CN108990909B - Method for trapping bradysia odoriphaga live insects and feeding bradysia odoriphaga live insects indoors in large quantities - Google Patents
Method for trapping bradysia odoriphaga live insects and feeding bradysia odoriphaga live insects indoors in large quantities Download PDFInfo
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- CN108990909B CN108990909B CN201810519888.8A CN201810519888A CN108990909B CN 108990909 B CN108990909 B CN 108990909B CN 201810519888 A CN201810519888 A CN 201810519888A CN 108990909 B CN108990909 B CN 108990909B
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- A—HUMAN NECESSITIES
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- A23K10/00—Animal feeding-stuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/90—Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
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Abstract
The invention discloses a method for trapping and indoor mass feeding of bradysia odoriphaga live insects, and belongs to the technical field of agricultural scientific research and insect breeding. The method for trapping live insects of the invention is to utilize a method of burying fat pork and pleurotus geesteranus in a small flowerpot to induce female bradysia odoriphaga to lay eggs, and a large number of larvae are obtained by hatching eggs and are used as initial test insects. Simple and easy to operate, and can easily obtain a large amount of initial test insects. According to the feeding method, peat soil is used as a feeding environment, pleurotus geesteranus is used as a feed, the feed does not need to be replaced and excrement does not need to be cleaned in a growth period, labor and time are saved, test materials are easy to obtain, the cost is low, the operation is simple, various test observations are convenient to conduct, the close-relative propagation of offspring can be avoided, and the population can not be degraded due to the close-relative propagation in the perennial feeding. The survival rate of each insect state is very high, and more than 80 percent of eggs can develop into imagoes. Therefore, the invention can trap a large amount of larvae as an initial insect source in a short time and realize annual mass rearing of the bradysia odoriphaga.
Description
Technical Field
The invention belongs to the technical field of agricultural scientific research and insect breeding, and particularly relates to a method for trapping and massively feeding bradysia odoriphaga live insects indoors.
Background
Bradysia odoriphaga (Bradysia sp.) is a diptera, Hyphomycota, Myxocyprinus superfamily, the largest genus of the family Occidomycidae, Bradysia winnnertz, 455 species are known worldwide and 131 species are recorded in our country. The bradysia odoriphaga mainly comprises underground young tissue of Liliaceae plant such as leek, mycelium and fruiting body primordium of wild and cultivated edible fungi, and young underground stem and leaf of greenhouse garden flower plant. The bradysia odoriphaga larvae which are harmful to the leeks mainly eat pseudostems, bulbs, leaf sheaths, young stems and tender buds of the leeks, so that underground parts of the leeks are rotted, overground parts of the leeks are thin, weak, withered and yellow, even the whole leeks die, the quality of the leeks is seriously affected, and the yield is greatly reduced. The bradysia odoriphaga which is harmful to edible fungi takes mushroom mycelium, stipe, pleopodium, fruiting body primordium and the like from larvae, damages culture materials of the strains, causes fungus degeneration and primordium disappearance, blackens and rots fungus bags, turns yellow and sticks damaged fruiting bodies, leads the fruiting bodies to lose commodity value, reduces the quality of the mushrooms and reduces the yield. When the bradysia odoriphaga is harmful to greenhouse phalaenopsis seedlings, larvae are harmful to young roots and young leaves at the base, the damaged leaves are yellow, withered and shed, and young roots are damaged to form rotten roots and hollow roots, so that the production of the phalaenopsis seedlings is severely restricted. When the tissue culture seedlings take roots outside bottles of the Chinese cypress and the sandalwood which are harmful to greenhouse gardens, the bradysia odoriphaga gnaws the epidermis and the phloem or the tender xylem of the underground stem of garden plants such as the unrerooted Chinese cypress, the sandalwood and the like through larvae, so that the seedlings cannot take roots, and the seedlings which are too tender are rotten and lodged; the seedlings with higher lignification degree are left with bald xylem (as shown in b in figure 1), the overground part is not laid down, the false appearance of the survival seedlings is caused, but the seedlings are withered and yellow finally, and the plant damage rate can reach more than 90% when the bradysia odoriphaga is serious, thereby bringing serious threat to the propagation of the seedlings.
In order to research the scientific and efficient technology for preventing and treating the bradysia odoriphaga, the life history, the life habit, the occurrence rule, the biological characteristics and the like of the bradysia odoriphaga need to be systematically researched, a large number of larvae with the same insect age are needed to be used for preventing and treating experiments, and the acquisition of experimental insects and mass feeding are the basis for researching the insects.
