CN103333813A - Compound microbial preparation and preparation method thereof - Google Patents
Compound microbial preparation and preparation method thereof Download PDFInfo
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- CN103333813A CN103333813A CN2012104064569A CN201210406456A CN103333813A CN 103333813 A CN103333813 A CN 103333813A CN 2012104064569 A CN2012104064569 A CN 2012104064569A CN 201210406456 A CN201210406456 A CN 201210406456A CN 103333813 A CN103333813 A CN 103333813A
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Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a compound microbial preparation and a preparation method thereof, and belongs to the fields of environmental protection and microbiology. The compound microbial preparation comprises EM bacteria, bacillus subtilis, beauveria bassiana and bacillus thuringiensis. According to the invention, the compound microbial preparation is prepared from complementary and antergic microbial strains. The compound microbial preparation has advantages of various kinds, reasonable compatibility, strong function, high economic benefit, and the like. The compound microbial preparation has good effects on biological deodorization and plant insect control.
Description
Technical field
The invention belongs to environmental protection and microorganism field, be specifically related to a kind of complex micro organism fungicide and preparation method thereof.
Background technology
1, stench is that a speciogenesis source is numerous, the sensation public hazards that contaminated area is wide.It not only makes the people feel bad and detests, and the many objectionable impuritiess in the foul smell directly are detrimental to health.For this reason, begin to cause gradually home and abroad environment scientific worker's attention and concern from the sixties in 20th century, and obtained many-sided achievement.Japan promulgated " odor prevention method " in 1971, and the U.S. promulgated " Clean Air sct " in 1971, and Germany stipulated frequency, the foul smell timing method of foul smell in 1993 in " instruction of relevant foul smell in the ambient atmosphere ".China starts late to the research of odor pollution, with reference to Japanese experience, in on January 15th, 1994 and on July 1st, 2003 implemented " odorant pollutant emission standard " respectively (GB1455421993) and " urban wastewater treatment firm pollutant emission standard " (GB1891822001) to odor concentration, and 8 kinds of odorous substances, that is: ammonia, Trimethylamine 99, sulfide, dimethyl sulfide, methyl disulfide, dithiocarbonic anhydride, vinylbenzene, and its measuring method also made concrete regulation.Repugnant substance is a lot, and at present the material that detects in the atmosphere can divide 8 classes, that is: 1, sulfide.2, aliphatic amide.3, hydro carbons.4, carbonyl compound.5, alcohols.6, phenols.7, lower fatty acid.8, indoles totally 147 kinds of gases.
Present stage, the treatment process of global odor pollution mainly contains gas washing method, ozone oxidation method, direct combustion method, absorption method, oxide treatment method, air oxidation process, masking method, air dilution method, acid-alkali washing method, biological chemistry decomposition method.And the reodorant that has in the market mainly contains chemical deodorizing agent and plant deodorant.Plant deodorant extraction process complexity, the production cost height; The chemical deodorizing agent also can cause secondary pollution except cost is higher.Therefore with low cost, technology is simple, uses the microbial deodorant of environmental protection to become the focus that the scientific research personnel pays close attention to and researches and develops.But all biological deodorants of Chu Xianing all exist can only degrade sulfide (as: hydrogen sulfide) and ammonia in the market, and there is defective in organism (total hydrocarbon) degradation capability.
2, the plant insect insect that mainly refers to harm the crops and products thereof and mite class etc.Kind is many, distribution is wide, breeding is fast, quantity is big, except the heavy losses that directly cause farm crop and products thereof, still propagates the media of Plant diseases.
The generation harm present situation of gardens disease and pest:
It is a kind of normal spontaneous phenomenon that the generation of gardens disease and pest exists, do not have set worm, not have tree not sick, the sick worm in gardens is an important component part of association in the landscape ecology system, is that natural enemy existence, the indispensable foodstuff ornamental plant of procreation have oneself's modification, self-compensating ability.
The many natural control action kou that also has the disease worm in the environment, green plants, disease and pest and natural enemy are in a kind of equilibrium state under the state usually.But urban ecology is fragile relatively, nature adjusting power is limited, city trees and shrubs has very strong dependency to surrounding environment, the healthy growth of city trees and shrubs is subjected to multiplely to influence because of rope, manage and protect unreasonably, the unusual change of weather, pollution and other artificial interference of urban air can break established equilibrium state at any time, bring out the serious generation of gardens disease and pest, and develop into natural disaster.The sick worm in gardens breaks out and causes disaster is a kind of common phenomenon of being not difficult, and is to be subjected to some undesirable elements influences and the result that causes the eubiosis to be destroyed.Therefore should be familiar with and treat the generation of gardens disease and pest objectively, follow its objective law, scientifically go to administer.The correlative study of China's gardens disease and pest is started late, and is just developed rapidly later on from 80 years, by generaI investigation, tentatively found out the kind of China's gardens disease and pest, distribute and the harm situation, grasped the pests occurrence rule of the sick worm in most of gardens, also found many control measures effectively.
