CN112471368A - Artificial feed for cowpea pod borer larvae and preparation method thereof - Google Patents
Artificial feed for cowpea pod borer larvae and preparation method thereof Download PDFInfo
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- A23K50/90—Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
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- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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Abstract
The invention belongs to the technical field of biological feed, and discloses an artificial feed for cowpea pod borer larvae and a preparation method thereof, wherein mung beans and wheat germ powder are placed in a 120 ℃ oven to be baked for 8 hours and then ground by a grinder; weighing agar, adding 1200mL of water, and boiling to completely melt the agar; according to the raw materials and the proportion of the artificial feed for the cowpea pod borer larvae, mung bean powder, wheat germ powder, yeast, cane sugar, methyl p-hydroxybenzoate, potassium sorbate, wechsler's salt, ascorbic acid, choline chloride and compound vitamin are sequentially added into boiled and melted agar water, stirred, uniformly mixed and poured into a clean container, and the container is placed into a refrigerator at 4 ℃ for storage after being cooled to room temperature. The artificial feed for the cowpod borer larvae provided by the invention is simple in components and easy to prepare, optimizes the preparation steps, can be used for continuously feeding the cowpod borers on a large scale indoors, and provides a reference basis for feeding other lepidoptera larvae.
Description
Technical Field
The invention belongs to the technical field of biological feed, and particularly relates to artificial feed for cowpod borer larvae and a preparation method thereof.
Background
The cowpea pod borers Maruca testulalis (Geyer) belong to Lepidoptera, the family of Bombycidae, commonly known as Diatraea viridis and Diatraea virgifera, and are important agricultural pests in bean crops. Is widely distributed worldwide, and occurs in most areas of china.
In recent years, along with global warming, changes of cultivation modes and cultivation systems, the population quantity of the cowpod borers is remarkably increased, and the main pest is that the pods are bored by larvae. The cowpod borers can occur for 2-10 generations all year round in China from north to south, especially in the south China (Shafen, 2007. study on the biological characteristics and artificial feeding method of the pod borers, Hunan: Master academic paper of Hunan agriculture university). The control of the cowpod borer is mainly dependent on chemical agents for prevention and treatment at present, because the larva of the cowpod borer has the property of boring, the chemical prevention and treatment can hardly achieve the ideal effect, the pesticide spraying frequency is increased and the dosage is increased at will by vegetable growers, so that the chemical prevention and treatment effect is reduced, the environment is seriously polluted, and the vegetable pesticide residue is over-standard (Jiguohui, 2001; Moyunfan, 2005). Therefore, how to effectively control the cowpod borers in the planting of the bean vegetables is an urgent problem to be solved. Meanwhile, the production of pollution-free vegetables puts more strict requirements and restrictions on chemical control. Therefore, a safe and effective control method is required.
Therefore, the artificial mass breeding of the cowpod borers is solved, and the provision of large-scale and regularly-developed cowpod borer larvae is the basis for researching the fields of cowpod borer ecology, biology, comprehensive prevention and control and the like. At present, 2 methods are mainly used for breeding indoor cowpea pod borers, firstly, the cowpea pod borers are bred by adopting natural hosts such as cowpea flowers and mung beans, but the natural hosts have long growth period, are easy to rot and need to be replaced frequently, and a large amount of manpower is consumed (Jinjiaxin, 2008); secondly, an artificial feed is adopted to research the formula of the artificial feed for the bean wild borers, although the problem of insect subculture can be solved by the method, the preparation method of the feed is too complicated and is unfavorable for operation, and formaldehyde needs to be added into the feed, so that the health of insect breeders is influenced (Wangbang, 2012).
In summary, the problems of the prior art are as follows: although the 2 common methods can also solve part of problems of experimental insects to some extent, the main problems are (1) that the breeding method is complex, the egg laying amount is low, and large-scale breeding is not facilitated; (2) the preparation process of the feed is complicated, the feed is easy to rot, and the feed needs to be replaced frequently. It is time-consuming and labor-consuming. As can be seen from the above, the existing methods all have different disadvantages and are difficult to meet the insect source requirements of various experiments for various insect states at various ages.
The difficulty of solving the technical problems is as follows: aiming at the problems, the invention provides the artificial feed which is simple in formula and easy to operate in the preparation process and is suitable for large-scale breeding of the cowpod borers, so that large-scale, regular and long-term breeding of standard insect sources is realized.
