CN109497353B - Application of graphene oxide in preparation of feed for lepidoptera and diptera insects - Google Patents

Application of graphene oxide in preparation of feed for lepidoptera and diptera insects Download PDF

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CN109497353B
CN109497353B CN201811430915.0A CN201811430915A CN109497353B CN 109497353 B CN109497353 B CN 109497353B CN 201811430915 A CN201811430915 A CN 201811430915A CN 109497353 B CN109497353 B CN 109497353B
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graphene oxide
feed
larvae
generation
agar
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CN109497353A (en
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王秀平
解海翠
李婧实
沈江洁
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Hebei Normal University of Science and Technology
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/90Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/30Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/105Aliphatic or alicyclic compounds
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/163Sugars; Polysaccharides
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/20Inorganic substances, e.g. oligoelements

Abstract

The invention discloses an application of graphene oxide in preparation of feeds for lepidoptera and diptera insects. Wherein the mass-to-volume ratio (mu g/mL) of the graphene oxide to the feed for lepidoptera and diptera insects is 500: 1-2000:1. The feed is prepared by the following method: preparing a graphene oxide solution, preparing a culture medium, and dissolving the culture medium in the graphene oxide solution to prepare the graphene oxide-based feed for the diptera insects. The graphene oxide-based lepidoptera and diptera insect feed can shorten the growth period of insects, improve the survival rate of larvae, increase the weight of the larvae and pupae, and greatly reduce the time and production cost in the practical application process.

Description

Application of graphene oxide in preparation of feed for lepidoptera and diptera insects
Technical Field
The invention belongs to the technical field of plant protection, and particularly relates to application of graphene oxide in preparation of feeds for lepidoptera and diptera insects.
Background
With the development of science and technology and the progress of nanotechnology, nanomaterials have shown wide application prospects in various fields. Graphene oxide is a novel two-dimensional carbon nanomaterial, has excellent characteristics of electricity, mechanics, optics and the like, and has a wide application prospect in the fields of biomedicine, sensing, environmental science, engineering science and the like. In recent years, the application of graphene oxide in the agricultural field has attracted wide interest of various scholars, for example, graphene oxide can load agricultural chemicals to achieve the functions of slow release and targeting, so that the utilization rate of the agricultural chemicals is improved, and the side effect on non-target organisms is reduced. However, the application of graphene oxide added to insect feed as an insect growth promoter has been reported.
The artificial mass propagation of insects is not only one of the important means for modern pest control research, but also an important way for obtaining natural product resources. The natural feed is used for feeding tested insects, and the aim of feeding the tested insects in large quantities is difficult to achieve due to the limitation of seasonality and the fact that the feed needs to be replaced for many times. The semi-artificial feed has the characteristics of no limitation of seasons, difficult mildewing, deterioration and drying, little or no change of feed in the feeding process and the like, is very convenient to manage, and makes large-scale industrialized production possible. However, the existing artificial feed has the problems of long feeding period, low insect individual survival and death rate, poor physical constitution of individual insects and the like, and causes certain trouble to the scale feeding of insects.
Disclosure of Invention
In view of the above, the invention provides an application of graphene oxide in preparing feeds for lepidoptera and diptera insects.
In order to solve the technical problems, the invention discloses application of graphene oxide in preparation of feeds for lepidoptera and diptera insects.
Optionally, the mass to volume ratio (μ g/mL) of graphene oxide to the feed for lepidopteran and dipteran insects is 500: 1-2000:1.
Optionally, the method comprises the following steps:
step 1, preparing a graphene oxide solution: dissolving graphene oxide in ionic water and carrying out ultrasonic treatment;
step 2, preparing a culture medium: dissolving corn flour in water, and blending into paste for later use; dissolving agar in water, boiling, stirring and melting; adding white sugar after agar is fully melted, stirring to fully dissolve, and stirring to fully dissolve; pouring the pasty corn flour into sugared agar water, stirring and boiling into paste; adding propionic acid and stirring; slightly cooling, adding yeast powder, stirring with a glass rod to mix well, adding purified water to constant volume, and preparing to obtain a culture medium;
step 3, preparing the graphene oxide-based feed for the diptera insects: and (3) dissolving a culture medium in the graphene oxide solution prepared in the step (1) to prepare the graphene oxide-based feed for the diptera insects.
Optionally, the mass-to-volume ratio (mg/ml) of the graphene oxide to the ionized water in the step 1 is 100:30-400: 30.
Optionally, the mass ratio of corn meal to water is 10: 38.
Optionally, the mass ratio of agar to water is 1: 20; the mass ratio of the white sugar to the agar is 9: 1.
Optionally, the volume-to-mass ratio (ml/g) of propionic acid to corn meal is 1: 20.