Garden plants is of a great variety, and its disease and pest is varied especially, it is reported that China has various gardens disease kind more than 5600,8265 kinds of garden pest insects, and the disease of wherein causing disaster worm is hundreds of.Gardens disease and pest hazard approach is various, and for example insect can sting food root and seedling, nibbles blade, eat into food limb, the tender tip, blade, sucks juice and lays eggs harm etc.; Disease can endanger root, stem, leaf, flower or whole plant, produces yellow, spot, rots, symptom such as deformity, and can form stain, mould layer, meal etc., thereby cause plant strain growth weak, even dead, ornamental value reduces, and influences city look and feel and urban ecological environment.Gardens disease and pest regionality is very strong, different provinces and cities, different areas can cause existence such as pest species distribution, hazard rating, pests occurrence rule than big difference because of difference such as envrionment conditions, green plants are formed, control basis, can not act accordingly and copy word by word, the dominant population of the sick worm in gardens, different areas has larger difference.In a word, disease and pest harm in gardens is very serious, and pest damage obviously overweights disease.
Current, urbanization process is constantly accelerated, and urban afforestation is extremely paid attention to, and various greenings such as park, community, sight spot are flourish, and urban forests fraction of coverage and per capita green area sustainable growth are built ecology homestead energetically and become each city to pursue jointly.Continuous expansion along with the greening scale.The diversity of the introduction of new greening species and greening layout, originally ornamental plant disease and pest harm of a great variety, that dispersion takes place is more serious, and kind constantly increases, and has presented many new situation, disease and pest rises one after another, and the new problem layer goes out constantly.
But a little less than the gardens plant protection relative thin, lag behind the greening developing steps, especially system investigation and the correlative study at gardens, Northwest of Shandong disease and pest still is in blank, kind distribution and pests occurrence rule to the gardens disease and pest can not be grasped, relevant less investment, the insect pest preventing and controlling strategy is unreasonable, measure is incapable, and the serious phenomenon that exists greening to separate with plant protection, and a lot of afforestation project units are busy with engineering construction.Be negligent of pest management, the blind import species.Caused the part disease and pest propagation, spread, ornamental plant is suffering serious harm, has had a strong impact on the effect of city look and feel and greening work.Therefore, at the concrete actual correlation study research of carrying out in my ground, thoroughly find out main green plants pest species and form and the harm level, grasp the pests occurrence rule of main kind, in order to instruct control very urgent.
The processing of plant pest now mainly is divided into:
1. cultural control.
2. chemical prevention, the human hazardous and noxious substances of using those insect to be produced lethal action, these hazardous and noxious substances mostly are that the human chemical means that passes through synthesizes, famous DDT is exactly wherein a kind of, people to it significant insecticidal action filled with exultation in disastrous effect also following, owing to be that the synthetic ecotope is difficult to its metabolic degradation, retain in vivo with in the soil for a long time, and biology caused huge injury, environmental pollution is serious, HUMAN HEALTH is made a big impact, also cause secondary pollution simultaneously, people also more and more recognize a lot of drawbacks of chemical process now.
3. utilize other useful animals pest controls, except predatism and parasitic enemy insect, also have birds, batrachia and other animal, the development of controlling number of pest is had great role.
4. biological control at first is to control worm with worm, utilizes the natural enemy insect pest control to be called with worm and controls worm, comprising the utilization of beneficial mite.Secondly control worm with biological bacteria, control worm with bacterium and be exactly pathogenic micro-organism (fungi, bacterium, the virus etc.) pest control that utilizes insect, wherein most widely used with bacterium and fungi.Therefore using the microorganism insecticide of environmental protection is the trend of scientific development.
Summary of the invention
The technical problem to be solved in the present invention be how to provide a kind of can deodorizing again can insect protected complex micro organism fungicide, solve single, functional poor, the shortcoming that economic benefit is not high of existing microbial inoculum kind.
