104 results about "Silkworm larvae" patented technology

The invention relates to a novel method for breeding cordyceps militaris by utilizing silkworm larvae. The method comprises the steps of taking live matured silkworm as the host, vaccinating cordyceps militaris strains through an injection method, and promoting silkworm to sleep, and the vaccination attainment rate is improved; after the method is adopted, the inactivation rate of the silkworm larvae reaches 90% to 100%, and the outgrowth rate of fruiting body reaches 85% to 100%; moreover, the water-solubility protein, polysaccharide, cordycepic acid, cordycepin and adenosine of the cordyceps militaris bred by the method are higher than those of cordyceps militaris bred by silkworm chrysalis in content. The novel method for breeding cordyceps militaris by utilizing silkworm larvae has the advantages of convenience in operation, low breeding cost, good experiment repeatability and wide application prospect, and is suitable for large-scale cultivation.

The invention relates to a cordyceps militaris fruiting body with silkworm larvae as hosts and a cultivation method of the cordyceps militaris fruiting body. The cultivation method includes the steps: disinfecting and washing the cordyceps militaris fruiting body, cutting the cordyceps militaris fruiting body into sections, and inoculating the sections to a solid culture medium for cultivation to obtain a bacterial mass; inoculating the bacterial mass to a primary liquid strain culture medium for cultivation to obtain a primary liquid strain; diluting the primary liquid strain with sterile water to obtain primary liquid strain diluent; mixing pearlite and vermiculite, mixing the disinfected mixture with the primary liquid strain diluent to obtain cordyceps cultivation soil mixture; wiping the silkworm larvae at four to five instar with alcohol, using a disposable syringe to suck the primary liquid strain, and injecting into the silkworm larvae; and inserting the inoculated silkworm larvae into the cordyceps cultivation soil mixture, maintaining heads of the larvae up, and harvesting after cultivation to obtain the cordyceps militaris fruiting body with the silkworm larvae as the hosts. The head of the cordyceps militaris fruiting body is more than 2cm in length, and content of effective ingredients is high.

A method for cultivating cordyceps militaris includes steps of (1) silkworm selection: 3rd-5thd silkworms at five years old are selected as hosts; (2) strain producing: cordyceps militaris strains are inoculated in fluid media for shake cultivating, and then liquid strains are obtained after the cordyceps militaris strains are filtered through a sterile gauze for strain ball removing; (3) nutrient soil producing: soil with 5-20% of water content is prepared and serves as nutrient soil after being sterilized at high template and high pressure; (4) inoculating: the silkworms are placed under ultraviolet for sterilizing for 20-50 min, and then 0.2-0.4ml liquid strains are injected to be inoculated into the silkworms; and (5) cultivating management: the inoculated silkworms are placed in the nutrient soil at 5-12 DEG C for light-shielded cultivating for 5-12 d, and then are transferred to be cultivated in the nutrient soil with light intensity to be 50-200 lx, temperature to be 21-23 DEG C, humidity to be 86-89% for 35-60 d to obtain the cordyceps militaris. The method has the advantages of being simple to operate, low in cost, high in infection rate and germination rate, good in stability and the like, is suitable for mass production and has a wide development prospect.

A silkworm breeding method includes the steps of breeding place selection and building, incubation of silkworm eggs, newly-hatched silkworm collection as well as spinning and cocooning management. The method further includes breeding of silkworms of 1-3 instar and breeding of silkworms of 4-5 instar. According to the silkworm breeding method, the breeding environment is strictly controlled in the growth and development stages of the silkworms, and due to the fact that silkworm larvae are poor in resistance and limited in disease resistance, the feed is carefully prepared, mulberry leaves are subjected to high-temperature steam treatment to reduce diseases caused by feed problems, and a certain amount of fermentation broth is added to the feed for feeding big silkworms of 4-5 instar to effectively increase feed intake of the silkworms and ensure yield and quality of silk.

