Method for artificial breeding and storage of the predatory crab spider, Misumenops asperatus

Abstract

The application discloses an artificial breeding and storage method of the farming and forestry pest predatory spider Zora spinimana represented by Malacosoma americanum. On the basis of obtaining the theoretical daily maximum predation amount of Zora spinimana to different instar Malacosoma americanum larvae, the artificial breeding and propagation of Zora spinimana are carried out by using the substitute prey of the oceanic worm, the yellow mealworm and the silkworm, and the suitable substitute prey size and the breeding condition are screened, and a low-temperature storage method is provided. The mass breeding of Zora spinimana can provide a stable research sample source for the Zora spinimana, and lays a solid foundation for in-depth research on the biological characteristics, behavior habits, genetic mechanisms and the like of the Zora spinimana. Further research on the storage method can ensure the long-term preservation and effective utilization of the Zora spinimana sample, maintains the physiological activity and genetic information of the Zora spinimana, and enables the Zora spinimana to continue to play a role in future research.

Description

Technical Field

[0001] This invention relates to a method for the artificial breeding and storage of the wavy crab spider, a predatory spider that feeds on agricultural and forestry pests, represented by the American white moth. Background Technology

[0002] Agricultural and forestry pests refer to various insects that harm the growth of crops and trees. They cause serious economic losses to agricultural and forestry production by feeding on plant tissues and spreading viruses or pathogens. Therefore, controlling agricultural and forestry pests is crucial to ensuring the safety and sustainable development of agricultural and forestry production. Biological control is an important method for controlling agricultural and forestry pests. It controls pest damage by utilizing the interrelationships between organisms, increasing the population of natural enemies, or introducing natural enemies. Spiders are important natural enemies of many agricultural and forestry pests, such as the fall armyworm, the American white moth, the green leafhopper, and the brown planthopper. They are characterized by their high consumption of live insects, large predation rates, excellent reproductive capacity, and strong resistance to adverse conditions, making them highly promising for biological control. Upadhyay et al. collected 58 relevant studies published between 1970 and 2017, covering cases of spider biological control worldwide. Using meta-analysis, they calculated the effect size (Hedge's d) and, through statistical models such as generalized least squares (GLS), concluded that spiders effectively controlled agricultural pests in 79% of the cases.

[0003] Wavy flower crab spider ( Xysticus croceus The *Crabweed undulatus* is a hunting spider widely distributed throughout China, characterized by its long lifespan, wide predatory range, strong predatory ability, and high resilience. It is an important predatory natural enemy of various agricultural and forestry pests, including the fall webworm, and possesses significant potential for biological control. Under natural conditions, the survival rate of young nymphs of the *Crabweed undulatus* is relatively low, facing threats such as predation by natural enemies and food shortages, resulting in a low survival rate. Often, less than 20% of individuals develop into adults, leading to low population densities and limited pest control. Therefore, artificial breeding research on the *Crabweed undulatus* is of great significance. Artificial breeding can mitigate natural risks, increase the survival rate of nymphs, and facilitate propagation. Simultaneously, leveraging the *Crabweed undulatus*'s excellent predatory abilities, new biological control agents can be developed, thereby reducing the use of chemical pesticides and protecting the ecological environment. Summary of the Invention

[0004] Currently, domestic research on the wavy-striped crab spider relies solely on identification after field collection, confirming its predatory nature as a natural enemy of agricultural and forestry pests. There is a lack of systematic research, and studies often depend on field collection, which is limited by various natural factors such as season, weather, and geographical location, leading to uncertainty and instability in sample acquisition. Although some scholars have conducted research on the life history, biological characteristics, and predatory responses of the related spider, the saddle-shaped crab spider, there are no standardized procedures for artificial breeding. Existing research on spiders often uses fruit flies or houseflies for artificial breeding. These two alternative prey species are difficult to breed, have relatively long breeding cycles, high feeding costs, and uniform body size, causing significant inconvenience for the management of spiders at different ages. Furthermore, due to physiological structure and habits, the wavy-striped crab spider and other spiders cannot effectively consume artificial feed. In addition, nymphs are relatively fragile due to incomplete physiological development, making their breeding and management a major challenge in artificial breeding. Therefore, exploring a feasible artificial breeding technique for the wavy-striped crab spider is of great significance.

[0005] Large-scale breeding of the wavy-striped crab spider can provide a stable source of research samples, laying a solid foundation for in-depth research on its biological characteristics, behavioral habits, and genetic mechanisms. Further research on its storage methods can ensure the long-term preservation and effective use of the crab spider samples, maintain the physiological activity and genetic information of the wavy-striped crab spider, and enable it to continue to play a role in future research.