Since eggs and pupae are difficult to find, sources of bradysia odoriphaga test insects are typically collected from larvae and adults. The larva collection is generally directly from the larva which is harmful on the occurring tunicate strain or the larva in the soil near the tunicate strain, because the bradysia odoriphaga larva is disturbed to quickly transfer hidden habits and has small larva bodies, if the occurrence of a not very serious plot is not very serious, the search for more initial larvae is very time-consuming and labor-consuming. Adult collection generally adopts the trematode, and the trematode inhales the adult and can cause the adult to be hurt, problem such as normal reproduction, because female adult flight ability is weaker, catches well, male adult flight ability is stronger, then difficult to catch with the trematode, and the trematode that is fit for this small insect also is difficult to purchase, self-control trematode more exists the difficult problem that lacks preparation material and preparation technique, and how the adult that the trematode inhales places in the environment that is fit for its existence and lay eggs and does not escape also the problem that faces. The late eye funguses occur in greenhouse forest flowers, one of the seedlings has 1 to several larvae which gnaw on the underground stem, so that the seedlings can not root, the emergence rate is seriously influenced, the larvae are transferred into harm or dispersed into soil to eat humus after gnawing one of the seedlings, therefore, the difficulty in searching more initial larvae in a greenhouse is higher, the seedlings are pulled out when insect sources are searched, sometimes misjudgment is carried out, the rooted seedlings are pulled out, artificial damage is caused to the seedlings, and the survival rate of the seedlings is further reduced.
The existing bradysia odoriphaga breeding technology is characterized in that fresh host feed is used for feeding, the feed needs to be replaced every day or every other day, or an edible fungus hypha culture medium needs to be prepared for feeding, the technical difficulty is high, or food residues and excrement need to be removed when the feed is artificially synthesized for feeding, so that the problems of complex operation, time consumption, labor consumption and the like exist.
Disclosure of Invention
In order to overcome the defects of the collection and feeding method of the live bradysia odoriphaga in the prior art, the invention aims to provide a method for trapping the live bradysia odoriphaga and feeding the live bradysia odoriphaga indoors in a large scale.
The invention can trap a large number of larvae as initial insect sources in a short time and realize annual mass feeding of the bradysia odoriphaga, and has the advantages of easily obtained test materials, low cost, simple operation, labor and time conservation, high survival rate of each insect state, easy observation and the like.
The purpose of the invention is realized by the following technical scheme:
a method for trapping and breeding a large number of bradysia odoriphaga live insects indoors comprises the following steps:
(1) live insect trapping: filling peat soil into the small flower pots, burying fat pork and pleurotus geesteranus as baits, placing the small flower pots in a greenhouse where bradysia odoriphaga occurs, and inducing female bradysia odoriphaga adults to lay eggs; after eggs are hatched into larvae, drilling the larvae into the soil to eat pleurotus geesteranus and fat pork; after 10-15 days, moving the small flowerpot to an insect breeding room, placing the small flowerpot under a dissecting mirror, turning out fat pork, and moving a large number of larvae on the fat pork and in the soil nearby to a culture dish;
(2) the breeding method comprises the following steps: adding peat soil and pleurotus geesteranus into the culture dish, and feeding larvae to adults;
(3) placing a culture dish with imagoes in an imago testing device, opening a dish cover, and flying the imagoes in the imago testing device; placing other culture dishes with imagoes in the imago test device to enable the imagoes from different culture dishes to be crossed;
(4) capturing single female adults after mating in the adult test device into a new culture dish, and enabling the female adults to lay eggs after 2-3 days;
(5) after eggs are hatched into larvae, adding peat soil and pleurotus geesteranus, culturing for 8-12 days, placing a culture dish under a dissecting mirror, poking soil blocks with tweezers, and searching for pupae;
(6) transferring the single pupa to a new culture dish, adding a proper amount of peat soil, covering handkerchief paper with a certain size and humidity on the peat soil, and allowing the pupa to emerge into adults within 1-3 d;
(7) male and female offspring from non-identical female adults are placed in the same adult test device to copulate, and close-relative propagation is avoided;
(8) capturing the copulated female imagoes to a culture dish, adding peat soil and pleurotus geesteranus after the female imagoes lay eggs, and repeating the feeding process to realize the purpose of feeding and breeding a large number of bradysia odoriphaga annually.
Preferably, the peat soil in the step (1) is smashed and then placed in a drying oven at the temperature of 60-80 ℃ for drying for 24-48 h, and tap water is added to prepare the peat soil with the relative humidity of 60-65%.
Preferably, the fat pork in the step (1) is raw fat pork and is cut into 1.0-1.5 cm3And (4) small blocks for standby.
Preferably, the depth of embedding fat pork and pleurotus geesteranus in the step (1) is 0.5-1.0 cm.
Preferably, the peat soil used in the steps (2), (5), (6) and (8) has a relative humidity of 60-65%, is placed in a tissue culture bottle with a diameter of 7cm and a height of 10cm, is placed in an autoclave, and is subjected to autoclaving at 121 ℃ and a temperature of 1.1kg/cm2Sterilizing under pressure for 40 min.
Preferably, the pleurotus geesteranus is pleurotus geesteranus pileus with the diameter of 2-3 cm or pleurotus geesteranus stipes with the length of 2-3 cm;
preferably, the pleurotus geesteranus is subpackaged into 9 x 13-12 x 18cm self-sealing bags, the self-sealing bags are placed indoors for 2-3 days, and released water is poured out.
Preferably, the feeding temperature of the insect feeding room is 25 +/-0.5 ℃, the relative humidity is 50-70%, and natural illumination is performed.
Adult test device, including being used for adult activity mating wide-mouth container 1 and wrapping up the white cotton 2 of 1 open end of wide-mouth container, the open end of white cotton 2 is provided with and is tied tightrope 3.
