CN111264473B - Method for breeding Exorista bigelovii by utilizing galleria mellonella - Google Patents

Method for breeding Exorista bigelovii by utilizing galleria mellonella Download PDF

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CN111264473B
CN111264473B CN202010098937.2A CN202010098937A CN111264473B CN 111264473 B CN111264473 B CN 111264473B CN 202010098937 A CN202010098937 A CN 202010098937A CN 111264473 B CN111264473 B CN 111264473B
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fly
transparent plastic
cage
larvae
disposable transparent
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CN111264473A (en
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蒋学建
黄华艳
罗辑
常明山
邹东霞
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Guangxi Zhuang Autonomous Region Forestry Research Institute
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Guangxi Zhuang Autonomous Region Forestry Research Institute
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00Rearing or breeding animals, not otherwise provided for; New breeds of animals
    • A01K67/033Rearing or breeding invertebrates; New breeds of invertebrates

Abstract

A method for breeding Exorista civilis Roem by using Selaginella furcifera comprises putting 60 pupae of Exorista civilis Roem in a breeding cage. The larva of the galleria mellonella treated by 75% alcohol and hot water of 60 ℃ is placed in a culture dish and put into a rearing cage. The culture dish is taken out every 2h, and the larvae of the greater wax moth on which the parasitic flies lay eggs are transferred into disposable plastic cups, wherein 50 heads are placed in each cup. After 15-20 days, taking the fly pupae out of the disposable transparent plastic cup and putting the fly pupae into a new disposable transparent plastic cup, wherein the number of the fly pupae in each disposable transparent plastic cup is 60. When the parasitic flies eclosion occurs, the disposable transparent plastic cup is moved into a new fly collecting cage, and a new round of breeding is started. The host greater wax moth larva used in the method is easier to obtain, the operation is simple, and the cost of feeding the Japanese chastetree flies is low.

