Feed for larvae of spodoptera frugiperda and preparation method of feed
By using a compound feed formula composed of corn flour, yeast powder, etc. and a specific fermentation method, the problems of short shelf life and high deformity rate of fall armyworm larvae feed were solved, and the feed effect of high survival rate and long shelf life was achieved.
Patent Information
- Application Number
- CN202511064038.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-31
- Publication Date
- 2025-10-03
AI Technical Summary
The existing feed for fall armyworm larvae has shortcomings in terms of shelf life and anti-corrosion properties, and the survival rate and health rate are low when raised at high density, which makes them prone to deformities and affects experimental research.
The compound feed formula composed of corn flour, yeast powder, agar, lecithin, cholesterol, trehalose, Bacillus cereus fermentation products and preservatives, combined with specific fermentation and preparation methods, can improve the pupation rate and emergence rate, reduce the deformity rate and extend the shelf life.
It improves the pupation and emergence rates of fall armyworm larvae, reduces the deformity rate, extends the shelf life of feed, and ensures the survival rate and health status of high-density breeding.
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of experimental insect breeding, and particularly relates to a feed for fall armyworm larvae and a preparation method thereof. Background Art
[0002] Fall armyworm (scientific name: Spodoptera frugiperda ), also known as fall armyworm, false armyworm or fall armyworm, belongs to the order Lepidoptera ( Lepidoptera ) Noctuidae ( Noctuidae ) Spodoptera ( Spodoptera The fall armyworm (Spodoptera frugiperda) is a global migratory pest native to tropical and subtropical regions of the Americas. It possesses exceptionally high reproductive capacity, migratory ability, and destructive power. It can harm over 350 plant species across 76 families, with a preference for gramineous crops such as corn, rice, wheat, and sugarcane, as well as soybeans, cotton, and fruit trees. Under suitable temperatures, a generation (egg → larva → pupa → adult) completes in approximately 30 days. Female moths can lay over 1,000 eggs in a single session, and adults can migrate 100 kilometers in a single night. Larvae feed on leaves, forming transparent "window-like" spots in younger larvae. Advanced larvae create irregular holes and notches, and can even bore into heart leaves and stems, resulting in "heart death" or even bare stems in corn plants. Given the destructive power and widespread nature of the fall armyworm, scientific research is necessary for its prevention. This research requires standardized breeding of experimental fall armyworms, primarily through the development of feed for fall armyworm larvae.
[0003] Research on feed for fall armyworm larvae has evolved from single plant raw materials to complex nutritional formulas. Many studies have shown that corn-based feed performs outstandingly in promoting development and reproduction. By comparing the effects of different feeds on the growth and development of fall armyworm, it was found that the "corn-artificial feed" combination can shorten the development period to 27.78 days, which is significantly better than single artificial feed or castor leaves. The fully artificial feed is based on corn flour, soybean flour, and wheat germ flour, with the addition of special ingredients such as maggot powder and spirulina, which significantly improves the egg-laying rate and the emergence rate. The semi-artificial feed uses a composite system of soybean flour, corn flour, and wheat flour. By adding lecithin and linalool to stimulate feeding, the larval survival rate is stabilized at 88%, and the biological parameters are maintained stable for 40 consecutive generations of cultivation.
[0004] Most artificial feeds can only be stored for about one day at room temperature, and even at low temperatures of 0°C, they can only maintain a shelf life of about one month. Feeds are prone to mold or bacteria growth, causing deterioration and affecting larval survival rates. Although some formulas have added preservatives such as potassium sorbate and methylparaben, the long-term preservation effect is still not ideal. When reared at high densities, larvae cannibalize each other due to competition for feed or space, resulting in a decrease in survival rate. Group rearing can easily lead to cross-infection of pathogenic microorganisms. Although antibiotics such as streptomycin are added, long-term use may induce drug resistance. Deformity problems that occur during the rearing of fall armyworm larvae (such as deformed pupae, developmental abnormalities, etc.) are one of the important factors affecting their large-scale rearing and experimental research. Summary of the Invention
[0005] The present invention aims to provide a feed for fall armyworm larvae and a preparation method thereof.
[0006] A feed for fall armyworm larvae comprises the following components in parts by weight: 100-200 parts of corn flour, 20-60 parts of yeast powder, 10-30 parts of agar, 3-5 parts of lecithin, 0.1-0.3 parts of cholesterol, 0.5-1.5 parts of trehalose, 1-4 parts of vitamins, 2-8 parts of Bacillus cereus fermentation product, 5-10 parts of corn leaf powder, 5-10 parts of preservatives, and 800-1200 parts of water.
[0007] The vitamins include the following components in parts by weight: 1-3 parts of vitamin C, 0.01-0.05 parts of nicotinamide, 0.01-0.03 parts of calcium pantothenate, 0.01-0.1 parts of vitamin B1, 0.01-0.03 parts of vitamin B2, 0.01-0.03 parts of vitamin B6, 0.01-0.05 parts of folic acid, 0.01-0.03 parts of biotin, and 0.002-0.006 parts of vitamin B12.