The significance of solving the technical problems is as follows: the research of large-scale, regular and long-term artificial feed and breeding technology suitable for the cowpod borers aims to improve the laying rate of adults and the survival rate of larvae and realize long-term subculture, and is an important basis for researching the ecology, biology, comprehensive prevention and control technology and production application of the cowpod borers.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides an artificial feed for cowpod borer larvae and a preparation method thereof.
The artificial feed for the cowpod borer larvae is prepared from, by mass, 10-13% of mung bean powder, 1.4-3% of wheat germ powder, 0.8-3% of yeast, 1-2% of sucrose, 1-3% of agar, 0.65-2.2% of methyl p-hydroxybenzoate, 0.3-1% of potassium sorbate, 0.3-1% of wecker salt, 0.2-1% of ascorbic acid, 0.3-1% of choline chloride, 0.05-0.5% of compound vitamin and 70-79% of distilled water.
The invention also aims to provide a preparation method of the artificial feed for the larvae of the cowpod borers, which comprises the following steps:
step one, placing mung bean and wheat germ powder in an oven for baking, and crushing by using a crusher after baking;
step two, weighing agar, adding water and boiling to completely melt the agar;
step three, according to the raw materials and the proportion of the artificial feed for the cowpea pod borer larvae, mung bean powder, wheat germ powder, yeast, sucrose, methyl p-hydroxybenzoate, potassium sorbate, wechsler's salt, ascorbic acid, choline chloride and compound vitamin are sequentially added into boiled and melted agar water and are continuously stirred;
step four, pouring the mixture obtained in the step three into a clean container after the mixture is uniformly mixed, and cooling the mixture; and cooling to room temperature to obtain artificial feed for breeding cowpod borer larva in large scale, and storing in refrigerator.
Further, the mung bean and wheat germ powder are placed in a 120 ℃ oven to be baked, and then are crushed by a crusher after being baked.
Further, the mung bean and wheat germ powder are placed in an oven to be baked for 8 hours in the first step.
Further, agar is weighed in the second step, and 1200mL of water is added for boiling.
And further, cooling to room temperature in the fourth step to obtain the artificial feed for breeding the cowpod borer larvae in a large scale, and storing in a refrigerator at 4 ℃ for later use.
The invention also aims to provide an artificial large-scale breeding method of the test population of the cowpod borer by applying the artificial feed for the larvae of the cowpod borer, which comprises the following steps:
(1) feeding the cowpod borer larvae with a sterilized 24-hole cell culture plate, putting prepared artificial feed accounting for 1/3 in each hole, and inoculating 1 initial-hatched cowpod borer larva into each hole;
(2) covering the cell culture plate with a layer of blow-molded paper, covering the cell culture plate, and binding rubber bands;
(3) and (5) placing the cowpea pod borer in a light incubator for culture, and observing and recording survival conditions of the cowpea pod borer larvae every day.
Further, the cowpod borer larvae fed with the feed are fed by a sterilized 24-hole cell culture plate, prepared artificial feed occupying the volume of the holes 1/3 is put into each hole, and 1 initial-hatched cowpod borer larva is inoculated into each hole.
Further, the culture conditions of the illumination incubator are as follows: the temperature was 26 ℃. + -. 2 ℃ and the relative humidity was 40%.
Further, the photoperiod in the illumination incubator is 14L: 10D.
In summary, the advantages and positive effects of the invention are: the artificial feed for the cowpod borer larvae provided by the invention is simple in components and easy to prepare, the preparation steps are optimized, the cowpod borers can be continuously fed indoors in a large scale, and meanwhile, a reference basis is provided for feeding other lepidoptera larvae. The feed has good palatability to cowpea pod borer larva by using mung bean, yeast, embryo and sucrose as main components, and simultaneously, the feed is added with potassium sorbate and methyl p-hydroxybenzoate commonly used in many food industries, so that the antiseptic effect can be improved, the ascorbic acid can be protected, and the situation of rotting and mildew of artificial feed in the larva feeding process can be avoided.
