CN1969653A - Antivirus microbe additive for silkworm and preparation method thereof - Google Patents
Antivirus microbe additive for silkworm and preparation method thereof Download PDFInfo
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- CN1969653A CN1969653A CNA2006101249799A CN200610124979A CN1969653A CN 1969653 A CN1969653 A CN 1969653A CN A2006101249799 A CNA2006101249799 A CN A2006101249799A CN 200610124979 A CN200610124979 A CN 200610124979A CN 1969653 A CN1969653 A CN 1969653A
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Abstract
The invention provides an antiviral microbiological additive for domestic silkworm, which is prepared from bacillus licheniformis SHK3, Bacillus licheniformis SHK3 through fermenting with a docket number of CCTCC No: 206115. The invention also discloses the process for preparing the antiviral microbiological additive for domestic silkworm, including aqueous agent and powdered agent.
Description
Technical field
The present invention relates to a kind of antivirus microbe additive for silkworm.The invention still further relates to the preparation method of this microbe additive.
Background technology
Silk is produced, and in existing more than the 5500 year history of China, is the traditional specialty industries of China, also is a kind of special goods that can monopolize the international market in China's export abroad trade.To the expanding economy of silkworm industry, governments at all levels are all paid much attention to, and scientific and technological circle breed silkworms to science and also carried out continuous research and discussion, and at mulberry, silkworm breeding, aspects such as flacherie control have obtained very big progress.But in the sericulture process, still often outbursts such as viral, bacteriosis that silkworm is infected through infection of digestive canal or through foreign object and enteron aisle disease, and there is not the good recipe of control, have a strong impact on silkworm and survive growth, be placed on small straw bundles to spin cocoons, cocoon and cocoon yield and silk quality, also affect silkworm raiser and silk export processing economic benefit of enterprises.
Be engaged in that silkworm is already studied or the practice of breeding silkworms is all known: bacteriosis is easily controlled, and viral disease is difficult anti-.Silkworm is in case morbidity, treat lately, leans on chemotherapy, and it produces little effect, particularly viral disease: as disease of digestive tract, hematologic disease, past medical help at all.As seen, the drug therapy after the silkworm morbidity is passive, and the also inaccessible reduction disease rate of dying increases cocoon yield, improves the purpose of silk quality.
Therefore, pressed in this area and can initiatively effectively strengthen silkworm antibody, anti-bacteria and anti-virus reduces the disease rate of dying, and increases the amount of cocoon production, improves the silkworm of silk quality and uses complex microorganism additive.
Summary of the invention
The objective of the invention is to overcome above-mentioned existing disadvantages of background technology part, and a kind of antivirus microbe additive for silkworm is provided.
Another object of the present invention is to provide the preparation method of this antivirus microbe additive for silkworm.
The objective of the invention is to reach by following measure: antivirus microbe additive for silkworm, it is by bacillus licheniformis SHK3, Bacillus licheniformis SHK3, forms through fermentation; Preserving number is CCTCC NO:M206115, and preservation date is on October 31st, 2006.
Bacillus licheniformis SHK3, Bacillus licheniformis SHK3, preserving number are the bacterial classification probation report of CCTCC NO:M206115
Bacterial classification source: Wuhan Nongda Biotechnology Co., Ltd.
Testing result:
| Test item | The result | Test item | The result |
| Gram’s staining cell shape cell dia＞1.0 μ m forms endospore sporulation sporangiocyst and expands the circular parasporal crystal catalase of gemma oxidizing ferment anaerobic growth VP test VP＜pH6 VP＞pH7 methyl red test | + shaft-like-+inferior end life---+-++-- | Produce acid: D-Glucose D-wood sugar Arabinose PEARLITOL 25C utilizes the glucose aerogenesis to utilize 50 ℃ of growths of citrate nitrate reduction pH, 5.7 growth 7%NaCl growth Starch Hydrolysis gelatin liquefaction hydrolysed casein urea | + + + + - + + + + + + + + - |
Name: Bacillus licheniformis SHK3, bacillus licheniformis SHK3
The difference of the feature of microorganism: bacterium colony is more flat, circular, small circle, light brown appears in neat in edge, wrinkle, coarse, central authorities.
