efficient longhorn beetle propagation techniques. Summary of the Invention

[0008] In view of the above problems, the purpose of this invention is to provide an artificial feed for the larvae of the Asian longhorn beetle (Sclerotium glabripennis). Another purpose of this invention is to provide a method suitable for indoor subculture rearing of the Asian longhorn beetle.

[0009] As a first aspect of the present invention, the present invention provides an artificial feed for the larvae of the longhorn beetle, *Aegilops glabripennis*.

[0010] The artificial feed comprises the following raw materials in parts by weight: 40.0-60.0 parts potato starch, 40.0-60.0 parts sucrose, 100.0-140.0 parts soybean flour, 25.0-40.0 parts casein, 1.5-4.0 parts cholesterol, 7.0-12.0 parts Wiener salt, 70.0-130.0 parts brewer's yeast, 250.0-450.0 parts microcrystalline cellulose, 60.0-90.0 parts agar powder, 3.0-6.0 parts citric acid, 3.0-6.0 parts sorbic acid, 3.0-6.0 parts methyl 4-hydroxybenzoate, 10.0-18.0 parts soybean oil, 1200.0-2000.0 parts distilled water, 4.0-8.0 parts ascorbic acid, and 1.5-6.0 parts propionic acid.

[0011] Preferably, the artificial feed comprises the following raw materials in parts by weight: 50.0 parts potato starch, 50.0 parts sucrose, 120.0 parts soybean flour, 30.0 parts casein, 3.0 parts cholesterol, 9.0 parts Wiegand salt, 90.0 parts brewer's yeast, 390.0 parts microcrystalline cellulose, 70.0 parts agar powder, 4.0 parts citric acid, 5.0 parts sorbic acid, 5.0 parts methyl 4-hydroxybenzoate, 14.0 parts soybean oil, 1550.0 parts distilled water, 6.0 parts ascorbic acid, and 4.0 parts propionic acid.

[0012] Preferably, the method for preparing the artificial feed includes the following steps:

[0013] Step A1: Weigh the raw materials according to the specified mass fractions. Add all raw materials except ascorbic acid and propionic acid to a beaker and mix thoroughly. Seal the beaker opening with aluminum foil. Place the beaker containing the mixture in an autoclave and sterilize at 121°C for 20 minutes to obtain a sterilized mixture.

[0014] Step A2: Transfer the sterilized mixture to a clean bench and let it cool to about 70°C. Add ascorbic acid and propionic acid and mix thoroughly to obtain artificial feed for the larvae of the longhorn beetle.

[0015] As a second aspect of the present invention, the present invention provides a method for raising the longhorn beetle with scaly shoulders, comprising the following steps:

[0016] Step B1: In mid-to-late July, collect host wood segments damaged by the longhorn beetle, dissect and collect young larvae under the bark, and use them as the first generation of insects for the longhorn beetle rearing.

[0017] Step B2, following steps A1 and A2, in a clean bench, dispense the artificial feed for the larvae of the longhorn beetle (A. globosa) into 60ml covered transparent plastic cups (4.0cm base diameter, 5.5cm mouth diameter, 3.5cm height), filling the cups to about 2 / 3 full. After the feed cools to room temperature, use sterile tweezers to make a small hole in the center of the feed. Then, insert a single longhorn beetle larva into the hole, cover the cup, and make five 0.5mm holes in the lid to ensure ventilation. Place the plastic cup containing the feed and the longhorn beetle in an artificial climate chamber at 25°C, RH=55%, L:D=0:24h for 8 weeks, without changing the feed.

[0018] Step B3: Remove the longhorn beetle larvae from Step B2 after 8 weeks of rearing. Following steps A1 and A2, in a clean bench, portion the artificial feed for the *Acer glabripennis* larvae into 60ml covered transparent plastic cups (4.0cm base diameter, 5.5cm mouth diameter, 3.5cm height), filling the cups to about 2 / 3 full. After the feed cools to room temperature, use sterile tweezers to make a small hole in the center of the feed. Insert one longhorn beetle larva into the hole, cover the cup, and poke five 0.5mm holes in the lid for ventilation. Place the plastic cup containing the feed and the longhorn beetle in an artificial climate chamber at 25°C, RH=55%, L:D=0:24h, and continue rearing for 4 weeks without changing the feed.