Technical scheme of the present invention is: a kind of complex micro organism fungicide, it comprises EM bacterium, subtilis, muscardine, Bacillus thuringiensis.
Further, described complex micro organism fungicide is mixed by the fermented liquid of EM bacterium, subtilis, muscardine, Bacillus thuringiensis.
Further, the total viable count of effective bacterium in the described complex micro organism fungicide is no less than 20 * 10
8Cfu/ml, the quantity of each effective bacterium is no less than 0.01 hundred million/ml.
The preparation method of complex micro organism fungicide is: the fermented liquid for preparing EM bacterium, subtilis, muscardine, Bacillus thuringiensis at first respectively;
Above-mentioned fermented liquid is got product by following weight proportion mixing: 4～8 parts of EM fermented liquids, 3 parts of fermentation of bacillus subtilis liquid, 1 part of fermentation of white muscardine fungi liquid, 1 part of Bacillus thuringiensis.
Further, the preparation method of described EM fermented liquid is:
With EM bacterial classification 10 grams, 0.1 kilogram in brown sugar (brown sugar dissolves with hot water earlier), add 1 kilogram water.Annotate: (add the red pool or white sugar after earlier water being heated to 100 ° of C, then continue heating 5 minutes.Add the EM bacterial classification when being cooled to 40 ℃), sealed fermenting (35～37 ℃) 3～5 days can be exitted once on the 3rd day, opened vessel port and smelt and have sour-sweet flavor namely to activate success, used as liquid spawn.
The liquid spawn that activation is good added second order fermentation jar sealed fermenting (35～37 ℃) 3～5 days, inoculum size 10%.The proportioning of second order fermentation liquid: brown sugar 10%, VITAMIN 0.3%, amino acid 0.1%; The finished product standard: smell is sour-sweet, pH value=4.0～6.0, viable bacteria content 〉=10,000,000,000/ml.
The preparation method of described fermentation of bacillus subtilis liquid is:
Subtilis is seeded in the substratum, and culture medium prescription is dextrose anhydrous 3.50 g/L, peptone 0.83 g/L, yeast extract paste 0.50 g/L, potassium primary phosphate 0.35 g/L and calcium carbonate 0.25 g/L, and 34 ± 2 ℃ of temperature are cultivated 16 h.
The preparation method of described fermentation of white muscardine fungi liquid is:
Muscardine is seeded in the substratum, and culture medium prescription is (g/L): corn steep liquor: 2, and sucrose: 3, KH
2PO
4: 0.226%, MgSO
47H
2O:0.0123%, FeSO
47H
20:0.0023%, ZnSO
4: 0.002%, CaCO
3: 0.4, potato juice: 20, glucose: 20, potassium primary phosphate: 3, agar: 15, peptone: 5, yeast extract paste: 5, pH6.Fermentation condition is: at constant temperature 25 degree, vibration 140r/min cultivates 96h, inoculum size 5%.
Described Bacillus thuringiensis preparation of fermentation liquid method is:
Bacillus thuringiensis is seeded in the substratum, and culture medium prescription is (g/L): peptone: 5, and extractum carnis: 30, NaCl: 5, agar: 15, W-Gum, glucose, sucrose are carbon source: 10, soybean cake powder: 1.5, yeast powder: 0.4, K
2HPO
4: 0.03, MgSO
47H
2O:0.02, CaCO
3: 0.04, ZnSO
4: 0.02, pH 7.0-7.2.
Fermentation condition is: initial pH 7.2, and autoclaving 20min. is 1% inoculation bacterial classification with volume fraction, places 28 ℃, carries out fermentation culture in the shaking table of 200 r/min; Behind 92 h gemma content in the fermented liquid is measured.
The application of complex micro organism fungicide of the present invention on biological deodorizing or botanical insect-proof.
The EM microbial inoculum
EM bacterium (Effective Microorganisms) is made up of about 80 kinds of microorganisms, and the EM bacterium is studied successfully according to professor by the ratio of Japanese Ryukyu university is good nineteen sixty-eight, puts goods on the market the eighties.The EM bacterium is a kind of little life bacteria preparation that a microorganism is composited surplus 6 genus 56 of photosynthetic bacterium, milk-acid bacteria, yeast and actinomycetes.The mechanism of action is the competition that forms EM bacterium and pathogenic micro-organism contention nutrition, because EM bacterium survival and reproduction very easily in soil, so can comparatively fast and stably occupy the ecologic niche in the soil, form the advantage group of useful microbial bacteria, thus the breeding of control pathogenic micro-organism and to the invasion and attack of crop.