[0008] The preparation method of the Bacillus cereus fermentation product is as follows: (1) activating Bacillus cereus: picking a single colony of Bacillus cereus cultured by streaking and placing it in LB broth culture medium, and culturing it on a shaker at 33-38°C and 100-300r / min for 12-36h to obtain an activated bacterial liquid; (2) adding 2-4% activated bacterial liquid prepared in step (1) to a fermentation buffer solution with a pH of 7 and a chicken feather content of 2-5% by weight; maintaining the fermentation temperature at 33-38°C and the fermentation time for 4-8d; (3) pouring the fermented liquid into two 100mL centrifuge tubes, balancing them, placing them opposite to each other in a centrifuge, centrifuging them at 2000-4000r / min for 8-12min, taking the supernatant, and vacuum freeze-drying and then crushing them.
[0009] The Bacillus cereus is Bacillus cereus ( Bacillus cereus) YSQ08, purchased from China Center for Type Culture Collection, with the strain collection number being CCTCC No. M2012322.
[0010] The fermentation buffer comprises: 0.1-0.8 g / L sodium chloride, 0.1-0.5 g / L calcium chloride, 0.1-0.5 g / L magnesium chloride, 1-3 g / L dipotassium hydrogen phosphate, and 0.5-1.5 g / L potassium dihydrogen phosphate.
[0011] The LB broth culture medium is prepared by mixing 8-12 g of tryptone, 3-7 g of yeast extract, and 3-7 g of NaCl, adding distilled water to make up to 1000 mL, and sterilizing by autoclaving at 121° C. for 15-25 min.
[0012] The preservative is one or more of sodium benzoate, potassium sorbate, propylparaben, monomethyl fumarate, ethyl p-hydroxybenzoate, and dimethyl fumarate.
[0013] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 20-40 minutes, heating to dissolve, then adding corn flour, stirring evenly, heating to boiling, and keeping warm for 5-10 minutes; (2) cooling to 60-70°C, adding corn leaf powder, lecithin, cholesterol, and trehalose, stirring evenly, and keeping warm for 3-5 minutes; (3) cooling to 35-50°C, adding yeast powder, vitamins, Bacillus cereus fermentation product, and preservatives, stirring evenly, and cooling to room temperature to prepare the feed.
[0014] The heating temperature in step (1) is 80-95°C.
[0015] The present invention has the beneficial effects of feeding fall armyworms with feed prepared in the present invention, which has high pupation and emergence rates, low deformity rates, and a long shelf life. The combined use of cholesterol and trehalose in the feed improves the pupation and emergence rates of fall armyworms, the combination of a Bacillus cereus fermentation product and corn leaf meal reduces the deformity rate, and the combination of monomethyl fumarate and ethyl parahydroxybenzoate improves the feed's antiseptic properties and extends its shelf life. DETAILED DESCRIPTION
[0016] To facilitate understanding of the present invention, the present invention will be described more fully below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive. Example 1
[0017] A feed for fall armyworm larvae comprises the following components in parts by weight: 150 parts of corn flour, 40 parts of yeast powder, 20 parts of agar, 4 parts of lecithin, 0.2 parts of cholesterol, 0.8 parts of trehalose, 2.194 parts of vitamins, 6 parts of Bacillus cereus fermentation product, 8 parts of corn leaf powder, 8 parts of a preservative, and 1000 parts of water; the vitamins comprise the following components in parts by weight: 2 parts of vitamin C, 0.03 parts of nicotinamide, 0.02 parts of calcium pantothenate, 0.05 parts of vitamin B1, 0.02 parts of vitamin B2, 0.02 parts of vitamin B6, 0.03 parts of folic acid, 0.02 parts of biotin, and 0.004 parts of vitamin B12; and the preservative is a mixture of monomethyl fumarate and ethyl parahydroxybenzoate in a mass ratio of 1:1.
[0018] The preparation method of the Bacillus cereus fermentation product is as follows: (1) Activation of Bacillus cereus: picking a single colony of Bacillus cereus that has been cultured by streaking is placed in LB broth culture medium, and cultured on a shaker at 37°C and 200 r / min for 24 hours to obtain an activated bacterial solution; the Bacillus cereus is Bacillus cereus ( Bacillus cereus ) YSQ08, purchased from China Center for Type Culture Collection, with a strain collection number of CCTCC No. M2012322; the LB broth culture medium was prepared as follows: 10 g of tryptone, 5 g of yeast extract, and 5 g of NaCl were mixed, the volume was fixed to 1000 mL with distilled water, and the mixture was sterilized at 121°C for 20 min; (2) the 3% activated bacterial solution prepared in step (1) was added to a fermentation buffer solution with a pH of 7 and a chicken feather content of 3%; the fermentation temperature was maintained at 37°C, and the fermentation time was 6 d; the fermentation buffer solution was composed of: 0.5 g / L sodium chloride, 0.3 g / L calcium chloride, 0.3 g / L magnesium chloride, 2 g / L potassium dihydrogen phosphate, and 1 g / L potassium dihydrogen phosphate; (3) the fermented liquid was poured into two 100 mL centrifuge tubes, balanced, and placed oppositely in a centrifuge, centrifuged at 3000 r for 10 min, the supernatant was taken, and the mixture was freeze-dried in vacuum and then crushed.