[0019] Step B4: Remove the longhorn beetle larvae from Step B3 after 4 weeks of rearing. Following steps A1 and A2, in a clean bench, portion the artificial feed for the *Acer glabripennis* larvae into 60ml covered transparent plastic cups (4.0cm base diameter, 5.5cm mouth diameter, 3.5cm height), filling the cups to about 2 / 3 full. After the feed cools to room temperature, use sterile tweezers to make a small hole in the center of the feed. Insert one longhorn beetle larva into the hole, cover the cup, and poke five 0.5mm holes in the lid for ventilation. Place the plastic cup containing the feed and the longhorn beetle in an artificial climate chamber at 25°C, RH=55%, L:D=0:24h, and continue rearing for 4 weeks without changing the feed.

[0020] Step B5: Place the longhorn beetle larvae, which have been raised for 4 weeks in step B4, along with their feed cups, in an artificial climate chamber at 5°C, RH=55%, L:D=0:24h, and store them at low temperature for 6 weeks.

[0021] Step B6: Transfer each larva of the longhorn beetle (Aegilops glabripennis) stored at low temperature in Step B5 to a 60ml transparent plastic cup with a lid (4.0cm base diameter, 5.5cm mouth diameter, 3.5cm height). Cover the cup with the lid, making five 0.5mm holes to ensure ventilation. Place the plastic cup containing the longhorn beetle larvae in an artificial climate chamber at 25°C, RH=55%, L:D=0:24h. After 20 days, begin daily checks on the pupation and emergence of the longhorn beetles.

[0022] Preferably, to achieve subculture, the following steps are also included:

[0023] Step B7: After the *Aegilops glabripennis* begins to emerge, prepare a lidded transparent plastic bottle (20.0cm base diameter, 15.0cm mouth diameter, 30.0cm height) with a volume of approximately 8L as a rearing box for the adults. Introduce 10 pairs of newly emerged male and female adults into the box. Place fresh *Acer negundo* branches (20.0cm long, 1.0-2.0cm diameter) into the box to supplement their nutrition. Prepare a rectangular wooden block (15.0cm long, 10.0cm wide, 3.0cm high), wrapping the surface of the block evenly with medical gauze twice and securing it with staples. Place the wooden block in the rearing box as a mating and oviposition substrate for the adults. Cover the bottle with the lid, making 10 0.5cm holes in the lid to ensure ventilation. Replace the branches in the rearing box every 2 days. Place the rearing box in an artificial climate chamber at 25°C, RH=55%, L:D=16:8h.

[0024] Step B8: After the *Avicennia martensii* begins laying eggs, remove the wooden block from the rearing box every 4 days. Use a brush to remove the eggs from the surface of the gauze and the seams. Then, put the wooden block back into the rearing box for the *Avicennia martensii* to continue laying eggs. Place the removed eggs in a sterile petri dish and place the dish in an artificial climate chamber at 25°C, RH=55%, L:D=16:8h until the eggs hatch. The newly hatched larvae can be fed artificial feed to achieve the propagation of *Avicennia martensii*.

[0025] The present invention has at least the following advantages and beneficial effects: (1) The raw materials of the artificial feed for larvae of the longhorn beetle of the present invention are entirely composed of commercially available chemical agents, completely eliminating the dependence on host plants, and the feed production is not limited by region, year, or season; (2) The raw material composition of the artificial feed for larvae of the longhorn beetle of the present invention is controllable, and the quality of the artificial feed produced therefrom is uniform between different batches, and the larvae of the longhorn beetle of the longhorn beetle fed with it have uniform development progress and quality. Experiments show that the survival rate of larvae of the longhorn beetle of the longhorn beetle is high, the culture cycle is shortened, and the breeding efficiency is high when the artificial feed for larvae of the longhorn beetle ...

[0026] The method for propagating the *Avicennia marina* (a type of longhorn beetle) according to this invention is simple to operate. Only three feed changes are needed throughout the larval stage, shortening the larval rearing period to no more than 200 days, significantly reducing the time compared to over 500 days in the natural environment. During the adult stage, only thin branches of the host plant are needed to supplement its nutrition; fresh host wood is not required for egg laying, and larvae do not need to be dissected from the wood again for propagation, greatly reducing the workload and cost of propagation.

