How to raise bat moth larvae

Abstract

The present invention relates to a method for raising bat moth larvae. During the growth stage of the bat moth larvae, juvenile hormone is applied to the bat moth larvae, and the concentration range is 25-1000ng / μL. Raising the bat moth larvae by the method of the present invention has good effects on the weight, survival rate, infection rate and dead insect formation rate of the single bat moth larvae.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, relates to a method for raising bat moth larvae, and specifically relates to a method for improving the weight, infection rate and dead insect formation rate of bat moth larvae. Background Art

[0002] Ophiocordyceps sinensis (Berk.)GH Sunget et al. is a precious traditional Chinese medicine produced in my country. It is mainly distributed in Tibet, Qinghai, Sichuan, Yunnan and Gansu. It belongs to the genus Hepialus of the family Hepialidae in the order Lepidoptera. It is an entomogenous ascomycete formed after the larvae are parasitized by Hirsutella sinensis. It belongs to the phylum Ascomycota, class Ascomycetes, subclass Sordariomycetidae, order Hypocreales, family Clavicipitaceae, genus Cordyceps (Frey) Link. of the kingdom Fungi. The growth conditions of Ophiocordyceps sinensis are very harsh and are closely related to altitude, vegetation and climate.

[0003] The bat moth has a life cycle of 4 or more years, with the larval stage lasting more than 3 years. The adult stage is shorter, about 25 to 28 days, and the individual survival period is generally 3 to 4 days. Cordyceps fungi invade the host bat moth larvae in July and August every year, making full use of the insect body's nutrition to reproduce hyphae, and even filling the entire body cavity. The infected larvae burrow into the soil to hibernate. Around October, when the ground temperature is 2 to 9 degrees Celsius, the larvae die and become dead insects.

[0004] In recent years, more and more scholars or research institutions have carried out a lot of research on the breeding of bat moth larvae in low-altitude areas in order to successfully breed Cordyceps sinensis and industrialize it. The survival rate, individual weight, infection rate and dead insect formation rate are all key research objects. Summary of the invention

[0005] In order to solve the problems existing in the prior art, the present invention provides a method for raising bat moth larvae, which not only improves the survival rate of the larvae and the weight of a single larvae, but also improves the subsequent infection rate and dead insect formation rate.

[0006] Specifically, the present invention provides a method for raising bat moth larvae, wherein juvenile hormone is applied to the bat moth larvae during their growth stage, with a concentration range of 25-1000 ng / μL.

[0007] In some embodiments, the concentration of the juvenile hormone ranges from 50-1000 ng / μL.

[0008] In some embodiments, the juvenile hormone comprises Altozar ZR-512, Methoprene ZR-515, Kinoprene ZR-777 or JH-286 farmplant.

[0009] In some embodiments, the growth stage is the 3rd and / or 4th age stage.

[0010] In some embodiments, the application is by spraying until the insect body is wetted.

[0011] In some embodiments, the applying means spraying the larvae once every 2-4 days at the 3rd instar stage until each larva is wet, and the concentration of the juvenile hormone ranges from 200-500 ng / μL.

[0012] In some embodiments, the applying refers to spraying the larvae once a day during the 4th instar stage, during the first 60-90 days, until each larvae is wet, and the concentration of the juvenile hormone ranges from 500-1000 ng / μL.

[0013] In some embodiments, the method of raising bat moth larvae comprises:

[0014] The third-instar bat moth larvae are raised under the conditions of a temperature of 12-15°C, a soil relative humidity of 35-40RH%, and an air relative humidity of 70-75RH%, and are fed with carrots cut into strips and Polygonum villosa (1:1) as feed; the larvae are sprayed with juvenile hormone once every 3 days until each larva is moistened.

[0015] In some embodiments, a method for raising bat moth larvae comprises:

[0016] The 4th instar bat moth larvae are raised under the conditions of a temperature of 10-12°C, a soil relative humidity of 35-40RH%, and an air relative humidity of 70-75RH%, and are fed with carrots cut into strips and Polygonum villosa (1:2) as feed; in the first 60-90 days, the larvae are sprayed with juvenile hormone once a day until each larva is moistened.