[0019] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 30 minutes, heating at 85°C to dissolve, then adding corn flour, stirring evenly, heating to boiling, and keeping warm for 6 minutes; (2) cooling to 65°C, adding corn leaf powder, lecithin, cholesterol, and trehalose, stirring evenly, and keeping warm for 4 minutes; (3) cooling to 40°C, adding yeast powder, vitamins, Bacillus cereus fermentation product, and preservatives, stirring evenly, and cooling to room temperature to prepare the feed. Example 2
[0020] A feed for fall armyworm larvae comprises the following components in parts by weight: 120 parts of corn flour, 25 parts of yeast powder, 12 parts of agar, 3 parts of lecithin, 0.1 part of cholesterol, 0.5 part of trehalose, 1.092 parts of vitamins, 3 parts of Bacillus cereus fermentation product, 5 parts of corn leaf powder, 5 parts of preservatives, and 800 parts of water; the vitamins comprise the following components in parts by weight: 1 part of vitamin C, 0.01 part of nicotinamide, 0.01 part of calcium pantothenate, 0.02 part of vitamin B1, 0.01 part of vitamin B2, 0.01 part of vitamin B6, 0.02 part of folic acid, 0.01 part of biotin, and 0.002 part of vitamin B12; and the preservative is a mixture of potassium sorbate and propylparaben in a mass ratio of 2:1.
[0021] The preparation method of the Bacillus cereus fermentation product is as follows: (1) Activation of Bacillus cereus: picking a single colony of Bacillus cereus that has been cultured by streaking is placed in LB broth medium, and cultured on a shaker at 36°C and 150 r / min for 18 hours to obtain an activated bacterial solution; the Bacillus cereus is Bacillus cereus ( Bacillus cereus ) YSQ08, purchased from China Center for Type Culture Collection, with a strain collection number of CCTCC No. M2012322; the LB broth culture medium was prepared as follows: 8 g of tryptone, 3 g of yeast extract, and 3 g of NaCl were mixed, the volume was fixed to 1000 mL with distilled water, and the mixture was sterilized at 121°C for 15 min; (2) the 2% activated bacterial solution prepared in step (1) was added to a fermentation buffer solution with a pH of 7 and a chicken feather content of 2%; the fermentation temperature was maintained at 36°C, and the fermentation time was 5 d; the fermentation buffer solution was composed of: 0.2 g / L sodium chloride, 0.1 g / L calcium chloride, 0.1 g / L magnesium chloride, 1 g / L potassium dihydrogen phosphate, and 0.5 g / L potassium dihydrogen phosphate; (3) the fermented liquid was poured into two 100 mL centrifuge tubes, balanced, and placed oppositely in a centrifuge, centrifuged at 2000 r for 12 min, the supernatant was taken, and the mixture was freeze-dried in vacuum and then crushed.
[0022] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 20 minutes, heating at 80°C to dissolve, then adding corn flour, stirring evenly, heating to boiling, and keeping warm for 10 minutes; (2) cooling to 60°C, adding corn leaf powder, lecithin, cholesterol, and trehalose, stirring evenly, and keeping warm for 5 minutes; (3) cooling to 35°C, adding yeast powder, vitamins, Bacillus cereus fermentation product, and preservatives, stirring evenly, and cooling to room temperature to prepare the feed. Example 3
[0023] A feed for fall armyworm larvae comprises the following components in parts by weight: 180 parts of corn flour, 50 parts of yeast powder, 30 parts of agar, 5 parts of lecithin, 0.3 parts of cholesterol, 1.5 parts of trehalose, 3.326 parts of vitamins, 6 parts of Bacillus cereus fermentation product, 9 parts of corn leaf powder, 8 parts of preservatives, and 1200 parts of water; the vitamins comprise the following components in parts by weight: 3 parts of vitamin C, 0.05 parts of nicotinamide, 0.03 parts of calcium pantothenate, 0.1 parts of vitamin B1, 0.03 parts of vitamin B2, 0.03 parts of vitamin B6, 0.05 parts of folic acid, 0.03 parts of biotin, and 0.006 parts of vitamin B12; and the preservative is a mixture of sodium benzoate and dimethyl fumarate in a mass ratio of 3:1.