[0027] By using the artificial feed and generation method of the present invention, the *Anoplophora glabripennis* can be continuously bred with test insects of different life stages and larval stages, providing a host for the breeding of obligate natural enemies of the *Anoplophora glabripennis*. Detailed Implementation

[0028] The present invention will be further described below with reference to the embodiments.

[0029] Unless otherwise specified, the methods used in the embodiments of this invention are all general techniques in the field, and all equipment and raw materials are commonly used products in this industry and can be purchased from the market.

[0030] Example 1

[0031] The artificial feed for the larvae of the longhorn beetle in this embodiment is composed of the following raw materials in parts by weight: 50.0g potato starch, 50.0g sucrose, 120.0g soybean flour, 30.0g casein, 3.0g cholesterol, 9.0g Wiener salt, 90.0g brewer's yeast, 390.0g microcrystalline cellulose, 70.0g agar powder, 4.0g citric acid, 5.0g sorbic acid, 5.0g methyl 4-hydroxybenzoate, 14.0g soybean oil, 1550.0ml distilled water, 6.0g ascorbic acid, and 4.0ml propionic acid.

[0032] The method for preparing artificial feed in this embodiment includes the following steps:

[0033] (1) Weigh the raw materials according to the weight proportions, add the remaining raw materials except ascorbic acid and propionic acid into the beaker, and mix thoroughly. Seal the mouth of the beaker with tin foil and sterilize it in a high-pressure autoclave at 121°C for 20 minutes to obtain mixture A.

[0034] (2) Transfer the mixture A to a clean bench, wait for it to cool to 70°C, add ascorbic acid and propionic acid, and mix thoroughly to obtain artificial feed for the larvae of the longhorn beetle.

[0035] (3) In the clean bench, the artificial feed is divided into the larval feed box. When the feed is cooled to room temperature, it can be used to feed the larvae of the longhorn beetle.

[0036] Example 2

[0037] The artificial feed for the larvae of the longhorn beetle in this embodiment is composed of the following raw materials in parts by weight: 40.0g potato starch, 40.0g sucrose, 100.0g soybean flour, 25.0g casein, 1.5g cholesterol, 7.0g Wiener salt, 70.0g brewer's yeast, 250.0g microcrystalline cellulose, 60.0g agar powder, 3.0g citric acid, 3.0g sorbic acid, 3.0g methyl 4-hydroxybenzoate, 10.0g soybean oil, 1200.0ml distilled water, 4.0g ascorbic acid, and 1.5ml propionic acid.

[0038] The method for preparing the artificial feed in this embodiment is exactly the same as that in Example 1, and will not be repeated here.

[0039] Example 3

[0040] The artificial feed for the larvae of the longhorn beetle in this embodiment is composed of the following raw materials in parts by weight: 60.0g potato starch, 60.0g sucrose, 140.0g soybean flour, 40.0g casein, 4.0g cholesterol, 12.0g Wiener salt, 130.0g brewer's yeast, 450.0g microcrystalline cellulose, 90.0g agar powder, 6.0g citric acid, 6.0g sorbic acid, 6.0g methyl 4-hydroxybenzoate, 18.0g soybean oil, 2000.0ml distilled water, 8.0g ascorbic acid, and 6.0ml propionic acid.

[0041] The method for preparing the artificial feed in this embodiment is exactly the same as that in Example 1, and will not be repeated here.

[0042] Comparative Example 1

[0043] The artificial feed in this comparative example consists of the following raw materials in parts by weight:

[0044] Potato starch 50.0g, sucrose 50.0g, soybean flour 120.0g, casein 10.0g, cholesterol 3.0g, Wiegand salt 9.0g, brewer's yeast 90.0g, microcrystalline cellulose 390.0g, agar powder 70.0g, citric acid 4.0g, sorbic acid 5.0g, methyl 4-hydroxybenzoate 5.0g, soybean oil 14.0g, distilled water 1550.0ml, ascorbic acid 6.0g, propionic acid 4.0ml.

[0045] The preparation method of the artificial feed in this comparative example is completely the same as that in Example 1, and will not be repeated here.