The EM microbial inoculum mainly contains following several big flora and forms:
1, photosynthetic bacteria group: the photosynthetic bacteria group in the EM bacterium liquid (aerobic and anaerobism) belongs to the autotrophy microorganism, it can utilize solar thermal energy that soil accepts or be the energy with the ultraviolet ray, hydrogen sulfide in the soil and the hydrogen in the hydrocarbon polymer are separated, the change objectionable impurities is innoxious substance, and be matrix with secretory product, organism, obnoxious flavour (hydrogen sulfide etc.) and the carbonic acid gas of plant root, nitrogen etc., synthetic carbohydrate, amino acid, vitamins, nitrogen compound and physiologically active substance etc. are rich soil and the main force that promotes animal and plant growth.The metabolic substd of photosynthetic bacteria or directly absorbed by plant perhaps becomes the nutrient of other microbial reproduction, if photosynthetic bacterium can breed, other beneficial microorganism also can be bred.
2, lactobacillus: milk-acid bacteria (anaerobic type), it produces lactic acid based on the materials such as carbohydrate that picked-up photosynthetic bacterium yeast produces.Lactic acid has very strong sterilizing ability, can effectively suppress the harmful microbe activity, and organic rapid corrupt the decomposition.Milk-acid bacteria can make under the normality not labile xylogen and Mierocrystalline cellulose etc. become to decompose easily, and eliminate all drawbacks that undecomposed organism produces, in the vital role that organic matter fermentation decomposes performance shock brigade, the organic substance that it will not become thoroughly decomposed changes into the effective nutrient of animals and plants.Milk-acid bacteria can also effectively suppress the pathogenic bacterium propagation that continuous cropping obstacle produces.
3, yeast flora: yeast (aerobic type) utilizes amino acid, carbohydrate and other organic substance, by fermentation, produces the fissional sensitization material of promotion.Yeast provides important nutrition guarantee for the production that the effective microbe that promotes other is bred needed matrix (food) in EM bacterium group army.In addition, the single cell protein of yeast production is the indispensable available nutrient of animal.
4, Gram positive actinomycetes group (good gas).It obtains amino acid, nitrogen etc. as matrix from photosynthetic bacterium, produce various antibiotics, VITAMIN and enzyme, can directly suppress pathogenic bacteria.It obtains harmful mould and the needed matrix of bacterial multiplication in advance, thereby suppresses their propagation, and creates the living environment of other beneficial microorganism propagation.The mixed clean bacterium effect of actinomycetes and photosynthetic bacterium is more much bigger than the lethality of actinomycetes individual combat.It has Degradation to the material that difficulty is decomposed as xylogen, Mierocrystalline cellulose, chitin etc., and easy passive plant absorbing, strengthens animals and plants to resistibility and the immunizing power of various diseases.Actinomycetes also can promote vinelandii and VA mycorhiza bacterium propagation.
5, the thread flora (anaerobies) of fermentation system.The Eurotium that uses during with fermentation alcohol is main body, and it can coexist with other microorganisms, and especially the generation to ester in the soil has good result.Because the alcohol generative power is strong, can prevents the generation of maggot and other insects, and can eliminate stench.
Therefore, each quasi-microorganism is all being brought into play important effect separately, central role is that photosynthetic bacterium and lactobacillus acidophilus are taken as the leading factor, its synthesis capability is supporting the activity of other microorganisms, also utilize simultaneously the material of other microorganisms, form the relation of coexistence and co-prosperity, guarantee that EM bacterium liquid status is stable, multiple functional, given play to the powerful energy that group army fights.The major function of EM bacterium liquid is to bring up optimum ecology.As long as use appropriate, it will with the rapid combination of optimum strength of place of arrival, produce antioxidant, remove oxidizing substance, eliminate corrupt, suppress pathogenic bacteria, formation is suitable for the good environment of animal and plant growth, and simultaneously, it also produces a large amount of benefit materials that easily absorb for animals and plants, as amino acid, organic acid, Polysaccharides, various VITAMIN, various biochemical enzyme, somatomedin, microbiotic and antiviral substance etc., improve vegeto-animal immunologic function.