Optionally, the mass ratio of the yeast powder to the corn flour is 7: 100.
optionally, the ratio of the total volume after volume fixing to the volume of the corn flour (ml/g) is 17: 2.
Optionally, the volume ratio of the culture medium in step 3 to the ionized water in step 1 is 170: 30.
Compared with the prior art, the invention can obtain the following technical effects:
the graphene oxide-based lepidoptera and diptera insect feed can shorten the growth period of larvae of insects, improve the survival rate of the larvae, increase the weight of the larvae and pupae, and greatly reduce the time and production cost in the practical application process.
Of course, it is not necessary for any one product in which the invention is practiced to achieve all of the above-described technical effects simultaneously.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention and not to limit the invention. In the drawings:
FIG. 1 is a graph showing the effect of different concentrations of graphene oxide on the progression of corn borer larvae;
FIG. 2 is the effect of graphene oxide of different concentrations on the pupal calendar phase of corn borer according to the present invention;
FIG. 3 is a graph showing the effect of different concentrations of graphene oxide on the duration of adult corn borers according to the present invention;
FIG. 4 is the effect of different concentrations of graphene oxide on oviposition of corn borer according to the present invention;
FIG. 5 is a graph of the effect of different concentrations of graphene oxide on the hatchability of ostrinia nubilalis eggs according to the present invention;
FIG. 6 is the survival rate of Asiatic corn borer larvae at different GO concentrations according to the present invention;
FIG. 7 is the pupation rate of Asiatic corn borer at different GO concentrations of the present invention;
FIG. 8 is the emergence rate of Asiatic corn borer at different GO concentrations of the present invention;
FIG. 9 is the larval weight of Asiatic corn borer at different GO concentrations of the present invention;
FIG. 10 is pupal weight of Asiatic corn borer at different GO concentrations of the present invention.
Detailed Description
The following embodiments are described in detail with reference to the accompanying drawings, so that how to implement the technical features of the present invention to solve the technical problems and achieve the technical effects can be fully understood and implemented.
The invention discloses an application of graphene oxide in preparation of feeds for lepidoptera and diptera insects.
The mass-to-volume ratio (mu g/mL) of the graphene oxide to the feed for lepidoptera and diptera insects is 500: 1-2000:1.
The invention also discloses a preparation method of the graphene oxide-based feed for diptera insects, which comprises the following steps:
step 1, preparing a graphene oxide solution: dissolving graphene oxide in ionized water and carrying out ultrasonic treatment, wherein the mass-volume ratio (mg/ml) of the graphene oxide to the ionized water is 100:30-400: 30;
step 2, preparing a culture medium: dissolving corn flour in water according to the mass ratio of 10:38, and blending into paste for later use; dissolving agar in water according to the mass ratio of 1:20, boiling, stirring and melting; adding 135g of white sugar after the agar is sufficiently melted, stirring to sufficiently dissolve the agar, wherein the mass ratio of the white sugar to the agar is 9:1, and stirring to sufficiently dissolve the agar; pouring the pasty corn flour into sugared agar water, stirring and boiling into paste; adding propionic acid and stirring, wherein the volume-to-mass ratio (ml/g) of the propionic acid to the corn flour is 1: 20; slightly cooling, and adding yeast powder, wherein the mass ratio of the yeast powder to the corn flour is 7: 100, stirring by using a glass rod, uniformly mixing, adding purified water to a constant volume, and preparing a culture medium, wherein the volume-to-mass ratio (ml/g) of the total volume to the corn flour is 17: 2;
step 3, preparing the graphene oxide-based feed for the diptera insects: and (3) dissolving a culture medium in the graphene oxide solution prepared in the step (1), wherein the volume ratio of the culture medium to the ionized water in the step (1) is 170:30, and preparing the graphene oxide-based feed for the diptera insects.
Example 1
Dissolving graphene oxide in ionized water and carrying out ultrasonic treatment, wherein the mass-volume ratio (mg/ml) of the graphene oxide to the ionized water is 100: 30; dissolving corn flour in water according to the mass ratio of 10:38, and blending into paste for later use; dissolving agar in water according to the mass ratio of 1:20, boiling, stirring and melting; adding 135g of white sugar after the agar is sufficiently melted, stirring to sufficiently dissolve the agar, wherein the mass ratio of the white sugar to the agar is 9:1, and stirring to sufficiently dissolve the agar; pouring the pasty corn flour into sugared agar water, stirring and boiling into paste; adding propionic acid and stirring, wherein the volume-to-mass ratio (ml/g) of the propionic acid to the corn flour is 1: 20; slightly cooling, and adding yeast powder, wherein the mass ratio of the yeast powder to the corn flour is 7: 100, stirring by using a glass rod, uniformly mixing, adding purified water to a constant volume, and preparing a culture medium, wherein the volume-to-mass ratio (ml/g) of the total volume to the corn flour is 17: 2; and (3) dissolving a culture medium in the graphene oxide solution prepared in the step (1), wherein the volume ratio of the culture medium to the ionized water in the step (1) is 170:30, and preparing the graphene oxide-based feed for the diptera insects.
