Method for artificial rearing of callosobruchus maculatus larvae and rearing device

Abstract

The application discloses a kind of artificial feeding method and feeding device of Callosobruchus maculatus larva.The application method is specifically as follows: after the seed embryo powder of legume seed is sifted through 50-70 mesh screen, it is mixed into group with pure water at the ratio of 1g:0.4-0.6mL, the surface of feed is shaped, and then a plurality of feeding grooves are inserted on the surface of feed, one Callosobruchus maculatus larva is picked into each feeding groove, and the Callosobruchus maculatus larva is cultured to eclosion under suitable environment.The application device comprises a feeding dish, a pressing plate for feed shaping and a punch for inserting a plurality of feeding grooves.The application method realizes artificial feeding of Callosobruchus maculatus larva to adult stage, has high survival rate, and is suitable for large-scale breeding.The application device can simply and quickly make Callosobruchus maculatus larva feed.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of insect rearing and the field of invasion biology, and particularly relates to a method and device for artificial rearing of Callosobruchus maculatus larvae. BACKGROUND

[0002] Callosobruchus maculatus (Fabricius) is an insect of the family Bruchidae in the order Coleoptera, and belongs to the genus Callosobruchus. The insects of the genus Callosobruchus, including Callosobruchus maculatus, are harmful quarantine organisms. Callosobruchus maculatus is a pest of the genus Callosobruchus that is widely distributed and has a relatively serious degree of harm. As a seed storage pest of legume crops, Callosobruchus maculatus mainly feeds on legume seeds in the form of a larva in a latent stage, thereby reducing the germination rate of the legume seeds and causing quality loss. Moreover, due to the strong reproductive capacity of Callosobruchus maculatus, the seeds can be completely bored in a few months, and the potential harm is huge.

[0003] Due to the habit of Callosobruchus maculatus larvae of hiding in legume seeds, and the insufficient biological research methods for Callosobruchus maculatus, it is difficult for the larvae to continue to feed after being peeled off from the legume seeds, and it is difficult to complete the generation development. The traditional rearing method of Callosobruchus maculatus larvae uses beans and must be reared from eggs to adults, which has a low rearing efficiency and is not conducive to the research on the larvae and subsequent development stages of Callosobruchus maculatus. SUMMARY

[0004] In view of the deficiencies of the prior art, the present application provides an artificial rearing method and a rearing device for Callosobruchus maculatus larvae. The artificial rearing method provided by the present application enables the Callosobruchus maculatus larvae peeled off from legume seeds to continue to develop to the adult stage, realizes artificial feed culture of Callosobruchus maculatus larvae, and has low rearing cost, high rearing efficiency of Callosobruchus maculatus larvae, and realizes scale rearing of Callosobruchus maculatus larvae.

[0005] To achieve the above-mentioned purpose, the technical solution adopted by the present application is as follows:

[0006] I. An artificial rearing method for Callosobruchus maculatus larvae

[0007] The artificial rearing method comprises the following steps:

[0008] 1) mixing a legume seed embryo powder with water to obtain a doughy feed, placing the doughy feed in a rearing dish, shaping the doughy feed into a cake shape, and inserting a plurality of holes on the surface of the cake-shaped feed to form a plurality of rearing grooves for accommodating Callosobruchus maculatus larvae;

[0009] In the step 1), the legume seed embryo powder is mixed with water after being sieved through a 50-70 mesh sieve.

[0010] In the step 1), the mass-volume ratio of the embryo powder to water is 1g: 0.4-0.6mL.

[0011] Specifically, the step 1) is that the embryo powder of peeled legume seeds is mixed with water and kneaded into a dough to obtain a doughy feed, then the doughy feed is placed in a feeding dish, the doughy feed is pressed by a pressing plate to shape the doughy feed into a cake, and after standing at room temperature for 1-2h, a puncher is used to uniformly insert a plurality of holes on the surface of the cake-shaped feed to form a plurality of feeding grooves.

[0012] In the step 1), the inner diameter of the feeding groove is 2-3mm, the depth of the feeding groove is 1.5-2mm, and the Callosobruchus maculatus larvae are collected at 12-14d after oviposition.

[0013] In the step 1), the ratio of the number of the feeding grooves to the area of the feed surface is 30-40: 39-41cm 2 .