The bacillus subtilis microbial inoculum
Subtilis is a kind of of bacillus.0.7～0.8 * 2～3 microns of individual cells, uniform coloring.No pod membrane, peritrichous can move.Gram-positive microorganism, 0.6～0.9 * 1.0～1.5 microns of gemma, oval to column, be positioned at thalline central authorities or inclined to one side slightly, gemma forms the back thalline and does not expand.The bacterium colony surface irregularity is opaque, dirty white or little yellow, and when growing in the liquid medium within, the normal wrinkle mould that forms.Aerophil.Available protein, multiple sugar and starch decompose tryptophane and form indoles.
1. the subtilyne that produces in the subtilis thalli growth process, polymyxin, nystatin, linear gramicidins isoreactivity material, these active substances have the obvious suppression effect to the conditioned pathogen of pathogenic bacterium or autogenous infection.
2. subtilis consumes the free oxygen in the environment rapidly, causes the enteron aisle hypoxemia, promotes useful anerobe growth, and produces lactic acid etc.
Organic acidClass reduces the enteron aisle pH value, suppresses other pathogenic bacterium growth indirectly.
3. stimulate growing of animal immune organ, activate T, bone-marrow-derived lymphocyte, improve immunoglobulin (Ig) and antibody horizontal, strengthen cellular immunization and humoral immune function, improve herd immunity.
4. enzymes such as the synthetic α-Dian Fenmei of subtilis thalline self, proteolytic enzyme, lipase, cellulase, in digestive tube with animal body in digestive enzymes together play a role.
5. can synthesise vitamins multiple vitamin B group such as B1, B2, B6, nicotinic acid, improve the activity of the plain and scavenger cell of animal body internal interference.
The muscardine agent
Muscardine (Beauveria Vuillemin) belongs to hyphomycetes, Moniliales, Moniliaceae, Beauveria, being a kind of insect pathogenic fungus of broad spectrum, is the preparation that is fermented, is processed into by the stiff Pseudomonas of insect pathogenic fungus imperfect fungi Moniliales clump spore actions and spoken parts in classical Chinese drama.
Multiple harmful insect is had parasitics, pathogenic strong, adaptability is strong.Can control insect population (insect) quickly and efficiently, not injure other natural enemy insects and beneficial organism simultaneously, with respect to the traditional chemical agricultural chemicals of high residue, high toxicity, high resistance, strong advantage be arranged.
Main by the insect cuticle contact infection, secondly also can be through digestive tube and respiratory tract infection.The approach that infects is different because of the difference of the kind of insect, worm attitude, envrionment conditions etc.
Scanning electron microscope or fluorescent dye are observed and are found, the of short duration growth on the cotton bollworm larvae body wall of the high toxic bacterial strain of beauveria bassiana namely forms the invasion structure, and low toxic bacterial strain produces elongated stolon at the larva body wall, and these mycelia can absorb nutrition in pest body, finally cause insect death.
The muscardine agent is the lower toxicity microbial pesticide, and people, animal are not had pathogenic effects, belongs to weak allergen, does not have " three cause " problem.Said preparation is nontoxic to people and animals, to fruit tree safety.
The bacillus thuringiensis bacterial classification
Bacillus thuringiensis (Bacillus thuringiensis) belongs to the Bacillaceae bacterium, a kind of gram positive bacterium that can produce parasporal crystal and gemma, some subspecies can also produce α and βWai Dusu, and Bacillus thuringiensis is current production rate maximum, the widest biotic pesticide of use.Its main active ingredient is a kind of or several insecticidal crystal proteins (insecticidal crystal proteins, ICPs), claim delta-endotoxin again, to insects such as lepidopteran, Coleoptera, Diptera, Hymenoptera, Homopteras, and animals and plants nematode, acarian constant pitch main drive thing have specific cytotoxicity, and to nontarget organism safety.
The insecticide active substance of bacillus thuringiensis mainly contains two kinds, i.e. crystal and spore. and crystal is parent toxin again, and it is a kind of protein.Complete crystal is not had an activity to insect, have only when crystal get by insect the food enter midgut after, under midgut liquid caustic soda condition, dissolve, resolve into littler molecule through proteolytic enzyme again--the toxicity peptide, be only the material that really has virulence, the toxicity peptide destroys the insect midgut cell, the enteron aisle alkali content haemocoele that bleeds, spore and thalline enter body cavity by ruined intestines wall, and growth and breeding within it, cause septicemia at last and death.
Another kind of situation, proteotoxin are destroyed the intestines wall and are caused the stomach paralysis, and insect is under hungry and disorganization's dual function and death.But the acid-basicity of not all insect stomach is identical, the insect that Here it is has to the bacillus thuringiensis sensitivity have to then insensitive reason place.
