Efficient propagation method for parasitic natural enemy products of housefly pupae

Abstract

The invention discloses an efficient propagation method for parasitic natural enemy products of housefly pupae, and relates to the technical field of animal husbandry pest biological control, and the method specifically comprises the following steps: S1, breeding host housefly pupae; s2, breeding a bee species of the pseudoalda persicae; s3, breeding the species of the commission minulescens; and S4, large-scale breeding of the mixed biocontrol product. According to the method disclosed by the invention, the Aada-like periodonia with stronger reproductive capacity and the terracotta figure bees with stronger field adaptive capacity are selected, and the propagation capacity and the pest control capacity of natural enemy products are improved through an artificial propagation mode of mixed bee inoculation and twice bee inoculation treatment.

Description

Technical Field

[0001] The invention relates to the technical field of biological control of livestock pests, and in particular to a method for efficiently propagating a parasitic natural enemy product of housefly pupae. Background Art

[0002] In integrated pest management, as the negative problems brought by chemical control become more prominent, discovering and utilizing new natural enemy resources for integrated pest management is an effective way to solve the above problems. The release and application of biological control products with high control efficiency can not only effectively reduce the frequency and amount of chemical pesticide use, but also greatly improve the safety of agricultural products. Parasitic natural enemies are important resources for biological pest control. Many parasitic wasps have been deeply developed and applied in the field of integrated pest management, such as Trichogramma spp. Trichogramma Aphidida Aphelinidae Aphidida Aphidiidae , Chrysanthemumidae Pteromalidae Numerous parasitic natural enemies of other families and genera play a good preventive and control role in pest control.

[0003] Compared with the exploration and utilization of parasitic natural enemy resources of agricultural and forestry pests, the biological control of sanitary pests in my country has made slow progress. In recent years, with the increase in the proportion of meat and eggs in the dietary structure, livestock and poultry breeding in my country has also developed rapidly, and flies, as an important companion pest of livestock and poultry breeding, are also causing more and more damage year by year. At present, chemical control measures are used to control flies in livestock and poultry farms in my country, but the development of pest resistance and non-point source pollution caused by the application of pesticides pose serious safety hazards to the healthy and safe production of meat and eggs. Therefore, the exploration and application of fly natural enemy resources and the development of fly biological control in this field can not only achieve sustainable control of fly pests in livestock and poultry farms, but also reduce the risk of pesticide residues in meat and egg products and improve product quality.

[0004] Chrysops adana Muscidifurax similadanacus and Aglaonema Spalangia endiusBoth belong to the family Chrysomelidae. These two parasitic wasps can be collected in the survey of natural enemy resources of housefly pupae in cattle farms in many places in northern my country, and the occurrence rate is relatively high. The natural population has a certain control effect on cattle farm flies, and is a potentially dominant parasitic natural enemy. The adana-like intestinal margin wasp is a new record species in my country in 2018. Its original host is housefly pupae; the fly pupa servant wasp is a widely distributed species in the world and has a good control effect on many kinds of flies. Although the occurrence rate of the fly pupa servant wasp in the wild is higher than that of the adana-like intestinal margin wasp, its reproductive capacity is significantly lower than that of the adana-like intestinal margin wasp. Therefore, if the two parasitic wasps can be used in combination for livestock and poultry fly control, it can not only ensure the high adaptability of the fly pupa servant wasp, but also reduce the cost of biological control products through the high reproductive efficiency of the adana-like intestinal margin wasp, which is very meaningful for the control of fly pests in livestock and poultry farms. However, there is currently no public literature reporting on the joint breeding technology of the two parasitic wasps, and it is unknown what kind of joint breeding method can reduce the interspecific competition between the two bees and thereby improve the reproductive efficiency of the parasitic wasps. Summary of the invention

[0005] In view of the above problems, the present invention provides a method for efficiently propagating a parasitic natural enemy product of housefly pupae.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is as follows: The present invention provides a method for efficiently propagating a parasitic natural enemy product of housefly pupae, which specifically comprises the following steps: S1: Reproduction of host housefly pupae; S2: breeding of Adana spp. S3: breeding of fly servant bees; S4: Mass propagation of mixed biological control products.

