A method for increasing the reproductive capacity of the carpenter bee by water bath processing of tenebrio molitor pupae

Abstract

The present application relates to a method for increasing the reproductive capacity of Sirex noctilio by water bath treatment of Tenebrio molitor pupae, which comprises preparing a series of necessary materials and equipment, including Tenebrio molitor larvae, culture dishes, absorbent paper, glass pipettes, etc., and then feeding Tenebrio molitor under suitable conditions until pupation. Next, select Tenebrio molitor pupae that have not been hatched for more than 48 hours and have similar quality, and then perform constant temperature water bath treatment at 30 DEG C for 3 minutes to improve their physiological state. The treated Tenebrio molitor pupae are used as parasitic hosts of Sirex noctilio, and the method effectively increases the parasitization rate and success rate of Sirex noctilio on the alternative host Tenebrio molitor pupae, provides high-quality parasitic wasp resources for biological control, helps to reduce the use of chemical pesticides, protects the ecological environment, and realizes the sustainable development of agriculture and forestry.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of artificial feeding of insects, in particular to a method for increasing the reproductive capacity of Sclerodermus alternatus by water bath treatment of yellow mealworm pupae. BACKGROUND

[0002] More than 500 million pine trees have been killed by pine wood nematode disease, and more than 300,000 hm of pine forests have been destroyed 2 , causing economic losses of over 100 billion yuan. Pine wood nematode disease is caused by the pathogen Bursaphelenchus xylophilus, but the pine wood nematode cannot spread and infect pine trees by itself, but mainly relies on the medium insect Monochamus alternatus for transmission and spread. Monochamus alternatus is not only the main medium insect of pine wood nematode disease, but also a serious and widely distributed bark borer of pine trees

[0003] Biological control is a safer and more environmentally friendly control method than chemical control, and has a long-lasting control effect. Chemical agents are difficult to use at the right time when controlling Monochamus alternatus, and have high costs and cause serious pollution, which can cause irreversible damage to the environment. Biological control, especially the use of natural enemy insects, has a better effect. Since natural enemy insects can search for and kill pests independently, and can sustainably control the population size of pests, biological control is one of the best methods for controlling Monochamus alternatus.

[0004] Sclerodermus alternatusi belongs to the order Hymenoptera, the family Bethylidae, and the genus Sclerodermus, and is a parasitic wasp that parasitizes Monochamus alternatus larvae discovered in Yunnan in 2008. This wasp is an ectoparasite of Monochamus alternatus larvae, and the parasitization rate of Monochamus alternatus larvae can reach more than 80%. The individual is large, the sex ratio is high, the lifespan is long, and the ability to attack pests, search for hosts, and reproduce is strong. In addition to killing Monochamus alternatus larvae through oviposition parasitization, the female wasp can also kill several host larvae during the supplementary nutrition stage. Many studies have explored the effects of yellow mealworm pupae Tenebrio molitor on the reproduction of other bethylid wasps under different treatments, and the reproduction of Sclerodermus alternatus using different hosts, but there are few reports on the use of yellow mealworm pupae to reproduce Sclerodermus alternatus. Therefore, a method for increasing the reproductive capacity of Sclerodermus alternatus by water bath treatment of yellow mealworm pupae is needed. SUMMARY

[0005] The present application provides a method for increasing the reproductive capacity of Sclerodermus alternatus by water bath treatment of yellow mealworm pupae.

[0006] To solve the above-mentioned technical problems, the present invention is achieved through the following technical solution: a method for increasing the reproductive capacity of *Symplocos pulvinata* wasps by water bath treatment of mealworm pupae, comprising the following steps:

[0007] Step 1: Material preparation: Prepare mealworm larvae, 90mm petri dishes, absorbent paper, 5ml glass finger tubes, constant temperature water bath, medical absorbent cotton rolls, storage box, wheat bran, stereo microscope, electronic balance, weighing paper, and artificial climate chamber.

[0008] The mealworm larvae were sourced from Shuyi Feed Raw Material Factory; the 90mm petri dishes were from Jiangsu Huaou Glass Co., Ltd.; the absorbent paper was from Zhengzhou Lingze Instrument Equipment Co., Ltd.; the 5ml glass finger tubes were from Huaou Glass Co., Ltd.; the constant temperature water bath was from Jintan Jincheng Guosheng Experimental Instrument Factory; the medical absorbent cotton rolls, storage boxes, wheat bran, and stereomicroscope were from Ningbo Sunny Instrument Co., Ltd.; the electronic balance was from Sartorius Scientific Instruments (Beijing) Co., Ltd.; and the weighing paper and artificial climate chamber were from Ningbo Plant Co., Ltd.

