CN107018997B - A method of fly spray is prepared using fly agaric - Google Patents
A method of fly spray is prepared using fly agaric Download PDFInfo
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- CN107018997B CN107018997B CN201710407021.9A CN201710407021A CN107018997B CN 107018997 B CN107018997 B CN 107018997B CN 201710407021 A CN201710407021 A CN 201710407021A CN 107018997 B CN107018997 B CN 107018997B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/002—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits
- A01N25/006—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits insecticidal
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- General Health & Medical Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Engineering & Computer Science (AREA)
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- Dentistry (AREA)
- Agronomy & Crop Science (AREA)
- Environmental Sciences (AREA)
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- Insects & Arthropods (AREA)
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Abstract
The present invention provides a kind of method for preparing fly spray using fly agaric, belongs to technical field of biological control, and shake-flask seed culture, seed tank culture, fermented and cultured, fermentation material including fly agaric are extracted.It is aided with increasing as matrix using extract-batrachotoxin alcohol and lures ingredient, is spray-dried, pelletizing forming and etc..The present invention mainly studies water-soluble substances in fly agaric fermentation material-batrachotoxin alcohol extraction process, and increasing is aided with as matrix using batrachotoxin alcohol, ingredient is lured to compound, with strong to fly virulence, lethal time is short, palatability is strong, and drug resistance is low, to safety of human and livestock's property height, the advantages that without generation " three wastes " pollution, is suitable for the extensive fly eradication work in the places such as family, hotel, school, hospital, office building, livestock and poultry farm, warehouse.
Description
Technical field
The present invention discloses a kind of method for preparing fly spray using fly agaric, especially relates to a kind of using macro fungi
The method for preparing fly spray belongs to biological control pest technical field.
Background technique
Currently, there is many shortcomings when killing fly for traditional fly spray, if toxicity is excessive, insect killing effect is good, but
There is security risk to people and animals；And toxicity is low small to safety of human and livestock's hidden danger, then it is poor to fly insect killing effect, there is insect killing effect with
Contradiction between safety.Therefore there is an urgent need for a kind of novel technical solutions in the prior art to solve the problems, such as this.
Fly agaric Amanita muscaria (L:Fr.) Lam. strain of the present invention belongs to Basidiomycetes, agaric
Mesh, amanita phalloides section are distributed across a kind of macro fungi of severe toxicity of Forest in Changbai Mountain Forest Region, summer and autumn all living creatures or scattered in Betulaceae, shell
On the ground, toxic component is batrachotoxin alcohol to the mixed forest that bucket section, pinaceae plant form, and is 5- hydroxy-n-dimethyltryptamine Yin
Diindyl derivative (belongs to alkaline matter).
Summary of the invention
The technical problems to be solved by the present invention are: a kind of method for preparing fly spray using fly agaric is provided, for solving
Fly spray effect certainly used at present is poor, and there is technical issues that environmental pollution is serious.
A kind of method preparing fly spray using fly agaric of the present invention, it is characterized in that the following steps are included:
1, fly agaric inclined-plane parent species are chosen and carry out shake-flask seed culture, high pressure sterilization pressure 1kg/cm3 keeps 30min, cold
But it being inoculated with afterwards, inoculum concentration is that the ratio between inclined-plane and shaking flask are 1:6, and cultivation temperature is 24~26 DEG C, 170~200 turns of rotary shaker/
Minute, incubation time is 160~240h, 20% or more bacterium ball weight in wet base；
Culture medium prescription used are as follows: 2 grams of peptone, 0.1 gram of yeast extract, 1.5 grams of maltose, 5 grams of yellow bean sprout, di(2-ethylhexyl)phosphate
0.15 gram of hydrogen potassium, 0.075 gram of magnesium sulfate, 0.001 gram of vitamin B1, pH value 6.5, water 1000ml；
2, seed tank culture: inoculum concentration is 5%~10%, and temperature is 24~26 DEG C, and mixing speed is 150~180r/ min,
Ventilatory capacity is 1:0.5v/v. min per minute, and tank presses 0.3 X 105Pa is cultivated 2~3 days；
Used medium formula are as follows: 50 grams of wheat bran, 2 grams of peptone, 20 grams of glucose, 1 gram of potassium dihydrogen phosphate, magnesium sulfate
0.3 gram, water 1000ml；
3, fermented and cultured: inoculum concentration is 5%~10%, and temperature is 24~26 DEG C, and mixing speed is 150~180r/ min, is led to
Tolerance is 1:0.5v/v. min per minute, is cultivated 5~6 days；Fermentation liquid retrogradation, mycelia start self-dissolving, and pH value is down to 4.5, residual sugar
Amount is down to 0.3%, and as fermentation termination when adenosine content is begun to decline puts tank leaching mycelium；
Used medium formula are as follows: 20 grams of sucrose, 5 grams of peptone, 20 grams of glucose, 20 grams of beancake powder, potassium dihydrogen phosphate
1.5 grams, 0.75 gram of magnesium sulfate, 1 gram of defoaming agent (soya-bean oil), water 1000ml；
4, the fermentation material by step 3) containing mycelium and fermentation liquid is divided into mycelium and filtrate, mycelium by plate compression
The amount of doubling water is heated to 80 DEG C, keeps the temperature 1h, and vacuum filtration removes residue, filtrate merges with extracting solution, is concentrated into relative density
1:1, as frog bacterium concentrate；
5, according to the volume ratio 1:4 of frog bacterium concentrate and water, 90 DEG C are injected into the frog bacterium concentrate of step 4)
Clear water, frog bacterium concentrate are immersed in 90 DEG C of thermostatted waters 2 hours, and toxin-frog mykol of fly agaric is dissolved in water, and filtering obtains
Batrachotoxin alcohol solution I and filter residue I；
6,90 DEG C of clear water with the medium volume of step 5) are injected into the filter residue of step 5) I, are kept for 2 hours, are filtered, and are obtained
Batrachotoxin alcohol solution II and filter residue II.