[0007] Furthermore, in the step S1, milk powder is used as feed, and adult houseflies are raised in insect cages, with 800 to 1000 adult flies in each cage, and wheat bran with a water content of 65% is used as a housefly egg-laying matrix and water source, and the egg-laying matrix is ​​replaced and milk powder is supplemented every day; housefly eggs are taken out and developed to initial eclosion, and the larvae are transferred to a plastic basin, and wheat bran with a water content of 65% is provided as larval feed, with 10,000 to 12,000 larvae in each basin, and the housefly pupae are washed out of the waste within 1 to 2 days after the larvae begin to pupate, and dried (the rearing conditions of adult houseflies: 26°C to 29°C, 55% to 70% RH, 16:8 L:D).

[0008] Furthermore, in step S2, the housefly pupae in step S1 are used to breed the species of the Adana pseudo-Galox spp., a plastic bottle is used as a breeding container, and the bee inoculation treatment is carried out in a dark room at a ratio of female bees: housefly pupae (host pupae) = 1:40. After 48 hours of inoculation, the bee species are removed, and the parasitic bees are developed and set aside.

[0009] Furthermore, in step S3, the housefly pupae in step S1 are used to breed fly pupa and small bee species, and a plastic bottle is used as a breeding container. The bee inoculation process is carried out in a dark room at a ratio of female bees: housefly pupae (host pupae) = 1:20. After 48 hours of inoculation, the bee species are removed and the parasitic bees are developed and set aside.

[0010] Furthermore, in the step S4, a mixed inoculation method is adopted, and the Adana intestinal margin golden bee and the fly pupa bee that are bred in steps S2 and S3 and emerge on the same day are mixed in a ratio of 1:1, and inoculated with the housefly pupae bred in step S1 in a ratio of female bees: housefly pupae (host pupae) = 1:40. After 48 hours of inoculation, the parasitic bee species are screened out and recovered, and the recovered parasitic bees are inoculated for a second time in a ratio of female bees: housefly pupae (host pupae) = 1:40. After 48 hours of inoculation, the bee species are removed, and the parasitic pupae obtained twice are developed separately. When the parasitic bees develop to the early pupa stage, they are stored as biological control products for standby use.

[0011] Furthermore, the length×width×height of the insect breeding cage=30cm×30cm×30cm; the length×width×height of the plastic basin=30cm×30cm×30cm.

[0012] Furthermore, the diameter×height of the plastic bottle=8cm×15cm; the environmental conditions for bee inoculation and development are both temperature 25±1°C and relative humidity 60±5%; the lighting conditions for bee inoculation are L:D=24:0; and the lighting conditions for development are L:D=16:8.

[0013] Furthermore, the diameter×height of the plastic bottle=8cm×15cm; the environmental conditions for bee inoculation and development are both temperature 25±1°C and relative humidity 60±5%; the lighting conditions for bee inoculation are L:D=24:0; and the lighting conditions for development are L:D=16:8.

[0014] Compared with the prior art, the present invention has the following beneficial effects: In the present invention, the Adana intestinal margin golden bee with stronger reproductive ability and the fly pupa figurine bee with stronger field adaptability are selected, and the artificial propagation method of mixed bee inoculation and two bee inoculation treatments is used to improve the propagation ability and pest control ability of the natural enemy product and reduce the production cost of the natural enemy product, specifically: (1) The mixed inoculation treatment can significantly improve the reproductive capacity of the Adana wasp, while the reproductive capacity of the fly pupa wasp did not change significantly, which generally improved the propagation efficiency of natural enemy products; (2) The choice of the two-time bee-grafting method maximizes the utilization rate of bee species and reduces the production cost of natural enemy products; (3) In the mixed bee inoculation treatment, the number of parasites of Adana wasp was high, and the fly pupa wasp had stronger environmental adaptability, which made the biological control product have stronger pest control ability; (4) The interspecific competition induced by the mixed breeding of Adana and Aglaonema promoted the improvement of the overall reproductive capacity of parasitic wasps. This conclusion provides a technical reference for the research related to parasitic natural enemies. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 This is a survey on the parasitism rate of two parasitic wasps (Adana intestinal margin golden wasp and fly pupa figurine wasp) in the test cattle farm in 2022 and 2023; Figure 2 is the daily egg-laying amount of the Adana intestinal margin golden wasp and the fly pupa servant wasp of the present invention; Figure 3 is the daily emergence rate of Adana intestinal margin golden wasp and fly pupa servant wasp; Figure 4 The first bee inoculation treatment under the two bee inoculation methods (mixed bee inoculation and single bee inoculation) of the present invention has an effect on the parasitic amount of Acropora adana and Acropora pupae; Figure 5 The invention discloses an effect of the second bee inoculation treatment on the parasitic amount of Acropora adana and Acropora pupae under the two bee inoculation methods (mixed bee inoculation and single bee inoculation) of the present invention.