The preparation method of antivirus microbe additive for silkworm, it comprises the steps:
1. bacterial classification is cultivated: bacillus licheniformis SHK3, Bacillus lichenifomisSHK3 bacterial classification are transplanted on the nutrient agar slant medium, are placed 36 ℃ of incubators, cultivated 24 hours, make the bacillus licheniformis rejuvenation;
2. shaking table is cultivated: get in bottles of 500 milliliters conical flasks of above-mentioned slant medium bacterial classification inoculation to 5, adorn 120 milliliters of nutrient broth mediums in every bottle of conical flask, every bottle graft kind 3 rings, moving to shaking table after the inoculation cultivated 18-24 hour, temperature is 36 ℃, be seeded to 50 liters of seed fermentation jars then and cultivate, adorn 35 liters of nutrient solutions in the fermentation tank;
3. 50 liters of seed fermentation jars are cultivated: make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 30min, when waiting to be chilled to 37 ℃, in ventilation than being 0.4-2: 1, rotating speed is 300-500r/min, temperature is under 35-37 ℃ the condition, to ferment 18-24 hour;
4. 1 ton of fermentation tank culture: in adorn 700 liters of nutrient solutions, make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 35min, when waiting to be chilled to 37 ℃, the kind daughter bacteria of cultivating in above-mentioned 50 liters of fermentation tanks is inoculated in 1 ton of fermentation tank, ferment; In ventilation than for 0.4-1: 1, rotating speed is 100-350r/min, temperature is under 35-37 ℃ the condition, fermentation time 36-48 hour; Transfer pH value 3.5-5, be the antivirus microbe additive for silkworm aqua.
In last this technical scheme, the culture medium composition in the described fermentation tank is: yeast extract 0.2%, peptone 1%, glucose 1%, sucrose 1%, calcium chloride 0.2%, sodium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, all the other are water.
In last this technical scheme, it also comprises the steps: to add carrier in the microbe additive aqua, again through being mixed into the solid, powdery thing, is antiviral bacteria composition solid pharmaceutical preparation after drying.
In last this technical scheme, described is to be carrier with dry starch, adds dry starch 1.25 weight portions with zymotic fluid 1 weight portion and adds under constantly stirring, and is stirred into graininess, and drying to water content gets final product between 10-12% again.
The present invention is the compositions such as metabolite that utilize antibiotic, antiviral microorganism and secretion thereof, can strengthen silkworm body immunity, stops virus to suck and viral duplicating, thereby reaches antibiotic, antiviral purpose.In addition, the multiple digestive ferment of microorganism secretion is cellulase, hemicellulase, and protease and amylase etc. can promote that silkworm digests and assimilates, and improve the utilization ratio of mulberry leaf, increase the weight of silk cocoon.Simultaneously, microbe additive is the silk improver of pure natural, and is without any side effects to silkworm and people.Therefore, the present invention is that collection is antibiotic, antiviral, enhancing silkworm immunity, improves the silk cocoon quality, is numerous silkworm raisers' increasing both production and income, is a kind of pure natural disease-resistant microorganism additive of silk processing enterprise synergy.
The specific embodiment
Embodiment 1
The preparation method of silkworm anti-virus type complex microorganism additive (aqua), it comprises the steps:
1. bacterial classification is cultivated: bacillus licheniformis SHK3, Bacillus licheniformisSHK3 bacterial classification are transplanted on the nutrient agar slant medium, are placed 36 ℃ of incubators, cultivated 24 hours, make the bacillus licheniformis rejuvenation;
2. shaking table is cultivated: get in bottles of 500 milliliters conical flasks of above-mentioned slant medium bacterial classification inoculation to 5, adorn 120 milliliters of nutrient broth mediums in every bottle of conical flask, every bottle graft kind 3 rings, moving to shaking table after the inoculation cultivated 18 hours, temperature is 36 ℃, be seeded to 50 liters of seed fermentation jars then and cultivate, adorn 35 liters of nutrient solutions in the fermentation tank;
3. 50 liters of seed fermentation jars are cultivated: make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 30min, when waiting to be chilled to 37 ℃, than being 0.4: 1, rotating speed is 300r/min in ventilation, and temperature is under 35 ℃ the condition, to ferment 18 hours;
4. 1 ton of fermentation tank culture: in adorn 700 liters of nutrient solutions, make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 35min, when waiting to be chilled to 37 ℃, the kind daughter bacteria of cultivating in above-mentioned 50 liters of fermentation tanks is inoculated in 1 ton of fermentation tank, ferment; Than being 0.4: 1, rotating speed is 100r/min in ventilation, and temperature is under 35 ℃ the condition, fermentation time 36 hours; Transfer pH value 3.5, be the antivirus microbe additive for silkworm aqua.