[0017] In addition, the present invention also provides the use of juvenile hormone in improving the weight, infection rate or dead insect formation rate of bat moth larvae.

[0018] Advantages of the bat moth larvae breeding method of the present invention:

[0019] 1. Increased the weight of a single worm.

[0020] 2. Improved the survival rate of bat moth larvae.

[0021] 3. The infection rate and dead insect formation rate of bat moth larvae were increased.

[0022] Detailed description

[0023] The invention provides a method for raising bat moth larvae. During the growth stage of the bat moth larvae, juvenile hormone is applied to the bat moth larvae, and the concentration range of the juvenile hormone is 25-1000 ng / μL.

[0024] The larvae of the bat moth are the basis for the growth of Cordyceps sinensis. In taxonomy, they belong to the family Hepialidae, the genus Hepiaua, and the Hepialusoberthur. Currently, through investigation, we know that there are more than 20 similar populations. Due to different growth areas, the species are also different. The bat moth larvae described in the present invention are the larvae of the Cordyceps sinensis.

[0025] The growth stages of bat moth larvae are divided by the number of times they molt, and each molt is one instar. Bat moth larvae generally molt 5-7 times.

[0026] The term "applying" as used in the present invention may be in various ways, such as spraying, applying, briefly soaking, and the like.

[0027] In some embodiments, the concentration of the juvenile hormone ranges from 50-1000 ng / μL.

[0028] In some embodiments, the concentration of the juvenile hormone ranges from 200-1000 ng / μL.

[0029] In some embodiments, the concentration of the juvenile hormone ranges from 200-500 ng / μL.

[0030] In some embodiments, the concentration of the juvenile hormone ranges from 500-1000 ng / μL.

[0031] In some embodiments, the applying means that at the 3rd instar stage, the concentration of the juvenile hormone is in the range of 200-500 ng / μL;

[0032] In some embodiments, the applying means that at the 4th instar stage, the concentration of the juvenile hormone is in the range of 500-1000 ng / μL.

[0033] In some embodiments, the applying means that at the 3rd instar stage, the concentration of the juvenile hormone is in the range of 200-500 ng / μL; at the 4th instar stage, the concentration of the juvenile hormone is in the range of 500-1000 ng / μL.

[0034] In some embodiments, the applying means that at the 3rd age stage, the concentration range of the juvenile hormone is 200 ng / μL;

[0035] In some embodiments, the applying means that at the 4th instar stage, the concentration range of the juvenile hormone is 500 ng / μL.

[0036] In some embodiments, the applying means that at the 3rd instar stage, the concentration range of the juvenile hormone is 200 ng / μL; at the 4th instar stage, the concentration range of the juvenile hormone is 500 ng / μL.

[0037] In some embodiments, the juvenile hormone comprises Altozar ZR-512, Methoprene ZR-515, Kinoprene ZR-777 or JH-286 farmplant.

[0038] In some embodiments, the juvenile hormone is kinoprene ZR-777.

[0039] In some embodiments, the growth stage is the 3rd and / or 4th age stage.

[0040] “And / or” means the relationship of “and” or the relationship of “or”.

[0041] In some embodiments, the growth stage is the 3rd or 4th instar stage.

[0042] In some embodiments, the growth stage is the 3rd and 4th instar stages.

[0043] In some embodiments, the application is by spraying until the insect bodies are wetted.

[0044] The term "wet" used in the present invention means that there is a water layer on the surface of the insect body, but there is no water flow, which does not affect the crawling and feeding of the larvae.

[0045] In some embodiments, the application refers to spraying the larvae once every 2-4 days at the 3rd instar stage until every larvae is wet; preferably, spraying once every 3 days.

[0046] In some embodiments, the application is at the 3rd instar stage, spraying the larvae once every 3 days until each worm body is wet, with a concentration of 200-500 ng / μL;

[0047] Spraying once every N days (N is a positive integer) means spraying again after N-1 days. For example, spraying once every 3 days means spraying on the first day and then spraying again after 2 days, that is, spraying on the 4th day.