[0016] Note: Figure 1 , Figure 4 and Figure 5 Different lowercase letters (a, b) indicate that the data were subjected to analysis of variance and there were significant differences at the 0.05 level. DETAILED DESCRIPTION

[0017] In order to make the purpose and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0018] The instruments, reagents, materials, etc. involved in the following examples, unless otherwise specified, are all conventional instruments, reagents, materials, etc. in the prior art and can be obtained through regular commercial channels. The experimental methods, detection methods, etc. involved in the following examples, unless otherwise specified, are all conventional experimental methods, detection methods, etc. in the prior art.

[0019] This example was carried out in a dairy farm in Yinchuan, Ningxia Hui Autonomous Region and in the laboratory of the Institute of Biological Control of Jilin Agricultural University, and the implementation period was 2022-2023.

[0020] Example 1 Investigation of parasitic natural enemy resources of flies in cattle farms From May to October in 2022 and 2023, respectively, regular surveys on the parasitic natural enemy resources of housefly pupae were conducted at the second dairy farm of Ningxia Nongken Dairy Co., Ltd., the third dairy farm of Ningxia Helan Mountain Dairy Original Breeding Co., Ltd., and the fifth dairy farm of Ningxia Nongken Dairy Co., Ltd. The specific method is: according to the occurrence site of fly pupae, pupae are collected regularly at fixed points by manual excavation on the 15th of each month (the survey period is postponed in case of rainy days), the area of ​​each point is set at 1m × 1m, the number of pupae collected at each point is 50-100 pupae, and a total of 2,000 pupae are collected from each farm. The collected fly pupae are brought back to the room for development. After the fly pupae emerge, the adult flies and pupal shells are removed, and the parasitic pupae are placed under laboratory conditions for development. After all the parasitic bees emerge, they are immersed in 75% alcohol, observed under a stereo microscope and an ultra-depth microscope, the number of parasitic bees is counted, and the parasitic bees are morphologically identified. The environmental conditions were temperature 25 ± 1 °C, relative humidity 60 ± 5%, and photoperiod L:D = 16:8.

[0021] In 2022 and 2023, a survey of the resources of natural enemies of housefly pupae was conducted in fixed cattle farms in Yinchuan area for two consecutive years. The results showed that the Adana intestinal margin golden wasp and the fly pupa figurine wasp were the two groups with the highest occurrence. The average annual collection volume of these two parasitic wasps accounted for 2.4% of the total collection volume. Figure 1 . As can be seen from the figure, the parasitism rates of Adana intestinal margin golden wasp and fly pupa wasp were 26% and 11% respectively in 2022; and 21% and 9% respectively in 2023. The results show that the wild populations of both parasitic wasps have a high occurrence rate, and the natural populations have a good control ability over the occurrence of pests. They are the dominant natural enemy groups for fly pest control in cattle farms and have the potential to be developed into biological control products. The occurrence rate of fly pupa wasp in the wild is higher, indicating that it has a stronger ability to adapt to the wild and has a better control ability over the occurrence of insect pests.