[0006] Therefore, the purpose of this invention is to provide a method for the artificial breeding and storage of the natural enemy spider, the wavy crab spider, laying a foundation for the use of this natural enemy in biological control.

[0007] This invention provides a predatory wavy flower crab spider ( Xysticus croceus The feeding and storage methods of the [product name] are characterized by including the following steps:

[0008] S1. Collect wavy-spotted crab spiders in the wild using the shaking or observation method;

[0009] S2. Bring the collected spiders back indoors and place them in a spider rearing box. Keep them at a temperature of 25±2℃ and a humidity of 60%. Overwintering spiders collected in the wild at temperatures below 0℃ should be placed in a 15±2℃ incubator for 24 hours to warm up before being reared.

[0010] S3. Feeding with alternative prey;

[0011] S4. Five days after the spiders develop into adults, place the male and female adult spiders in a breeding device at a ratio of 1:1 to allow them to mate. After mating, remove the male spider and keep the female. About three weeks after mating, the female spider lays eggs and exhibits egg-guarding behavior. The egg masses hatch 8-14 days after laying the eggs.

[0012] S5. Place newly hatched spiderlings in a spider rearing box or centrifuge tube and feed them with substitute prey larvae until they reach the 5th instar. Then, raise them according to the method in S3.

[0013] Furthermore, the method further includes the following steps:

[0014] S6. Place 4-year-old or older wavy crab spiders in a refrigerator at 4±1℃ and maintain a relative humidity of 50±10% for 4-6 months.

[0015] In a specific implementation, data was fitted using the Holling II type disk equation to establish a predatory functional response model of adult wavy crab spiders to fall webworms of different instars. The theoretical maximum predation count of adult wavy crab spiders against 2nd, 3rd, 4th, and 5th instar fall webworm larvae was found to be 9.056, 6.674, 5.502, and 2.514, respectively.

[0016] In a specific implementation, the spider rearing box in S2 ( Figure 1A The spider rearing box consists of a lower box and an upper lid. The bottom is circular with a diameter of 5cm and a height of 3cm. The upper lid has a circular opening with a diameter of 1cm, and a metal mesh with a mesh count of 3000 is attached to the opening. In a preferred embodiment, small damp cotton balls or a 1cm layer of clean coconut fiber with 1mL of water are placed inside the spider rearing box, along with small pieces of bark for hiding places.

[0017] In a specific implementation, the substitute prey in S3 consists of mealworm larvae, sea worm larvae, and silkworm larvae. Different sizes and species of substitute prey are selected based on the spider's age and the rearing equipment.

[0018] In a specific implementation, the breeding device in S4 ( Figure 1B The container consists of a lower box and an upper lid, and is a cuboid measuring 10×10×8cm. The upper lid has two 1cm diameter circular openings, each with a 3000-mesh metal mesh. The bottom of the lower box is lined with 3cm of coconut fiber, and fresh leaves are placed on top of the coconut fiber. In a preferred embodiment, broad-leaved leaves such as mulberry or poplar leaves are used, and these are replaced every two days.

[0019] In a specific implementation, in step S5, after adding coconut fiber and water or a wet cotton ball to the spider rearing box or centrifuge tube, the young nymph spiders are placed in for rearing.

[0020] In a preferred embodiment, a spider rearing box is filled with coconut fiber and water to maintain humidity, and young nymphs are raised individually.

[0021] In a specific implementation, the dormant device in S6 ( Figure 1CThe box consists of a black outer shell 3-1, a box body 3-2, and a lid 3-3. The bottom is a circle with a diameter of 5cm and a height of 3cm. The box body 3-2 contains 1cm of coconut soil 3-4 and 1mL of water. Several pieces of bark 3-5 are placed on the coconut soil. The lid 3-3 has 10 small holes 3-6 with a diameter of 0.1cm.