The method for breeding the accurate cowpod borer experimental population with the uniform insect stages provided by the invention overcomes the defects of the prior art. Experiments show that the feed based on the mung beans, the yeast, the germs and the cane sugar as main components has good palatability to cowpod borer larvae, and the feed can realize the artificial continuous feeding of the cowpod borers and provide insect source guarantee for indoor research. After a proper amount of choline chloride and some preservatives are added, the quality of the feed is ensured, the feeding mildew condition is avoided, the feeding and the oviposition can be promoted, and the technical support is provided for the indoor artificial mass propagation of the cowpod borers.
The prior art cannot realize batch and subculture of the cowpod borer test insects by the formula mainly because of the following reasons: (1) some components in the feed are not enough to meet the growth requirement of the cowpod borers, although the insects are lepidoptera insects, host plants of various insects are different, required nutrient components are different, and main hosts of the cowpod borers are leguminous crops, so that the formula selects mung bean flour which is a main host material as one of main raw materials, and is different from other insects (Chenzong 250351983; panhong jade 1991; Chen jin 2000; Lvfei 2007; Dachang, 2016). (2) The feed is not beneficial to the feeding of the low-age larvae, the survival rate of the low-age larvae is low, the low-age larvae are related to insufficient nutritional ingredients in the feed formula and overhigh water content (Xiaofen, 2008). (3) Although the subculture can be completed, the death rate of the larvae is high, the mortality rate is related to the ratio of feed preservatives, the feed is easy to generate bacteria, the insects are difficult to grow, the death rate of the larvae is too high, and the formula preparation process is complicated, so that the mass production is not facilitated (Wang Pan, 2012). The invention takes the green bean powder and the wheat germ powder as main raw materials, increases the proportion of the green bean powder to the wheat germ powder as the main raw materials, reduces the water content, removes formaldehyde which is unfavorable for people, uses the methyl p-hydroxybenzoate and the potassium sorbate as preservatives, and increases the proportion of the ascorbic acid to effectively ensure the bacteriostatic effect of the feed. The preparation process of the feed is simplified. The feed formula and the preparation process are suitable for group breeding of the young larvae (1-3 instars), and the young larvae can be fed singly after the larvae grow to 4 instars. This can greatly save the feeding time and also provide convenience for large-scale feeding.
Drawings
Fig. 1 is a flow chart of a preparation method of the artificial feed for the cowpod borer larvae, which is provided by the embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Aiming at the problems in the prior art, the invention provides an artificial feed for cowpod borer larvae and a preparation method thereof, and the invention is described in detail with reference to the accompanying drawings.
The artificial feed for the cowpod borer larvae provided by the embodiment of the invention comprises, by mass, 10-13% of mung bean flour, 1.4-3% of wheat germ flour, 0.8-3% of yeast (Angel yeast ordered by a reagent company), 1-2% of sucrose, 1-3% of agar, 0.65-2.2% of methyl p-hydroxybenzoate, 0.3-1% of potassium sorbate, 0.3-1% of Welsh's salt, 0.2-1% of ascorbic acid, 0.3-1% of choline chloride, 0.05-0.5% of vitamin complex and 70-79% of distilled water.
As shown in fig. 1, the preparation method of the artificial feed for the cowpod borer larvae provided by the embodiment of the invention comprises the following steps:
s101, placing the mung bean and wheat germ powder in a 120 ℃ oven for baking for 8 hours, baking, and crushing by using a crusher.
S102, weighing agar, adding 1200mL of water, boiling, and completely melting the agar.
S103, according to the raw materials and the proportion of the artificial feed for the cowpea pod borer larvae, mung bean powder, wheat germ powder, yeast, sucrose, methyl p-hydroxybenzoate, potassium sorbate, wechsler' S salt, ascorbic acid, choline chloride and vitamin complex are sequentially added into boiled and melted agar water and are continuously stirred.
And S104, S103, pouring the mixture into a clean container after uniformly mixing, and cooling. Cooling to room temperature to obtain artificial feed for breeding cowpod borer larva in large scale, and storing in 4 deg.C refrigerator.
The artificial large-scale breeding method of the cowpod borer test population provided by the embodiment of the invention comprises the following steps:
(1) feeding the cowpod borer larvae with a sterilized 24-hole cell culture plate, putting the prepared appropriate amount of artificial feed accounting for about 1/3 in each hole, and inoculating 1 initial-hatched cowpod borer larva in each hole.
(2) Covering the cell culture plate with a layer of blow-molded paper, covering the cell culture plate, and binding rubber bands.