Culture medium composition in the described fermentation tank is: yeast extract 0.2%, peptone 1%, glucose 1%, sucrose 1%, calcium chloride 0.2%, sodium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, all the other are water.
Embodiment 2
The preparation method of silkworm anti-virus type complex microorganism additive (aqua), it comprises the steps:
1. bacterial classification is cultivated: bacillus licheniformis SHK3, Bacillus licheniformisSHK3 bacterial classification are transplanted on the nutrient agar slant medium, are placed 36 ℃ of incubators, cultivated 24 hours, make the bacillus licheniformis rejuvenation;
2. shaking table is cultivated: get in bottles of 500 milliliters conical flasks of above-mentioned slant medium bacterial classification inoculation to 5, adorn 120 milliliters of nutrient broth mediums in every bottle of conical flask, every bottle graft kind 3 rings, moving to shaking table after the inoculation cultivated 24 hours, temperature is 36 ℃, be seeded to 50 liters of seed fermentation jars then and cultivate, adorn 35 liters of nutrient solutions in the fermentation tank;
3. 50 liters of seed fermentation jars are cultivated: make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 30min, when waiting to be chilled to 37 ℃, than being 2: 1, rotating speed is 500r/min in ventilation, and temperature is under 37 ℃ the condition, to ferment 24 hours;
4. 1 ton of fermentation tank culture: in adorn 700 liters of nutrient solutions, make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 35min, when waiting to be chilled to 37 ℃, the kind daughter bacteria of cultivating in above-mentioned 50 liters of fermentation tanks is inoculated in 1 ton of fermentation tank, ferment; Than being 1: 1, rotating speed is 350r/min in ventilation, and temperature is under 37 ℃ the condition, fermentation time 48 hours; Transfer pH value 5, be the antivirus microbe additive for silkworm aqua.
Culture medium composition in the described fermentation tank is: yeast extract 0.2%, peptone 1%, glucose 1%, sucrose 1%, calcium chloride 0.2%, sodium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, all the other are water.
Embodiment 3
The preparation method of silkworm anti-virus type complex microorganism additive (aqua), it comprises the steps:
1. bacterial classification is cultivated: bacillus licheniformis SHK3, Bacillus licheniformisSHK3 bacterial classification are transplanted on the nutrient agar slant medium, are placed 36 ℃ of incubators, cultivated 24 hours, make the bacillus licheniformis rejuvenation;
2. shaking table is cultivated: get in bottles of 500 milliliters conical flasks of above-mentioned slant medium bacterial classification inoculation to 5, adorn 120 milliliters of nutrient broth mediums in every bottle of conical flask, every bottle graft kind 3 rings, moving to shaking table after the inoculation cultivated 21 hours, temperature is 36 ℃, be seeded to 50 liters of seed fermentation jars then and cultivate, adorn 35 liters of nutrient solutions in the fermentation tank;
3. 50 liters of seed fermentation jars are cultivated: make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 30min, when waiting to be chilled to 37 ℃, than being 1.2: 1, rotating speed is 400r/min in ventilation, and temperature is under 36 ℃ the condition, to ferment 21 hours;
4. 1 ton of fermentation tank culture: in adorn 700 liters of nutrient solutions, make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 35min, when waiting to be chilled to 37 ℃, the kind daughter bacteria of cultivating in above-mentioned 50 liters of fermentation tanks is inoculated in 1 ton of fermentation tank, ferment; In ventilation than for 0.4-1: 1, rotating speed is 225r/min, temperature is under 36 ℃ the condition, fermentation time 42 hours; Transfer pH value 4, be the antivirus microbe additive for silkworm aqua.
Culture medium composition in the described fermentation tank is: yeast extract 0.2%, peptone 1%, glucose 1%, sucrose 1%, calcium chloride 0.2%, sodium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, all the other are water.