The open end of the wide-mouth container 1 is connected with one end of the white cotton cloth 2 through a staple.
The wide-mouth container 1 is a wide-mouth cylindrical transparent plastic container with a bottom; the edible oil tank is formed by refitting a 5L edible oil tank, a bottle opening and a bottle neck part of the oil tank are cut off, and the rest cylindrical part is used as a wide-mouth container 1 of the device.
The wide-mouth container 1 is 13-14-20-21 cm long and wide.
The length and the width of the white cotton cloth 2 are 52-56-40-42 cm; the length of the white cotton cloth 2 is similar to the circumference of the opening end of the wide-mouth container 1.
The length of the fastening rope 3 is 70-75 cm.
The fastening rope 3 is a white thread rope.
The method specifically comprises the following steps:
(1) preparing a culture dish for feeding: and (3) paving qualitative filter paper with the diameter of 9cm on the bottom layer of a disposable plastic culture dish with the diameter of 9cm, adding tap water to the filter paper by using a 10-15 cm long rubber dropper, and sucking open water on the filter paper in the culture dish by using other dry filter paper for later use.
(2) Preparing peat soil for feeding: smashing imported peat soil purchased in the flower market in the amount required by the experiment, adding tap water to prepare peat soil with the relative humidity of 60-65%, filling the peat soil into a tissue culture bottle with the diameter of 7cm and the height of 10cm, filling 3/4 of the volume of the tissue culture bottle, covering the tissue culture bottle, putting the tissue culture bottle into an autoclave, and sterilizing at the temperature of 121 ℃ and 1.1kg/cm2Sterilizing under pressure for 40min to kill various microorganisms in peat soil, such as bacteria, fungi, nematodes, mites, etc., and standing at room temperature.
(3) Preparing bait and feed for trapping insects and raising: the method comprises the steps of buying fat pork in quantity required by experiments on farmer markets, and cutting the fat pork into 1.0-1.5 cm in a pest breeding room3The small blocks are placed in a refrigerator at the temperature of 18 ℃ below zero for freezing storage, and are taken out for thawing for later use. The method comprises the steps of purchasing pleurotus geesteranus in the quantity required by experiments on farmer markets, subpackaging the pleurotus geesteranus into self-sealing bags of 9 x 13-12 x 18cm in an insect breeding room (feed is not suitable to be exposed in the air so as to prevent some parasitic fly insects from spawning and breeding in food and polluting the food), placing the pleurotus geesteranus for 2-3 days at room temperature to enable moisture in the pleurotus geesteranus to be naturally released, pouring off moisture in the bags, sealing, placing the bags in a refrigerator at-18 ℃ for freezing, taking out the pleurotus geesteranus before use, and using the pleurotus geesteranus after thawing.
(4) Making an adult test device: cutting off the bottle mouth part of the edible oil barrel with the volume of 5L, and leaving a wide-mouth container part with the length, width and height of about 13-14, 20-21 cm. And sewing a piece of white cotton cloth with the length and width of 52-56-40-42 cm into a cylinder, wherein the circumference of the cylinder is equal to that of the wide-mouth container and is about 52-56 cm, one end of the cylinder is sleeved on the open end of the wide-mouth container and is connected together by a staple, the other end of the white cotton cloth cylinder is threaded with a piece of white thread rope which can be pulled tightly, a movable opening is formed, and the adult test device is completed.
(5) Live insect trapping method
The imports purchased in the flower marketThe peat soil is smashed according to the quantity required by the test, the peat soil is placed in a drying box at the temperature of 60-80 ℃ and dried for 24-48 hours to kill various micro organisms in the peat soil, the peat soil is taken out and added with tap water to prepare the peat soil with the relative humidity of 60-65%, the peat soil is placed in small plastic flowerpots with the upper caliber of 9cm, the lower caliber of 6cm and the height of 8cm, the number of the flowerpots is determined according to the test requirement, each flowerpot is filled with the peat soil with the relative humidity of 60-65% of 5/6 volume, and the peat soil is compacted. The center of the flowerpot is embedded with 1.0-1.5 cm3A small square of raw fat pork and a piece of Pleurotus geesteranus pileus with a diameter of 2-3 cm or Pleurotus geesteranus stipe with a length of 2-3 cm, embedded to a depth of 0.5-1.0 cm. The flowerpot is placed in a greenhouse where the late-eyed funguses occur, the late-eyed funguses female adults can lay eggs in soil seams of the flowerpot, and the eggs are hatched into larvae and then are drilled into the soil to eat the fat pork and the mushrooms. Get back the insect room with the flowerpot after 10 ~ 15d, during according to soil humidity drench water management in the flowerpot, keep soil relative humidity at 60 ~ 65%. Taking back the flowerpot of the insect breeding room, directly placing the flowerpot under an anatomical lens for microscopic examination, turning out fat pork by using forceps, wherein a large number of 3-4-year-old late eye mushroom mosquito larvae are in motion on the fat pork and in soil near the fat pork, hundreds of larvae can be found in the flowerpot with large trapping amount, and gently clamping the larvae out of the culture dish prepared in the step (1) by using the forceps. In order to prevent clamping the larvae, a small amount of peat soil near the larvae can be clamped out together. Each culture dish can hold about 30-50 larvae.