[0046] Comparative Example 2

[0047] The artificial ingredient in this comparative example is composed of the following raw materials in parts by weight: 50.0g potato starch, 50.0g sucrose, 120.0g soybean flour, 30.0g casein, 3.0g cholesterol, 9.0g Wiener salt, 90.0g brewer's yeast, 390.0g microcrystalline cellulose, 100.0g agar powder, 4.0g citric acid, 5.0g sorbic acid, 5.0g methyl 4-hydroxybenzoate, 14.0g soybean oil, 1550.0ml distilled water, 6.0g ascorbic acid, and 4.0ml propionic acid.

[0048] The preparation method of the artificial feed in this comparative example is completely the same as that in Example 1, and will not be repeated here.

[0049] Comparative Example 3

[0050] The artificial feed in this comparative example is composed of the following ingredients in parts by weight: 50.0g potato starch, 50.0g sucrose, 120.0g soybean flour, 30.0g casein, 3.0g cholesterol, 9.0g Wiener salt, 90.0g brewer's yeast, 390.0g microcrystalline cellulose, 70.0g agar powder, 4.0g citric acid, 5.0g sorbic acid, 5.0g methyl 4-hydroxybenzoate, 14.0g soybean oil, 1550.0ml distilled water, and 6.0g ascorbic acid.

[0051] The preparation method of the artificial feed in this comparative example is completely the same as that in Example 1, and will not be repeated here.

[0052] Larvae of the *Aegilops glabripennis* were reared using the artificial feeds prepared in Examples 1-3 and Comparative Examples 1-3, respectively. One newly hatched *Aegilops glabripennis* larva was placed in each larval feed box, with a total of 100 larvae inoculated for each feed. The rearing period was 8 weeks, constituting the first cycle. The artificial feeds described in Examples 1-3 and Comparative Examples 1-3 were then prepared again, and the larvae that survived the first cycle were reared for 4 weeks, constituting the second cycle. The artificial feeds described in Examples 1-3 and Comparative Examples 1-3 were then prepared again, and the larvae that survived the second cycle were reared for 4 weeks, constituting the third cycle, after which the larval rearing was completed. The number of surviving larvae and their weight were recorded at the end of each rearing cycle. The rearing equipment was an artificial climate chamber, with conditions set at 25°C, RH=55%, and L:D=0:24h. Specific experimental results are shown in Table 1.

[0053] Table 1. Analysis of the effects of artificial feed formulations on larvae of the longhorn beetle (Anoplophora glabra).

[0054]

[0055] Note: The data listed in Table 1 are mean ± standard error. The same letter in the same column indicates no significant difference, and different letters in the same column indicate significant difference.

[0056] As shown in Table 1, Comparative Example 1 reduced the proportion of casein. The results showed that although the number of surviving longhorn beetles in Comparative Example 1 was not significantly different from that in the Example 1, the beetles' weight at each feeding cycle and their final weight were significantly lower than in the Example 1 (the smaller the beetle's weight, the weaker its vitality after molting). This indicates that casein plays a crucial role in the weight gain of juvenile longhorn beetles.

[0057] The results showed that the survival rate of *Anoplophora glabripennis* larvae fed using Examples 1-3 was significantly higher than that of Comparative Examples 2 and 3. The artificial feed in Comparative Example 2, compared to Example 1, had increased agar powder content, which increased feed stickiness, affecting beetle activity and significantly reducing the survival rate and body weight of the beetles fed with it (for example, 100 parts by weight of agar powder resulted in excessive stickiness, negatively impacting beetle survival). The artificial feed in Comparative Example 3, compared to Example 1, removed propionic acid, causing mold growth during beetle rearing, greatly affecting beetle survival and resulting in lower body weight.

[0058] In summary, the relative contents of agar, casein, and propionic acid in the artificial feed for larvae of the longhorn beetle are key factors in determining larval rearing efficiency.

[0059] Example 4

[0060] The method for breeding and propagating the longhorn beetle with sclerotium brevicornu in this embodiment includes the following steps:

[0061] (1) Steps for preparing artificial feed for larvae: The raw materials and preparation methods are the same as in Example 1.