Embodiment 4
Aqua among the foregoing description 1 or embodiment 2 or the embodiment 3 is added dry starch, adding dry starch 1.25 weight portions with zymotic fluid 1 weight portion adds under constantly stirring and is stirred into graininess, again drying to water content between 10-12%, be solid pharmaceutical preparation.Application example and using method:
Disease-resistant type silkworm is used for silkworm with microbe additive and freshens food, using method: 1 milliliter of this agent (aqua or pulvis) is poured in the bottle of cleaning, add 8 milliliters to 10 milliliters cold boiling water (or pure water) again, fully shake the back placement and soak 5-8 hour (generally speaking, in the time of 32-34 ℃, soaked 5 hours, and in the time of 26-28 ℃, soaked 8 hours.If temperature is low excessively, then soak prolongation, the temperature height, then soak time shortens relatively), spray mulberry leaf with miniaturised nebuliser, copy while spraying, make the mulberry leaf west be stained with this agent solution, be advisable not drip and feed silkworm.Since three length of times, freshen food every other day once, till being placed on small straw bundles to spin cocoons.Aqua after general 1 jin of immersion can spray 10 jin of mulberry leaf, and is edible once for a silkworm (the 22000-30000 head is generally 25000).
The disease-resistant type silkworm of the present invention is half a year with the shelf-life of complex microorganism additive aqua, and the shelf-life of solid powder is 1 year.
Use this agent to the growing of silkworm, the prevention flacherie, the contrast effect of cocoon yield is table 1, table 2, table 3.
The present invention can prevent the bacillary and viral disease of silkworm, thereby substitutes the traditional chemical medicine gradually, reaches the bacillary and viral disease of prevention silkworm, increases silk cocoon weight, improves silk quality, shortens breeding cycle, saves mulberry leaf, increases economic efficiency.
Table 1 anti-virus type silkworm is with the prevention of damage by disease effect of complex microorganism additive to silkworm
| The test order | Experiment batch (I) | Experiment batch (II) | |
| Test period | 3-August 25 August | 17-September 8 August | |
| Handle | Project | Number, the incidence of disease | Number, the incidence of disease |
| Complex microorganism additive 1: 8 | For an examination number (head) | 60 | 60 |
| A morbidity number (head) | 0 | 1 | |
| Incidence of disease % | 0 | 1.67 | |
| Composite microbial | For an examination number (head) | 60 | 60 |
| A morbidity number (head) | 0 | 0 | |
| Thing additive 1: 10 | Incidence of disease % | 0 | 0 |
| Contrast (clear water) | For an examination number (head) | 60 | ?60 |
| A morbidity number (head) | 15 | ?21 | |
| Incidence of disease % | 25 | ?35 |
Table 2 anti-virus type silkworm grows to silkworm with complex microorganism additive and the cocoon quality influence
The disease-resistant type complex microorganism of table 3 flows back to the output effect to silkworm
Analyze as can be known by statistics: after adopting anti-virus type silkworm of the present invention to breed silkworms with microbe additive following effect is arranged: (1) adds food additive of the present invention can save mulberry leaf 15%; (2) increase cocoon yield 12.5%; (3) improve the disease-resistant rate 18% of silkworm; (4) shorten feeding time 10 hours; (5) improve 1 of cocoon quality and wait the group level.Thereby improve the economic benefit about 15% at sericulture family.
Claims (5)
1, antivirus microbe additive for silkworm is characterized in that it is by bacillus licheniformis SHK3, Bacillus licheniformis SHK3, forms through fermentation; Preserving number is CCTCCNO:M206115.