Method for mass indoor breeding
(6) And (3) placing the larvae lured to be collected in the step (5) in the culture dish prepared in the step (1) and then immediately adding the peat soil prepared in the step (2) to the volume of the culture dish 3/4, placing 2-3 pleurotus geesteranus prepared in the step (3) on the peat soil, covering the dish cover, sleeving the dish cover and the dish bottom together by using a rubber band to prevent the dish cover from being opened inadvertently, and when imagoes occur, enabling the imagoes to fly out and escape. A plurality of culture dishes can be sleeved together to save culture space.
(7) After the larvae are raised for more than 10 days, the adult larvae can be seen in the culture dish. And (3) moving the culture dish with the imagoes to an imago test device, opening a dish cover, flying the imagoes to the imago test device, and tightening the imagoes by a white line to prevent the imagoes from escaping. Adults in different culture dishes can be placed in the same adult test device, and tens of adults can be placed in one adult test device. And (3) fully mating the male and female adults in the adult test device, capturing the single female adult after mating into the culture dish prepared in the step (1) after 30-60 min, and sleeving the dish bottom and the dish cover with a rubber band to wait for the female adult to lay eggs without any operation.
(8) And after 2-3 days, the female adults can lay eggs, the egg laying duration of the female adults is 4-8 hours, and the eggs die quickly after the eggs are laid. The female adults lay eggs on folds formed by the filter paper and the dish wall of the culture dish (as shown in fig. 7), or lay the eggs directly on the filter paper close to the dish wall, wherein the eggs are mostly laid by more than two or even dozens of eggs, and the eggs are rarely scattered by a single egg. The amount of oviposition can now be examined under a dissecting scope. At this time, depending on the humidity of the filter paper, a few drops of tap water were dropped onto the filter paper without eggs by a dropper, so as to maintain the appropriate humidity in the petri dish, and the eggs were allowed to normally hatch. The eggs are hatched into small larvae after about 3 days, the number of the unhatched eggs can be investigated under a dissecting mirror at the moment, and the egg hatchability is calculated. According to the egg laying amount investigation of 15 female adults, the egg laying amount of 1 female bradysia odoriphaga adults is 46-192 grains, the average is 120.67 grains, and the egg hatchability is 61.5-98.1%.
(9) After eggs are hatched into small larvae, adding 3/4-volume peat soil prepared in the step (2) into a culture dish, placing 2-3 pleurotus geesteranus prepared in the step (3) on the peat soil, covering a dish cover, and sleeving the dish cover and the dish bottom together by using a rubber band. Before the adult emergence period, the dish cover can be opened and placed under a dissecting mirror, and then the soil block and the pleurotus geesteranus are turned over by using forceps to observe the development conditions of the larvae and the pupae.
(10) In order to prevent the close-relative mating propagation of the offspring of the same female adult in the same dish, the pupae need to be fed separately in the pupal stage. Under the condition that the culture temperature is 25 +/-0.5 ℃, the pupa growing period is about 16-20 days after the female adult spawns, at the moment, the culture dish is placed under a dissecting mirror, the soil block is poked by using tweezers, the pupa can be found in the middle of the soil block, and at the moment, the male and female pupas can be identified. Because the pupae are very small, the soil at the tail part of the pupae needs to be completely removed for identifying the male and female pupaes, and the pupae can be injured in the removing process, so that the male and female pupaes do not need to be identifiedIn the case, the pupae and the soil near the body can be taken out to the culture dish prepared in the step (1), because the pupae needs high humidity for development and is not enough, the pupae skin can be dried, and the pupae can not be eclosized, the body of the pupae needs to be covered with the peat soil prepared in the step (2) with the length, the width and the height of about 3-4, 0.8-1.0 cm, and then 3-4 cm is used2The small piece of wet handkerchief paper is covered on peat soil to increase the culturing humidity of pupa, then the dish cover is covered, and the dish bottom and the dish cover are sleeved by a rubber band. Pupae emerge into adults after 1-3 days, and the adults can move in the culture dish. The method can make eclosion rate of pupa reach more than 95%. At this time, the culture dish was placed under a dissecting mirror without opening the dish lid to identify the male and female adult insects. And (4) putting male and female adults from different dishes into the adult test device prepared in the step (4) for copulation. Through experimental observation, the female and male adults begin to copulate in the adult test device within 10 seconds at the fastest speed and within 4 minutes at the slowest speed, the copulation time is 2-35 min, and after the copulation is completed, the female adult is captured into the culture dish prepared in the step (1) and the female adult is waited to lay eggs. The male adults have multiple copulation habits, and the male adults left in the adult test device can continue to copulate with other female adults.
The culture process is repeated to realize the purpose of breeding a large amount of bradysia odoriphaga yearly.
The feeding process is carried out in a constant-temperature insect feeding room, the feeding temperature is 25 +/-0.5 ℃, the relative humidity is 50-70%, and natural illumination is carried out.
Compared with the prior art, the invention has the following advantages and effects:
(1) the method for trapping live insects of the invention is to utilize a method of burying fat pork and pleurotus geesteranus in a small flowerpot to induce female bradysia odoriphaga to lay eggs, and a large number of larvae are obtained by hatching eggs and are used as initial test insects. Simple and easy to operate, and can easily obtain a large amount of initial test insects.