[0022] Example 2 Comparison of reproductive characteristics between Aglaonema pseudoadana and Aglaonema pupae (1) Reproduction of housefly pupae and reproduction of two parasitic bees on housefly pupae Housefly Musca domestica :The housefly pupae were collected from Yinchuan in September 2022, and the collected housefly pupae were brought back to the laboratory for propagation. The specific breeding of host housefly pupae is as follows: using milk powder as feed, raising housefly adults in insect cages of length × width × height = 30cm × 30cm × 30cm, with 800~1000 adult flies per cage, using wheat bran with a water content of 65% as the housefly egg-laying matrix and water source, replacing the egg-laying matrix and supplementing milk powder every day; taking out the housefly eggs and developing them to initial eclosion, transferring the larvae to a plastic basin of length × width × height = 30cm × 30cm × 30cm, and providing wheat bran with a water content of 65% as larval feed, with 10,000~12,000 larvae per basin, washing the housefly pupae from the waste within 1~2 days after the larvae begin to pupate, and drying them for later use.

[0023] Parasitic wasps: Both Adana-like intestinal margin golden wasp and fly pupa servant wasp were collected from Yinchuan area in September 2022, and identified, expanded and conserved indoors at the Institute of Biological Control of Jilin Agricultural University. The above-mentioned housefly pupae are used as breeding hosts for the development of bee species of Adana-like intestinal margin golden wasp and fly pupa servant wasp. The specific method is: using a plastic bottle with a diameter × height = 8cm × 15cm as a breeding container, the newly emerged and fully mated Adana-like intestinal margin golden wasp is placed in a dark room at a ratio of female bees: housefly pupae = 1:40, and the newly emerged and fully mated fly pupa servant wasp is placed in a ratio of female bees: housefly pupae = 1:20. After 48 hours of bee placement, the bee species are removed, the parasitic wasps are developed, and the parasitic wasps are pupated to count the single female parasitism, emergence rate, and female ratio of the two parasitic wasps. The environmental conditions for bee inoculation and development were both temperature (25±1)℃ and relative humidity (60±5)%; the light conditions for bee inoculation were L:D=24:0; the light conditions for development were L:D=16:8.

[0024] The results showed that both parasitic wasps had good reproductive characteristics on housefly pupae. The statistical analysis of reproductive parameters such as the parasitism load per female, emergence rate and female ratio showed that the above indicators were significantly higher than those of the housefly pupa servant wasps (see Table 1). Specifically, the parasitism load per female of the housefly was 24.9 in 48 hours, while that of the housefly pupa servant wasps was 13.6; the emergence rate of the housefly was 99.3% and that of the housefly pupa servant wasps was 84.4%; the offspring of the housefly and the housefly were mainly female, with female ratios of 0.72 and 0.64, respectively.

[0025] Table 1 Reproductive characteristics of two parasitic wasps on housefly pupae

[0026] Note: Different lowercase letters (a, b) indicate that there are significant differences in the reproductive parameters of the two parasitic wasps at the 0.05 level when variance analysis is performed.

[0027] (2) Comparison of reproductive characteristics of two parasitic wasps The two parasitic wasps that had just emerged were fully mated in the insect tank and then introduced into the test tubes, with one pair in each tube (♀:♂=1:1), and fresh housefly pupae (40 Adana intestinal margins of the golden bee and 20 housefly pupae of the housefly servant bee) were provided. The test tubes were sealed with gauze, and the host pupae were replaced every 24 hours. When the female bee died, no host was provided. The parasitic pupae were placed under the original conditions for development, and the daily egg production and emergence rate of the two parasitic wasps were recorded. Each bee species was a treatment, and each treatment was repeated 20 times. The experimental environment conditions were (25±1)℃, humidity RH (60±5)%, and a photoperiod of 14L:10D in an artificial climate chamber.

[0028] The results showed that by observing the daily egg-laying amount of the two parasitic wasps for 10 days, it was found that the egg-laying amount of the Adana-like intestinal margin wasp first increased and then decreased with the increase of age. The peak of egg-laying occurred on the 5th day after eclosion, and the average daily egg-laying amount in 10 days was 12-30 eggs. There was no significant difference in the egg-laying amount of the fly pupa servant wasp in the first 4 days after eclosion, and it began to decline after the 5th day. The average daily egg-laying amount in 10 days was 2-9 eggs. The egg-laying amount of the Adana-like intestinal margin wasp at each tested age was significantly higher than that of the fly pupa servant wasp ( Figure 2 ); The emergence rate of the tested day-old Adana wasp was 95%-99%, which was significantly higher than the 80%-92% of the fly pupa wasp ( Figure 3 ). The results showed that the reproductive efficiency of the Adana wasp was significantly higher than that of the fly pupa wasp.