Description

Method for breeding Exorista bigelovii by utilizing galleria mellonella
Technical Field
The invention belongs to the technical field of biological control, relates to a breeding method of natural enemy insects, and particularly relates to a method for breeding Exorista civilis Roem by utilizing galleria mellonella.
Background
The Exorista japonica is a large-egg parasitic fly, a female fly lays an egg which does not complete embryo development on the body surface of a host, the larva is hatched and then drilled into the body cavity of a host larva to live in a compatible parasitic life, finally the host is drilled out in the pupal stage of the host, and pupation is beside the killed host pupal, so that the biological control effect is achieved. The host of the parasitoid is wide, more than 30 pests such as silkworm, pine moth, fall webworm, armyworm and prodenia litura are natural hosts, and the parasitic rate can reach more than 90 percent at most. Therefore, the parasitoid has wide application prospect in the aspect of preventing and controlling agricultural and forestry pests.
At present, experts successfully feed the Exorista civilis with hosts such as Trichoplusia agnata, armyworm, beet armyworm, meadow moth and the like. In order to continue to reduce the breeding cost of the Exorista civilis Rond, it is necessary to select intermediate hosts that can be more easily bred. Therefore, the inventors expanded the intermediate host selection range of the Exorista civilis into its non-natural hosts. Experiments find that the parasitic rate of the Exorista parva Hemsl to the larvae of the wax moth which is not treated by the technology reaches more than 21 percent, so the wax moth becomes the research target of the people. The greater wax moth belongs to Lepidoptera Lepidopetera, Pyralidae and Gallerineae, which are important large pests of honeybee nest and are called nest worms. In 2013, the inventor breaks through the indoor large-scale propagation technology of the greater wax moth and obtains patent grant (ZL 201310416520.6) in 2017. By analyzing the nutrient components of the greater wax moth (Jiangjian greater wax moth nutrient component analysis and evaluation, Guangxi forestry science, 2016,45(04): 452-), the penaeid beetle has the components of crude protein, crude fat and the like which are higher than the content of the natural host silkworm of the Japanese migratory fly under the condition of the same weight, and can be more suitable to be used as an intermediate host of the Japanese migratory fly for feeding the Japanese migratory fly. The activity of the larvae of the greater wax moth imposes the habit of spinning, and experiments show that the larvae of the greater wax moth find a hidden place to spin and wrap themselves before the parasitic flies lay eggs, so that the host parasitic rate is low. The inventor finds that the activity and the silking ability of the greater wax moth larvae can be obviously reduced after the larvae of the greater wax moth are treated by alcohol and hot water through repeated experiments in order to ensure the survival of the greater wax moth, the parasitic rate of the greater wax moth larvae by the Japanese rice fly larvae is increased from 21 percent to more than 60 percent under the conditions of the temperature of 20-30 ℃ and the humidity of 75-90 percent, and also finds the degeneration phenomenon of the rice fly after more than ten generations of breeding, thereby forming the invention.
Disclosure of Invention
The invention aims to provide a simple and effective breeding method for breeding the Exorista civilis Rond by utilizing the greater wax moth.
The invention is realized by the following steps:
a method for breeding Exorista civilis Roem by using Chilo suppressalis comprises placing pupae of Exorista civilis Roem (60 + -1) in a breeding cage, eliminating weak and malformed Exorista civilis Roem imagoes after eclosion, and ensuring that the number of Exorista civilis Roem in each cage is 50 + -1, and the ratio of male to female is 2-3: 2; putting 20% of honey water into the cage for supplementing nutrition for the adult parasitic flies, putting the processed hosts in a culture dish, and putting the hosts into a feeding cage; taking out the culture dish after the parasitic flies lay eggs, observing under a dissecting mirror, transferring host galleria mellonella larvae on which the parasitic flies lay eggs into a container, supplementing the hosts again in the culture dish, continuously putting the host galleria mellonella into a breeding cage for breeding, and intensively placing the container for collecting the hosts to be laid, so that measures for preventing the harm of ants are taken; after raising and breeding for a period of time, taking out the fly pupae from the container, intensively placing the fly pupae into a new container, removing weak pupae, wherein the number of the fly pupae in each container is 60 +/-1, and sealing the container by roll paper; when the parasitic flies eclosion occurs, the container is moved into a new fly collecting cage, and a new round of breeding is started.
Further, the host is robust caenorhabditis elegans aged larva with the body length of more than 3 cm.
Further, the processing method of the processed host comprises the following steps: soaking larva of greater wax moth in 75% alcohol for 2min, taking out, soaking in 60 deg.C hot water for 10 s, taking out, and air drying.
Furthermore, the timing of transferring the larvae of the galleria mellonella into the rearing cage is that the time for taking out the larvae of the galleria mellonella each time and collecting the fly eggs is 2 hours after the larvae of the galleria mellonella move into the rearing cage for 3 days.
Furthermore, the container is a disposable transparent plastic cup, the application method comprises the steps of filling a crease on the bottom of the cup to flatten the roll paper before the oviposition host is moved in, covering a layer of same paper after 10 +/-1 heads of the egg is moved in, flattening tips of the paper, moving into the egg-laid greater wax moth, covering a layer of same paper, flattening tips of the paper until the number of the egg-laid greater wax moth larvae in each disposable transparent plastic cup reaches 50 +/-1, covering the mouth of the disposable transparent plastic cup with a section of roll paper, fixing the roll paper by a rubber band, and finally recording information such as date and the like on the paper by a marker pen.
Further, the period of taking the fly pupae out of the container is after the larva of the galleria mellonella, which is laid eggs, is moved into the disposable transparent plastic cup for 15-20 days.
The invention has the technical advantages and beneficial effects that:
1. in order to reduce the breeding cost of the Exorista civilis Rond, the technology screens the greater wax moth from the non-natural host of the Exorista civilis Rond, creatively uses simple treatment measures to treat the larvae of the greater wax moth, reduces the movement capacity and the spinning capacity of the greater wax moth, enables the larvae of the greater wax moth to become the best intermediate host of the Exorista civilis Rond at present, and enables the parasitism rate of the Exorista civilis Rond on the greater wax moth to be increased from 21% to over 60%.
2. The breeding feed for the host greater wax moth larvae used by the technology is easy to obtain, the breeding period is short, the breeding method is simple, no plant diseases and insect pests are generated in the breeding process, the survival rate is high, and the breeding cost is low. At present, the larvae of the wax moth have been commercialized and sold on the market, about 0.1 yuan/head, and a small amount of silkworms, beet armyworms and the like are sold on the market as commodities with the prices of at least 1 yuan/head and 5 yuan/head respectively.
3. The technology uses the larvae of the galleria mellonella hubner which has strong hunger-dependent ability, and the larvae of the galleria mellonella hubner treated by alcohol and hot water can continuously survive for more than 1 month without pupation, thereby avoiding the reduction of parasitic effect caused by the falling of the eggs of the gypsophila japonica caused by host molting.
4. The technique for breeding the Exorista civilis Rond has simple operation steps and high breeding efficiency.
Detailed Description
In the following, the embodiments of the present invention are described in further detail, and the examples are only one of the preferred embodiments of the present invention, and general changes and substitutions by those skilled in the art within the technical scope of the present invention should be included in the protection scope of the present invention.
The feed materials used in the following examples are commercially available unless otherwise specified; the greater wax moth is obtained from the breeding center of natural enemies of pests in Guangxi forestry; the seed source of the Japanese Pistis Chaetoceros is collected from the Bombycis Mori.
Example 1
(1) Collection of seed sources
Collecting mature larvae of Bombycis Mori, placing into fly collecting cage, collecting adult Bombycis Mori, transferring into new fly collecting cage as seed source, and removing weak and abnormal fly. 50 heads of the Japanese soldier fly were transplanted into each cage, and the ratio of male to female was 2-3: 2. 20% of honey water is put into the cages for supplementing nutrition for adults.
(2) Inoculation of
After the parasitoid fly is moved into a rearing cage for 3 days, 50 old larvae of the galleria mellonella treated by 75% alcohol and hot water of 60 ℃ are placed in a culture dish and put into the rearing cage, and parasitoid fly eggs are collected every 2 hours.
(3) Collection of fly-catching eggs
Taking out the culture dish, observing under a dissecting mirror, transferring the larvae of the galleria mellonella which have spawned by the parasitic flies into disposable transparent plastic cups, filling a fold on the bottom of the cup before transferring into the disposable transparent plastic cups, flattening the roll paper, covering a layer of the same paper after transferring into 10 heads, flattening tips, transferring into the galleria mellonella which has spawned, covering a layer of the same paper again, flattening the tips until 50 galleria mellonella larvae are transferred into each disposable transparent plastic cup. The mouth of the disposable transparent plastic cup is covered by a section of roll paper and fixed by a rubber band. Finally, information such as date is recorded on the paper with a marker pen. And (4) replenishing the larvae of the galleria mellonella in the culture dish and then continuously putting the larvae into the rearing cage.
(4) Management of fly-killing eggs in pupa stage
The disposable transparent plastic cups are placed in a centralized way, and measures for preventing the harm of ants are taken. And after 15-20 days, taking the fly pupae out of the cup, intensively placing the fly pupae into a new disposable transparent plastic cup, removing weak and small pupae, wherein the number of the fly pupae in each disposable transparent plastic cup is 60, and the disposable transparent plastic cup is sealed by roll paper.
(5) Management of adults
Observing the eclosion condition of the fly pupae every day, moving the plastic cup into a new rearing cage when the fly pupae eclosion occurs, and opening the disposable transparent plastic cup opening. Arranging the new rearing cage according to the method, placing the treated larva of the galleria mellonella 3d later, taking out the disposable transparent plastic cup after all the pupae of the parasitifer in the rearing cage are eclosized, and starting a new round of rearing and breeding.
The temperature and humidity condition test of the method comprises the following steps:
the influence of temperature and humidity factors on the propagation of the fly is researched by adopting a quantitative control method, and other factors are unchanged when the influence of a certain factor is researched. The following experiments were performed in 5 replicates per level. The constant factors are: the number of the parasitic flies per cage is 50, the number of the parasitic flies per larva of the greater wax moth is 1, and the lighting is 10h under the condition of 4O W headlights per day. The factors varied were:
temperature: 5 levels at 35 deg.C, 30 deg.C, 25 deg.C, 20 deg.C, 15 deg.C;
relative humidity: 90%, 75%, 60%, 45%, 30% 5 levels.
The test results are shown in tables 1 and 2. Test results show that the temperature of 25-30 ℃ is the optimal temperature range for the egg laying of the bactrocera dorsalis, the temperature of 20-30 ℃ is the optimal temperature range for the parasitism hosts of the bactrocera dorsalis, and the temperature of 25-30 ℃ is the optimal temperature range for pupation of larvae of the bactrocera dorsalis; the optimal humidity for spawning is 75%, 75-90% is the optimal humidity range of the host parasitized by the bactrocera dorsalis eggs, and 60-90% is the optimal humidity range of the pupation of the bactrocera dorsalis. According to the conclusion, the temperature is 25-30 ℃, the relative humidity is 75-90% which is the best temperature and humidity range of the parasitic effect of the Japan Exploris lucidi, and according to the climate conditions in Guangxi, the process adopts the conditions of 30 ℃ and 75% of relative humidity, so that the propagation cost can be greatly reduced.