[0009] Step 2: Mealworm rearing: In the rearing room at 25℃, use a 25cm×16cm×7cm storage box to fill with an appropriate amount of wheat bran to raise mealworms, and add a small amount of fruit peels as needed to supplement moisture, and wait for them to pupate;

[0010] Step 3: Selection of mealworm pupae: After the mature mealworm larvae molt and pupate, select pupae that have emerged within 48 hours and are of similar quality and place them in a petri dish.

[0011] Step 4: Place the culture dish containing the mealworm pupae in a 30℃ constant temperature water bath for 3 minutes, then remove it and place it on absorbent paper to stand at room temperature until it is naturally dried. Then, use tweezers to take the mealworm pupae and place them into a glass finger tube.

[0012] Step 5: Parasitic wasp rearing: Take pine beetle swollen-legged wasps that have emerged 3 days ago and have mated from an artificial climate chamber, and introduce them at a wasp-to-insect ratio of 3:1. Put one treated pupa into each tube, and introduce 3 female wasps with a paintbrush. Seal the tube with cotton plugs and place it in an artificial climate chamber with a temperature of 27±0.5℃, a humidity of 65%±5%, and a photoperiod of 14L:10D for rearing.

[0013] By treating the pupae with a constant temperature water bath at 30°C, the physiological activities inside the pupae can be stimulated, which may include promoting the synthesis or transformation of certain nutrients and improving their surface microenvironment, making the pupae more suitable as a parasitic host for the pine sawyer beetle.

[0014] The internal structure and nutritional state of the yellow mealworm pupae treated by water bath are more conducive to the parasitization and reproduction of the Sirex noctilio.

[0015] The use of an artificial climate box to provide a stable and suitable breeding environment for the parasitoid can ensure its reproduction in the best state, thereby increasing the reproductive capacity of the Sirex noctilio.

[0016] The beneficial effects of the present application are:

[0017] 1. The artificial breeding method of the Sirex noctilio of the present application is simple and easy to operate, and is particularly suitable for indoor breeding. Even if the relevant staff do not have relevant professional knowledge, they can learn and operate independently in a short period of time. The incubator used in the present application can be adjusted in size according to the experimental requirements and control requirements, and a worm breeding room can also be established, which occupies less space and has lower breeding cost. The method of the present application can mass-produce the Sirex noctilio. By softening the epidermis of the yellow mealworm pupae, the parasitization rate and success rate of the Sirex noctilio on the alternative host yellow mealworm pupae are improved, and the number of offspring of the Sirex noctilio is also increased, thereby saving the breeding cost and providing a large amount of insect source to reduce the ecological environmental impact of the pine wood nematode and the Sirex noctilio on the pine forest. DETAILED DESCRIPTION

[0018] The technical solutions in the embodiments of the present application will be described below in conjunction with the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor are within the scope of protection of the present application.

[0019] A method for increasing the reproductive capacity of *Symplocos pulmonata* wasps through water bath treatment of mealworm pupae includes the following steps: Step 1: Material preparation: Prepare mealworm larvae, 90mm culture dishes, absorbent paper, 1.5ml glass finger tubes, a constant temperature water bath, medical absorbent cotton rolls, storage boxes, wheat bran, a stereomicroscope, an electronic balance, weighing paper, and an artificial climate chamber; Step 2: Mealworm rearing: Under 25℃ conditions in the rearing room, use a 25cm×16cm×7cm storage box filled with an appropriate amount of wheat bran to rear the mealworms, and add a small amount of fruit peels as needed to supplement moisture, waiting for them to pupate; Step 3: Selection of mealworm pupae: After the mature mealworm larvae molt and pupate, select... Step 4: Place the pupae containing mealworm pupae in a 30℃ constant temperature water bath for 3 minutes, then remove them and place them on absorbent paper to stand at room temperature until they are naturally dried. Use tweezers to take the mealworm pupae and place them into glass finger tubes. Step 5: Parasitic wasp rearing: Take the pine brown longhorn beetle swollen-legged wasps that have emerged 3 days ago and have mated in an artificial climate chamber. Introduce the wasps at a ratio of 3:1, placing one treated pupa in each tube. Use a brush to introduce 3 female wasps. Seal the tubes with cotton plugs and place them in an artificial climate chamber with a temperature of 27±0.5℃, a humidity of 65%±5%, and a photoperiod of 14L:10D.