Example 2
Dissolving graphene oxide in ionized water and carrying out ultrasonic treatment, wherein the mass-volume ratio (mg/ml) of the graphene oxide to the ionized water is 200: 30; dissolving corn flour in water according to the mass ratio of 10:38, and blending into paste for later use; dissolving agar in water according to the mass ratio of 1:20, boiling, stirring and melting; adding 135g of white sugar after the agar is sufficiently melted, stirring to sufficiently dissolve the agar, wherein the mass ratio of the white sugar to the agar is 9:1, and stirring to sufficiently dissolve the agar; pouring the pasty corn flour into sugared agar water, stirring and boiling into paste; adding propionic acid and stirring, wherein the volume-to-mass ratio (ml/g) of the propionic acid to the corn flour is 1: 20; slightly cooling, and adding yeast powder, wherein the mass ratio of the yeast powder to the corn flour is 7: 100, stirring by using a glass rod, uniformly mixing, adding purified water to a constant volume, and preparing a culture medium, wherein the volume-to-mass ratio (ml/g) of the total volume to the corn flour is 17: 2; and (3) dissolving a culture medium in the graphene oxide solution prepared in the step (1), wherein the volume ratio of the culture medium to the ionized water in the step (1) is 170:30, and preparing the graphene oxide-based feed for the diptera insects.
Example 3
Dissolving graphene oxide in ionized water and carrying out ultrasonic treatment, wherein the mass-to-volume ratio (mg/ml) of the graphene oxide to the ionized water is 400: 30; dissolving corn flour in water according to the mass ratio of 10:38, and blending into paste for later use; dissolving agar in water according to the mass ratio of 1:20, boiling, stirring and melting; adding 135g of white sugar after the agar is sufficiently melted, stirring to sufficiently dissolve the agar, wherein the mass ratio of the white sugar to the agar is 9:1, and stirring to sufficiently dissolve the agar; pouring the pasty corn flour into sugared agar water, stirring and boiling into paste; adding propionic acid and stirring, wherein the volume-to-mass ratio (ml/g) of the propionic acid to the corn flour is 1: 20; slightly cooling, and adding yeast powder, wherein the mass ratio of the yeast powder to the corn flour is 7: 100, stirring by using a glass rod, uniformly mixing, adding purified water to a constant volume, and preparing a culture medium, wherein the volume-to-mass ratio (ml/g) of the total volume to the corn flour is 17: 2; and (3) dissolving a culture medium in the graphene oxide solution prepared in the step (1), wherein the volume ratio of the culture medium to the ionized water in the step (1) is 170:30, and preparing the graphene oxide-based feed for the diptera insects.
Comparative example 1
Adding 30ml of deionized water into 1 200ml beaker; dissolving corn flour in water according to the mass ratio of 10:38, and blending into paste for later use; dissolving agar in water according to the mass ratio of 1:20, boiling, stirring and melting; adding 135g of white sugar after the agar is sufficiently melted, stirring to sufficiently dissolve the agar, wherein the mass ratio of the white sugar to the agar is 9:1, and stirring to sufficiently dissolve the agar; pouring the pasty corn flour into sugared agar water, stirring and boiling into paste; adding propionic acid and stirring, wherein the volume-to-mass ratio (ml/g) of the propionic acid to the corn flour is 1: 20; slightly cooling, and adding yeast powder, wherein the mass ratio of the yeast powder to the corn flour is 7: 100, stirring by using a glass rod, uniformly mixing, adding purified water to a constant volume, and preparing a culture medium, wherein the volume-to-mass ratio (ml/g) of the total volume to the corn flour is 17: 2; dissolving a culture medium in the ionized water in the step 1, wherein the volume ratio of the culture medium to the ionized water in the step 1 is 170:30, and preparing the graphene oxide-based feed for the diptera insects.