Claims (2)

1. A method for breeding the Exorista civilis Roem by utilizing the wax moth is characterized in that: putting 60 +/-1 heads of pupae of the Japanese soldier fly pupae on the same day into a rearing cage, eliminating weak and malformed adult soldier fly pupae after eclosion, and ensuring that the number of the soldier fly pupae in each cage is 50 +/-1, and the male-female ratio is 2-3: 2; putting 20% of honey water into the cage for supplementing nutrition for the adult parasitic flies, putting the processed hosts in a culture dish, and putting the hosts into a feeding cage; taking out the culture dish after the parasitic flies lay eggs, observing under a dissecting mirror, transferring host galleria mellonella larvae on which the parasitic flies lay eggs into a container, supplementing the hosts again in the culture dish, continuously putting the host galleria mellonella into a breeding cage for breeding, and intensively placing the container for collecting the hosts to be laid, so that measures for preventing the harm of ants are taken; after raising and breeding for a period of time, taking out the fly pupae from the container, intensively placing the fly pupae into a new container, removing weak pupae, wherein the number of the fly pupae in each container is 60 +/-1, and sealing the container by roll paper; when the parasitic flies eclosion occurs, the container is moved into a new fly collecting cage, and a new round of breeding is started;
the host is robust wax moth mature larva with the body length of more than 3 cm;
the processing method of the processed host comprises the following steps: soaking larva of galleria mellonella in 75% alcohol for 2min, taking out, soaking in 60 deg.C hot water for 10 s, taking out, and air drying;
the timing of transferring the larvae of the galleria mellonella into the rearing cage is that the time for taking out the larvae of the galleria mellonella each time and collecting the fly eggs is 2 hours after the larvae of the galleria mellonella move into the rearing cage for 3 days;
the container is a disposable transparent plastic cup, the application method comprises the steps of filling a section of fold and flattening roll paper at the bottom of the cup before the oviposition host is moved in, covering a layer of same paper after 10 +/-1 heads are moved in, flattening tips, moving into the oviposition greater wax moth, covering a layer of same paper, flattening tips until the number of the larvae of the greater wax moth moved in each disposable transparent plastic cup reaches 50 +/-1, covering the mouth of the disposable transparent plastic cup with a section of roll paper, fixing the disposable transparent plastic cup with a rubber band, and finally recording date information on the paper with a marker pen;
the time for taking out the fly pupae from the container is after the larva of the galleria mellonella, which is laid eggs, is moved into the disposable transparent plastic cup for 15-20 days.
2. The method for propagating the rice fly larvae as claimed in claim 1, wherein the method comprises the steps of: the operation steps are as follows:
(1) collecting seed sources: placing collected mature larvae of the Bombycis Mori into a fly collecting cage, collecting adult Bombycis Mori, transferring into a new fly collecting cage as seed source, removing weak, small and malformed fly, transferring 50 heads of the Bombycis Mori into each cage with male-female ratio of 3:2, and placing 20% honey water into the cages for supplementing nutrition for the adult;
(2) inoculation: after the parasitic flies are moved into a rearing cage for 3 days, 50 old larvae of the greater wax moth treated by 75% alcohol and 60 ℃ hot water are placed in a culture dish and put into the rearing cage, and fly-killing eggs are collected every 2 hours;
(3) collecting the fly-killing eggs: taking out the culture dish, observing under a dissecting mirror, transferring the larvae of the galleria mellonella which have spawned by the parasitic flies into disposable transparent plastic cups, filling a crease on the bottom of each disposable transparent plastic cup before the larvae are transferred into the disposable transparent plastic cups, flattening the paper rolls, covering a layer of the same paper after 10 heads are transferred, flattening the tips of the paper rolls, transferring the eggs of the galleria mellonella which have spawned into the disposable transparent plastic cups, covering a layer of the same paper, flattening the tips of the paper rolls until 50 galleria mellonella larvae are transferred into each disposable transparent plastic cup, covering the mouths of the disposable transparent plastic cups with a section of the paper rolls, fixing the paper rolls by rubber bands, recording date information on the paper by marker pens, replenishing the galleria mellonella larvae in the culture dish again, and then continuously putting the galleria mellonella into a rearing cage;
(4) managing the fly-killing eggs in the pupal stage: collectively placing the disposable transparent plastic cups, taking measures for preventing ants from harming, after 15-20d, taking the fly pupae out of the cups, collectively placing the fly pupae into a new disposable transparent plastic cup, removing weak and small pupae, wherein the number of the fly pupae in each disposable transparent plastic cup is 60, and the disposable transparent plastic cups are sealed by roll paper;
(5) adult management: observing eclosion condition of the fly pupae every day, when the fly pupae eclosion occurs, moving the disposable transparent plastic cup into a new rearing cage, opening the mouth of the disposable transparent plastic cup, arranging the new rearing cage according to the method in the step (1),
3d, placing the treated larvae of the galleria mellonella, taking out the disposable transparent plastic cup after all the parasitic flies in the rearing cage are eclosized, and starting a new round of rearing and breeding;
the method comprises the following steps of: the illumination is 10h of headlamp illumination of 4O W each day; the temperature is 30 ℃; the relative humidity was 75%.
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