[0048] In some embodiments, the method for raising bat moth larvae includes: raising 3rd instar bat moth larvae at a temperature of 12-15°C, a soil relative humidity of 35-40RH%, and an air relative humidity of 70-75RH%, and feeding them with carrots cut into strips and Polygonum villosa (1:1) as feed; spraying the larvae with juvenile hormone once every 3 days, until each larva is moistened.

[0049] In some embodiments, the application refers to spraying the larvae once a day during the first 60-90 days of the 4th instar stage until each larva is wet. Preferably, spraying is performed once a day during the first 60, 70, 80 or 90 days. Preferably, spraying is performed once a day during the first 60 days.

[0050] The first N days (N is a positive integer) refer to the N days starting from the time the bat moth larvae molt; for example, in the 4th instar stage, the first 60 days refer to the 60 days starting from the time the bat moth larvae molt from the 3rd instar to the 4th instar.

[0051] In some embodiments, the application is at the 4th instar stage, during the first 60-90 days, by spraying the larvae once a day until each worm body is wet, with a concentration ranging from 500-1000 ng / μL.

[0052] In some embodiments, the method for raising bat moth larvae includes: raising 4th-instar bat moth larvae at a temperature of 10-12°C, a soil relative humidity of 35-40RH%, and an air relative humidity of 70-75RH%, and feeding them with carrots cut into strips and Polygonum villosa (1:2) as feed; for the first 60-90 days, spraying the larvae with juvenile hormone once a day, until each larva is moistened.

[0053] In some embodiments, the application is performed at the 3rd instar stage by spraying the larvae once every 3 days until each worm body is wet; at the 4th instar stage, during the first 60-90 days, the application is performed once a day by spraying the larvae until each worm body is wet.

[0054] In some embodiments, the application is at the 3rd instar stage, spraying the larvae once every 3 days until each worm body is wet, with a concentration of 200 ng / μL; at the 4th instar stage, during the first 60-90 days, spraying the larvae once a day until each worm body is wet, with a concentration of 500 ng / μL.

[0055] The present invention relates to temperature, humidity, concentration. Although specific numerical values ​​are given, they include a difference of at least ±20%, such as 10 degrees, which refers to 8-12 degrees. In some embodiments, it includes a difference of at least ±10%, such as 10 degrees, which refers to 9-11 degrees. In some embodiments, it includes a difference of at least ±5%. In some embodiments, it includes a difference of at least ±4%. In some embodiments, it includes a difference of at least ±3%. In some embodiments, it includes a difference of at least ±2%. In some embodiments, it includes a difference of at least ±1%.

[0056] In addition, the present invention also provides the use of juvenile hormone in improving the weight, infection rate or dead insect formation rate of bat moth larvae.

[0057] In addition, the present invention also provides a method for using juvenile hormone to improve the weight, infection rate or dead insect formation rate of bat moth larvae. DETAILED DESCRIPTION

[0058] The embodiments of the present invention are described in detail below, wherein the same or similar reference numerals throughout represent the same or similar elements or elements with the same or similar functions. The following embodiments are exemplary and are intended to be used to explain the present invention, and should not be construed as limiting the present invention.

[0059] Example 1

[0060] 1-A. Fifty 3rd-instar bat moth larvae were raised under the conditions of 12-15°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum bulbil (1:1) were fed as feed. Juvenile hormone (Kinoprene ZR-777, concentration 50ng / μL) was sprayed on the larvae once every 3 days, and the number of deaths was counted based on the wetness of each larvae, and they were weighed before molting.

[0061] 1-B. 50 3rd-instar bat moth larvae were raised under the conditions of 12-15°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum bulbil (1:1) were fed as feed. Juvenile hormone (Kinoprene ZR-777, concentration 200ng / μL) was sprayed on the larvae once every 3 days, and the number of deaths was counted based on the wetness of each larvae, and they were weighed before molting.

[0062] 1-C. 50 3rd-instar bat moth larvae were raised under the conditions of 12-15°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum bulbil (1:1) were fed as feed. Juvenile hormone (Kinoprene ZR-777, concentration 3000ng / μL) was sprayed on the larvae once every 3 days, and the number of deaths was counted based on the wetness of each larvae, and they were weighed before molting.