The technical effects of the invention are illustrated below with reference to specific experimental data:
firstly, the influence of graphene oxide on the growth duration and the fertility of corn borers:
one) experimental method:
1. and (3) measuring the growth period, egg laying amount and hatching rate of the corn borers:
the newly hatched larvae of the Asiatic corn borers are respectively filled in prepared artificial feed (the feed is cut into small pieces of 5 x 5 cm), 250 insects are inoculated in each box of feed, and the insect inoculation date is recorded. Culturing the inoculated feed in a climatic chamber (greenhouse condition: temperature: 28 deg.C, relative humidity 75%, and light cycle L: D: 12), observing every day until pupation, and recording pupation date (larval stage) of each larva; picking pupae from the feed every day after the larva pupates, putting the pupae into a 15mL centrifuge tube, putting a cotton ball at the tube opening for wetting, collecting the pupae in an artificial climate box until the pupae emerge as adult insects, and recording the date (pupa calendar period) when each pupae emerges as adult insects; after the adult insects eclosion, putting the male and female pairs in a paper cup for breeding until the adult insects die, and observing and recording the death date (the calendar of the adult insects) of each adult insect; and (3) selecting 30 pairs of adult insects in the peak period for each treatment, performing oviposition test, repeating for three times, and recording the number of oviposits and the hatchability (oviposition amount and hatchability).
2. The data processing and analyzing method comprises the following steps: all assay determinations were completely randomized, with three replicates per treatment. And (3) statistically analyzing the experimental data by adopting DPS software and an Excel table to obtain the average value +/-SE (standard error), wherein the p value is less than 0.05 (or less than 0.01), and the statistical significance is determined by adopting the t test.
II) experimental results:
1. influence of graphene oxide with different concentrations on corn borer larva in later stage
As can be seen from FIG. 1, in the F0 generation, the larvae of the ostrinia nubilalis raised on the feed without graphene oxide had a period of 20 days, the larvae of the ostrinia nubilalis raised on the feed with a graphene oxide concentration of 500. mu.g/mL (example 1, the same below) had a period of 18.33 days, the larvae of the ostrinia nubilalis raised on the feed with a graphene oxide concentration of 1000. mu.g/mL (example 2, the same below) had a period of 17.17 days, and the larvae of the ostrinia nubilalis raised on the feed with a graphene oxide concentration of 2000. mu.g/mL (example 3, the same below) had a period of 16.33 days; in the F1 generation, the larva of the corn borer bred by the feed without the graphene oxide is 18.5 days, the larva of the corn borer bred by the feed with the graphene oxide concentration of 500 mug/mL is 18.17 days, the larva of the corn borer bred by the feed with the graphene oxide concentration of 1000 mug/mL is 16.17 days, and the larva of the corn borer bred by the feed with the graphene oxide concentration of 2000 mug/mL is 15.17 days; in the F2 generation, the larvae of the corn borer bred by the feed without the graphene oxide are 16.67 days old, the larvae of the corn borer bred by the feed with the graphene oxide concentration of 500 mug/mL are 13.67 days old, the larvae of the corn borer bred by the feed with the graphene oxide concentration of 1000 mug/mL are 13 days old, and the larvae of the corn borer bred by the feed with the graphene oxide concentration of 2000 mug/mL are 11.83 days old. As can be seen from table 1, there were very significant differences in the course of the raised asian corn borer larvae at 1-fold, 2-fold and 4-fold increase in graphene oxide concentration, respectively, compared to the concentration of the control (p < 0.01). The results show that the graphene oxide with different concentrations has obvious influence on the larval stage of the ostrinia nubilalis, and the graphene oxide can shorten the larval stage and has a dose effect.
2. The influence of the graphene oxide with different concentrations on the pupa calendar of the corn borer:
as can be seen from fig. 2, in the F0 generation, the pupal period of the ostrinia nubilalis bred by the feed without adding graphene oxide is 5.13 days, the pupal period of the ostrinia nubilalis bred by the feed with the graphene oxide concentration of 500 μ g/mL is 5.50 days, the pupal period of the ostrinia nubilalis bred by the feed with the graphene oxide concentration of 1000 μ g/mL is 5.61 days, the pupal period of the ostrinia nubilalis bred by the feed with the graphene oxide concentration of 2000 μ g/mL is 5.82 days, the pupal period of the ostrinia nubilalis bred by the feed with the graphene oxide concentration of 500 μ g/mL in the F1 generation is 6.51 days, the pupal period of the ostrinia nubilalis bred by the feed with the graphene oxide concentration of 500 μ g/mL is 6.30 days, the pupal period of the feed with the graphene oxide concentration of 1000 μ g/mL is 6.54 days, and the pupal period of the feed with the graphene oxide concentration of 2000 μ g/mL is 6.51 days; in the F2 generation, the pupa duration of the corn borer bred by the feed without the graphene oxide is 6.27 days, the pupa duration of the corn borer bred by the feed with the graphene oxide concentration of 500 mug/mL is 6.6 days, the pupa duration of the corn borer bred by the feed with the graphene oxide concentration of 1000 mug/mL is 6 days, and the pupa duration of the corn borer bred by the feed with the graphene oxide concentration of 2000 mug/mL is 6.27 days. As can be seen from Table 1, the graphene oxide concentration is increased by 1 time, 2 times and 4 times respectively compared with the control concentration, and the pupa calendar period of the Asiatic corn borers bred under the conditions has no significant difference (p > 0.05). From the above results, it can be seen that the graphene oxide with different concentrations has no significant effect on the pupal calendar period of the ostrinia nubilalis.