[0022] This invention studies the predatory functional response of the wavy-striped crab spider to fall webworm larvae of different instars, determining that it conforms to the Holling type II predatory functional response model. When the density of fall webworm larvae is low, the predation amount of adult wavy-striped crab spiders increases with increasing prey density. The theoretical maximum predation amount of 2nd, 3rd, 4th, and 5th instar fall webworm larvae by adult wavy-striped crab spiders is calculated to be 9.056, 6.674, 5.502, and 2.514 larvae, respectively. Based on the predatory functional response data, a search effect model was further established. As the prey density increases, the search effect of adult wavy-striped crab spiders on fall webworm larvae gradually decreases. Through research on the feeding behavior of wavy-striped crab spiders of different instars to different substitute prey sizes, it was determined that 1 body length is suitable for feeding 2nd-4th instar nymphs, while 1.5 body length is suitable for 4th instar and above. A study of the feeding behavior of *Crabapple spp.* spiders at different instars with varying body sizes and prey species revealed that for prey of 1 times their body length, younger nymphs preferred to feed on larvae of crickets and silkworms, while their predation on yellow beetle larvae gradually increased with age. For prey of 1.5 times their body length, younger nymphs again preferred cricket larvae and yellow beetle larvae, while their predation on silkworm larvae gradually increased with age. A study on the survival rate of younger nymphs under different rearing conditions showed that using spider rearing boxes and moist coconut coir for humidity control yielded the best results for individual rearing. This invention further provides a low-temperature storage method for *Crabapple spp.* spiders, increasing the management and control of artificial breeding, reducing the breeding rate during non-productive stages, and enabling the control of the consistency of predator and prey release times during the production and breeding stages. Attached Figure Description

[0023] Figure 1A Spider breeding box.

[0024] Figure 1B Breeding equipment.

[0025] Figure 1C Feeding equipment.

[0026] Figure 2 The predatory function response and search effect of the wavy crab spider to larvae of the fall webworm at different instars.

[0027] Figure 3Selection of alternative prey size by wavy-flowered crab spiders at different ages.

[0028] Figure 4 The selection of alternative prey of different sizes and species by wavy crab spiders at different ages.

[0029] Figure 5 . Care conditions for young nymphs of the wavy crab spider.

[0030] Figure 6 The wavy-spotted crab spider feeds on the larvae of the yellow powder beetle.

[0031] Figure 7 Egg-guarding behavior of the wavy-flowered crab spider.

[0032] Figure 8 2-year-old nymphs of the wavy-flowered crab spider.

[0033] Figure 9 The nymphs of the wavy crab spider prey on the larvae of the fall webworm.

[0034] Figure 10 Male and female adult crab spiders. Detailed Implementation

[0035] A predatory wavy-striped crab spider ( Xysticus croceus The feeding and storage methods of the [product name] are characterized by including the following steps:

[0036] S1. Collect wavy-spotted crab spiders in the wild using the shaking or observation method;

[0037] S2. Bring the collected spiders back indoors and place them in a spider rearing box. Keep them at a temperature of 25±2℃ and a humidity of 60%. Overwintering spiders collected in the wild at temperatures below 0℃ should be placed in a 15±2℃ incubator for 24 hours to warm up before being reared.

[0038] S3. Feeding with alternative prey;

[0039] S4. Five days after the spiders develop into adults, place the male and female adult spiders in a 1:1 ratio in a breeding device to allow them to mate. After mating, remove the male spider and keep the female. About three weeks after mating, the female spider lays eggs and exhibits egg-guarding behavior. The egg masses hatch 8-14 days after laying the eggs.

[0040] S5. Place newly hatched spiderlings in a spider rearing box or centrifuge tube and feed them with substitute prey larvae until they reach the 5th instar. Then, raise them according to the method in S3.

[0041] Furthermore, the method further includes the following steps:

[0042] S6. Place 4-year-old or older wavy crab spiders in a refrigerator at 4±1℃ and maintain a relative humidity of 50±10% for 4-6 months.

[0043] From June to September, the shaking-drop method is used to collect spiders from trees such as mulberry and elm. The specific procedure is as follows: A 1-square-meter (m²) white netting (or gauze, plastic sheeting) is laid flat on the ground below the side branches of the target tree. The branches are gently shaken to create vibrations, causing the wavy-flowered crab spiders to fall from the leaves onto the netting, or they are allowed to slowly descend using hanging threads. The collected spiders are placed in pre-prepared centrifuge tubes, the tube openings are sealed with absorbent cotton, and the spiders are taken back to the tree. From November to March of the following year, targeted searches and collection can be conducted in overwintering sites such as under the bark at the base of trees, in fallen leaves, and in crevices around rocks.

[0044] The collected spiders were brought back indoors to a spider rearing box. Figure 1A Spiders are kept separately in the enclosure. The spider enclosure consists of a lower box 1-1 and an upper box cover 1-2. The bottom is a circle with a diameter of 5cm and a height of 3cm. The upper box cover 1-2 has a circular opening with a diameter of 1cm, and a metal mesh 1-3 with a mesh count of 3000 is attached to the circular opening.