The invention discloses a method for expressing recombinant canine Alpha interferon with silkworm. Silkworm baculovirus serves as a carrier and recombinant virus containing a canine Alpha interferon gene is structured. Silkworm serves as a host of the recombinant baculovirus, and the recombinant canine Alpha interferon containing biological activity is expressed. The method solves the defects that complicated renaturation operations are needed in expressing the canine Alpha interferon in colon bacillus, the plasmid transformants in yeast expression are unstable and the production cost of mammal culture cells is too high, most of the recombinant canine Alpha interferon expressed by silkworm larvae is secreted into haemolymph, which is very good for the quick extraction of protein. Experimental results show that the method for using the recombinant baculovirus to produce recombinant canine Alpha interferon with silkworm is feasible. The largely expression of the recombinant canine Alpha interferon opens up broad prospects for the application of the Alpha interferon to medical and curing fields.

An aseptic culture method for silkworm larvae includes disinfecting silkworm ova one day before they ar incubated by immersing in formaldehyde solution for 10-15 min and then in alcohol for 1-4 min, drying in air, inculating to obtain larvae and culturing.

The invention discloses a method for earthing-cultivating silkworm insect grass by simulating an ecological environment. The method comprises the following steps: (1) selecting a strain host; (2) enlarging preparation solution of cultivated strain liquid; (3) preprocessing and postprocessing inoculation; (4) preparing an embedding substrate; and (5) management of earthing-cultivating: inoculatingthe strains optimized in the step (2) in a fluid nutrient medium added with 5-8 wt% of silkworm chrysalis meal, after enlarging the cultivation, inoculating in healthy silkworm larvae in the step (1)for infecting and rigidifying, and embedding infected and rigidified silkworm bodies to be developed in the step (3) in a transparent plastic box filled with the plant ash embedding substrate; after sporophore is grown to be matured, taking out the silkworm bodies from the plant ash embedding substrate, and cleaning up plant ash adhered to the periphery of the silkworm bodies, to obtain the matured silkworm insect grass. The method has the characteristics of low sundry fungus contamination rate, high infection rigidifying rate, high production rate, good stability of the obtained silkworm insect grass, higher yield, low cost, and full and regular appearance of the silkworm insect grass, is suitable for the large-scale production, and has the extensive development prospect.

The invention relates to a method for cultivating silkworm larvae into silkworm cordyceps by simulating a natural ecological environment. The method comprises the process that the silkworm larvae of 4-5 years are firstly selected, the temperature, the humidity, the illumination (light) intensity and the like are controlled under a specific environment after the silkworm larvae are inoculated with cordyceps militaris spores, and the temperature, the humidity, the light and the like are regulated and controlled along with the cultivation progress and the development situation of hyphae. The process comprises the following steps: a, breeding: strain bodies are selected for each generation of strains through sexual propagation; b, open inoculation: a tray is used for breeding and infrared rays are used for sterilization; and c, the cordyceps are kept fresh under original ecology of a living body. The method disclosed by the invention has the advantages that the strain bodies are selected for each generation of strains through the sexual propagation, so that the yield of the cordyceps is high after inoculation, the cordyceps have high quality, and the output of the cordyceps is guaranteed; infrared ray sterilization measures in open inoculation are adopted to greatly promote the popularization of large-scale production in a silkworm cordycep industry, and the production cost is low; and the cordyceps are kept fresh under the original ecology of the living body, and under the original ecology condition of the living body, an essence extracting effect is better.

The present invention provides a method for producing a selenium-enriched feed additive. The method consists of the following steps: (1) selenium-enriched yeasts, plant stalks, fruits, vegetables, mulberry leaves and wheat brans are mixed, and the mixture is fermented to obtain fermentation products; (2) the fermentation products are used to feed grass-eating fly larvae, tenebrio molitor larvae or silkworm larvae to obtain chicken edible pupae; (3) the chicken edible pupas are added into a chicken basic feed to feed chickens and chicken manure produced by the chicken is collected; (4) chicken feathers of the chicken manure are removed, the feather removed chicken manure is dehydrated, and the dehydrated chicken manure is used for feeding grass-eating fly larvae, tenebrio molitor larvae or silkworm larvae to obtain feed pupae; and (5) the feed pupae is separated, the separated feed pupae is dried in a conventional manner, the dried feed pupae is crushed, and the crushed feed pupae is granulated to obtain the selenium-enriched feed additive. The present invention also provides a use of the selenium-enriched feed additive for feeding fish.