3. Influence of graphene oxide with different concentrations on adult corn borers in calendar period
As can be seen from FIG. 3, in the F0 generation, the adult age of the ostrinia nubilalis bred by the feed without the graphene oxide is 4.11 days, the adult age of the ostrinia nubilalis bred by the feed with the graphene oxide concentration of 500 mug/mL is 4.33 days, the adult age of the ostrinia nubilalis bred by the feed with the graphene oxide concentration of 1000 mug/mL is 4.08 days, and the adult age of 2000 mug/mL is 4.07 days; in the F1 generation, the adult corn borers bred by the feed without the graphene oxide is 3.75 days old, the adult corn borers bred by the feed with the graphene oxide concentration of 500 mug/mL is 3.47 days old, the adult corn borers bred by the feed with the graphene oxide concentration of 1000 mug/mL is 3.71 days old, and the adult corn borers bred by the feed with the graphene oxide concentration of 2000 mug/mL is 3.90 days old; in the F2 generation, the adult corn borers bred by the feed without the graphene oxide is 3.63 days old, the adult corn borers bred by the feed with the graphene oxide concentration of 500 mug/mL is 3.9 days old, the adult corn borers bred by the feed with the graphene oxide concentration of 1000 mug/mL is 3.87 days old, and the adult corn borers bred by the feed with the graphene oxide concentration of 2000 mug/mL is 3.77 days old. As can be seen from Table 1, the graphene oxide concentration is increased by 1 time, 2 times and 4 times respectively compared with the control concentration, and the adult stage of the Asian corn borers bred under the conditions has no significant difference (p > 0.05). From the above results, it can be seen that the graphene oxide with different concentrations has no significant effect on the adult duration of the ostrinia nubilalis.
TABLE 1 analysis of variance of graphene oxide with different concentrations in development stages of larvae, pupae and adults of Asiatic corn borer
Figure BDA0001882693890000081
4. Influence of graphene oxide with different concentrations on oviposition amount of corn borer
As can be seen from FIG. 4, in the F0 generation, the feed without added graphene oxide gave rise to 134.8 eggs of ostrinia nubilalis, the feed with added graphene oxide concentration of 500. mu.g/mL gave rise to 166.02 eggs of ostrinia nubilalis, the feed with added graphene oxide concentration of 1000. mu.g/mL gave rise to 183.31 eggs, and the feed with added graphene oxide concentration of 2000. mu.g/mL gave rise to 158.13 eggs; in the F1 generation, the feed without the added graphene oxide can feed the ostrinia nubilalis with the egg laying amount of 122.33, the feed with the added graphene oxide concentration of 500 mug/mL can feed the ostrinia nubilalis with the egg laying amount of 129, the feed with the added graphene oxide concentration of 1000 mug/mL of 134 and the feed with the added graphene oxide concentration of 2000 mug/mL of 126.33; in the F2 generation, the feed without the added graphene oxide feed has the feed bred corn borer oviposition amount of 95.87, the feed with the added graphene oxide concentration of 500 mug/mL has the feed bred corn borer oviposition amount of 97.84, the feed with the added graphene oxide concentration of 1000 mug/mL has 106.43, and the feed with the added graphene oxide concentration of 2000 mug/mL has 108.77. As can be seen from Table 2, the graphene oxide concentration is increased by 1 time, and the egg laying amount of the Asian corn borers raised by 2 times and 4 times respectively has no significant difference (p > 0.05) compared with the control concentration. From the above results, it can be seen that the different concentrations of graphene oxide have no significant effect on the oviposition amount of the ostrinia nubilalis.