[0045] Place small, damp cotton balls or a 1cm layer of clean coconut fiber with 1mL of water inside the rearing box, and add small pieces of bark for hiding places. Rear the spiders at a temperature of 25±2℃ and a humidity of 60%. For overwintering spiders collected in the wild when the temperature is below 0℃, first place them in a 15±2℃ incubator to warm them up, then transfer them to a 25±2℃ environment for rearing.

[0046] The physiological structure and hunting habits of the wavy-spotted crab spider present significant challenges to rearing it using artificial feed. Its chelicerae are less developed than those of other spiders like the tube-nesting spider and wolf spider, and both juvenile and adult spiders prefer live insects; artificial feed remains stationary, and the spiders will not consume it. Therefore, live insects are used as alternative prey, including mealworm larvae, silkworm larvae, and larvae of silkworms. Mealworms and silkworm larvae are easy to manage, reproduce rapidly, and are low-cost; multiple larvae of different instars can coexist at the same time, allowing for the acquisition of larvae of varying sizes. Silkworms develop quickly, and their eggs can be stored in a refrigerator for immediate hatching and feeding; spiders also prefer to feed on silkworm larvae. Therefore, using silkworm larvae, mealworms, and silkworms as alternative prey is simple, low-cost, and effective. To ensure rearing success, the appropriate age of the alternative prey should be selected based on the spider's age, and the size of the alternative prey should ideally be between 1 and 2 times the spider's body length. If the substitute prey is too small, the spider will not be willing to eat it, and the number of substitute prey to feed will need to be increased, increasing the workload; if the prey is too large, the spider will not dare to eat it or it will cause harm to the spider.

[0047] Female wavy crab spiders have a pre-egg-laying period, therefore they need to wait a few days after their last molt before mating. Male spiders undergo a dramatic color change after reaching adulthood, with their forelimbs turning black. One male and one female spider, both over 7 days old and fully grown, are selected and transferred to a breeding facility. Figure 1B The female spider is allowed to mate in the breeding device. Fresh leaves, such as mulberry or poplar leaves, are used in the breeding device to help her build an egg chamber; these leaves are changed every two days. The breeding device (…) Figure 1B It consists of a lower box body 2-1 and an upper box cover 2-2, and is a cuboid with dimensions of 10×10×8cm. The upper box cover 2-2 has two circular openings with a diameter of 1cm, and a metal mesh 2-3 is attached to the circular openings. The metal mesh 2-3 has a mesh count of 3000.

[0048] Before laying eggs, the female spider constructs an egg chamber. If building on a leaf, she uses silk to glue the edges of the leaf together, forming a cylindrical or conical egg chamber; if there are no leaves, she builds the egg chamber at the junction of the top and sides of the box. Egg laying usually occurs at night. The female spider first spins silk to create a mat-like structure, then lays the eggs on this mat. The egg mass is piled up. After laying the eggs, the female spider spins silk again to create an upper mat that completely covers the egg mass. The upper and lower mats together form an egg sac to protect the egg mass from external environmental disturbances. The average number of eggs laid is 124.1 (n=5), with a minimum of 68 and a maximum of 162. The egg sac is surrounded by messy silk, and the female spider guards it, waiting for the nymphs to hatch. No feeding is required during this period. Under conditions of 25±2℃ and 60% humidity, nymphs hatch and emerge after 8-14 days, with an average hatching rate of 94.19% (n=5).

[0049] In the first instar stage, the spiderlings rely entirely on the nutrients stored in the eggs for growth and development, requiring no external foraging activity. Upon reaching the second instar, the spiderlings emerge from the egg sac and disperse naturally using gossamer threads. Third instar spiderlings exhibit cannibalistic behavior; therefore, after emerging from the egg sac, they should be collected and housed individually in a spider rearing box or centrifuge tube. Because young spiderlings are fragile, they should not be brushed directly. Instead, use feathers or strips of paper to collect them from the bottom, then place them in a spider rearing box or centrifuge tube, keeping them moist with coconut fiber and water or damp cotton balls, and housed individually. Before the third instar, maintain a high humidity level in the rearing environment, keeping it at 70% ± 10%, while avoiding condensation that could interfere with the spiderlings' activities. The second to fourth instar nymphs of the wavy crab spider have the highest mortality risk and require meticulous management. After the fourth instar, the spiderlings' predatory abilities and resilience significantly improve; under artificial rearing conditions, over 95% can successfully develop into adults.