[0029] Example 3 Effect of mixed inoculation on the total amount of eggs laid by two parasitic wasps The newly emerged Adana intestinal edge golden bee and fly pupa servant bee were obtained from Example 2. The two parasitic bees that emerged on the same day were mixed in a mixed bee infusion method at a ratio of 1:1 for female bees, and the bee infusion treatment was carried out at a ratio of 1:40 for female bees: housefly pupae. After 48 hours, the adult bees were screened out, the bee species were recovered, and the second bee infusion treatment was carried out at a ratio of 1:40 for female bees: housefly pupae. The bee species were removed 48 hours after the bee infusion, and the parasitic pupae obtained twice were developed to emergence respectively. Each bee infusion was a treatment, and 20 replicates were set for each treatment. The number of parasites and the number of emergence of the two bees in each treatment were counted. The experimental environment conditions were an artificial climate chamber with (25±1)℃, humidity RH (60±5)%, and a photoperiod of 14L:10D.

[0030] The results showed that the use of two kinds of parasitic wasps mixed inoculation treatment, the two inoculations can significantly increase the parasitic amount and total parasitic amount of Adana intestinal edge golden wasp. The parasitic amount after the first inoculation treatment was as follows: Figure 4 . As can be seen from the figure, when treated with bees alone, the parasitic amount of the Adana-like intestinal margin golden wasp was 22.0 grains / female bee, while when treated with mixed bees, the parasitic amount increased to 32.2 grains / female bee, and the parasitic amount increased by 46.4%; the two bee treatments had no effect on the parasitic amount of the fly pupa wasp; the total parasitic amount in the single bee treatment (1 Adana-like intestinal margin golden wasp + 1 fly pupa wasp) was 34.2 grains, and the total parasitic amount in the mixed bee treatment (1 Adana-like intestinal margin golden wasp + 1 fly pupa wasp) was 41.25 grains, and the parasitic amount increased by 20.6%. The parasitic amount after the second bee treatment is as follows Figure 5As shown in the figure, in the second bee inoculation treatment, the two parasitic wasps still maintained a high parasitic amount. When treated with bees alone, the parasitic amount of the second bee inoculation of the Adana intestinal margin golden wasp was 20.81 grains / female bee, and when treated with mixed bees, its parasitic amount was 27.45 grains / female, which increased by 31.9%; the parasitic amount of the fly pupa figurine wasp in the two bee inoculation treatments was 10.22 grains / female and 9.43 grains / female, respectively, with no significant change; in addition, compared with the single bee inoculation treatment, the total parasitic amount of the mixed bee inoculation treatment was also significantly increased.

[0031] Based on the results of Examples 1-3, both the Adana-like intestinal margin golden bee and the fly pupa servant bee have a high occurrence rate in cattle farms, and the fly pupa servant bee accounts for a higher proportion, indicating that the fly pupa servant bee has a higher ability to adapt to the wild, but the Adana-like intestinal margin golden bee has a significantly higher reproductive capacity than that of the fly pupa servant bee. Comprehensively evaluating the reproduction cost and control effect, the Adana-like intestinal margin golden bee has more potential to be developed into a biological control product; the mixed inoculation treatment can significantly increase the reproduction of the Adana-like intestinal margin golden bee, while the reproduction of the fly pupa servant bee has no significant change.

[0032] Normally, when two bee species compete, the reproductive capacity of one will increase while the other will decrease. However, in the present invention, through mixed bee inoculation, the reproductive capacity of the fly pupa servant wasp with strong field adaptability and weak reproductive capacity is not reduced during the breeding process, and the reproductive capacity of the Adana intestinal margin golden wasp with weak field adaptability and strong reproductive capacity is significantly improved on the original basis, that is, the interspecific competition induced by mixed bee inoculation promotes the significant improvement of the total reproductive capacity of parasitic wasps. For natural enemy products, its expansion efficiency is greatly improved. However, the improvement of the expansion efficiency is not at the expense of the reproductive capacity and field adaptability of a certain bee species, but on the basis of maintaining the original efficiency, the total reproductive efficiency is additionally improved, achieving an expansion effect of 1+1>2, which is very different from the prior art.