[0014] 2) The pre-collected Callosobruchus maculatus larvae are picked into the feeding grooves;

[0015] 3) The feeding dish with the picked Callosobruchus maculatus larvae is placed in a suitable environment for cultivation until the Callosobruchus maculatus larvae emerge;

[0016] In the step 3), the suitable environment is specifically: temperature of 28-32℃, humidity of 60-65% RH;

[0017] In the step 3), after 6-8d of cultivation, the Callosobruchus maculatus larvae start to emerge.

[0018] II. A feeding device for Callosobruchus maculatus larvae

[0019] The feeding device comprises a feeding dish, a pressing plate and a puncher; the pressing plate is used to shape the feed, so that the doughy feed placed in the feeding dish is pressed into a cake shape and the surface of the cake-shaped feed is flat; the puncher is used to insert a plurality of feeding grooves;

[0020] A plurality of through holes are uniformly provided on the pressing plate, and a plurality of punch columns are uniformly arranged on the bottom surface of the puncher; the number and arrangement position of the punch columns are the same as those of the through holes on the pressing plate, and the arrangement positions are vertically aligned one by one; when the puncher is pressed from top to bottom, the bottom end of each punch column penetrates through the corresponding through hole and inserts into the cake-shaped feed below the pressing plate.

[0021] The ratio of the number of the through holes / punch columns to the area of the feed surface is 30-40: 39-41cm 2 .

[0022] The feeding dish is made of transparent plastic material.

[0023] The beneficial effects of the present application are as follows:

[0024] 1. The method takes the larva stage of Callosobruchus maculatus as the feeding starting point, and is a supplement to the feeding method of Callosobruchus maculatus, so that the feeding method of Callosobruchus maculatus is more flexible, and the technical problem that the larva of Callosobruchus maculatus is difficult to continue to feed after being peeled off from the legume seeds and is difficult to complete the generation development is solved, so that the larva of Callosobruchus maculatus peeled off from the legume seeds can continue to feed and normally complete the generation development.

[0025] 2. The raw material cost and feeding cost of the method are low, the feeding efficiency of the larva of Callosobruchus maculatus is high, the survival rate is high, and the large-scale feeding of the larva of Callosobruchus maculatus is realized.

[0026] 3. The device simulates the natural form and living habit of the larva of Callosobruchus maculatus, ensures that the feeding process conforms to the natural growth state, and through the design of quick punching, the dependence on professional knowledge and experience is reduced, so that the artificial feeding method is more easy to popularize and operate.

[0027] 4. The device uniformly forms feeding grooves of the same size on the surface of the feed, so that the growth environment of all the larvae of Callosobruchus maculatus is kept uniform, and thus an effective way is provided for studying the biological characteristics and physiological and biochemical characteristic changes presented by the larva of Callosobruchus maculatus and its subsequent development stages. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 It is a structural schematic view of the feeding device in the present application;

[0029] Figure 2 It is a photo schematic view of starting to feed the larva of Callosobruchus maculatus by the artificial feeding method in the present application;

[0030] Figure 3 It is a photo schematic view of starting to pupate into adult insects of Callosobruchus maculatus fed by the artificial feeding method in the present application.

[0031] Among them, 1, feeding dish, 2, pressing plate, 3, punch. DETAILED DESCRIPTION

[0032] The present application will be further described in detail below in combination with the drawings and specific embodiments.

[0033] The first aspect of the present application provides an artificial feeding method of Callosobruchus maculatus larvae according to the living characteristics of the larvae in legume seeds. The artificial feeding method of the present application simulates the morphology of legume seed embryos by shaping the feed, creates grooves in the feed to simulate the larval feeding channels formed in legume seeds, and controls the moisture of the feed to enhance the adaptability of the larvae to the feed, thereby achieving the continuous cultivation of Callosobruchus maculatus larvae to the adult stage.

[0034] The specific steps of the artificial feeding method of the present application are as follows:

[0035] 1) After mixing the embryo powder of peeled legume seeds with water, a doughy feed is obtained. The doughy feed is placed in a feeding dish 1, shaped into a cake, and a plurality of holes that can accommodate Callosobruchus maculatus larvae are inserted into the upper surface of the obtained cake-shaped feed. Each hole serves as a feeding groove, and a plurality of feeding grooves are obtained.