[0050] The wavy crab spider exhibits hibernation in low-temperature environments, with 1st-4th instar nymphs having an extremely low survival rate under these conditions. Therefore, 5th instar and older nymphs, as well as adult spiders, are selected to induce hibernation for storage. First, the spiders to be stored are fed to a satiated state using yellow powdery beetle larvae matching their growth stage. Then, they are transferred to the hibernation device. Figure 1C In this process, the spiders are placed in a 15°C incubator for one day and then transferred to a 4°C refrigerator. After one day of low-temperature induction, the spiders enter a dormant state. The survival rate of dormant spiders varies depending on their age, but generally, this method can preserve them for about 4-6 months. The dormancy device (…) Figure 1C The box consists of a black outer shell 3-1, a box body 3-2, and a lid 3-3. The bottom is a circle with a diameter of 5cm and a height of 3cm. The box body 3-2 contains 1cm of coconut soil 3-4 and 1mL of water. Several pieces of bark 3-5 are placed on the coconut soil. The lid 3-3 has 10 small holes 3-6 with a diameter of 0.1cm.

[0051] The following describes in detail the breeding and storage methods of the wavy crab spider provided by the present invention, with reference to specific embodiments. Example 1

[0052] This embodiment studies the functional response of the wavy-striped crab spider to the predation of the fall webworm. Fall webworm larvae of different instars and at different density gradients were placed in spider rearing boxes. The experimental setup and statistical results are shown in Table 1.

[0053] Table 1. Predation density settings for American white moths at different ages and average daily predation of the wavy-striped crab spider.

[0054]

[0055] Then, one adult *Crabappa undulata* spider that had been starved for 24 hours was introduced into the rearing box. Each treatment was repeated 5 times. The spider rearing box was placed under conditions of 25±1℃ and 60%±10% humidity. After 24 hours, the number of spiders preying on *Pteris vittata* larvae was counted. The Holling II type disc equation Na=aNT / (1+aT) was used. h N) Fits the experimental data to the equation, where a represents the instantaneous attack rate of the spherical civet, reflecting the predator's attack capability against its prey; T h The treatment time represents the time required for the wavy-spotted crab spider to prey on a fall webworm larva; Na represents the daily prey amount of the wavy-spotted crab spider per unit time; N represents the prey density; and T represents the experimental time, which is 1 day in this example. By fitting the experimental data, we obtained the predatory functional response equation of adult wavy-spotted crab spiders to fall webworm larvae of different instars, and calculated the theoretical maximum prey amount and search effect of the wavy-spotted crab spider. The search effect equation is S=a / (1+aT) hN), the results are shown in Table 2:

[0056] Table 2. Parameters of the predatory function response of adult *Crabweed undulatus* spiders to larvae of different instars of *Pteris vittata*.

[0057]

[0058] The results showed that the predation amount (Na) of adult *Symplocos wavyis* spiders on fall webworm larvae increased with increasing prey density (N) and decreased with increasing fall webworm instar. The theoretical maximum predation amount of 2nd, 3rd, 4th, and 5th instar fall webworm larvae by adult *Symplocos wavyis* spiders was 9.056, 6.674, 5.502, and 2.514, respectively. To verify the accuracy of the fitted equation, a chi-square test was performed on the results. The test results showed that the chi-square test was satisfactory. 2 All less than χ (0.05，42) =9.49, corresponding to p-values ​​greater than 0.992, which is significant at a level greater than 0.05. Therefore, it can be confirmed that the predatory functional response of adult *Crassula ovata* spiders to *Pteryx fallis* larvae conforms to the Holling type II disk equation.

[0059] When the density of fall webworm larvae is low, the predation rate of adult *Crassula ovata* spiders increases with increasing prey density; however, once the prey density reaches a certain level, due to limitations in the spiders' processing capacity or satiety levels, the predation rate tends to stabilize and no longer increases significantly with further increases in prey density. Based on predation response data, a search effect model was further established, consisting of... Figure 2 It is evident that the searching effect of adult *Symplocos rubrotinctum* spiders on fall webworm larvae gradually decreases as prey density increases. These findings provide theoretical support for alternative prey feeding methods in spider reproduction.

[0060] Example 2

[0061] This embodiment describes the screening of alternative prey for the wavy-spotted crab spider. Ten species of native and alternative prey for the wavy-spotted crab spider were obtained through field collection and purchase. The spiders were continuously fed with these native and alternative prey, and the ten prey species were artificially bred. The experimental results are shown in Table 3.