(2) According to the feeding method, peat soil is used as a feeding environment, pleurotus geesteranus is used as a feed, the feed does not need to be replaced and excrement does not need to be cleaned in a growth period, labor and time are saved, test materials are easy to obtain, the cost is low, the operation is simple, various test observations are convenient to conduct, the close-relative propagation of offspring can be avoided, and the population can not be degraded due to the close-relative propagation in the perennial feeding. The survival rate of each insect state is very high, and more than 80 percent of eggs can develop into imagoes.
Drawings
FIG. 1 shows the subterranean stem (a) of the bradysia odoriphaga larva, pinus massoniana, and the xylem (b) remaining after gnawing.
FIG. 2 shows a feed for Pleurotus geesteranus.
FIG. 3 shows that fat pork and Pleurotus geesteranus are added into a small flowerpot to induce female adults to lay eggs.
FIG. 4 shows a tissue culture bottle sterilized by adding peat soil with suitable humidity under high temperature and high pressure.
FIG. 5 is a petri dish with filter paper laid at an appropriate humidity.
FIG. 6 shows the culture dish containing peat soil, Pleurotus geesteranus feed (a) and multiple culture dishes nested together (b).
FIG. 7 shows the slow eye mushroom laying eggs on the folds formed by the filter paper and the culture dish wall.
FIG. 8 shows how the larvae eat Pleurotus geesteranus feed.
FIG. 9 is pupae of the bradysia odoriphaga.
FIG. 10 is the rearing of mosquito pupae of Armillaria esculenta.
FIG. 11 shows an adult test device (a) and an adult test device (b) that is fastened during an adult mating test.
FIG. 12 is a schematic view of an adult test device.
FIG. 13 shows the cross-breeding of male and female adults in the adult test device.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
Example 1
(1) Preparing a culture dish for feeding: a qualitative filter paper with the diameter of 9cm is laid on the bottom layer of a disposable plastic culture dish with the diameter of 9cm, tap water is added onto the filter paper through a 10-15 cm long rubber head dropper, and the clear water on the filter paper in the culture dish is sucked dry through another piece of dry filter paper for later use (as shown in figure 5).
(2) Preparing peat soil for feeding: general flower marketSmashing the purchased imported peat soil in the amount required by the test, adding tap water to prepare peat soil with the relative humidity of 60-65%, filling the peat soil into a tissue culture bottle with the diameter of 7cm and the height of 10cm, filling 3/4 of the volume of the tissue culture bottle, covering the tissue culture bottle with a cover, putting the tissue culture bottle into an autoclave, and sterilizing at the temperature of 121 ℃ and the pressure of 1.1kg/cm2Sterilizing under pressure for 40min, taking out, and standing at room temperature (as shown in FIG. 4).
(3) Preparing bait and feed for trapping insects and raising: the method comprises the steps of buying fat pork in quantity required by experiments on farmer markets, and cutting the fat pork into 1.0-1.5 cm in a pest breeding room3The small blocks are placed in a refrigerator at the temperature of 18 ℃ below zero for freezing storage, and are taken out for thawing for later use. Buying the pleurotus geesteranus required by the experiment on the farmer market, subpackaging the pleurotus geesteranus into 9 x 13-12 x 18cm self-sealing bags in a worm breeding chamber, placing the bags at room temperature for 2-3 days to naturally release water in the pleurotus geesteranus, pouring the water in the bags, sealing the bags, placing the bags in a refrigerator at-18 ℃ for freezing, taking out the bags before use, and using the bags after thawing (as shown in figure 2).
(4) Making an adult test device: adult test device, its schematic diagram is shown in fig. 12, including being used for adult activity mating wide-mouth container 1 and wrapping up the white cotton 2 of wide-mouth container open end, the open end of white cotton is provided with and is tied rope 3. The open end of the wide-mouth container 1 is connected with one end of the white cotton cloth 2 through a staple. The wide-mouth container 1 is a wide-mouth cylindrical transparent plastic container with a bottom; the edible oil tank is formed by refitting a 5L edible oil tank, a bottle opening and a bottle neck part of the oil tank are cut off, and the rest cylindrical part is used as a wide-mouth container 1 of the device. The wide-mouth container 1 is 13-14-20-21 cm long and wide. The length and width of the white cotton cloth 2 are 52-56-40-42 cm, the white cotton cloth is sewn into a cylindrical shape, and one end of the cylindrical shape is bound with the opening end of the wide-mouth container 1 through a staple. The length of the fastening rope 3 is 70-75 cm. The fastening rope 3 is a white thread rope.
The adult test devices thus produced and the adult test devices fastened at the time of adult mating test are shown in fig. 11 a and 11 b.