5. Influence of graphene oxide with different concentrations on hatchability of ostrinia nubilalis eggs
As can be seen from FIG. 5, in the F0 generation, the hatchability of the ostrinia nubilalis eggs bred by the feed without adding graphene oxide is 65.33%, the hatchability of the ostrinia nubilalis eggs bred by the feed with the graphene oxide concentration of 500 mug/mL is 66.48%, the hatchability of the ostrinia nubilalis eggs bred by the feed with the graphene oxide concentration of 1000 mug/mL is 90.39%, and the hatchability of the ostrinia nubilalis eggs bred by the feed with the graphene oxide concentration of 2000 mug/mL is 94.15%; in the F1 generation, the hatchability of the ostrinia nubilalis eggs fed by the feed without the graphene oxide is 89.7 percent, the hatchability of the ostrinia nubilalis eggs fed by the feed with the graphene oxide concentration of 500 mug/mL is 88.18 percent, the hatchability of the ostrinia nubilalis eggs fed by the feed with the graphene oxide concentration of 1000 mug/mL is 91.23 percent, and the hatchability of the ostrinia nubilalis eggs fed by the feed with the graphene oxide concentration of 2000 mug/mL is 91.4 percent; in the F2 generation, the hatchability of the ostrinia nubilalis eggs fed by the feed without the graphene oxide is 92.56%, the hatchability of the ostrinia nubilalis eggs fed by the feed with the graphene oxide concentration of 500 mug/mL is 94.78%, the hatchability of the ostrinia nubilalis eggs fed by the feed with the graphene oxide concentration of 1000 mug/mL is 97.4%, and the hatchability of the ostrinia nubilalis eggs fed by the feed with the graphene oxide concentration of 2000 mug/mL is 97.87%. As can be seen from table 2, the hatchability of the ostrinia nubilalis eggs can be increased compared to the concentration of the control, but the difference is not significant at some generations.
TABLE 2 analysis of variance of graphene oxide on oviposition and hatchability of ostrinia furnacalis across generations
Figure BDA0001882693890000091
Third), the research results of the invention show that the graphene oxide with different concentrations has obvious influence on the larval stage of the Asiatic corn borer. With the increase of the concentration of the graphene oxide, the development period of the larvae of the Asiatic corn borer F0-F2 generation is remarkably shortened. The graphene oxide has no obvious influence on the pupal stage, the adult stage, the egg laying amount and the hatching rate of the corn borer.
Secondly, the growth and development of the Asiatic corn borers are influenced by the graphene oxide:
one) test method:
1. and (3) test treatment:
the assay sets up 3 GO concentration treatments, 2000. mu.g/mL, 1000. mu.g/mL and 500. mu.g/mL. Three GO with different concentrations are added in the process of preparing the artificial feed for the ostrinia nubilalis, and normal feed without the GO is used as a control, and each treatment is repeated for three times. The Asiatic corn borer larva breeding container is a sterilized plastic box with the length of 15cm, the width of 15cm and the height of 10cm, a round hole with the diameter of 4cm is arranged on a box cover, a gauze with 70 meshes of stainless steel is used for sealing the round hole, the Asiatic corn borer larva breeding container is placed in an artificial climate box for breeding, and the temperature in the box is (28 +/-1) DEG C; relative humidity (70 ± 10)%; the illumination period L and D are 12h to 12h, (L, 6: 00-18: 00; D, 18: 00-6: 00).
2. Determination of survival rate of Asiatic corn borers:
and (3) inoculating the primary larvae of the ostrinia furnacalis hatched on the same day into the prepared artificial feed, recording the inoculation date of 250 larvae in each box, feeding the ostrinia furnacalis in an artificial climate box, and observing the growth and development conditions of the ostrinia furnacalis every day. Selecting all larvae of each box and placing the larvae in newly configured feed when the larvae are from 5 years before pupation, recording the total number of the larvae, and calculating the survival rate (%) of the larvae; after all the larvae pupate, recording the pupation number, and calculating the pupation rate (%); randomly selecting 5 pupae from each box, observing the eclosion condition, and calculating the eclosion rate (%).
3. Determination of the body weight of Asiatic corn borer individuals:
selecting 15 larvae (at 5 ages before pupation) with the same size from each box, weighing the larvae, and recording the original data; after pupation of larvae (weighing pupae weight on day 2 after pupation), selecting 15 pupaes with the same size from each box, weighing the pupae weight, recording original data, respectively putting the pupae into a 15mL centrifuge tube for single-head feeding every day, marking with a marker pen, putting into a cotton ball, dipping water, moisturizing, and putting into a magnetic disk in an artificial climate box.
4. Statistical analysis
All assay determinations were completely randomized, with three replicates per treatment. And (3) carrying out statistical analysis on the experimental data by adopting DPS software and an Excel table, and carrying out variance analysis on the difference significance among treatments by adopting a Duncar new repolarization method.