[0036] Preferably, the embryo powder of peeled legume seeds is sieved through a 50-70 mesh sieve before being mixed with water. The purpose of sieving is to ensure that the feed has a relatively uniform texture. In specific implementation, the mesh number of the sieve can be selected as 50, 60, or 70, and is preferably 70. The feed prepared using the embryo powder that can pass through a 70 mesh sieve has a water retention rate that is relatively most suitable for the development of the larvae, so that the feed can maintain the shape after shaping throughout the entire cultivation process without cracking or deformation, and can maintain a humidity similar to that of the legume embryo environment, thereby best ensuring the adaptability of the larvae to the feed.

[0037] Preferably, the mass-volume ratio of the embryo powder to water is 1g:0.4-0.6mL. Under this preferred mass-volume ratio, the obtained feed is not only easier to shape and does not stick to the larvae, but also has the highest similarity to the legume embryo environment after drying, which is most conducive to the adaptation and feeding of the larvae. In specific implementation, the mass-volume ratio of the embryo powder to water is most preferably 1g:0.5mL.

[0038] Preferably, each feeding groove has the same inner diameter and depth and does not completely penetrate the cake-shaped feed, so as to ensure that there is a certain gap between the bottom of the feeding groove and the bottom of the feeding dish 1.

[0039] Preferably, the inner diameter of each feeding groove is 2-3mm, and the depth of the feeding groove is 1.5-2mm. The method of the present application simulates the morphology of legume seed embryos by creating grooves in the feed to simulate the larval feeding channels formed in legume seeds.

[0040] Preferably, the ratio of the number of feeding grooves to the area of the feed surface is 30-40:39-41cm 2 , preferably 30-40:40cm 2, to improve the feeding density of larvae as much as possible under the premise of ensuring that the larvae have sufficient nutrition to complete the generation development, improve the feed utilization rate, and reduce the feeding cost.

[0041] Preferably, the legume seeds include, but are not limited to, one or a combination of cowpea, adzuki bean, mung bean and chickpea. Preferably, the cowpea.

[0042] As an optional embodiment of the present application, step 1) is specifically: a certain amount of clean legume crop seeds are prepared in advance, soaked for 1 h, peeled, and placed in a 65℃ oven to dry the moisture; the dried legume seed embryo is ground into powder with a flour mill, and the peeled legume seed embryo powder is mixed with water after being sieved through a 70-mesh sieve, and kneaded into a dough until the feed becomes a dough without obvious powder, to obtain a dough feed; then the dough feed is placed in a feeding dish 1, and a pressing plate 2 is placed above the dough feed, and the pressing plate 2 is pressed downward to extrude the feed, so that the feed is fully attached to the dish wall and the surface is flat; then the feeding dish 1 containing the pressing plate 2 and the cake-shaped feed is placed in an open state in an indoor environment or a ventilated environment for 1-2 h, at which time the surface of the feed is pressed gently with fingers without sticking feeling; the puncher 3 is placed on the pressing plate 2, the position of the puncher 3 is adjusted so that the punching columns on the bottom surface of the puncher 3 and the number and arrangement position of the through holes on the pressing plate 2 are the same and are vertically aligned one by one, and then the puncher 3 is pressed downward so that each punching column is inserted into the cake-shaped feed below the corresponding through hole, and then a plurality of holes are uniformly inserted into the upper surface of the cake-shaped feed, and a plurality of feeding grooves are obtained.

[0043] 2) According to the method of picking one four-stripe bean weevil larva into each feeding groove, the pre-collected four-stripe bean weevil larvae are picked into the feeding grooves.

[0044] Preferably, the four-stripe bean weevil larvae are collected at 12-14 days after oviposition.

[0045] 3) The feeding dish 1 with the picked four-stripe bean weevil larvae is placed in a suitable environment and cultured until the four-stripe bean weevil larvae are eclosed into adults.

[0046] Usually, after 6-8 days of culture, four-stripe bean weevil larvae begin to eclose.

[0047] Preferably, the suitable environment refers to a culture box, and the parameters of the culture box are set as: total darkness, temperature of 30±2℃, humidity of 60-65% RH, and open culture. No additional water is needed to be added to the surface of the feed to maintain humidity during the feeding process. The environment is very dry when the larvae are naturally fed with bean seeds, and the open culture of the artificial feed using the method of the present application can approach the natural feeding state.