7, the batrachotoxin alcohol solution II of the batrachotoxin alcohol solution I of step 5) and step 6) is mixed, it is dense is put into decompression
Compression apparatus, the pressure intensity parameter that vacuum concentration equipment is arranged is 60~65K pa, and heating temperature is 85~90 DEG C, is concentrated into original solution
The 5~10% of volume obtain batrachotoxin alcohol concentrate；
8, the filter residue of step 6) II is Tumblied Dry to water content less than 23%；
9) take fry dulcet maize flour with drying after filter residue II mixed by quality 1:1, be ground by pulverizer
White sugar is added into hybrid particles less than the hybrid particles of 200 mesh in diameter, and the mass ratio of hybrid particles and white sugar is 25:1, obtains
Obtain spice；
10, the batrachotoxin alcohol concentrate atomizing of step 4) is sprayed on the spice of step 9), every kilogram of spice sprinkling
1000ml batrachotoxin alcohol concentrate, and 0.01% sesame oil is added, stirring 10 times or more, drying machine drying to water content 55~60%；
11, spice of the step 10 after dry is put into pelletizing forming equipment, pelletizing forming, then fly spray preparation finishes.
The positive effect of the present invention is:
By the extraction process for the mycelium water solubility toxic agent substance that fly agaric is obtained through submerged fermentation, and with batrachotoxin alcohol
Being aided with increasing for matrix lures ingredient to compound, have it is strong to fly virulence, lethal time is short, and palatability is strong, and drug resistance is low, to people
Raise it is highly-safe, without generation " three wastes " pollution the advantages that, be suitable for family, hotel, school, hospital, office building, livestock and poultry cultivation
The extensive fly eradication work in the places such as field, warehouse.It is strong that the fly agaric species strain of artificial culture does not jeopardize the mankind in enforcement of regulations
Health does not carry animals and plants cause of disease, free from environmental pollution.The passage of fly agaric strain 18 more than generation is not degenerated, and ageing resistance, steady is belonged to
Qualitative and metabolic capability is stronger, yield and the higher strain of yield.
Specific embodiment
By following embodiment further illustrate description the present invention, do not limit the invention in any way, without departing substantially from
Under the premise of technical solution of the invention, easy to accomplish any of those of ordinary skill in the art made for the present invention changes
Dynamic or change is fallen within scope of the presently claimed invention.