[0024] The preparation method of the Bacillus cereus fermentation product is as follows: (1) Activation of Bacillus cereus: picking a single colony of Bacillus cereus that has been cultured by streaking is placed in LB broth culture medium, and cultured on a shaker at 35°C and 300 r / min for 36 hours to obtain an activated bacterial solution; the Bacillus cereus is Bacillus cereus ( Bacillus cereus ) YSQ08, purchased from China Center for Type Culture Collection, with a strain collection number of CCTCC No. M2012322; the LB broth culture medium was prepared as follows: 12 g of tryptone, 6 g of yeast extract, and 6 g of NaCl were mixed, the volume was fixed to 1000 mL with distilled water, and the mixture was sterilized at 121°C for 25 min; (2) the 4% activated bacterial solution prepared in step (1) was added to a fermentation buffer solution with a pH of 7 and a chicken feather content of 4%; the fermentation temperature was maintained at 35°C, and the fermentation time was 8 d; the fermentation buffer solution was composed of: 0.8 g / L sodium chloride, 0.5 g / L calcium chloride, 0.5 g / L magnesium chloride, 3 g / L potassium dihydrogen phosphate, and 1.5 g / L potassium dihydrogen phosphate; (3) the fermented liquid was poured into two 100 mL centrifuge tubes, balanced, and placed oppositely in a centrifuge, centrifuged at 4000 r for 8 min, the supernatant was taken, and the mixture was freeze-dried in vacuum and then crushed.
[0025] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 40 minutes, heating at 90°C to dissolve, then adding corn flour, stirring evenly, heating to boiling, and keeping warm for 10 minutes; (2) cooling to 70°C, adding corn leaf powder, lecithin, cholesterol, and trehalose, stirring evenly, and keeping warm for 5 minutes; (3) cooling to 50°C, adding yeast powder, vitamins, Bacillus cereus fermentation product, and preservatives, stirring evenly, and cooling to room temperature to prepare the feed.
[0026] Comparative Example 1: A feed for fall armyworm larvae, comprising the following components in parts by weight: 150 parts of corn flour, 40 parts of yeast powder, 20 parts of agar, 4 parts of lecithin, 1 part of cholesterol, 2.194 parts of vitamins, 6 parts of Bacillus cereus fermentation product, 8 parts of corn leaf powder, 8 parts of preservative, and 1000 parts of water; the vitamins include the following components in parts by weight: 2 parts of vitamin C, 0.03 parts of nicotinamide, 0.02 parts of calcium pantothenate, 0.05 parts of vitamin B1, 0.02 parts of vitamin B2, 0.02 parts of vitamin B6, 0.03 parts of folic acid, 0.02 parts of biotin, and 0.004 parts of vitamin B12; the preservative is a mixture of monomethyl fumarate and ethyl parahydroxybenzoate in a mass ratio of 1:1.
[0027] The preparation method of the Bacillus cereus fermentation product is as follows: (1) Activation of Bacillus cereus: picking a single colony of Bacillus cereus that has been cultured by streaking is placed in LB broth culture medium, and cultured on a shaker at 37°C and 200 r / min for 24 hours to obtain an activated bacterial solution; the Bacillus cereus is Bacillus cereus ( Bacillus cereus ) YSQ08, purchased from China Center for Type Culture Collection, with a strain collection number of CCTCC No. M2012322; the LB broth culture medium was prepared as follows: 10 g of tryptone, 5 g of yeast extract, and 5 g of NaCl were mixed, the volume was fixed to 1000 mL with distilled water, and the mixture was sterilized at 121°C for 20 min; (2) the 3% activated bacterial solution prepared in step (1) was added to a fermentation buffer solution with a pH of 7 and a chicken feather content of 3%; the fermentation temperature was maintained at 37°C, and the fermentation time was 6 d; the fermentation buffer solution was composed of: 0.5 g / L sodium chloride, 0.3 g / L calcium chloride, 0.3 g / L magnesium chloride, 2 g / L potassium dihydrogen phosphate, and 1 g / L potassium dihydrogen phosphate; (3) the fermented liquid was poured into two 100 mL centrifuge tubes, balanced, and placed oppositely in a centrifuge, centrifuged at 3000 r for 10 min, the supernatant was taken, and the mixture was freeze-dried in vacuum and then crushed.
[0028] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 30 minutes, heating at 85°C to dissolve, then adding corn flour, stirring evenly, heating to boiling, and keeping warm for 6 minutes; (2) cooling to 65°C, adding corn leaf powder, lecithin, and cholesterol, stirring evenly, and keeping warm for 4 minutes; (3) cooling to 40°C, adding yeast powder, vitamins, Bacillus cereus fermentation product, and preservatives, stirring evenly, and cooling to room temperature to prepare the feed.