[0063] 1-D. 50 3rd-instar bat moth larvae were raised at a temperature of 12-15°C, a soil relative humidity of 35-40 RH%, and an air relative humidity of 70-75 RH%, and fed with carrots cut into strips and Polygonum villosa (1:1) as feed. The number of deaths was counted and weighed before molting.

[0064] The results are shown in Table 1:

[0065] Table 1

[0066] Group Deaths Survival rate Average weight 1-A 12 76％ 0.45 1-B 11 78％ 0.58 1-C 18 64％ 0.44 1-D 16 68％ 0.35

[0067] Conclusion: From the above table, we can see that low concentrations of juvenile hormone have a positive effect on the survival rate and body weight of bat moth larvae.

[0068] Example 2

[0069] 2-A. 50 4-instar bat moth larvae obtained in 1-B of Example 1 were placed under the conditions of 10-12°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum bulbil (1:2) were added as feed. The larvae were sprayed with juvenile hormone (Kinoprene ZR-777, concentration 500ng / μL) once a day until each larva was wetted. After 90 days of spraying, the larvae were no longer sprayed and grew naturally. The number of deaths was counted and weighed before molting.

[0070] 2-B. 50 4th-instar bat moth larvae obtained in 1-B of Example 1 were raised under the conditions of 10-12°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum villosa (1:2) were added as feed. The larvae were sprayed with juvenile hormone (Kinoprene ZR-777, concentration 500ng / μL) once a day until each larva was wetted. After 60 days of spraying, the larvae were no longer sprayed and grew naturally. The number of deaths was counted and weighed before molting.

[0071] 2-C. 50 4th-instar bat moth larvae obtained in 1-B of Example 1 were raised under the conditions of 10-12°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum villosa (1:2) were added as feed. The larvae were sprayed with juvenile hormone (Kinoprene ZR-777, concentration 500ng / μL) once a day, and the number of deaths was counted based on the wetness of each larva, and they were weighed before molting.

[0072] 2-D. 50 4th-instar bat moth larvae obtained in 1-B of Example 1 were raised under the conditions of 10-12°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and were fed with carrots cut into strips and Polygonum villosa (1:2) as feed. They were allowed to grow naturally, the number of deaths was counted, and they were weighed before molting.

[0073] 2-E. 50 4th-instar bat moth larvae obtained in 1-D of Example 1 were placed in a temperature of 10-12°C, a soil relative humidity of 35-40 RH%, and an air relative humidity of 70-75 RH%, and fed with carrots cut into strips and Polygonum villosa (1:2) as feed. They were allowed to grow naturally, the number of deaths was counted, and they were weighed before molting.

[0074] 2-F. 50 4th-instar bat moth larvae obtained in 1-D of Example 1 were raised under the conditions of 10-12°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum villosa (1:2) were added as feed. The larvae were sprayed with juvenile hormone (Kinoprene ZR-777, concentration 500ng / μL) once a day until each larva was wetted. After 60 days of spraying, the larvae were no longer sprayed and grew naturally. The number of deaths was counted and weighed before molting.

[0075] Table 2

[0076]

[0077]

[0078] Conclusion: From the above table, we can see that spraying juvenile hormone for a certain period of time is beneficial to survival and weight gain.

[0079] Example 3

[0080] Preparation of bacterial feed: In a sterile clean area, extend the fruiting body of Cordyceps sinensis into a pre-cultured qualified plate, with the body of Cordyceps sinensis exposed outside the plate, and cover the body with sterile gauze containing sterile water to keep it moist, cover the dish, and allow the ascospores to naturally eject onto the surface of the culture medium of the plate. The ejection time is not less than 7 days, and the light is exposed for 12 hours per day with a light intensity of 300Lx. The plate containing the strain is cultured in a program-controlled mold incubator to form colonies at a culture temperature of 15°C for 50 days. The colonies are purified and cultured 2-3 times, and the purified colonies are placed in a liquid culture medium containing 10 5 spores, and spray the liquid culture medium onto the feed to form a bacterial feed (carrots cut into strips + Polygonum villosa (1:2)).

[0081] Prepare the soil matrix containing bacteria: spray the infection solution of ascospores (60 / mL) into the soil matrix.