[0020] The equipment used in the following embodiments is as follows:

[0021] 1. Insect rearing room: Includes the rearing room for the Asian longhorn beetle, the area of ​​which varies depending on the scale, usually 15-20m². 2 An air conditioner is installed in the insect rearing room to control the temperature, and a microcomputer time control switch is installed to automatically control the lights in the insect rearing room.

[0022] 2. Mealworm rearing box: 25cm×16cm×7cm, which can be enlarged as needed.

[0023] 3. Water bath treatment: 30℃, 3 min

[0024] 4. Bee-to-insect ratio: 3:1

[0025] Example 1: After mature mealworm larvae molt and pupate, several mealworm pupae of similar size were selected and placed in a petri dish. Each larva was heated in a 30°C water bath for 3 minutes, then removed and placed on absorbent paper to air dry at room temperature. After air drying, the pupae were placed into glass finger tubes using tweezers. Three-day-old *Symplocos pulmonata* wasps, which had emerged from the incubator, were removed and introduced into the incubator at a wasp-to-larvae ratio of 3:1. The egg-laying time and number of *Symplocos pulmonata* wasps, egg hatching time and number, pupation time and number, and emergence time and number were observed. After complete emergence, the total number of offspring wasps and the number of individual female and male offspring were counted.

[0026] Example two: After the old matured larvae of yellow mealworms molted and pupated, several yellow mealworms with similar body size were selected and placed in a culture dish. After being heated in a 35℃ constant temperature water bath for 9 minutes, they were taken out and placed on an absorbent paper at room temperature for natural drying. After drying, the yellow mealworms were taken out with a pair of tweezers and placed in a glass test tube. Three-day-old Sirex noctilio wasp larvae in a constant temperature box were taken out, and the wasp larvae were matched according to the ratio of 3:1. The time and number of oviposition, the time and number of egg hatching, the time and number of pupation, and the time and number of eclosion were recorded. After complete eclosion, the total number of offspring wasp larvae, and the number of female and male offspring wasp larvae were counted.

[0027] Table: 30℃ water bath feeding Sirex noctilio wasp larvae

[0028]

[0029] Although the embodiments of the present application have been shown and described, it is to be understood that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present application, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for increasing the reproductive capacity of *Symplocos pulmonata* wasps through water bath treatment of mealworm pupae, characterized in that... Includes the following steps: Step 1: Material preparation: Prepare mealworm larvae, petri dishes, absorbent paper, glass finger tubes, constant temperature water bath, medical absorbent cotton rolls, storage box, wheat bran, stereo microscope, electronic balance, weighing paper, and artificial climate chamber; Step 2: Mealworm rearing: In the rearing room at 25℃, use a storage box to load an appropriate amount of wheat bran to raise mealworms, and add a small amount of fruit peels as needed to supplement moisture, and wait for them to pupate; Step 3: Selection of mealworm pupae: After the mature mealworm larvae molt and pupate, select pupae that have emerged within 48 hours and are of similar quality and place them in a petri dish. Step 4: Place the culture dish containing the mealworm pupae in a 30℃ constant temperature water bath for 3 minutes, then remove it and place it on absorbent paper to stand at room temperature until it is naturally dried. Then, use tweezers to take the mealworm pupae and place them into a glass finger tube. Step 5: Parasitic wasp rearing: Take pine beetle swollen-legged wasps that have emerged 3 days ago and have mated from an artificial climate chamber, and introduce them at a wasp-insect ratio of 3:

1. Put 1 treated pupa and 3 female wasps into each tube, seal the tube, and then put it into an artificial climate chamber with a temperature of 27±0.5℃, a humidity of 65%±5%, and a photoperiod of 14L:10D for rearing. The petri dish is set to 90mm in diameter, and the glass finger tube is set to 5ml in volume. The storage box measures 25cm × 16cm × 7cm and is used to hold an appropriate amount of wheat bran to feed mealworms. In step five, three female bees are inserted into a glass finger tube using a brush, and the tube is sealed with a cotton plug.

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