[0062] Table 3. Selection of alternative prey for the wavy-spotted crab spider.

[0063]

[0064] The results showed that aphids, leafhoppers, and leaf beetles require fresh foliage as food. Under artificial breeding conditions, the timely supply of fresh foliage cannot be guaranteed, or the cost of fresh foliage is too high, making stable production impossible. During feeding, aphids have fragile body walls that are difficult to separate from foliage, and leafhoppers are highly mobile, making separate feeding in individual boxes difficult. The breeding and management of vestigial-winged fruit flies and housefly larvae is difficult, and the larvae cannot feed spiders; only the adults can be used as prey. Fruit flies are easily contaminated with genes by normal fruit flies in the environment during breeding, resulting in a large number of winged fruit flies in the offspring, which can fly like houseflies, making separate feeding in individual boxes difficult. The artificial breeding and management of fall webworms is difficult, with high environmental requirements and complex operations. Furthermore, they can only be collected from the wild, making stable production impossible. Silkworms can be purchased at any time and are relatively inexpensive. They are also native prey, and their eggs can be refrigerated and hatched as needed, with a rapid growth rate, making them a relatively ideal prey. Yellow-faced beetle larvae and sea worm larvae mostly live underground in nature, while the wavy-striped crab spider is an arboreal spider, not their native prey. Furthermore, they are readily available, with a stable and low-cost supply; artificial breeding is simple, and larvae of different instars and sizes can coexist, facilitating feeding spiders of different ages and making the process convenient; their bodies are rich in nutrients, allowing the spiders to molt and develop normally, resulting in good feeding efficiency, making them ideal alternative prey. Therefore, silkworm larvae, yellow-faced beetle larvae, and sea worm larvae were selected as alternative prey.

[0065] Example 3

[0066] This embodiment studies the selection of alternative prey size by the wavy-spotted crab spider. The specific experimental method is as follows: One yellow powdery beetle larva each with a length of 0.5, 1, 1.5, 2, and 3 times the spider's body length were placed in a spider rearing box. Then, wavy-spotted crab spider nymphs and adults of different instars, after being starved for 24 hours, were added, with 20 replicates per group. The spider rearing boxes were placed under conditions of 25±1℃ and 60%±10% humidity. The body size of the yellow powdery beetle larvae first consumed by the wavy-spotted crab spider at each instar was recorded and analyzed. The experimental results are as follows: Figure 3 As shown.

[0067] The results showed that during the early nymphal stage, spiders preferred to feed on prey 0.5-1.5 times their body length, while after reaching the 4th instar, they preferred to prey 1-2 times their body length. Therefore, during the 2nd-4th instar nymphal stage, prey of 1 times their body length was used as a substitute for food, and during the 4th instar and above stage, prey of 1.5 times their body length was used as a substitute for food.

[0068] Example 4

[0069] This embodiment studies the selection of alternative prey species by the wavy-spotted crab spider. The specific experimental method is as follows: Two each of *Echinochloa crus-galli* larvae, *Pterocarya spp.* larvae, and *Bombyx mori* larvae (each one times the spider's body length) were placed in a spider rearing box. Since *Echinochloa crus-galli* larvae are relatively small, and their mature larvae are shorter than 7th instar nymphs and adults, three each of *Pterocarya spp.* larvae and *Bombyx mori* larvae were placed in each box after a 24-hour starvation period, with 20 replicates per group. The spider rearing boxes were placed at a temperature of 25±1℃ and a humidity of 60%±10%, and the types of alternative prey first consumed by the wavy-spotted crab spiders at each instar were recorded and analyzed. The experimental results are as follows: Figure 4 As shown in Figure a.

[0070] The results showed that young spider nymphs preferred to prey on silkworm larvae and beetle larvae, while their predation on yellow beetle larvae gradually increased with age. This is because the forelimbs and chelicerae of young spider nymphs are not fully developed, resulting in poor predation ability. The body walls of young silkworm and beetle larvae are thin, making them easier for the spider nymphs to pierce and consume. As the spider matures, its predation ability gradually increases, allowing it to more easily pierce the epidermis of yellow beetle larvae, thus increasing its predation tendency on them.