[0062] (2) Larval rearing steps: One newly hatched larva of the longhorn beetle *Aegilops glabripennis* was introduced into each feed box, for a total of 200 larvae. The larvae were reared for 8 weeks, constituting the first cycle. Artificial feed was then prepared again, and the larvae that survived the first cycle were introduced into the feed. The larvae were reared for 4 weeks, constituting the second cycle. Artificial feed was then prepared again, and the larvae that survived the second cycle were introduced into the feed. The larval rearing was then completed. The rearing equipment was an artificial climate chamber, with all conditions set at 25°C, RH=55%, and L:D=0:24h.

[0063] (3) Larval storage steps: After the larvae have been reared, place them in an artificial climate chamber at 5°C, RH=55%, L:D=0:24h and store them at low temperature for 6 weeks.

[0064] (4) Larval pupation and emergence steps: After refrigeration, the larvae were placed in an artificial climate chamber at 25°C, RH=55%, L:D=0:24h, and their pupation and emergence progress was observed and recorded.

[0065] (5) Adult rearing steps: Place 10 pairs of newly emerged male and female adults into an adult rearing box. Place maple branches in the box to supplement their nutrition, and place wood blocks wrapped with gauze as mating and egg-laying substrate. Replace the branches in the box every 2 days. Place the rearing box in an artificial climate chamber at 25°C, RH=55%, L:D=16:8h. A total of 3 rearing boxes were set up, with a total of 30 pairs of adults.

[0066] (6) Egg collection procedure: Every 4 days, remove the wooden block from the rearing box, use a brush to remove the eggs from the surface of the gauze and the seams, and place them in a sterile glass petri dish. Then, put the wooden block back into the rearing box for the longhorn beetle to continue laying eggs. Continue to collect eggs until all the females in the rearing box die, and record the number of eggs.

[0067] (7) Collection of larvae for propagation: The culture dishes containing the eggs of the Asian longhorn beetle were placed in an artificial climate chamber at 25°C, RH=55%, L:D=16:8h, and the hatching rate of the eggs was recorded. The newly hatched larvae obtained can be fed with artificial feed to achieve propagation of the Asian longhorn beetle.

[0068] Example 5

[0069] The method for breeding and propagating the longhorn beetle with sclerotium pedunculata in this embodiment includes the following steps:

[0070] (1) Preparation of artificial feed for larvae: The raw materials and preparation method are the same as in Example 1.

[0071] (2) Larval rearing steps: Same as the larval rearing steps described in Example 4.

[0072] (3) Larval storage steps: After the larvae have been reared, place them in an artificial climate chamber at 10°C, RH=55%, L:D=0:24h and store them at low temperature for 6 weeks.

[0073] (4) Larval pupation and emergence steps: Same as the larval pupation and emergence steps described in Example 4.

[0074] (5) Adult insect rearing steps: Same as the adult insect rearing steps described in Example 4.

[0075] (6) Egg collection steps: Same as the egg collection steps described in Example 4.

[0076] (7) Steps for collecting subculture larvae: Same as the steps for collecting subculture larvae described in Example 4.

[0077] Comparative Example 4

[0078] The propagation and rearing method of the *Aegilops glabripennis* in this comparative example includes the following steps:

[0079] (1) Preparation of artificial feed for larvae: The raw materials and preparation method are the same as in Example 1.

[0080] (2) Larval rearing steps: Same as the larval rearing steps described in Example 4.

[0081] (3) Larval storage steps: After the larvae have been reared, they are not refrigerated. Instead, they are stored in an artificial climate chamber at 25°C, RH=55%, L:D=0:24h for 6 weeks.

[0082] (4) Larval pupation and emergence steps: Same as the larval pupation and emergence steps described in Example 4.

[0083] (5) Adult insect rearing steps: Same as the adult insect rearing steps described in Example 4.

[0084] (6) Egg collection steps: Same as the egg collection steps described in Example 4.

[0085] (7) Steps for collecting subculture larvae: Same as the steps for collecting subculture larvae described in Example 4.

[0086] Comparative Example 5

[0087] The comparative method for breeding and propagating the longhorn beetle with sclerotium pedunculata includes the following steps:

[0088] (1) Preparation of artificial feed for larvae: The raw materials and preparation method are the same as in Example 1.

[0089] (2) Larval rearing steps: Same as the larval rearing steps described in Example 4.

[0090] (3) Larval storage steps: Same as the larval storage steps described in Example 4.