Secondly, result and analysis:
one) Effect of different GO concentrations on survival Rate of Asiatic corn borer
1. Influence of different GO concentrations on survival rate of Asiatic corn borer larvae
As shown in fig. 6, in the F0 generation, the survival rate of larvae was 40.53% in the control group, and the survival rates of larvae were 42.53%, 48.13% and 51.20% at GO concentrations of 500 μ g/mL, 1000 μ g/mL and 2000 μ g/mL, respectively, as can be seen from table 3, there was a very significant difference in the improvement of asian corn borer larvae survival rate with increasing GO concentration compared to the control group concentration (p < 0.01); larvae survival in the F1 generation was 33.60% for the control group and 38.67%, 42.40% and 48.67% for GO concentrations of 500, 1000 and 2000 μ g/mL, respectively, as also seen in table 3, significantly increased with increasing GO concentration compared to the control group concentration (p < 0.01); in the F2 generation, the survival rate of the larvae in the control group was 38.40%, and the survival rates of the larvae were 41.73%, 42.00% and 44.00% at GO concentrations of 500. mu.g/mL, 1000. mu.g/mL and 2000. mu.g/mL, respectively, as can be seen from Table 3, GO increased the survival rate of larvae of corn borer, but the difference was not significant (p > 0.05), compared to the control group concentration.
2. Influence of different GO concentrations on pupation rate of Asiatic corn borer
As shown in fig. 7, in the F0 generation, the pupation rate of the control group was 32.80%, and the pupation rates were 40.80%, 43.87% and 45.87% at GO concentrations of 500 μ g/mL, 1000 μ g/mL and 2000 μ g/mL, respectively, and as can be seen from table 3, there was a significant difference in the increase in the pupation rate of asian corn borer with the increase in GO concentration as compared to the control group concentration (p < 0.05); in the F1 generation, the pupation rate of the control group was 28.27%, the pupation rate of GO was 29.07%, 33.33% and 37.33% at concentrations of 500. mu.g/mL, 1000. mu.g/mL and 2000. mu.g/mL, respectively, and it can be seen from Table 3 that the increase in GO concentration had no significant effect on the increase in pupation rate (p > 0.05) compared to the control group concentration; in the F2 generation, the pupation rates of the control group were 33.60%, when the GO concentrations were 500 μ g/mL, 1000 μ g/mL and 2000 μ g/mL, the pupation rates were 37.47%, 34.53% and 36.53%, respectively, and it can be seen from Table 3 that GO has no significant effect on the pupation rate of Asian corn borer (p > 0.05) compared with the control group concentration.
3. Effect of different GO concentrations on emergence rate of asian corn borer:
FIG. 8 shows that in the F0 generation, the eclosion rate of the control group was 87.91%, the eclosion rate was 85.54%, 81.84% and 88.00% at GO concentrations of 500 μ g/mL, 1000 μ g/mL and 2000 μ g/mL, respectively, and it can be seen from Table 3 that the increase in GO concentration had no significant effect on the eclosion rate of Asian corn borer (p > 0.05) compared to the control group concentration; in the F1 generation, the eclosion rate of the control group was 93.40%, and the eclosion rates were 92.33%, 89.19% and 88.67% at GO concentrations of 500, 1000 and 2000 μ g/mL, respectively, and it can also be seen from Table 3 that an increase in GO concentration had a very significant effect on the increase in the eclosion rate compared to the control group concentration (p < 0.01); in the F2 generation, the control group had an emergence rate of 91.69% and the GO concentrations were 93.69%, 94.11% and 96.27% at 500. mu.g/mL, 1000. mu.g/mL and 2000. mu.g/mL, respectively. As can be seen from Table 3, GO has a certain promoting effect (p < 0.05) on the increase of the feathering rate compared with the control group concentration.
TABLE 3 analysis of variance of survival rate, pupation rate and emergence rate of larvae of Ostrinia furnacalis Guenee in each generation under different GO concentrations
Figure BDA0001882693890000121
II) Effect of different GO concentrations on the body weight of Asiatic corn borers
1. Effect of different GO concentrations on Asiatic corn borer larva weight
FIG. 9 shows that in the F0 generation, the larvae weight was 0.0617g, 0.0706g and 0.0814g for the control group with a weight of 0.0519g, GO concentration of 500. mu.g/mL, 1000. mu.g/mL and 2000. mu.g/mL; in the F1 generation, when the weight of the larvae of the control group is 0.0550g and the GO concentration is 500 mug/mL, 1000 mug/mL and 2000 mug/mL, the weight of the larvae is 0.0733g, 0, 0784g and 0.0905g respectively; in the F2 generation, when the weight of the larvae of the control group is 0.0508g, and the GO concentration is 500. mu.g/mL, 1000. mu.g/mL and 2000. mu.g/mL, the weight of the larvae is 0.0557g, 0.0618g and 0.0674g, respectively. As can be seen from table 4, there was a very significant difference in the body weight of the larvae of the asian corn borer reared at 1-fold, 2-fold and 4-fold increase in GO concentration, respectively, compared to the control group concentration (p < 0.01). From the above results, it can be seen that GO has a significant promoting effect on the increase in the body weight of larvae of ostrinia nubilalis.