A method of utilizing cordyceps militaris to inoculate castor silkworm 5-year larvae to cultivate silkworm cordyceps includes following steps: (1), selecting a cordyceps militaris host; (2), performing enlarged culture on a strain liquid preparation solution; (3), treating host castor silkworm after inoculating; (4), performing cultivation management on silkworm cordyceps fruiting bodies: inoculating a cordyceps militaris bevel mother strain onto a solid flat plate culture medium for activation, inoculating into a liquid culture medium, inoculating into healthy castor silkworm larvae after enlarged culture for infection ossification, and putting infected and ossified silkworm without bacteria spawning into a transparent plastic box after going through sterile treatment for culture management to obtain silkworm cordyceps. The advantage that castor silkworm larvae are big in size, soft in body and rich in nutrition is fully utilized to cultivate the fruiting bodies; infectious bacteria contamination rate is low, infection ossification rate and grass emerging rate are high, and obtained castor silkworm cordyceps is high in stability and fruiting body yield, rich in active ingredient content, suitable for large-scale production and wide in development prospect.

The invention provides a factory-like production method of silkworm cordyceps. The method is characterized by comprising the steps as follows: separating fruiting bodies of cordyceps militaris, sterilizing and washing the fruiting bodies, putting the fruiting bodies on a slant culture medium, and placing the slant culture medium in an incubator for culture; placing a PDA (potato dextrose agar) culture medium in a shake flask, placing white mycelia in the shake flask for shake culture, inoculating the white mycelia on a rice culture medium, after the white mycelia grow over the rice culture medium, taking out the mycelia, adding a glucose liquid, mashing the mycelia, and obtaining a nutrition fungus suspension as an inoculation fluid; stabbing and injecting pleopod parts of silkworm larvae with a medical injector sucked with the inoculation liquid to obtain the inoculated silkworm larvae, obliquely placing the silkworm larvae on the sterilized rice culture medium, performing light-shielding variable-temperature culture on the silkworm larvae firstly, and performing light culture after white hyphae grow over the larvae; and collecting the silkworm cordyceps. The factory-like production method is simple in technology, the mass production can be realized, and the silkworm cordyceps with shapes similar to natural cordyceps sinensis and medium values basically identical with the cordyceps sinensis can be produced.

The invention provides a method for identifying sex for silkworm larvae. It comprises: extracting DNA from silkworm larvae, testing nucleic acid by using spectrophotometer and agarose gel electrophoresis; designing inducer by using gentic sequence relative to factor deciding the silkworm larvae sex, carrying out PCR amplification; agarose gel electrophoresis for PCR amplification; identifying the sex according to result got from electrophoresis. The method is easy to operate and can increase the producing benefit for owners.

The invention relates to the field of gene engineering, in particular to a method for preparing human interleukin 28A by a silkworm bioreactor. Recombined silkworm Bombyx mori recombinant baculovirus with a human interleukin 28A (hIL-28A) gene is obtained by an gene engineering method and is used for inoculating silkworm larvae or pupae, so that the human interleukin 28A is expressed with high efficiency in the silkworm larvae or pupae. Because the invention uses the silkworm bioreactor to produce the recombinant human interleukin 28A (hIL-28A), silkworms can be edible or medicinal, the silkworm larvae and pupae expressing the hIL-28A can be directly freeze-dried, and an expression product does not need to be purified and can be directly used as an oral medicine, so that the invention has the advantages of simple preparation process, low cost, convenient use of the oral medicine and the like and can also reduce some side effects of an injection medicament form.

The invention relates to Chinese herbal medicine external medicine powder, in particular to mask powder for removing female face stains and achieving beauty maintaining and youth keeping functions through Chinese herbal medicine. The mask powder is composed of 150-180 g of radix angelica dahurica, 30-45 g of American ginseng, 15-35 g of stigma croci, 50-70 g of tribulus terrestris, 25-40 g of silkworm larvae, 15-30 g of cordyceps flowers, 50-75 g of monthly roses, 30-45 g of pearls, 30-45 of talc, 10-20 g of saponin, 30-50 of mung beans, 50-70 g of roses, 20-35 of excrementum passeris, 30-50 of peach blossoms, 25-40 g of ampelopsis japonica, 25-40 of radix paeoniae alba, 30-45 of raw rhizoma atractylodis macrocephalae, 25-40 g of bletilla striata, 20-35 g of semen pharbitidis, 30-45 g of rhizoma typhonii, and 100-150 g of white poria. When used, the mask powder is ground into 120-mesh fine powder, a proper amount of honey is added, the mixture is stirred to be in a pasty shape and is evenly smeared to the face, after about 90 minutes, the mask powder is completely removed with warm water, skin whitening, acne removing and stain removing effects are obvious, and obvious curative effects are achieved for brown stains, butterfly stains, erythema, age stains and the like. Due to reasonable application of the collateral dredging transportation and transformation function of the Chinese herbal medicine, skin cells can absorb effective components in the powder in short time, skin restoration is accelerated, and therefore the effects of resisting aging, removing the stains, whitening skin and maintaining beauty are achieved. It is proved through a large number of clinical implementation cases that no side effect is generated when the mask powder is used, and high stability is achieved.