Embodiment 1
1, the fly agaric inclined-plane parent species (Am-A) for choosing artificial culture carry out shake-flask seed culture, culture medium prescription used
Are as follows: peptone 2%, 0.1 gram of yeast extract, 1.5 grams of maltose, 5 grams of yellow bean sprout, 0.15 gram of potassium dihydrogen phosphate, 0.075 gram of magnesium sulfate,
Before 0.001 gram of vitamin B1, pH value 6.5(disappear).High pressure sterilization pressure 1kg/cm3 keeps 30min, is inoculated with after cooling, inoculum concentration
Be 1:6 for the ratio between inclined-plane and shaking flask, i.e. the shaking flask (loading amount 100ml) that an inclined-plane meets 6 250ml, cultivation temperature be 24~
26 DEG C, 170~200 revs/min of rotary shaker, incubation time is 160~240h, and 20 grams of bacterium ball weight in wet base or more can be shaking flask
Seed is spare；
2, seed tank culture: used medium formula are as follows: 50 grams of wheat bran, 2 grams of peptone, 20 grams of glucose, biphosphate
1 gram of potassium, 0.3 gram of magnesium sulfate, water 1000ml；Inoculum concentration be 5%~10%, temperature be 24~26 DEG C, mixing speed be 150~
180r/ min, ventilatory capacity are 1:0.5v/v. min per minute, and tank presses 0.3 X 105Pa is cultivated 2~3 days；
3, fermented and cultured: used medium formula are as follows: 20 grams of sucrose, 5 grams of peptone, 20 grams of glucose, 20 grams of beancake powder,
1.5 grams of potassium dihydrogen phosphate, 0.75 gram of magnesium sulfate, 1 gram of defoaming agent (soya-bean oil), water 1000ml；Inoculum concentration is 5%~10%, and temperature is
24~26 DEG C, mixing speed is 150~180r/ min, and ventilatory capacity is 1:0.5v/v. min per minute, is cultivated 5~6 days；At this time
Fermentation liquid retrogradation, mycelia start self-dissolving, and pH value is down to 4.5 or so, and residual sugar amount is down to 0.3% or so, when adenosine content is begun to decline
As fermentation termination can put tank leaching mycelium；
4, the fermentation material containing mycelium and fermentation liquid is divided into mycelium and filtrate by plate compression, mycelium doubles
Amount water is heated to 80 DEG C, keeps the temperature 1h, and vacuum filtration removes residue, filtrate merges with extracting solution, is concentrated into relative density 1:1, i.e.,
For frog bacterium concentrate；
5, according to the volume ratio 1:4 of frog bacterium concentrate and water, 90 DEG C of clear water, frog are injected into frog bacterium concentrate
Bacterium concentrate is immersed in 90 DEG C of thermostatted waters 2 hours, and frog verticillium toxin-frog mykol is dissolved in water, and filtering obtains batrachotoxin alcohol water
Solution I and filter residue I；
6,90 DEG C of clear water with the medium volume of step 5 are injected into filter residue I, are kept for 2 hours, and filtering obtains batrachotoxin alcohol
Aqueous solution II and filter residue II；
7, aqueous solution I and aqueous solution II are mixed, is put into vacuum concentration equipment, the pressure intensity parameter of vacuum concentration equipment is set
For 60~65K pa, heating temperature is 85~90 DEG C, is concentrated into the 5~10% of original solution volume, obtains batrachotoxin alcohol concentrate；
8, filter residue II is Tumblied Dry to water content less than 23%；
9, take fry dulcet maize flour with drying after filter residue II mixed by quality 1:1, be ground by pulverizer
White sugar is added into hybrid particles less than the hybrid particles of 200 mesh in diameter, and the mass ratio of hybrid particles and white sugar is 25:1, obtains
Obtain spice；
10, batrachotoxin alcohol concentrate atomizing is sprayed on spice, every kilogram of spice sprinkling 1000ml batrachotoxin is pure and strong
Contracting liquid, and 0.01 gram of sesame oil is added, stirring 10 times or more, drying machine drying to water content 55~60%；
11, spice of the step 10 after dry is put into pelletizing forming equipment, pelletizing forming, then fly spray preparation finishes.
Test example 1
1, water-soluble batrachotoxin alcohol toxic agent water extraction conditions selection
Under other identical conditions, different extraction temperatures (60,70,80,90,100 DEG C), extraction time are studied respectively
The single factor tests such as (1,2,3,4,5h), extraction time (1,2,3,4,5 time) and solid-liquid ratio (1:2,1:3,1:4,1:6) are to water-soluble clam
Toad poison alcohol toxic agent yield influences；
2, water-soluble batrachotoxin alcohol toxic agent water extracts orthogonal test
On the basis of single factor experiment, select extraction temperature, extraction time, extraction time and solid-liquid ratio for investigation factor,
Orthogonal test is carried out using orthogonal arrage L9(34), using water-soluble batrachotoxin alcohol toxic agent yield as inspection target, screens optimum extraction
Condition；
3, the infiltration, dissolution of water-soluble batrachotoxin alcohol toxic agent, diffusivity increase with temperature and are increased, the viscosity of solution with
Temperature is increased and is reduced.Therefore, Extracting temperature select 90 DEG C it is appropriate；
4, the influence test by extraction time to water-soluble batrachotoxin alcohol toxic agent yield, finally obtaining extraction time is 2h
It is advisable；
5, the influence test by extracting times to water-soluble batrachotoxin alcohol toxic agent yield, finally obtaining extracting times is 2
It is secondary to be advisable；
6, the influence test by solid-liquid ratio to water-soluble batrachotoxin alcohol toxic agent yield, comprehensively considers solid-liquid ratio finally with 1:
4 are advisable；
7, according to orthogonal experiments it is found that the primary sequence for influencing water-soluble batrachotoxin alcohol toxic agent extraction factor is followed successively by
Extraction temperature, extraction time, solid-liquid ratio, extracting times；
8, the water-soluble optimal extraction conditions of batrachotoxin alcohol toxic agent are solid-liquid ratio 1:4；Under 90 DEG C of water temperature conditions, extract 2 times,
It extracts 2 hours every time, it is easy to operate, it is easy to control；Under optimal extracting factor, water-soluble batrachotoxin alcohol toxic agent yield
Average out to 2.5%；
The products characteristics that the technologies such as present invention use, submerged fermentation, extraction, concentration, drying, molding, foundation are developed,
According to low cost, easily control, functional principle of guaranteeing the quality, rely on existing feasible replenishment of process, reasonable combination, optimum choice, foundation is most
Good process route.