2. Influence of different GO concentrations on pupa weight of asian corn borer:
as can be seen from fig. 10, in the F0 generation, the pupal weights of the control group were 0.0533g, and the GO concentrations were 500 μ g/mL, 1000 μ g/mL, and 2000 μ g/mL, the pupal weights were 0.0631g, 0.0686g, and 0.0739g, respectively, and as can be seen from table 4, there was a very significant difference in the increase in the pupal weight of the asian corn borer with the increase in the GO concentration as compared to the control group concentration (p < 0.01); in the F1 generation, when the pupal weight of the control group was 0.0495g and the GO concentration was 500. mu.g/mL, 1000. mu.g/mL and 2000. mu.g/mL, the pupal weights were 0.0557g, 0, 0645g and 0.0747g, respectively, and it can be seen from Table 4 that the increase in GO concentration had a very significant effect on the increase in pupal weight compared to the control group concentration (p < 0.01); in the F2 generation, when the pupa weight of the control group is 0.0631g, and the GO concentration is 500 mug/mL, 1000 mug/mL and 2000 mug/mL, the pupa weight is 0.0502g, 0.0538g and 0.0576g respectively, and it can be shown in Table 4 that GO has no significant effect on the pupa weight of Asian corn borer (p > 0.05) compared with the control group.
TABLE 2 analysis of variance of the weights of larvae and pupae of Asiatic corn borer in different generations under different GO concentrations
Figure BDA0001882693890000131
In summary, the graphene oxide is used as an external factor influencing insect growth, and within a certain concentration range, the graphene oxide can promote the weight increase of the Asiatic corn borers, improve the survival rate and pupation rate of the Asiatic corn borers, and influence but not obvious influence on the pupation weight and the eclosion rate.
While the foregoing description shows and describes several preferred embodiments of the invention, it is to be understood, as noted above, that the invention is not limited to the forms disclosed herein, but is not to be construed as excluding other embodiments and is capable of use in various other combinations, modifications, and environments and is capable of changes within the scope of the inventive concept as expressed herein, commensurate with the above teachings, or the skill or knowledge of the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (10)

1. Use of graphene oxide in the preparation of a feed for lepidoptera insects capable of improving the larvae of the F0 generation, the larvae survival rate of the F0 generation, the larvae weight of the F0 generation, the pupa weight of the F0 generation, the larvae of the F1 generation, the larvae survival rate of the F1 generation, the larvae weight of the F1 generation, the pupa weight of the F1 generation, the larvae of the F2 generation, the hatchability of the F2 generation and the larvae weight of the F2 generation, wherein the lepidoptera insects are the larvae of the F0, the F1 and the F2 generation of corn borers.
2. The use according to claim 1, wherein the mass to volume ratio of graphene oxide to lepidopteran insect feed, μ g/ml, is 500: 1-2000:1.
3. Use according to claim 1, characterized in that it comprises the following steps:
step 1, preparing a graphene oxide solution: dissolving graphene oxide in ionic water and carrying out ultrasonic treatment;
step 2, preparing a culture medium: dissolving corn flour in water, and blending into paste for later use; dissolving agar in water, boiling, stirring and melting; adding white sugar after the agar is fully melted, and stirring to fully dissolve the agar; pouring the pasty corn flour into sugared agar water, stirring and boiling into paste; adding propionic acid and stirring; slightly cooling, adding yeast powder, stirring with a glass rod to mix uniformly, adding purified water to a constant volume, and preparing to obtain a culture medium;
step 3, preparing the graphene oxide-based lepidoptera insect feed: and (3) dissolving a culture medium in the graphene oxide solution prepared in the step (1) to prepare the graphene oxide-based lepidoptera insect feed.
4. The use according to claim 3, wherein the mass-to-volume ratio of graphene oxide to ionized water in step 1, mg/ml, is 100:30-400: 30.
5. Use according to claim 3, wherein the mass ratio of corn meal to water is 10: 38.
6. Use according to claim 3, characterized in that the mass ratio of agar to water is 1: 20; the mass ratio of the white sugar to the agar is 9: 1.
7. The use according to claim 3, wherein the volume to mass ratio of propionic acid to corn meal, ml/g, is 1: 20.
8. The use of claim 3, wherein the mass ratio of yeast powder to corn flour is 7: 100.
9. the use of claim 3, wherein the ratio of the total volume after volume fixing to the volume of the corn flour, ml/g, is 17: 2.
10. The use according to claim 3, wherein the volume ratio of the culture medium in step 3 to the ionized water in step 1 is 170: 30.
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