The invention discloses a preparation method of an orally-taken vaccine for treatment of hemorrhage of a grass carp, which comprises the following steps of: (1) preparing a recombinant baculovirus BmNPV-IIVP6 with hemorrhagic virus structure protein VP6 genes of the grass carp in a genetic engineering method; (2) grafting the amplified recombinant baculovirus BmNPV-IIVP6 to a five-instar silkworm larvae or pupae; (4) collecting the silkworm larvae or pupae subjected to virus inoculation after 4 to 6 days, and preparing freeze-dried powder in a freeze-drying method; and (4) mixing the freeze-dried powder prepared in Step (3) and powdered fish feed, to obtain the orally-taken vaccine for treatment of hemorrhage of the grass carp. The content of the freeze-dried powder in the orally-taken vaccine for treatment of hemorrhage of the grass carp is 1-10 percent by weight. The vaccine can be directly used as an orally-taken vaccine, and has the advantages of simple preparation process, low cost, convenient use and dual functions of protein subunit vaccines and nucleic acid vaccines.

The invention discloses a determination method of the content of 1-deoxynojirimycin in silkworm larvae. The determination method comprises the following steps of: taking 0.05mol/L HCl as a solvent and swirling for 15-20 seconds at a room temperature; extracting for two times by using a step of ultrasonic treatment for 20-30 minutes to effectively extract DNJ (Deoxynojirimycin) from the silkworm larvae; marking the DNJ for 20-25 minutes at 20 DEG C by utilizing 5mmol/L 9-fluorenyl methylclhlorofonmate (FMOC-C1) to form a DNJ-FMOC complex with the fluorescence; and detecting by utilizing a reverse phase high performance liquid chromatography provided with a fluorescence detector and calculating the content of the DNJ according to a peak area. The determination method of the content of the DNJ in the silkworm larvae provided with the invention has the advantages of high precision, repeatability and high sample recovery rate and good stability, can be used for detecting the content of the DNJ in a lot of silkworm larva products, and has active social benefits and economic benefits for propelling pharmaceutical development of the silkworm larvae.

The invention relates to a method for constructing a BmNPV-domestic silkworm larva polygene expression system, which comprises the following steps: (1) an I-SceI expression cassette controlled by arabinose promoter (pBAD) is recombined into the genome of E.coli D H10B to obtain new recipient bacteria I-SceI DH10B; (2) a donor plasmid used for combined transformation is constructed; (3) a recombination site of cre/loxp replaces v-cath gene and chitin gene in the BmBacmid genome, and two I-SceI sites replace p70 gene in the BmBacmid to obtain a receptor I-sceI BmMultibac; (4) an exogenous gene is induced by the three methods of cre-loxp recombination, Tn7 recombination and combined transformation; (5) a recombined Bacmid DNA is prepared by a conventional method, and is used for transfecting insect cells or injected into the silkworm larva. The BmNPV-domestic silkworm larva polygene expression system lays a foundation for studying multimeric proteins, the interaction between proteins and preparing virus-like particles.

The invention discloses a traditional Chinese medicine preparation for treating bronchitis and asthma. The traditional Chinese medicine preparation is characterized by being prepared from the following raw material medicines in parts by weight: 6-15 parts of dried tangerine or orange peel, 5-10 parts of peucedanum praeruptorum, 9-12 parts of ark shell, 10-15 parts of liquorice, 6-9 parts of silkworm larvae, 5-10 parts of oyster plant inflorescence, 8-12 parts of honey-fried ephedra, 6-15 parts of rhizoma nelumbinis and 5-10 parts of gallnut. The traditional Chinese medicine preparation has notable curative effect on trachitis and asthma caused by insufficiency of both the spleen and the kidney and has the effects of clearing lung and eliminating phlegm as well as relieving cough and asthma, at the same time improves the functions of tonifying qi and kidney as well as soothing the liver and activating the blood, and is free of toxic or side effect as the materials are all traditional Chinese medicinal materials; the medicines are easy to obtain, the medicines are low in cost, and the burden of patients can be alleviated.