[0091] (4) Larval pupation and emergence steps: Same as the larval pupation and emergence steps described in Example 4.

[0092] (5) Adult insect rearing steps: Replace the branches that provide nutrition for the adults with weeping willow branches, and follow the same adult insect rearing steps as described in Example 4.

[0093] (6) Egg collection steps: Same as the egg collection steps described in Example 4.

[0094] (7) Steps for collecting subculture larvae: Same as the steps for collecting subculture larvae described in Example 4.

[0095] Comparative Example 6

[0096] The comparative method for breeding and propagating the longhorn beetle with sclerotium pedunculata includes the following steps:

[0097] (1) Preparation of artificial feed for larvae: The raw materials and preparation method are the same as in Example 1.

[0098] (2) Larval rearing steps: Same as the larval rearing steps described in Example 4.

[0099] (3) Larval storage steps: Same as the larval storage steps described in Example 4.

[0100] (4) Larval pupation and emergence steps: Same as the larval pupation and emergence steps described in Example 4.

[0101] (5) Adult insect rearing steps: Replace the branches that provide nutrition for the adults with arrow-shaped poplar branches, and follow the same adult insect rearing steps as described in Example 4.

[0102] (6) Egg collection steps: Same as the egg collection steps described in Example 4.

[0103] (7) Steps for collecting subculture larvae: Same as the steps for collecting subculture larvae described in Example 4.

[0104] The specific test results are shown in Table 2.

[0105] Table 2 Analysis of the rearing effects of the Brilliant-shouldered Star Beetle (A. chinensis) in generations

[0106]

[0107] Note: The data listed in Table 2 are mean ± standard error. The same letter in the same column indicates no significant difference, and different letters in the same column indicate significant difference.

[0108] As shown in Table 2, the pupation and emergence rates of the longhorn beetle larvae raised in Examples 4 and 5, as well as Comparative Examples 5 and 6, were significantly higher than those raised in Comparative Example 4 (which did not undergo low-temperature storage) after the low-temperature storage process. Furthermore, their generation time was also shorter than that of Comparative Example 4. Therefore, low-temperature storage can significantly improve the pupation and emergence progress and success rate of *A. glabra*. Meanwhile, even with low-temperature storage, the generation time of the longhorn beetles raised in Example 4 was significantly shorter than that in Example 5, indicating that a low-temperature storage temperature of 5°C is better than 10°C and more conducive to accelerating the rearing progress of *A. glabra*. In addition, the egg production of the adult longhorn beetles raised in Examples 4 and 5 was significantly higher than that in Comparative Examples 5 and 6, indicating that supplementing the nutrition of adults with *Acer negundo* significantly improves the egg production of *A. glabra*. The longhorn beetles raised in Comparative Example 4 not only had the lowest egg production but also a lower egg hatching rate, indicating that the low-temperature storage process before larval pupation also has a very positive effect on improving the reproductive fitness of adults.

[0109] In summary, the key to ensuring breeding efficiency in the reproduction of the Asian longhorn beetle lies in the low-temperature storage process after the completion of the larval rearing cycle and the supplementation of nutrition with Acer negundo during the adult stage.

[0110] The above description is merely a preferred embodiment of the invention and is not intended to limit the invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the invention should be included within the protection scope of the invention.