(3) Placing the mixture in a light incubator at 26 +/-2 ℃ and 40% relative humidity, wherein the photoperiod is 14L: 10D.
(4) The survival condition of the cowpod borer larvae is observed and recorded every day.
The present invention will be further described with reference to the following examples.
The raw materials and the proportion of the artificial feed for industrially breeding the cowpod borer larvae on a large scale are shown in table 1.
TABLE 1 raw materials and formulation of artificial feed for industrial mass rearing of cowpod borer larvae
| Composition (I) | Formulation 1 (%) | Formulation 2 (%) | Formulation 3 (%) |
| Agar-agar | 2 | 3 | 1.5 |
| Mung bean powder | 13 | 19 | 10 |
| Wheat germ powder | 2 | 1.4 | 3 |
| Yeast | 0.8 | 2 | 3 |
| Sucrose | 1 | 1.5 | 2 |
| P-hydroxybenzoic acid methyl ester | 0.65 | 1.2 | 2.2 |
| Potassium sorbate | 0.3 | 0.5 | 1 |
| Ascorbic acid | 0.2 | 1 | 0.5 |
| Choline chloride | 1 | 0.3 | 0.6 |
| Compound vitamin | 0.05 | 0.1 | 0.5 |
| Distilled water | 79 | 70 | 75.7 |
Example 1
100 heads of cowpea pod borer pupae are placed in a spawning cage, after the cowpea pod borer pupae are eclosized and spawned, the newly hatched larvae are used as a test insect source, are inoculated into the feed of the formula 1 in the table 1, and are placed in an insect breeding room for breeding, wherein the breeding conditions of the larvae are 26 +/-2 ℃ and the relative humidity is 40%.
Example 2
100 heads of cowpea pod borer pupae are placed in a spawning cage, after the cowpea pod borer pupae are eclosized and spawned, the newly hatched larvae are used as a test insect source, are inoculated into the feed of the formula 2 in the table 1, and are placed in an insect breeding room for breeding, wherein the breeding conditions of the larvae are 26 +/-2 ℃ and the relative humidity is 40%.
Example 3
100 heads of cowpea pod borer pupae are placed in a spawning cage, after the cowpea pod borer pupae are eclosized and spawned, the newly hatched larvae are used as a test insect source, are inoculated into the feed of the formula 3 in the table 1, and are placed in an insect breeding room for breeding, wherein the breeding conditions of the larvae are 26 +/-2 ℃ and the relative humidity is 40%.
Example 4: evaluation of feed Effect
The artificial feed for the cowpod borer larvae prepared by the 3 examples is used for breeding among insect breeding rooms, the larva breeding conditions are 26 +/-2 ℃, at least 20 generations of breeding are continuously carried out under the condition that the relative humidity is 40%, and the population degradation is not caused. As shown in Table 2, the survival rate of the larvae can reach more than 80% by the 3 formulas, which shows that the feed based on mung beans, yeast, germs and sucrose as main components has good palatability to the larvae of the cowpod borer, and the feed can realize the artificial continuous feeding of the cowpod borer and provide insect source guarantee for indoor research. After a proper amount of choline chloride and some preservatives are added, the quality of the feed is ensured, the feeding mildew condition is avoided, the feeding and the oviposition can be promoted, and the technical support is provided for the indoor artificial mass propagation of the cowpod borers.
TABLE 2 indoor artificial feeding metering evaluation of cowpod borer
| Formulation of | Hatchability% | Survival rate of 1 instar larva% | Larvae through the course (d)26 ℃ +/-2 DEG C | Weight of pupa |
| 1 | 82.2 | 80.2 | 12.65 | 40.7 |
| 2 | 80.8 | 81.3 | 13.45 | 41.2 |
| 3 | 80.1 | 82.6 | 13.54 | 40.3 |
Example 5: indoor artificial feeding generation life parameter table for cowpea pod borer
Test materials and methods:
a test insect source: collecting larvae with consistent insect age from a mung bean test base in agricultural academy of sciences in Hubei province, feeding fresh mung bean pods into an insect breeding box in a single-head mode, after pupation and eclosion and spawning of adults, taking newly hatched larvae as a tested insect source, and continuously feeding for 20 generations under the conditions that the indoor temperature is 26 ℃ and 2 ℃, the relative humidity is 70 ℃ and the photoperiod is 14L to 10D. Test feed: the artificial feed formula 1.