[0033] In summary, the mixed bee inoculation treatment of the present invention can not only reduce the production cost of biological control products, but also improve the pest control ability and breeding efficiency of parasitic natural enemy insects.

[0034] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for efficiently propagating a parasitic natural enemy product of housefly pupae, characterized in that: The specific steps include: S1: Reproduction of host housefly pupae; S2: breeding of Adana spp. S3: breeding of fly servant bees; S4: Mass propagation of mixed biological control products.

2. The method for efficiently propagating a parasitic natural enemy product of housefly pupae according to claim 1, characterized in that: In the step S1, milk powder is used as feed, and adult houseflies are raised in insect cages, with 800 to 1000 adult flies in each cage; wheat bran with a water content of 65% is used as a housefly egg-laying matrix and water source, and the egg-laying matrix is ​​replaced and milk powder is supplemented every day; housefly eggs are taken out and developed to initial eclosion, the larvae are transferred to a plastic basin, and wheat bran with a water content of 65% is provided as larval feed, with 10,000 to 12,000 larvae in each basin; and housefly pupae are washed out from waste materials within 1 to 2 days after the larvae begin to pupate, and dried.

3. The method for efficiently propagating a parasitic natural enemy product of housefly pupae according to claim 1, characterized in that: In step S2, the housefly pupae in step S1 are used to breed the species of the Adana intestinal margin hornbills, a plastic bottle is used as a breeding container, and the bee inoculation process is carried out in a dark room at a ratio of female bees to housefly pupae of 1:

40. After 48 hours of inoculation, the bee species are removed, and the parasitic bees are developed and set aside.

4. The method for efficiently propagating a parasitic natural enemy product of housefly pupae according to claim 1, characterized in that: In step S3, the housefly pupae in step S1 are used to breed fly pupa and small bee species, and a plastic bottle is used as a breeding container. The bee species are inoculated in a dark room at a ratio of female bees to housefly pupae of 1:

20. After 48 hours of inoculation, the bee species are removed and the parasitic bees are developed for later use.

5. The method for efficiently propagating a parasitic natural enemy product of housefly pupae according to claim 1, characterized in that: In the step S4, a mixed bee inoculation method is adopted, the Adana intestinal margin golden bee and the fly pupa bee bred in steps S2 and S3 and emerging on the same day are mixed in a ratio of 1:1, and inoculated with the housefly pupae bred in step S1 in a ratio of female bees: housefly pupae=1:40, and the parasitic bee species are screened out and recovered 48 hours after the bee inoculation, and the recovered parasitic bees are subjected to a second bee inoculation treatment in a ratio of female bees: housefly pupae=1:40, and the bee species are removed 48 hours after the bee inoculation, and the parasitic pupae obtained twice are developed separately, and when the parasitic bees develop to the early pupa stage, they are stored as biological control products for standby use.

6. The method for efficiently propagating a parasitic natural enemy product of housefly pupae according to claim 2, characterized in that: The insect cage has a length×width×height=30cm×30cm×30cm; the plastic basin has a length×width×height=30cm×30cm×30cm.

7. The method for efficiently propagating a parasitic natural enemy product of housefly pupae according to claim 3, characterized in that: The diameter×height of the plastic bottle is 8cm×15cm; the environmental conditions for bee inoculation and development are both 25±1°C and 60±5% relative humidity; the light conditions for bee inoculation are L:D=24:0; and the light conditions for development are L:D=16:

8.

8. The method for efficiently propagating a parasitic natural enemy product of housefly pupae according to claim 4, characterized in that: The diameter×height of the plastic bottle is 8cm×15cm; the environmental conditions for bee inoculation and development are both 25±1°C and 60±5% relative humidity; the light conditions for bee inoculation are L:D=24:0; and the light conditions for development are L:D=16:8.

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