Claims

1. A method for the propagation and rearing of the longhorn beetle, characterized in that, Includes the following steps: Step B1: Collect newly hatched larvae of the longhorn beetle (Anoplophora glabripennis) for use as the first generation of insects for artificial rearing. Step B2: The newly hatched larvae are transferred to a larval feed box containing artificial feed and raised in an artificial climate chamber for 6-10 weeks. Step B3: Transfer the reared longhorn beetle larvae to a larval feed box containing artificial feed and continue to raise them in an artificial climate chamber for 3-5 weeks. Step B4: Transfer the longhorn beetle larvae raised in step B3 to larval feed boxes containing artificial feed, and continue to raise them in an artificial climate chamber for 3-5 weeks. Step B5: Place the longhorn beetle larvae along with the feed box in a low-temperature artificial climate chamber and store them at low temperature for 5-7 weeks. Step B6: After refrigeration, the longhorn beetle larvae are transferred to an artificial climate chamber to pupate and emerge as adults. Step B7: Transfer the newly emerged male and female longhorn beetles in pairs to an adult rearing box. Place a branch of maple leaf in the box to supplement their nutrition, and place a wooden block covered with gauze as a substrate for mating and oviposition. Step B8: After the longhorn beetles mate and lay eggs, collect the eggs on the gauze into a sterile culture dish. After the eggs hatch into larvae, they are fed artificial feed for generation. In particular, no feed needs to be changed during each larval rearing cycle from steps B2 to B4; The artificial feed is composed of the following raw materials in parts by weight: 40.0-60.0 parts potato starch, 40.0-60.0 parts sucrose, 100.0-140.0 parts soybean flour, 25.0-40.0 parts casein, 1.5-4.0 parts cholesterol, 7.0-12.0 parts Wiener salt, 70.0-130.0 parts brewer's yeast, 250.0-450.0 parts microcrystalline cellulose, 60.0-90.0 parts agar powder, 3.0-6.0 parts citric acid, 3.0-6.0 parts sorbic acid, 3.0-6.0 parts methyl 4-hydroxybenzoate, 10.0-18.0 parts soybean oil, 1200.0-2000.0 parts distilled water, 4.0-8.0 parts ascorbic acid, and 1.5-6.0 parts propionic acid; The artificial feed is prepared by the following steps: Step A1: Weigh the raw materials according to the mass fractions, and thoroughly mix the remaining raw materials except ascorbic acid and propionic acid. Then, autoclave to obtain a sterilized mixture. Step A2: When the sterilized mixture cools to 65℃-75℃, add ascorbic acid and propionic acid, and stir thoroughly to obtain artificial feed for the larvae of the longhorn beetle.

2. The method for breeding and propagating the longhorn beetle *Aegilops glabripennis* according to claim 1, characterized in that... The artificial feed consists of the following raw materials in parts by weight: Potato starch 50.0 parts, sucrose 50.0 parts, soybean flour 120.0 parts, casein 30.0 parts, cholesterol 3.0 parts, Wiegand salt 9.0 parts, brewer's yeast 90.0 parts, microcrystalline cellulose 390.0 parts, agar powder 70.0 parts, citric acid 4.0 parts, sorbic acid 5.0 parts, methyl 4-hydroxybenzoate 5.0 parts, soybean oil 14.0 parts, distilled water 1550.0 parts, ascorbic acid 6.0 parts, propionic acid 4.0 parts.

3. The method for breeding and propagating the longhorn beetle as described in claim 1, characterized in that, The autoclaving conditions in step A1 are: moist heat sterilization at 121°C for 20 minutes.

4. The method for breeding and propagating the longhorn beetle *Aegilops glabripennis* according to any one of claims 1 to 3, characterized in that... Step B2: Feed for 8 weeks; Step B3: Feed for 4 weeks; Step B4: Feed for 4 weeks; Step B5: Store at low temperature for 6 weeks.

5. The method for breeding and propagating the longhorn beetle with sclerotium pedunculata according to claim 1, characterized in that: The twigs of the compound maple, which provide nutrition for the adults, are replaced every two days; the substrate for the adults to mate and lay eggs is made of wood blocks covered with gauze.

6. The method for breeding and propagating the longhorn beetle according to any one of claims 1 to 3, characterized in that: The larval feed box is a covered transparent plastic cup with a volume of 60ml, a bottom diameter of 4.0cm, a mouth diameter of 5.5cm, and a height of 3.5cm; the adult rearing box is a covered transparent plastic bottle with a volume of 8L, a bottom diameter of 20.0cm, a mouth diameter of 15.0cm, and a height of 30.0cm.

7. The method for breeding and propagating the longhorn beetle according to any one of claims 1 to 3, characterized in that: The larval rearing environment conditions were: 25℃, RH=55%, L:D=0:24h; the larval refrigeration environment conditions were: 5℃, RH=55%, L:D=0:24h; and the larval pupation and emergence environment conditions were: 25℃, RH=55%, L:D=0:24h.

8. The method for breeding and propagating the longhorn beetle with sclerotium pedunculata according to claim 7, characterized in that: The environmental conditions for adult rearing and egg hatching were: 25℃, RH=55%, L:D=16:8h.

Images