2, the preparation method of antivirus microbe additive for silkworm is characterized in that it comprises the steps:
1. bacterial classification is cultivated: bacillus licheniformis SHK3, Bacillus licheniformisSHK3 bacterial classification are transplanted on the nutrient agar slant medium, are placed 36 ℃ of incubators, cultivated 24 hours, make the bacillus licheniformis rejuvenation;
2. shaking table is cultivated: get in bottles of 500 milliliters conical flasks of above-mentioned slant medium bacterial classification inoculation to 5, adorn 120 milliliters of nutrient broth mediums in every bottle of conical flask, every bottle graft kind 3 rings, moving to shaking table after the inoculation cultivated 18-24 hour, temperature is 36 ℃, be seeded to 50 liters of seed fermentation jars then and cultivate, adorn 35 liters of nutrient solutions in the fermentation tank;
3. 50 liters of seed fermentation jars are cultivated: make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 30min, when waiting to be chilled to 37 ℃, in ventilation than being 0.4-2: 1, rotating speed is 300-500r/min, temperature is under 35-37 ℃ the condition, to ferment 18-24 hour;
4. 1 ton of fermentation tank culture: in adorn 700 liters of nutrient solutions, make defoamer with rape oil or soya-bean oil, with 121 ℃ of sterilization 35min, when waiting to be chilled to 37 ℃, the kind daughter bacteria of cultivating in above-mentioned 50 liters of fermentation tanks is inoculated in 1 ton of fermentation tank, ferment; In ventilation than for 0.4-1: 1, rotating speed is 100-350r/min, temperature is under 35-37 ℃ the condition, fermentation time 36-48 hour;
Transfer pH value 3.5-5, be the antivirus microbe additive for silkworm aqua.
3, the preparation method of a kind of antivirus microbe additive for silkworm according to claim 2 is characterized in that the culture medium composition in the described fermentation tank is: yeast extract 0.2%, peptone 1%, glucose 1%, sucrose 1%, calcium chloride 0.2%, sodium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, all the other are water.
4, the preparation method of a kind of antivirus microbe additive for silkworm according to claim 2, it is characterized in that it also comprises the steps: to add carrier in the microbe additive aqua, again through being mixed into the solid, powdery thing, be antiviral bacteria composition solid pharmaceutical preparation after drying.
5, the preparation method of a kind of antivirus microbe additive for silkworm according to claim 4, it is characterized in that described is to be carrier with dry starch, adding dry starch 1.25 weight portions with zymotic fluid 1 weight portion adds under constantly stirring, be stirred into graininess, drying to water content gets final product between 10-12% again.
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| CNA2006101249799A CN1969653A (en) | 2006-11-09 | 2006-11-09 | Antivirus microbe additive for silkworm and preparation method thereof |
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| CNA2006101249799A CN1969653A (en) | 2006-11-09 | 2006-11-09 | Antivirus microbe additive for silkworm and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102626184A (en) * | 2012-04-03 | 2012-08-08 | 浙江大学 | Feed additive for improving laying yield of female silkworm moths and preparation method thereof |
| CN106290716A (en) * | 2016-07-18 | 2017-01-04 | 广西壮族自治区蚕业技术推广总站 | Detect contaminated Folium Mori toxicity and the method for safety |
| CN108048372A (en) * | 2018-01-30 | 2018-05-18 | 西南大学 | A kind of bacillus pumilus and its application of prevention silkworm nuclear polyhedrosis virus infection |
| CN109336664A (en) * | 2018-11-29 | 2019-02-15 | 广西壮族自治区蚕业技术推广总站 | A kind of method of Silkworm Raise and the innoxious compost treatment of silkworm excrement |
| CN117530371A (en) * | 2023-12-15 | 2024-02-09 | 山东农业大学 | Silkworm artificial feed additive containing alkaline protease and preparation method and application thereof |
-
2006
- 2006-11-09 CN CNA2006101249799A patent/CN1969653A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102626184A (en) * | 2012-04-03 | 2012-08-08 | 浙江大学 | Feed additive for improving laying yield of female silkworm moths and preparation method thereof |
| CN102626184B (en) * | 2012-04-03 | 2013-06-05 | 浙江大学 | Feed additive for improving laying yield of female silkworm moths and preparation method thereof |
| CN106290716A (en) * | 2016-07-18 | 2017-01-04 | 广西壮族自治区蚕业技术推广总站 | Detect contaminated Folium Mori toxicity and the method for safety |
| CN108048372A (en) * | 2018-01-30 | 2018-05-18 | 西南大学 | A kind of bacillus pumilus and its application of prevention silkworm nuclear polyhedrosis virus infection |
| CN109336664A (en) * | 2018-11-29 | 2019-02-15 | 广西壮族自治区蚕业技术推广总站 | A kind of method of Silkworm Raise and the innoxious compost treatment of silkworm excrement |
| CN117530371A (en) * | 2023-12-15 | 2024-02-09 | 山东农业大学 | Silkworm artificial feed additive containing alkaline protease and preparation method and application thereof |
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