[0029] Comparative Example 2: A feed for fall armyworm larvae, comprising the following components in parts by weight: 150 parts of corn flour, 40 parts of yeast powder, 20 parts of agar, 4 parts of lecithin, 1 part of trehalose, 2.194 parts of vitamins, 6 parts of Bacillus cereus fermentation product, 8 parts of corn leaf powder, 8 parts of preservative, and 1000 parts of water; the vitamins include the following components in parts by weight: 2 parts of vitamin C, 0.03 parts of nicotinamide, 0.02 parts of calcium pantothenate, 0.05 parts of vitamin B1, 0.02 parts of vitamin B2, 0.02 parts of vitamin B6, 0.03 parts of folic acid, 0.02 parts of biotin, and 0.004 parts of vitamin B12; the preservative is a mixture of monomethyl fumarate and ethyl parahydroxybenzoate in a mass ratio of 1:1.
[0030] The preparation method of the Bacillus cereus fermentation product is as follows: (1) Activation of Bacillus cereus: picking a single colony of Bacillus cereus that has been cultured by streaking is placed in LB broth culture medium, and cultured on a shaker at 37°C and 200 r / min for 24 hours to obtain an activated bacterial solution; the Bacillus cereus is Bacillus cereus ( Bacillus cereus ) YSQ08, purchased from China Center for Type Culture Collection, with a strain collection number of CCTCC No. M2012322; the LB broth culture medium was prepared as follows: 10 g of tryptone, 5 g of yeast extract, and 5 g of NaCl were mixed, the volume was fixed to 1000 mL with distilled water, and the mixture was sterilized at 121°C for 20 min; (2) the 3% activated bacterial solution prepared in step (1) was added to a fermentation buffer solution with a pH of 7 and a chicken feather content of 3%; the fermentation temperature was maintained at 37°C, and the fermentation time was 6 d; the fermentation buffer solution was composed of: 0.5 g / L sodium chloride, 0.3 g / L calcium chloride, 0.3 g / L magnesium chloride, 2 g / L potassium dihydrogen phosphate, and 1 g / L potassium dihydrogen phosphate; (3) the fermented liquid was poured into two 100 mL centrifuge tubes, balanced, and placed oppositely in a centrifuge, centrifuged at 3000 r for 10 min, the supernatant was taken, and the mixture was freeze-dried in vacuum and then crushed.
[0031] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 30 minutes, heating at 85°C to dissolve, then adding corn flour, stirring evenly, heating to boiling, and keeping warm for 6 minutes; (2) cooling to 65°C, adding corn leaf powder, lecithin, and trehalose, stirring evenly, and keeping warm for 4 minutes; (3) cooling to 40°C, adding yeast powder, vitamins, Bacillus cereus fermentation product, and preservatives, stirring evenly, and cooling to room temperature to prepare the feed.
[0032] Comparative Example 3: A feed for the larvae of fall armyworm, comprising the following components in parts by weight: 150 parts of corn flour, 40 parts of yeast powder, 20 parts of agar, 4 parts of lecithin, 0.2 parts of cholesterol, 0.8 parts of trehalose, 2.194 parts of vitamins, 14 parts of Bacillus cereus fermentation product, 8 parts of preservative, and 1000 parts of water; the vitamins include the following components in parts by weight: 2 parts of vitamin C, 0.03 parts of nicotinamide, 0.02 parts of calcium pantothenate, 0.05 parts of vitamin B1, 0.02 parts of vitamin B2, 0.02 parts of vitamin B6, 0.03 parts of folic acid, 0.02 parts of biotin, and 0.004 parts of vitamin B12; the preservative is a mixture of monomethyl fumarate and ethyl parahydroxybenzoate in a mass ratio of 1:1.
[0033] The preparation method of the Bacillus cereus fermentation product is as follows: (1) Activation of Bacillus cereus: picking a single colony of Bacillus cereus that has been cultured by streaking is placed in LB broth culture medium, and cultured on a shaker at 37°C and 200 r / min for 24 hours to obtain an activated bacterial solution; the Bacillus cereus is Bacillus cereus ( Bacillus cereus ) YSQ08, purchased from China Center for Type Culture Collection, with a strain collection number of CCTCC No. M2012322; the LB broth culture medium was prepared as follows: 10 g of tryptone, 5 g of yeast extract, and 5 g of NaCl were mixed, the volume was fixed to 1000 mL with distilled water, and the mixture was sterilized at 121°C for 20 min; (2) the 3% activated bacterial solution prepared in step (1) was added to a fermentation buffer solution with a pH of 7 and a chicken feather content of 3%; the fermentation temperature was maintained at 37°C, and the fermentation time was 6 d; the fermentation buffer solution was composed of: 0.5 g / L sodium chloride, 0.3 g / L calcium chloride, 0.3 g / L magnesium chloride, 2 g / L potassium dihydrogen phosphate, and 1 g / L potassium dihydrogen phosphate; (3) the fermented liquid was poured into two 100 mL centrifuge tubes, balanced, and placed oppositely in a centrifuge, centrifuged at 3000 r for 10 min, the supernatant was taken, and the mixture was freeze-dried in vacuum and then crushed.