(5) Live insect trapping method
Purchase flowers on the marketSmashing the bought imported peat soil in an amount required by a test, placing the smashed peat soil in a drying box at 60-80 ℃, drying for 24-48 h, taking out, adding tap water to prepare peat soil with relative humidity of 60-65%, placing the peat soil into small plastic flowerpots with an upper caliber of 9cm, a lower caliber of 6cm and a height of 8cm, filling each flowerpot with peat soil with relative humidity of 60-65% in 5/6 volume according to the amount required by the test, and compacting. The center of the flowerpot is embedded with 1.0-1.5 cm3Burying a small square of raw fat pork and a piece of Pleurotus geesteranus pilus with a diameter of 2-3 cm or Pleurotus geesteranus stipe with a length of 2-3 cm into a depth of 0.5-1.0 cm (as shown in FIG. 3), placing the flowerpot in a greenhouse where delayed eye muscae volitantes occur, and taking the flowerpot back to the insect-raising room after 10-15 days. During the period, the relative humidity of the soil is kept between 60 and 65 percent according to the water sprinkling management of the soil humidity in the flowerpot. Directly placing the flowerpot of the insect-raising room under an anatomical lens for microscopic examination, turning over fat pork by using forceps, wherein a large amount of 3-4-year-old late-eyed muscae volitantes larvae are in motion on the fat pork and in soil near the fat pork. And (4) slightly clamping the larvae out of the culture dish prepared in the step (1) by using forceps. In order to prevent clamping the larvae, a small amount of peat soil near the larvae can be clamped out together. Each culture dish can hold about 30-50 larvae.
The purpose of putting fat pork and mushrooms into the flowerpot for trapping live insects is that the mushrooms are the best foodstuff which is favored by the bradysia odoriphaga to take food and has the most comprehensive nutrition and can ensure the normal development of the bradysia odoriphaga in life, if only mushroom baits are embedded in fruit soil, the mushrooms are taken to eat or decay in the soil quickly, and then larvae can move dispersedly and are not easy to find. After the fat pork is put in, the fat pork is not easy to rot and has attraction effect on the bradysia odoriphaga larvae, and after the bradysia odoriphaga eating the mushrooms, the bradysia odoriphaga will be attracted to the surroundings by the smell of the fat pork, so that the larvae are easy to find. If only fat pork is put in the flowerpot, the bradysia odoriphaga will develop badly and die due to the single nutrition of the fat pork. The fat pork and the mushrooms are combined to be used as baits, so that the respective defects are overcome, and the purpose of trapping a large number of well-developed live bradysia odoriphaga is achieved.
Method for mass indoor breeding
(6) And (3) placing the larvae trapped in the step (5) in the culture dish prepared in the step (1) and then immediately adding the peat soil prepared in the step (2) to the volume of the culture dish 3/4, placing 2-3 pieces of pleurotus geesteranus prepared in the step (3) on the peat soil, covering the dish cover, sleeving the dish cover and the dish bottom together by using a rubber band, and sleeving a plurality of culture dishes together to save the culture space (as shown in a in figure 6 and b in figure 6).
(7) After the larvae are raised for more than 10 days, the adult larvae can be seen in the culture dish. And (3) moving the culture dish with the imagoes to an imago test device, opening a dish cover, flying the imagoes to the imago test device, and tightening the imagoes by a white line to prevent the imagoes from escaping. Adults in different culture dishes can be placed in the same adult test device, and tens of adults can be placed in one adult test device. And (3) fully mating the male and female adults in the adult test device, capturing the single female adult after mating into the culture dish prepared in the step (1) after 30-60 min, and sleeving the dish bottom and the dish cover with a rubber band to wait for the female adult to lay eggs without any operation. Adult females were crossed in the adult test apparatus as shown in FIG. 13.
(8) After 2-3 days, the female adult can lay eggs (as shown in figure 7), and the eggs die soon after laying. The amount of oviposition can now be examined under a dissecting scope. Then, according to the humidity of the filter paper, a few drops of tap water are dropped on the filter paper without eggs by using a rubber head dropper so as to keep the proper humidity in the culture dish and ensure that the eggs are normally hatched. The eggs are hatched into small larvae after about 3 days, the number of the unhatched eggs can be investigated under a dissecting mirror at the moment, and the egg hatchability is calculated. According to the egg laying amount investigation of 15 female adults, the egg laying amount of 1 female bradysia odoriphaga adults is 46-192 grains, the average is 120.67 grains, and the egg hatchability is 61.5-98.1%.
(9) After eggs are hatched into small larvae, 3/4 volumes of peat soil prepared in the step (2) is added into a culture dish, 2-3 pleurotus geesteranus prepared in the step (3) are placed on the peat soil, a dish cover is covered, the dish cover and the dish bottom are sleeved together through a rubber band, and a larva breeding process is started (as shown in fig. 6 and 8).
(10) In order to prevent the close-relative mating propagation of the offspring of the same female adult in the same dish, the pupae need to be fed separately in the pupal stage. Under the condition that the culture temperature is 25 +/-0.5 ℃, the pupa growing period is about 16-20 days after the female adult spawns,at this point, the petri dish was placed under a dissecting scope, and the soil mass was removed with forceps, and pupae were found in the middle of the soil mass (as shown in fig. 9). In order to avoid pupae injury, the pupae and soil near the body are taken out to the culture dish prepared in the step (1) together by using tweezers, the upper cover of the pupae body is covered with peat soil prepared in the step (2) with the length, width and height being about 3-4, 0.8-1.0 cm, and then 3-4 cm is used2The small piece of wet handkerchief paper is covered on peat soil (as shown in figure 10) to increase the humidity of pupa culture, and then the dish cover is covered, and the bottom and the dish cover are sleeved with rubber bands. Pupae emerge into adults after 1-3 days, and the adults can move in the culture dish. At this time, the culture dish was placed under a dissecting mirror without opening the dish lid to identify the male and female adults. Male and female adults from different dishes are placed into the adult test device prepared in step (4) for copulation (as shown in FIG. 13). And (3) after mating is finished, catching the female imagoes into the culture dish prepared in the step (1) and waiting for the female imagoes to lay eggs. The male adult is left in the adult test device and can continue to copulate with other female adults.