Test example 2
Virulence test
1, henhouse is taken to lure the fly ovum grain of production, 3~7 days adult flies after receptacle is sprouted wings, every cage (35 × 35 × 40
It ㎝) 150 or so, raises daily with 10% brown sugar water cotton balls；
2,10 ㎎ of fly spray of the present invention is diluted to thin viscous be applied on paper to be placed on small porcelain dish, is put in small equipped with food 8 is starved
When fly small pupa cage in, every one disk of cage, in different time observe fly feeding and be poisoned to death fly number, start to be poisoned to death
The death rate of time, 24 hours.Temperature is 25~30 DEG C when experiment, air humidity 70~90%, and illumination is 500Lux or more；
3, fly spray virulence test as a result, fly spray of the present invention has kills fly effect well in Jing little cage.Fly feeding 2
Minute i.e. it is visible be poisoned to death, 24 hours death rates are in 100%, and 24 hours without anabiosis；
4, fly spray of the present invention is diluted to 1:500 solution directly to spray in fly body, fly 5 minutes i.e. visible death by poisoning
It dies, 24 hours death rates are 100%；
5, fly spray has many advantages, such as that poisoning is fast, no repellent phenomenon.
Test example 3
Select the Density of flies higher positions such as metope around brewery's vinasse pond, poor pond.
It takes 100g fly spray of the present invention to mix with 80ml water and mixes paste, and smearing metope (10~30 pieces/40 ㎡, 10 × 13
㎝/block)；
Density is 228/cage before fly eradication, makes 1 day dead fly number 1788 after medicine, and 2 days are 1468, and 7 days every cages are close after going out
Degree is 28；
Prove that fly spray of the present invention has immediate effect, holding effect, flies are without anabiosis.
Claims (1)
1. a kind of method for preparing fly spray using fly agaric, it is characterized in that the following steps are included:
1) fly agaric inclined-plane parent species are chosen and carry out shake-flask seed culture, high pressure sterilization pressure 1kg/cm3 keeps 30min, after cooling
Inoculation, inoculum concentration are the 250ml shaking flask that an inclined-plane connects that 6 loading amounts are 100ml, and cultivation temperature is 24~26 DEG C, are rotatably shaken
170~200 revs/min of bed, incubation time are 160~240h, 20% or more bacterium ball weight in wet base；
Culture medium prescription used are as follows: 2 grams of peptone, 0.1 gram of yeast extract, 1.5 grams of maltose, 5 grams of yellow bean sprout, potassium dihydrogen phosphate
0.15 gram, 0.075 gram of magnesium sulfate, 0.001 gram of vitamin B1, pH value 6.5, water 1000ml；
2) seed tank culture: inoculum concentration is 5%~10%, and temperature is 24~26 DEG C, and mixing speed is 150~180r/ min, ventilation
Amount is 1:0.5v/v. min per minute, and tank presses 0.3 X 105Pa is cultivated 2~3 days；
Used medium formula are as follows: 50 grams of wheat bran, 2 grams of peptone, 20 grams of glucose, 1 gram of potassium dihydrogen phosphate, 0.3 gram of magnesium sulfate,
Water 1000ml；
3) fermented and cultured: inoculum concentration is 5%~10%, and temperature is 24~26 DEG C, and mixing speed is 150~180r/ min, ventilatory capacity
For 1:0.5v/v. min per minute, cultivate 5~6 days；Fermentation liquid retrogradation, mycelia start self-dissolving, and pH value is down to 4.5, residual sugar amount drop
To 0.3%, as fermentation termination when adenosine content is begun to decline puts tank leaching mycelium；
Used medium formula are as follows: 20 grams of sucrose, 5 grams of peptone, 20 grams of glucose, 20 grams of beancake powder, potassium dihydrogen phosphate 1.5
Gram, 0.75 gram of magnesium sulfate, 1 gram of defoaming agent, water 1000ml；
4) fermentation material by step 3) containing mycelium and fermentation liquid is divided into mycelium and filtrate by plate compression, and mycelium adds one