[0034] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 30 minutes, heating at 85°C to dissolve, then adding corn starch, stirring evenly, heating to boiling, and keeping warm for 6 minutes; (2) cooling to 65°C, adding lecithin, cholesterol, and trehalose, stirring evenly, and keeping warm for 4 minutes; (3) cooling to 40°C, adding yeast powder, vitamins, Bacillus cereus fermentation product, and preservatives, stirring evenly, and cooling to room temperature to prepare the feed.
[0035] Comparative Example 4: A feed for fall armyworm larvae, comprising the following components in parts by weight: 150 parts of corn flour, 40 parts of yeast powder, 20 parts of agar, 4 parts of lecithin, 0.2 parts of cholesterol, 0.8 parts of trehalose, 2.194 parts of vitamins, 14 parts of corn leaf powder, 8 parts of preservatives, and 1000 parts of water; the vitamins include the following components in parts by weight: 2 parts of vitamin C, 0.03 parts of nicotinamide, 0.02 parts of calcium pantothenate, 0.05 parts of vitamin B1, 0.02 parts of vitamin B2, 0.02 parts of vitamin B6, 0.03 parts of folic acid, 0.02 parts of biotin, and 0.004 parts of vitamin B12; the preservative is a mixture of monomethyl fumarate and ethyl parahydroxybenzoate in a mass ratio of 1:1.
[0036] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 30 minutes, heating at 85°C to dissolve, then adding corn flour, stirring evenly, heating to boiling, and keeping warm for 6 minutes; (2) cooling to 65°C, adding corn leaf powder, lecithin, cholesterol, and trehalose, stirring evenly, and keeping warm for 4 minutes; (3) cooling to 40°C, adding yeast powder, vitamins, and preservatives, stirring evenly, and cooling to room temperature to prepare the feed.
[0037] Comparative Example 5: A feed for fall armyworm larvae, comprising the following components in parts by weight: 150 parts of corn flour, 40 parts of yeast powder, 20 parts of agar, 4 parts of lecithin, 0.2 parts of cholesterol, 0.8 parts of trehalose, 2.194 parts of vitamins, 6 parts of Bacillus cereus fermentation product, 8 parts of corn leaf powder, 8 parts of monomethyl fumarate, and 1000 parts of water; the vitamins include the following components in parts by weight: 2 parts of vitamin C, 0.03 parts of nicotinamide, 0.02 parts of calcium pantothenate, 0.05 parts of vitamin B1, 0.02 parts of vitamin B2, 0.02 parts of vitamin B6, 0.03 parts of folic acid, 0.02 parts of biotin, and 0.004 parts of vitamin B12.
[0038] The preparation method of the Bacillus cereus fermentation product is as follows: (1) Activation of Bacillus cereus: picking a single colony of Bacillus cereus that has been cultured by streaking is placed in LB broth culture medium, and cultured on a shaker at 37°C and 200 r / min for 24 hours to obtain an activated bacterial solution; the Bacillus cereus is Bacillus cereus ( Bacillus cereus) YSQ08, purchased from China Center for Type Culture Collection, with a strain collection number of CCTCC No. M2012322; the LB broth culture medium was prepared as follows: 10 g of tryptone, 5 g of yeast extract, and 5 g of NaCl were mixed, the volume was fixed to 1000 mL with distilled water, and the mixture was sterilized at 121°C for 20 min; (2) the 3% activated bacterial solution prepared in step (1) was added to a fermentation buffer solution with a pH of 7 and a chicken feather content of 3%; the fermentation temperature was maintained at 37°C, and the fermentation time was 6 d; the fermentation buffer solution was composed of: 0.5 g / L sodium chloride, 0.3 g / L calcium chloride, 0.3 g / L magnesium chloride, 2 g / L potassium dihydrogen phosphate, and 1 g / L potassium dihydrogen phosphate; (3) the fermented liquid was poured into two 100 mL centrifuge tubes, balanced, and placed oppositely in a centrifuge, centrifuged at 3000 r for 10 min, the supernatant was taken, and the mixture was freeze-dried in vacuum and then crushed.
[0039] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 30 minutes, heating at 85°C to dissolve, then adding corn flour, stirring evenly, heating to boiling, and keeping warm for 6 minutes; (2) cooling to 65°C, adding corn leaf powder, lecithin, cholesterol, and trehalose, stirring evenly, and keeping warm for 4 minutes; (3) cooling to 40°C, adding yeast powder, vitamins, Bacillus cereus fermentation product, and monomethyl fumarate, stirring evenly, and cooling to room temperature to prepare the feed.