The culture process is repeated to realize the purpose of breeding a large amount of bradysia odoriphaga yearly.
The feeding is carried out in a constant-temperature insect breeding room, the feeding temperature is 25 +/-0.5 ℃, the relative humidity is 50-70%, and the natural illumination is carried out.
Example 2
Preparing non-copulated male and female adults: the adult bradysia odoriphaga is small, and unlike large insects, female adults and male adults can be simply placed together for cross breeding and other experimental observations. The artificial breeding of the larvae is generally carried out in a culture dish with the diameter of 9cm, and offspring male and female adults from one female adult are generated in the same culture dish, so that the close-relative breeding is easily caused. When pupa stage occurs, single pupa is placed in a separate culture dish for breeding (as shown in figure 10), and pupa eclosion can obtain female adult or male adult which are not crossed.
The application method of the adult test device comprises the following steps: a pair of culture dishes for generating male and female adults are placed into a wide-mouth container 1 from the open end of a white cotton cloth 2 of an adult test device in sequence, the cover of the culture dishes is opened, the adults are placed into the adult test device, the culture dishes are taken out, and then the open end of the white cotton cloth 2 is fastened, so that the copulation of the male and female bradysia odoriphaga adults in the device can be observed (as shown in figure 13).
Capturing female adults after mating: after the sex adult mating is accomplished, will spread the culture dish that has moist filter paper and arrange wide-mouth container 1 in, one hand operation, open the dish lid about 30 degrees angles after, external force forces the sex adult to fly, and operation open-ended culture dish catches in the sex adult to the culture dish, covers the dish lid after catching, and outside shifting out adult test device, continue the cultivation of sex adult. The method for catching male adults is the same as that for catching female adults.
The method for determining the number of times of male adult cross-overs comprises the following steps: placing the culture dish containing the male imago which is obtained by breeding single pupae and has not crossed the tail into an imago testing device from the opening end of the white cotton cloth 2, opening the dish cover, placing the male imago into the wide-mouth container 1, taking out the culture dish from the opening end of the white cotton cloth 2, then placing the culture dish containing the female imago which is obtained by breeding single pupae and has not crossed the tail into the same imago testing device by the same method, and fastening the opening end of the white cotton cloth 2 by the fastening rope 3 to prevent the imago from escaping. At this time, it can be observed that the female adults flying into the wide-mouth container 1 have weak flight ability, generally only crawl along the wall of the wide-mouth container 1, fly occasionally, while the male adults have strong flight ability, generally fly in the wide-mouth container 1 continuously, the female adults can be found within 10 seconds to 5 minutes, the male adults curl the end of the abdomen towards the abdomen, are close to the end of the abdomen of the female adults, clamp the end of the abdomen of the female adults by a clasper, and then arrange a line to start to meet (as shown in fig. 13). And recording the time of the transaction. And after the mating is finished, the female adults are captured and moved out of the device by using a culture dish paved with filter paper with proper humidity. And then the female adult with the head not crossed with the tail is put in the female adult, and the rest is done in the same way until the male adult does not cross with the tail. The cross-over of the 4 male adults tested is shown in table 1. Table 1 shows that the male imagoes of the bradysia odoriphaga can be crossed 4-6 times, the crossing time is 2 '02' (2 minutes 02 seconds) to 35 '34' (35 minutes 34 seconds), and the crossing time is longer and longer as the crossing times of the male imagoes are increased.
TABLE 1 test of the number of crossovers of adult males
The method for determining the ratio of the adult female to the adult male of the single female generation comprises the following steps: the maximum egg laying of the female bradysia odoriphaga adults can reach as many as 200, the average egg laying is 120, the highest raising survival rate can reach more than 80 percent, and therefore dozens of adults can emerge every day in the peak period of adult emergence. The eclosion duration of the single-female offspring adults is 10-15 d, and the number of the adults generated per day is 1-50. The adult insect is in a peak period, and the investigation of the sex ratio is time-consuming and labor-consuming. The use of an adult test device simplifies and makes the test accurate. The method comprises the steps of moving a culture dish with imagoes from the open end of a white cotton cloth 2 of an imago test device to a wide-mouth container 1 every 1-2 days, opening a dish cover, flying the imagoes into the wide-mouth container 1, moving out the culture dish, and fastening the open end of the white cotton cloth 2 by a fastening rope 3. Due to the small volume of the device, the whole device with the live imagoes can be put into a refrigerator with the temperature of 18 ℃ below zero for freezing for 20min, and after being taken out, the dead imagoes are poured into a culture dish and identified under a dissecting mirror. The female-male ratio can be calculated by integrating the number of the female-male adults in the whole adult generation period, and the female-male ratio of the delayed muscae volitantes is 1.66: 1.