[0071] Example 5

[0072] This embodiment studies the selection of alternative prey species by the wavy-spotted crab spider. The specific experimental method is as follows: Two each of *Ichthyophthirius multifiliis* larvae, *Pterocarya spp.* larvae, and *Bombyx mori* larvae (each 1.5 times the spider's body length) were placed in a spider rearing box. Since *Ichthyophthirius multifiliis* larvae are relatively small, their mature larvae are less than 1.5 times the body length of 6th and 7th instar nymphs and adults; therefore, three each of *Pterocarya spp.* larvae and *Bombyx mori* larvae were placed in each rearing box. Then, wavy-spotted crab spider nymphs and adults of different instars were introduced after 24 hours of starvation treatment, with 20 replicates per group. The spider rearing boxes were placed under conditions of 25±1℃ and 60%±10% humidity, and the species of alternative prey first consumed by the wavy-spotted crab spiders at each instar were recorded and analyzed. The experimental results are as follows: Figure 4 As shown in b.

[0073] The results showed that young spider nymphs were more inclined to prey on silkworm larvae and yellow beetle larvae, while the predation tendency towards silkworm larvae gradually increased with the spider's age. This is because young spider nymphs feed by biting the head or tail of their prey and injecting venom. Silkworm larvae and yellow beetle larvae, which are 1.5 times their body length, are more slender than silkworm larvae, making it easier for the spider nymphs to bite their heads or tails. As the spider develops, its forelimbs and chelicerae become more developed, allowing it to restrict the prey's movement and bite the prey in the middle for feeding. Silkworm larvae have thinner body walls, making them easier to feed on, thus increasing the spider's predation tendency towards silkworm larvae.

[0074] Example 6

[0075] This embodiment studies the breeding and management of young nymphs of the wavy-spotted crab spider.

[0076] Treatment 1: Place the second-instar nymphs that have emerged from the egg sac into a spider rearing box. Add 1cm of coconut fiber to the bottom of the box, pour in 1mL of water, and add a small clump of moss (such as...). Figure 5 (a) The spider density is 1 spider / box. Add 2 silkworm larvae with a length of 1.5 times that of the spider every 2 days. Clean up the food residue from the previous feeding before the next feeding to prevent mold. Add 1mL of clean water every 5 days.

[0077] Treatment 2: Place the second-instar nymphs that have emerged from the egg sac into a spider rearing box. Add 1cm of coconut fiber to the bottom of the box, pour in 1mL of water, and add a small clump of moss (such as...). Figure 5 (a) The spider density is 5 spiders / box. Add 10 silkworm larvae (1.5 times the length of the spider) every 2 days. Clean up the food residue from the previous feeding before the next feeding to prevent mold. Add 1mL of clean water every 5 days.

[0078] Process 3: Place the second-instar nymphs that have emerged from the egg sac into a spider rearing box, and add small pieces of damp cotton balls to the bottom of the box. Figure 5 -b). The spider density is 1 spider / box. Every 2 days, add 2 yellow powder beetle larvae or sea worm larvae that are 1.5 times the length of the spider's body. Before the next feeding, clean up the food scraps from the previous feeding to prevent mold, and replace the wet cotton balls at the same time.

[0079] Process 4: Place the second-instar nymphs that have emerged from the egg sac into a spider rearing box, and add small pieces of damp cotton balls to the bottom of the box. Figure 5 -b). The spider density is 5 spiders per box. Every 2 days, add 10 yellow powder beetle larvae or sea worm larvae that are 1.5 times the length of the spider's body. Before the next feeding, clean up any leftover food to prevent mold growth and replace the wet cotton balls.

[0080] Process 5: Place the second-instar nymphs that have emerged from the egg sac into a 10mL centrifuge tube, seal the tube opening tightly with absorbent cotton, add 2cm of coconut fiber to the bottom of the tube, and then add 0.5mL of water. Figure 5 (c) The spider density is 1 spider / tube. Add 2 silkworm larvae with a length of 1.5 times that of the spider every 2 days. Clean up the food scraps from the previous feeding before the next feeding to prevent mold. Add 0.5 mL of clean water every 5 days.

[0081] Process 6: Place the second-instar nymphs that have emerged from the egg sac into a 10mL centrifuge tube, seal the tube opening tightly with absorbent cotton, and add a small piece of damp cotton ball to the bottom of the tube. Figure 5 (d). The spider density is 1 spider / tube. Add 2 yellow powder beetle larvae or sea worm larvae that are 1.5 times the length of the spider's body every 2 days. Before the next feeding, clean up the food scraps from the previous feeding to prevent mold, and replace the wet cotton balls at the same time.