Amount water is heated to 80 DEG C again, keeps the temperature 1h, and vacuum filtration removes residue, and filtrate merges with extracting solution, is concentrated into relative density 1:1,
As frog bacterium concentrate；
5) according to the volume ratio 1:4 of frog bacterium concentrate and water, 90 DEG C of clear water is injected into the frog bacterium concentrate of step 4),
Frog bacterium concentrate is immersed in 90 DEG C of thermostatted waters 2 hours, and toxin-frog mykol of fly agaric is dissolved in water, and filtering obtains frog
Malicious alcohol solution I and filter residue I；
6) 90 DEG C of clear water with the medium volume of step 5) are injected into the filter residue of step 5) I, are kept for 2 hours, and filtering obtains frog
Malicious alcohol solution II and filter residue II；
7) the batrachotoxin alcohol solution II of the batrachotoxin alcohol solution I of step 5) and step 6) is mixed, is put into reduced pressure dress
It sets, the pressure intensity parameter that vacuum concentration equipment is arranged is 60~65K pa, and heating temperature is 85~90 DEG C, is concentrated into original solution volume
5~10%, obtain batrachotoxin alcohol concentrate；
8) filter residue of step 6) II is Tumblied Dry to water content less than 23%；
9) take fry dulcet maize flour with drying after filter residue II mixed by quality 1:1, diameter is ground by pulverizer
Less than the hybrid particles of 200 mesh, it is added white sugar into hybrid particles, the mass ratio of hybrid particles and white sugar is 25:1, obtains medicine
Material；
10) the batrachotoxin alcohol concentrate atomizing of step 7) is sprayed on the spice of step 9), every kilogram of spice sprinkling
1000ml batrachotoxin alcohol concentrate, and 0.01 gram of sesame oil is added, stirring 10 times or more, drying machine drying to water content 55~60%；
11) spice of the step 10) after dry is put into pelletizing forming equipment, pelletizing forming to get.
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| CN201710407021.9A CN107018997B (en) | 2017-06-02 | 2017-06-02 | A method of fly spray is prepared using fly agaric |
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| CN201710407021.9A CN107018997B (en) | 2017-06-02 | 2017-06-02 | A method of fly spray is prepared using fly agaric |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101861796A (en) * | 2010-05-18 | 2010-10-20 | 四川大学 | Method for culturing amanita pantherina by using waste distillage after fermentation of coloured rice rich in trace elements |
| CN103627757A (en) * | 2013-12-02 | 2014-03-12 | 云南大学 | Method for improving toxin-producing capacity of small amanita pantherina mycelia |
| CN105961442A (en) * | 2016-05-16 | 2016-09-28 | 吉林省生物研究所 | Method for preparing rodenticide from lampteromycetes japonicus |
| CN106316631A (en) * | 2016-08-27 | 2017-01-11 | 广西新六合环保有限责任公司 | Leaf fertilizer for killing peanut aphids |
-
2017
- 2017-06-02 CN CN201710407021.9A patent/CN107018997B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101861796A (en) * | 2010-05-18 | 2010-10-20 | 四川大学 | Method for culturing amanita pantherina by using waste distillage after fermentation of coloured rice rich in trace elements |
| CN103627757A (en) * | 2013-12-02 | 2014-03-12 | 云南大学 | Method for improving toxin-producing capacity of small amanita pantherina mycelia |
| CN105961442A (en) * | 2016-05-16 | 2016-09-28 | 吉林省生物研究所 | Method for preparing rodenticide from lampteromycetes japonicus |
| CN106316631A (en) * | 2016-08-27 | 2017-01-11 | 广西新六合环保有限责任公司 | Leaf fertilizer for killing peanut aphids |
Non-Patent Citations (2)
| Title |
|---|
| Catching flies with Amanita muscaria:traditional recipes from Slovenia and their efficacy in the extraction of ibotenic acid;Mateja Lumpert,Samo Kreft;《Journal of Ethnopharmacology》;20160407;1-8 * |
| 毒蘑菇菌株及毒素粗提液对樟子松枯梢病菌生长的影响;祁金玉,宋瑞清;《林业科技》;20060525;第31卷(第3期);20-23 * |
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