[0082] 3-A. The 5th-instar bat moth larvae obtained in 2-B of Example 2 were raised under the conditions of 10-12°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum bulbil (1:2) were added as feed. The temperature was gradually increased by 15-16°C within 10 days. After continuing to raise for 10 days, the bat moth larvae were placed on a soil substrate containing bacteria, and the bacteria-containing feed was added. The temperature was 15-16°C, the soil relative humidity was 40-45RH%, and the air relative humidity was 70-75RH%. The infection rate at 3 months and the dead insect formation rate at 12 months were investigated.

[0083] 3-B. The 5th-instar bat moth larvae obtained in 2-D of Example 2 were raised under the conditions of 10-12°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum bulbil (1:2) were added as feed. The temperature was gradually increased by 15-16°C within 10 days. After continuing to raise for 10 days, the bat moth larvae were placed on a soil substrate containing bacteria, and the bacteria-containing feed was added. The temperature was 15-16°C, the soil relative humidity was 40-45RH%, and the air relative humidity was 70-75RH%. The infection rate at 3 months and the dead insect formation rate at 12 months were investigated.

[0084] 3-C. The 5th-instar bat moth larvae obtained in 2-E of Example 2 were raised under the conditions of 10-12°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum bulbil (1:2) were added as feed. The temperature was gradually increased by 15-16°C within 10 days. After continuing to raise for 10 days, the bat moth larvae were placed on a soil substrate containing bacteria, and the bacteria-containing feed was added. The temperature was 15-16°C, the soil relative humidity was 40-45RH%, and the air relative humidity was 70-75RH%. The infection rate at 3 months and the dead insect formation rate at 12 months were investigated.

[0085] 3-D. The 5th-instar bat moth larvae obtained in 2-F of Example 2 were raised under the conditions of 10-12°C, 35-40RH% soil relative humidity, and 70-75RH% air relative humidity, and carrots cut into strips + Polygonum bulbil (1:2) were added as feed. The temperature was gradually increased by 15-16°C within 10 days. After continuing to raise for 10 days, the bat moth larvae were placed on a soil substrate containing bacteria, and the bacteria-containing feed was added. The temperature was 15-16°C, the soil relative humidity was 40-45RH%, and the air relative humidity was 70-75RH%. The infection rate at 3 months and the dead insect formation rate at 12 months were investigated.

[0086] Table 3

[0087] Group Infection rate Dead insect formation rate 3-A 76％ 52％ 3-B 68％ 32％ 3-C 66％ 26％ 3-D 74％ 41％

[0088] Conclusion: Juvenile hormone can increase the infection rate and the formation rate of dead insects.

[0089] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0090] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A method for raising bat moth larvae, characterized in that: During the growth stage of the bat moth larvae, juvenile hormone is applied to them, wherein the juvenile hormone includes Altozar ZR-512, Methoprene ZR-515, Kinoprene ZR-777 or JH-286 farmoplant; The growth stage is the 3rd and / or 4th age stage; The application means spraying the larvae once every 2-4 days until each larva is moistened at the 3rd instar stage, and the concentration of the juvenile hormone ranges from 200-500 ng / μL; The application refers to spraying the larvae once a day during the 4th instar stage, during the first 60-90 days, until each larvae is wet, and the concentration of the juvenile hormone ranges from 500-1000 ng / μL.

2. The breeding method according to claim 1, characterized in that The application is by spraying until the insect bodies are wet.

3. The breeding method according to claim 1, characterized in that include: The third-instar bat moth larvae are raised under the conditions of a temperature of 12-15°C, a soil relative humidity of 35-40RH%, and an air relative humidity of 70-75RH%, and are fed with carrots cut into strips and Polygonum villosa (1:1) as feed; the larvae are sprayed with juvenile hormone once every 3 days until each larva is moistened.

4. The breeding method according to claim 1, characterized in that include: The 4th instar bat moth larvae are raised under the conditions of a temperature of 10-12°C, a soil relative humidity of 35-40RH%, and an air relative humidity of 70-75RH%, and are fed with carrots cut into strips and Polygonum villosa (1:2) as feed; in the first 60-90 days, the larvae are sprayed with juvenile hormone once a day until each larva is moistened.

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