The invention relates to the field of biological feed cultivation technology, particularly to a method for cultivating silkworms for biological feed of parent shrimp, comprising the steps of establishing standard silkworm breeding pond, integrally treating water in the silkworm pond, preparing the medium, incubation of the silkworm larvae, and cultivating the silkworm and detection of bacteria. The cultivation method for the silkworm and the the cultivated silkworm has the following advantages: health, no injury, hypertrophy and gloss, high survival rate. This solves the key technology of cultivation of silkworm and provide a new development path for safe and efficient cultivation of silkworms.Also, the cultivation method provides safe and reliable nutrition for cultivate the parent shrimp with better healthy and gonadal development. Moreover, the cultivation method provides a guarantee for producing the shrimp seedlings with high quality and promotes health and sustainable development of shrimp breeding industry.

A process for preparing the zymolase of agricultural organophosphorus chemical by use of silkwork bioactivator includes such steps as recombining said zymolase gene with baculovirus vector, infecting host silkworm, expressing of said zymolase, collecting the body fluid of young silkworm or pupa containing said zymolase, homogenizing, and preparing enzyme preparation.

The invention discloses an application of Ag nanoparticles to preparation of medicines for restraining nosema bombycis as a main pathogen of bombyx mori. The Ag nanoparticles are spherical, are 3-24nmin dimension and are 0.23nm in crystal lattice distance. Through Ag nanoparticle solution of different concentrations, the toxicity of bombyx mori larvae is tested, and the result shows that the weight, the cocoon shell rate and the oviposition number of the bombyx mori larvae between groups do not have significant differences, which indicates that the Ag nanoparticle solution having the concentration being 100 [mu]g/mL or below is safe to the bombyx mori larvae; a mechanism of Ag nanoparticles for resisting nosema bombycis is analyzed, then the inventor finds that the Ag nanoparticles can seriously destroy the structure of the nosema bombycis, when the Ag nanoparticles act with spore coats, the permeability is increased, so that some substances of DNA, proteins and the like are destroyed, the infestation capacity is greatly reduced, the pathogenic capacity of the nosema bombycis is reduced, and the disinfection effects of the nosema bombycis can be achieved; and the Ag nanoparticleshave important significance in controlling the nosema bombycis of the bombyx mori.

The invention discloses a fusion protein of CTB (Cellulose Tribenzoate), human insulin and glutamic acid decarboxylase 3p531 fragments and application of the fusion protein. The application of the fusion protein is in preparing a medicament for treating type I diabetes. Silkworm larvae or silkworm chrysalides infected by recombinant silkworm baculoviruses efficiently express the fusion protein which has higher activity than that of the fusion protein expressed by prokaryotes directly and higher yield than that of the fusion protein expressed by transgenic plants; a silkworm body has multiple natural protein protective agents, thereby having a protection function for expression products and facilitating the more stable expression products; the prepared fusion protein can be prepared into the medicament for treating the type I diabetes and relieving injection pains and infection threats for patients.

The invention discloses Bacillus bombyseptieus parasporal crystal toxin (Bb toxin) protein and a truncated segment thereof as well as a recombinant expression vector and an application of the Bacillus bombyseptieus parasporal crystal toxin protein. The amino acid sequence of the Bacillus bombyseptieus parasporal crystal toxin protein is shown by SEQ ID NO.4, and the truncated segment of the Bacillus bombyseptieus parasporal crystal toxin protein is shown by SEQ ID NO.8. The survival rate of domestic silkworms is obviously reduced (about 30%) after silkworm larvae are additionally fed with the truncated segment, which shows that the gene can be used for coding a toxic protein capable of causing lepidopteron, namely the domestic silkworms to death, and the Bacillus bombyseptieus parasporal crystal toxin protein can be used for preparing biological pesticides for resisting lepidoptera pests and has wide application prospect.