[0048] The second aspect of the present application provides a feeding device for the above-mentioned artificial feeding method of Callosobruchus maculatus larvae. The feeding device realizes rapid shaping of the feed and convenient preparation of the feeding groove through the cooperation of the through holes on the pressing plate 2 and the punching columns on the puncher 3. In addition, the through holes on the pressing plate 2 and the punching columns on the puncher 3 are carefully designed in terms of shape, size and distribution density, etc. to simulate the natural form and living habits of Callosobruchus maculatus larvae, ensuring the biological simulation of the feeding process. This design reduces the dependence on professional knowledge and experience, making the artificial feeding method more easy to popularize and operate.

[0049] As shown in Figure 1 The feeding device includes a feeding dish 1, a pressing plate 2 and a puncher 3; the feeding dish 1 is used as a container for culturing Callosobruchus maculatus larvae; the pressing plate 2 is used to press the mass of feed placed in the feeding dish 1 into a cake shape and make the surface of the cake-shaped feed flat; the puncher 3 is used to insert a plurality of holes on the surface of the cake-shaped feed to form a plurality of feeding grooves. The outer diameter of the pressing plate 2 is correspondingly arranged with the inner diameter of the feeding dish 1, so that the pressing plate 2 can be placed inside the feeding dish 1.

[0050] A plurality of through holes are uniformly arranged on the pressing plate 2, and a plurality of punching columns are uniformly arranged on the bottom surface of the puncher 3. The number and arrangement position of the punching columns are the same as those of the through holes on the pressing plate 2 and are vertically aligned. Each punching column is gap-fitted with the corresponding through hole, so that the punching column can be inserted into the corresponding through hole and reciprocally move up and down in the through hole.

[0051] When the puncher 3 is pressed from top to bottom, the bottom end of each punching column is inserted into the upper surface of the cake-shaped feed below the pressing plate 2 after passing through the corresponding through hole.

[0052] The punching column is columnar, and the difference between the axial length of the punching column and the thickness of the pressing plate 2 corresponds to the depth setting of the feeding groove, preferably 1.5-2 mm.

[0053] Preferably, the feeding dish 1 is made of transparent plastic material.

[0054] The present application will be further described below in conjunction with specific examples:

[0055] Example 1

[0056] In this example, cowpea seeds are used as the main component of the feed to feed Callosobruchus maculatus larvae to the adult form. The specific steps are as follows:

[0057] (1) Prepare 100 g of market-purchased cowpea seeds in a dish, add an appropriate amount of purified water, just slightly cover the surface of the cowpea seeds, soak for 1 h, then peel and dry in a 65℃ oven;

[0058] (2) using a household flour mill to grind the dried legume seeds to a powder, and then sieving the powder through a 70-mesh sieve to remove larger particles and ensure a uniform texture;

[0059] (3) 2.5 g of the sieved flower cowpea seed embryo powder was mixed with 1.25 mL of purified water at a ratio of 1 g of the flower cowpea seed embryo powder to 0.5 mL of purified water, and then stirred and kneaded until the feed was initially formed into a dough with no obvious powder on the surface;

[0060] (4) The dough-like feed was placed in the feeding dish 1, and the pressing plate 2 was used to press the feed to make it fully adhere to the dish wall and have a smooth surface;

[0061] In this embodiment, the feeding dish 1 is a circular culture dish with an inner diameter of 3.5 cm and a height of 2 cm, and the thickness of the cake-like feed is about 2-3 mm;

[0062] (5) The feeding dish 1, the feed, and the pressing plate 2 were placed in an indoor or ventilated environment for 1-2 h, and at this time, the surface of the feed was gently pressed with fingers without sticking feeling;

[0063] (6) The surface of the feed was vertically pressed using the puncher 3 to create feeding grooves that can accommodate four-vaned bean weevil larvae; the pressing plate 2 and the puncher 3 were removed from the feeding dish 1;

[0064] In this embodiment, the number of feeding grooves was 36 and 37, respectively;

[0065] (7) About 30 four-vaned bean weevil larvae at the age of 12-14 days after oviposition were selected and placed in the feeding grooves, with one four-vaned bean weevil larva in each feeding groove, and the feeding dish 1 containing the four-vaned bean weevil larvae was placed in an environment with a temperature of about 30°C and a humidity of 65% RH in the dark and open culture;

[0066] The photo of the four-vaned bean weevil larvae at the beginning of the artificial feeding method in the present application is shown in Figure 2 ;

[0067] (8) After about 7 days, some four-vaned bean weevil larvae began to pupate. The photo of the four-vaned bean weevil larvae that began to pupate into adults using the artificial feeding method in the present application is shown in Figure 3 .