[0040] Comparative Example 6: A feed for fall armyworm larvae, comprising the following components in parts by weight: 150 parts of corn flour, 40 parts of yeast powder, 20 parts of agar, 4 parts of lecithin, 0.2 parts of cholesterol, 0.8 parts of trehalose, 2.194 parts of vitamins, 6 parts of Bacillus cereus fermentation product, 8 parts of corn leaf powder, 8 parts of ethyl hydroxybenzoate, and 1000 parts of water; the vitamins include the following components in parts by weight: 2 parts of vitamin C, 0.03 parts of nicotinamide, 0.02 parts of calcium pantothenate, 0.05 parts of vitamin B1, 0.02 parts of vitamin B2, 0.02 parts of vitamin B6, 0.03 parts of folic acid, 0.02 parts of biotin, and 0.004 parts of vitamin B12.
[0041] The preparation method of the Bacillus cereus fermentation product is as follows: (1) Activation of Bacillus cereus: picking a single colony of Bacillus cereus that has been cultured by streaking is placed in LB broth culture medium, and cultured on a shaker at 37°C and 200 r / min for 24 hours to obtain an activated bacterial solution; the Bacillus cereus is Bacillus cereus ( Bacillus cereus) YSQ08, purchased from China Center for Type Culture Collection, with a strain collection number of CCTCC No. M2012322; the LB broth culture medium was prepared as follows: 10 g of tryptone, 5 g of yeast extract, and 5 g of NaCl were mixed, the volume was fixed to 1000 mL with distilled water, and the mixture was sterilized at 121°C for 20 min; (2) the 3% activated bacterial solution prepared in step (1) was added to a fermentation buffer solution with a pH of 7 and a chicken feather content of 3%; the fermentation temperature was maintained at 37°C, and the fermentation time was 6 d; the fermentation buffer solution was composed of: 0.5 g / L sodium chloride, 0.3 g / L calcium chloride, 0.3 g / L magnesium chloride, 2 g / L potassium dihydrogen phosphate, and 1 g / L potassium dihydrogen phosphate; (3) the fermented liquid was poured into two 100 mL centrifuge tubes, balanced, and placed oppositely in a centrifuge, centrifuged at 3000 r for 10 min, the supernatant was taken, and the mixture was freeze-dried in vacuum and then crushed.
[0042] The method for preparing feed for fall armyworm larvae is carried out according to the following steps: (1) adding agar into water, soaking for 30 minutes, heating at 85°C to dissolve, then adding corn flour, stirring evenly, heating to boiling, and keeping warm for 6 minutes; (2) cooling to 65°C, adding corn leaf powder, lecithin, cholesterol, and trehalose, stirring evenly, and keeping warm for 4 minutes; (3) cooling to 40°C, adding yeast powder, vitamins, Bacillus cereus fermentation product, and ethyl hydroxybenzoate, stirring evenly, and cooling to room temperature to prepare the feed.
[0043] Experimental Example: Rearing of Fall Armyworm: Male and female adults were placed in an insect cage at a ratio of 1:1 for mating and laying eggs; the adults were fed 10% honey water; the egg-laying paper was wetted and attached to the inner wall of the insect cage, the egg-laying paper was changed once a day, and the top of the insect cage was covered with gauze to seal it; the egg mass was cut from the egg-laying paper and placed in a small insect box, the box was covered with a layer of absorbent paper and then covered with a lid; after the larvae hatched, the larvae were raised to the third instar using the feed prepared in Examples 1-3 and Comparative Examples 1-4; the third instar larvae were placed in a single-hole insect box filled with feed, covered with a layer of absorbent paper and then covered with a lid, the feed was changed once in the middle, and the larvae were raised until they cocooned and pupated; the pupae were picked out with tweezers, and the pupae were spread flat on the insect cage with absorbent paper, and waited for the adult to emerge. The pupation rate, pupal emergence rate, and deformity rate of Spodoptera frugiperda larvae (deformities include deformities of pupae and adults after emergence) were statistically analyzed using SPSS 24.0 software. Measurement data were expressed as `x±(mean±standard deviation). The Kolmogorov-Smirnov test was used for data normality testing. For data that met the normal distribution, the t-test was used to compare the mean differences between the two groups. P<0.05 was considered statistically significant. The results are shown in Table 1: Table 1 Experimental group Pupation rate% Emergence rate% Deformity rate% Example 1 93.4±1.4 95.2±1.3 1.2±0.1 Example 2 92.1±2.1 94.3±1.5 1.3±0.2 Example 3 91.9±1.8 95.9±1.2 1.5±0.1 Comparative Example 1 82.3±1.1* 83.5±1.7* 1.6±0.3 Comparative Example 2 78.9±1.4* 84.2±1.6* 1.3±0.2 Comparative Example 3 91.1±0.8 93.3±1.1 5.2±0.4* Comparative Example 4 90.3±1.2 94.7±1.7 6.8±0.1* Note: * represents P < 0.05 compared with the group in Example 1.