[0082] Table 4. Survival rate of nymphs of the wavy-spotted crab spider under different rearing conditions

[0083]

[0084] The results are shown in Table 4. When using spider rearing boxes and moist coconut fiber to maintain humidity, individual rearing resulted in the highest survival rate. Second-instar nymphs exhibit cannibalistic behavior, therefore individual rearing was adopted. In practice, coconut fiber provided good humidity control, but yellow meal beetle larvae and silkworm larvae would burrow into the soil, necessitating the use of silkworm larvae for feeding. Using moist cotton balls minimized environmental disturbance, allowing the spiders to quickly locate prey and facilitating the removal of food residue. However, moist cotton balls dried quickly, were complex to handle, and water droplets could easily drip, affecting the nymphs' activity and potentially causing their death. Centrifuge tubes were difficult to operate and made it hard to remove food residue. Therefore, using spider rearing boxes and moist coconut fiber for humidity control yielded the best results for individual rearing of young nymphs.

[0085] Example 7

[0086] This embodiment studies the storage conditions of the wavy-spotted crab spider. Fifth-instar nymphs and adults were selected for storage. The spiders to be stored were fed with yellow powder beetles until satiated and then placed in a hibernation device. Figure 1C The hibernation device containing the spider was placed in a 15°C incubator for one day, then transferred to a 4°C refrigerator. After one day, the spider began to enter a hibernation state. Once the spider entered hibernation, it could continue to be stored in the 4°C refrigerator, with the humidity inside the container checked periodically to ensure it remained at 50% ± 10%.

[0087] Using this method, the dormancy rate can reach over 95%. During storage, regularly check the spiders' condition to ensure they are in a dormant state and undisturbed. If signs of awakening are observed or the storage environment changes, adjust the storage conditions promptly. Generally, this method can preserve spiders for approximately 4-6 months.

Claims

1. A predatory natural enemy, the wavy-striped crab spider ( Xysticus croceus The feeding method of ) is characterized by, Includes the following steps: S1. The collected wavy crab spiders were brought back indoors and placed in a spider rearing box, where they were kept at a temperature of 25±2℃ and a humidity of 60%. S2. Use one, two, or three of the following as alternative prey for feeding: worms, yellow powder beetles, and silkworms; S3. After developing into adult spiders, place male and female adult spiders in a breeding device at a 1:1 ratio and allow them to mate. After mating, remove the male spider and keep the female spider. From the time the female spider lays eggs after mating, the egg mass will hatch in 8-14 days. S4. Place newly hatched spiderlings in a spider rearing box or centrifuge tube and feed them with larvae that serve as prey as described in S2 until they reach the 5th instar. S5. Place the wavy crab spiders raised using the aforementioned method in a refrigerator at 4±1℃, maintain a relative humidity of 50±10%, and store for 4-6 months; In step S1, the wavy crab spiders are collected in the wild by shaking or observation. If the overwintering spiders are collected in the wild at a temperature below 0℃, they need to be placed in a 15±2℃ incubator for 24 hours to warm up before being raised. In step S1, the spider breeding box consists of a lower box body and an upper box cover. The bottom surface is a circle with a diameter of 5cm and a height of 3cm. The upper box cover has a circular opening with a diameter of 1cm, and a metal mesh with a mesh count of 3000 is attached to the circular opening. The spider rearing box contains small, damp cotton balls and small pieces of bark for hiding places; or the spider rearing box contains a 1cm layer of clean coconut fiber and 1mL of water, and small pieces of bark for hiding places. In step S3, the breeding device consists of a lower box and an upper box cover. It is a cuboid with dimensions of 10×10×8cm. The upper box cover has two circular openings with a diameter of 1cm, and a metal mesh with a mesh count of 3000 is attached to the circular openings. The bottom of the lower box is covered with 3cm thick coconut fiber, and fresh leaves are placed on the coconut fiber. Use mulberry leaves and poplar leaves, and change them every 2 days.

2. The method according to claim 1, characterized in that, In step S4, coconut fiber and water are added to the spider rearing box or centrifuge tube, or wet cotton balls are added, and then the young nymphs are placed in for rearing.

3. The method according to claim 1, characterized in that, In step S4, coconut fiber and water are added to a spider rearing box to maintain humidity, and young nymphs are raised separately.

4. The method according to claim 1, characterized in that, In step S5, when placing it in the refrigerator, it is placed in the hibernation device and then placed in the refrigerator. The hibernation device includes a black outer shell, which consists of a box body and a lid. The bottom is a circle with a diameter of 5cm and a height of 3cm. The box body contains 1cm of coconut fiber and 1mL of water. Several pieces of bark cover the coconut fiber. The lid has 10 small holes with a diameter of 0.1cm.

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