[0068] (9) The number of adults that pupated each day was counted until there were no new adults that pupated for three consecutive days, and the survival rate was calculated.

[0069] In this embodiment, the survival rate of the four-vaned bean weevil larvae that were fed using the method of the present application until pupation was 90% (63 / 70), and the pupation rate of the four-vaned bean weevil larvae fed using cowpea seeds in a natural state was about 85-95%.

[0070] In summary, the method of the present application is not only suitable for artificial cultivation of Callosobruchus maculatus larvae, but also supports large-scale breeding. In addition, the method of the present application also provides an effective way for studying the biological characteristics and physiological and biochemical characteristic changes of Callosobruchus maculatus larvae and subsequent development stages.

[0071] The application of the present application is not limited to the embodiments described. Any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present application, and therefore the protection scope of the present application should be limited by the scope defined by the claims.

Claims

1. A method for artificial rearing of Callosobruchus maculatus (Coleoptera: Chrysomelidae) larvae, characterized by: The artificial feeding method comprises the following steps: 1) After the seed embryo powder of peeled legume seeds is sieved through a 50-70 mesh sieve, the seed embryo powder is mixed with water to obtain a doughy feed, the doughy feed is placed in a feeding dish (1), the doughy feed is shaped into a cake, and after the cake-shaped feed is left to stand at room temperature for 1-2 h, a plurality of holes are inserted on the surface of the cake-shaped feed to form a plurality of feeding grooves; The mass / volume ratio of the seed embryo powder to water is 1 g:0.4-0.6 mL; the thickness of the cake-shaped feed is 2-3 mm; the inner diameter of the feeding groove is 2-3 mm, and the depth of the feeding groove is 1.5-2 mm; The feeding device comprises a feeding dish (1), a pressing plate (2), and a hole puncher (3); the pressing plate (2) is used for shaping the feed; the hole puncher (3) is used for inserting a plurality of feeding grooves; a plurality of through holes are formed in the pressing plate (2), and a plurality of hole punching columns are arranged on the bottom surface of the hole puncher (3); the number and arrangement positions of the hole punching columns are the same as those of the through holes and are aligned; when the hole puncher (3) is pressed from top to bottom, the bottom ends of the hole punching columns respectively pass through the corresponding through holes and are inserted into the cake-shaped feed below the pressing plate (2); The step 1) specifically comprises the following steps: the seed embryo powder of peeled legume seeds is mixed with water and kneaded into a doughy feed, the doughy feed is then placed in the feeding dish (1), the doughy feed is pressed by the pressing plate (2) to shape the doughy feed into a cake, and a plurality of holes are uniformly inserted on the surface of the cake-shaped feed by using the hole puncher (3) to form a plurality of feeding grooves; 2) The pre-collected Callosobruchus maculatus larvae are picked into the feeding grooves; The Callosobruchus maculatus larvae are collected at 12-14 d after oviposition; 3) The feeding dish in which the Callosobruchus maculatus larvae are picked is placed in a suitable environment, and is cultured until the Callosobruchus maculatus larvae are eclosed.

2. The method according to claim 1, wherein: In the step 1), the ratio of the number of the feeding troughs to the area of the surface of the cake feed is 30 to 40: 39 to 41 cm 2 .

3. The method according to claim 1, wherein the method is characterized by: In the step 3), the suitable environment is specifically as follows: the temperature is 28-32 ℃, and the humidity is 60-65% RH; in the step 3), the Callosobruchus maculatus larvae start to eclose after being cultured for 6-8 d.

4. The method for artificial rearing of Callosobruchus maculatus (Fabricius) larvae according to claim 1, characterized in that: The feeding dish (1) is made of transparent plastic.

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