The test method comprises the following steps: the test insects are raised according to the method, the test insects are continuously raised for 20 generations, and the incubation rate of the eggs, the survival rate of the larvae, the developmental stage of the larvae, the pupal weight, the service life of adults and the egg laying amount are counted in each generation. The results show that the life parameter difference among generations is very small, the population development is neat, and the results show that the cowpod borers bred by the artificial feed can completely meet the requirement of indoor subculture breeding, and a large number of test insects with neat and consistent growth and development can be obtained.
TABLE 3 indoor artificial feeding generation life parameter table for cowpod borer
Note that the data in the table are mean values and standard errors, and the same letters after the same row data indicate insignificant differences after multiple comparisons of new repolarization differences (P > 0.05).
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (10)
1. An artificial feed for cowpod borer larvae is characterized by comprising, by mass, 10-13% of mung bean powder, 1.4-3% of wheat germ powder, 0.8-3% of yeast, 1-2% of sucrose, 1-3% of agar, 0.65-2.2% of methyl p-hydroxybenzoate, 0.3-1% of potassium sorbate, 0.3-1% of weber's salt, 0.2-1% of ascorbic acid, 0.3-1% of choline chloride, 0.05-0.5% of vitamin complex and 70-79% of distilled water.
2. The method for preparing the artificial feed for the larvae of the cowpod borers according to claim 1, wherein the method for preparing the artificial feed for the larvae of the cowpod borers comprises the following steps:
step one, placing mung bean and wheat germ powder in an oven for baking, and crushing by using a crusher after baking;
step two, weighing agar, adding water and boiling to completely melt the agar;
step three, according to the raw materials and the proportion of the artificial feed for the cowpea pod borer larvae, mung bean powder, wheat germ powder, yeast, sucrose, methyl p-hydroxybenzoate, potassium sorbate, wechsler's salt, ascorbic acid, choline chloride and compound vitamin are sequentially added into boiled and melted agar water and are continuously stirred;
step four, pouring the mixture obtained in the step three into a clean container after the mixture is uniformly mixed, and cooling the mixture; and cooling to room temperature to obtain artificial feed for breeding cowpod borer larva in large scale, and storing in refrigerator.
3. The method for preparing an artificial feed for cowpea pod borer larvae, according to claim 2, wherein the first step is to bake the mung beans and the wheat germ powder in an oven at 120 ℃, bake the powder and then crush the powder by a crusher.
4. The method for preparing an artificial feed for cowpod borer larva according to claim 2, wherein the first step is to bake the mung bean and wheat germ powder in an oven for 8 hours.
5. The method of claim 2, wherein the step two comprises weighing agar, adding 1200 water, and boiling.
6. The method for preparing artificial feed for cowpod borer larva according to claim 2, wherein the fourth step is cooling to room temperature to obtain artificial feed for large scale rearing of cowpod borer larva, and storing in 4 deg.C refrigerator for further use.
7. A method for artificially feeding a test population of cowpod borers using the artificial feed for larvae of cowpod borers according to claim 1, wherein the method for artificially feeding a test population of cowpod borers comprises the following steps:
(1) feeding the cowpod borer larvae with a sterilized 24-hole cell culture plate, putting prepared artificial feed accounting for 1/3 in each hole, and inoculating 1 initial-hatched cowpod borer larva into each hole;
(2) covering the cell culture plate with a layer of blow-molded paper, covering the cell culture plate, and binding rubber bands;
(3) and (5) placing the cowpea pod borer in a light incubator for culture, and observing and recording survival conditions of the cowpea pod borer larvae every day.
8. The method of claim 7, wherein the fed cowpod borer larvae are fed using a sterilized 24-well cell culture plate, wherein the artificial feed prepared in the volume of 1/3 is placed in each well, and 1 initial hatching of the cowpod borer larvae is added to each well.
9. The method for artificially feeding a test population of cowpod borers according to claim 7, wherein the light incubator is under the following conditions: the temperature was 26 ℃. + -. 2 ℃ and the relative humidity was 40%.
10. The method of artificially feeding a test population of cowpod borers according to claim 6, wherein the photoperiod in the light incubator is 14L: 10D.
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