the above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A method for trapping and breeding a large number of bradysia odoriphaga live insects indoors is characterized by comprising the following steps:
(1) live insect trapping: filling peat soil into the small flower pots, burying fat pork and pleurotus geesteranus as baits, placing the small flower pots in a greenhouse where bradysia odoriphaga occurs, and inducing female bradysia odoriphaga adults to lay eggs; after eggs are hatched into larvae, drilling the larvae into the soil to eat pleurotus geesteranus and fat pork; after 10-15 days, moving the small flowerpot to an insect breeding room, placing the small flowerpot under a dissecting mirror, turning out fat pork, and moving a large number of larvae on the fat pork and in the soil nearby to a culture dish;
(2) the breeding method comprises the following steps: adding peat soil and pleurotus geesteranus into the culture dish, and feeding larvae to adults;
(3) placing a culture dish with imagoes in an imago testing device, opening a dish cover, and flying the imagoes in the imago testing device; placing other culture dishes with imagoes in the imago test device to enable the imagoes from different culture dishes to be crossed;
the adult test device comprises a wide-mouth container for the activity copulation of adults and white cotton cloth wrapping the open end of the wide-mouth container, wherein the open end of the white cotton cloth is provided with a tying rope;
(4) capturing single female adults after mating in the adult test device into a new culture dish, and enabling the female adults to lay eggs after 2-3 days;
(5) after eggs are hatched into larvae, adding peat soil and pleurotus geesteranus, culturing for 8-12 days, placing a culture dish under a dissecting mirror, poking soil blocks with tweezers, and searching for pupae;
(6) transferring the single pupa to a new culture dish, adding a proper amount of peat soil, covering handkerchief paper with a certain size and humidity on the peat soil, and allowing the pupa to emerge into adults within 1-3 d;
(7) male and female offspring from non-identical female adults are placed in the same adult test device to copulate, and close-relative propagation is avoided;
(8) capturing the copulated female imagoes to a culture dish, adding peat soil and pleurotus geesteranus after the female imagoes lay eggs, and repeating the feeding process to realize the purpose of feeding and breeding a large number of bradysia odoriphaga annually.
2. The method for trapping and mass-rearing of bradysia odoriphaga live insects according to claim 1, wherein:
and (2) mashing the peat soil in the step (1), drying at 60-80 ℃ for 24-48 h, and adding tap water to prepare the peat soil with a relative humidity of 60-65%.
3. The method for trapping and mass-rearing of bradysia odoriphaga live insects according to claim 1, wherein:
the fat pork in the step (1) is raw fat pork,cutting into 1.0-1.5 cm3And (4) small blocks for standby.
4. The method for trapping and mass-rearing of bradysia odoriphaga live insects according to claim 1, wherein:
the depth of embedding fat pork and pleurotus geesteranus in the step (1) is 0.5-1.0 cm.
5. The method for trapping and mass-rearing of bradysia odoriphaga live insects according to claim 1, wherein:
the peat soil used in the steps (2), (5), (6) and (8) has a relative humidity of 60-65%, is filled into a tissue culture bottle with a diameter of 7cm and a height of 10cm, and is cultured at a temperature of 121 ℃ and a concentration of 1.1kg/cm2Sterilizing under pressure for 40 min.
6. The method for trapping and mass-rearing of bradysia odoriphaga live insects according to claim 1, wherein:
the pleurotus geesteranus is pleurotus geesteranus pileus with the diameter of 2-3 cm or pleurotus geesteranus stipes with the length of 2-3 cm.
7. The method for trapping and mass-rearing of bradysia odoriphaga live insects according to claim 1, wherein:
the pleurotus geesteranus is subpackaged into self-sealing bags of 9 x 13-12 x 18cm, placed indoors for 2-3 d, and released water is poured out.
8. The method for trapping and mass-rearing of bradysia odoriphaga live insects according to claim 1, wherein:
the feeding temperature of the insect feeding room is 25 +/-0.5 ℃, the relative humidity is 50-70%, and natural illumination is performed.
9. The method for trapping and mass-rearing of bradysia odoriphaga live insects according to claim 1, wherein:
the opening end of the wide-mouth container is connected with one end of the white cotton cloth through a staple;
the wide-mouth container is a cylindrical transparent plastic container with a wide mouth and a bottom.
10. The method for trapping and mass-rearing of bradysia odoriphaga live insects according to claim 1, wherein:
the length, width and height of the wide-mouth container are 13-14, 20-21 cm;
the length and the width of the white cotton cloth are 52-56-40-42 cm;
the length of the tying rope is 70-75 cm;
the tightening rope is a white thread rope.
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| KR100421779B1 (en) * | 2001-04-16 | 2004-03-10 | 삼성에버랜드 주식회사 | Korean entomopathogenic nematode, steinernema carpocapsae gsn 1 and control method for insect |
| CN104798734A (en) * | 2015-05-19 | 2015-07-29 | 天津市植物保护研究所 | Culture medium and method for feeding predatory coenosia larvae indoors |
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