[0044] The mold-proofing effects of the feeds prepared in Examples 1-3 and Comparative Examples 5-6 were tested according to GB / T 13092-2006, "Determination of the Total Fungal Count in Feeds." During storage, the mold count in the samples was measured every 20 days. Statistical analysis was performed using SPSS 24.0 software. Data were expressed as x ± (mean ± standard deviation). The Kolmogorov-Smirnov test was used to verify normality. For data that conformed to a normal distribution, the t-test was used to compare mean differences between the two groups. P < 0.05 was considered statistically significant. The results are shown in Table 2: Table 2 Experimental group <![CDATA[Initial mold count (10 3 / g)]]> <![CDATA[Mold count on the 20th day (10 3 / g)]]> <![CDATA[Mold count on the 40th day (10 3 / g)]]> Example 1 1.1±0.1 3.2±0.3 5.7±0.2 Example 2 1.2±0.1 3.3±0.2 5.6±0.2 Example 3 1.1±0.2 3.9±0.2 5.9±0.1 Comparative Example 5 1.1±0.1 6.5±0.1* 11.2±0.3* Comparative Example 6 1.3±0.2 6.6±0.3* 9.8±0.4* Note: * represents P < 0.05 compared with the group in Example 1.
[0045] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.
Claims
1. A feed for fall armyworm larvae, characterized in that: The invention comprises the following components in parts by weight: 100-200 parts of corn flour, 20-60 parts of yeast powder, 10-30 parts of agar, 3-5 parts of lecithin, 0.1-0.3 parts of cholesterol, 0.5-1.5 parts of trehalose, 1-4 parts of vitamins, 2-8 parts of Bacillus cereus fermentation product, 5-10 parts of corn leaf powder, 5-10 parts of preservative and 800-1200 parts of water.
2. The feed for fall armyworm larvae according to claim 1, characterized in that The vitamins include the following components in parts by weight: 1-3 parts of vitamin C, 0.01-0.05 parts of nicotinamide, 0.01-0.03 parts of calcium pantothenate, 0.01-0.1 parts of vitamin B1, 0.01-0.03 parts of vitamin B2, 0.01-0.03 parts of vitamin B6, 0.01-0.05 parts of folic acid, 0.01-0.03 parts of biotin, and 0.002-0.006 parts of vitamin B12.
3. The feed for fall armyworm larvae according to claim 1, characterized in that The preparation method of the Bacillus cereus fermentation product is as follows: (1) activating Bacillus cereus: picking a single colony of Bacillus cereus cultured by streaking and placing it in LB broth culture medium, and culturing it on a shaker at 33-38°C and 100-300r / min for 12-36h to obtain an activated bacterial liquid; (2) adding 2-4% activated bacterial liquid prepared in step (1) to a fermentation buffer solution with a pH of 7 and a chicken feather content of 2-5% by weight; maintaining the fermentation temperature at 33-38°C and the fermentation time for 4-8d; (3) pouring the fermented liquid into two 100mL centrifuge tubes, balancing them, placing them opposite to each other in a centrifuge, centrifuging them at 2000-4000r / min for 8-12min, taking the supernatant, and vacuum freeze-drying and then crushing them.
4. The feed for fall armyworm larvae according to claim 3, characterized in that The Bacillus cereus is Bacillus cereus ( Bacillus cereus ) YSQ08, the deposit number is CCTCC No. M2012322.
5. The feed for fall armyworm larvae according to claim 3, characterized in that: The fermentation buffer comprises: 0.1-0.8 g / L sodium chloride, 0.1-0.5 g / L calcium chloride, 0.1-0.5 g / L magnesium chloride, 1-3 g / L dipotassium hydrogen phosphate, and 0.5-1.5 g / L potassium dihydrogen phosphate.
6. The feed for fall armyworm larvae according to claim 3, characterized in that: The LB broth culture medium is prepared by mixing 8-12 g of tryptone, 3-7 g of yeast extract, and 3-7 g of NaCl, adding distilled water to make up to 1000 mL, and sterilizing by autoclaving at 121° C. for 15-25 min.
7. The feed for fall armyworm larvae according to claim 1, characterized in that The preservative is one or more of sodium benzoate, potassium sorbate, propylparaben, monomethyl fumarate, ethyl p-hydroxybenzoate, and dimethyl fumarate.
8. The method for preparing feed for fall armyworm larvae according to claim 1, characterized in that: The process is as follows: (1) add agar to water, soak for 20-40 minutes, heat to dissolve, then add cornstarch, stir evenly, heat to boiling, and keep warm for 5-10 minutes; (2) cool to 60-70°C, add corn leaf powder, lecithin, cholesterol, and trehalose, stir evenly, and keep warm for 3-5 minutes; (3) cool to 35-50°C, add yeast powder, vitamins, Bacillus cereus fermentation product, and preservatives, stir evenly, and cool to room temperature to make it.
9. The method for preparing feed for fall armyworm larvae according to claim 8, characterized in that: The heating temperature in step (1) is 80-95°C.