The major advantage of bacillus thuringiensis (BT) sterilant:
1) the BT sterilant is to the person poultry safety, but will avoid contacting with eye and wound.
2) the BT sterilant can be used for a long time and directly cause the harvest season, and this is the difference with other sterilant.
3) can use continuously, when not required interbody spacer.
4) control of BT different strains has specificity, to beneficial insect and non-target pest safety.
5) birds and animal can eat the insect that is caused death by BT safely.
6) use the plant of BT and do not produce harm, environmentally safe reaches and does not does not kill and wound pest natural enemy.
7) do not produce resistance, but will note the excessive BT of using.
8) prevention effect is good.
9) easy to use.
The present invention compared with prior art has following advantage:
The present invention adopts the microbial strains of complementary antagonism to make complex micro organism fungicide, and this microbial inoculum has that kind is complete, compatibility is reasonable, functional strong, economic benefit advantages of higher.
Complex micro organism fungicide of the present invention all has good effect on biological deodorizing, botanical insect-proof.
Embodiment
A kind of complex micro organism fungicide, it comprises EM bacterium, subtilis, muscardine, Bacillus thuringiensis.
Further, described complex micro organism fungicide is mixed by the fermented liquid of EM bacterium, subtilis, muscardine, Bacillus thuringiensis.The total viable count of effective bacterium in the described complex micro organism fungicide is no less than 20 * 10
8Cfu/ml, the quantity of each effective bacterium is no less than 0.01 hundred million/ml.
Embodiment
The preparation method of complex micro organism fungicide is: the fermented liquid for preparing EM bacterium, subtilis, muscardine, Bacillus thuringiensis at first respectively; Then above-mentioned fermented liquid is mixed by following weight proportion and get product 4～8 parts of EM fermented liquids, 3 parts of fermentation of bacillus subtilis liquid, 1 part of fermentation of white muscardine fungi liquid, 1 part of Bacillus thuringiensis.
The preparation method of EM fermented liquid is:
With EM bacterial classification 10 grams, 0.1 kilogram in brown sugar (brown sugar dissolves with hot water earlier), add 1 kilogram water.Annotate: (add the red pool or white sugar after earlier water being heated to 100 ° of C, then continue heating 5 minutes.Add the EM bacterial classification when being cooled to 40 ℃), sealed fermenting (35～37 ℃) 3～5 days can be exitted once on the 3rd day, opened vessel port and smelt and have sour-sweet flavor namely to activate success, used as liquid spawn.
The liquid spawn that activation is good added second order fermentation jar sealed fermenting (35～37 ℃) 3～5 days, inoculum size 10%.The proportioning of second order fermentation liquid: brown sugar 10%, VITAMIN 0.3%, amino acid 0.1%; The finished product standard: smell is sour-sweet, pH value=4.0～6.0, viable bacteria content 〉=10,000,000,000/ml.
The preparation method of fermentation of bacillus subtilis liquid is:
Subtilis is seeded in the substratum, and culture medium prescription is dextrose anhydrous 3.50 g/L, peptone 0.83 g/L, yeast extract paste 0.50 g/L, potassium primary phosphate 0.35 g/L and calcium carbonate 0.25 g/L, and 34 ± 2 ℃ of temperature are cultivated 16 h.
The preparation method of fermentation of white muscardine fungi liquid is:
Muscardine is seeded in the substratum, and culture medium prescription is (g/L): corn steep liquor: 2, and sucrose: 3, KH
2PO
4: 0.226%, MgSO
47H
2O:0.0123%, FeSO
47H
20:0.0023%, ZnSO
4: 0.002%, CaCO
3: 0.4, potato juice: 20, glucose: 20, potassium primary phosphate: 3, agar: 15, peptone: 5, yeast extract paste: 5, pH6.Fermentation condition is: at constant temperature 25 degree, vibration 140r/min cultivates 96h, inoculum size 5%.
Bacillus thuringiensis preparation of fermentation liquid method is:
Bacillus thuringiensis is seeded in the substratum, and culture medium prescription is (g/L): peptone: 5, and extractum carnis: 30, NaCl: 5, agar: 15, W-Gum, glucose, sucrose are carbon source: 10, soybean cake powder: 1.5, yeast powder: 0.4, K
2HPO
4: 0.03, MgSO
47H
2O:0.02, CaCO
3: 0.04, ZnSO
4: 0.02, pH 7.0-7.2.
Fermentation condition is: initial pH7.2, and autoclaving 20min. is 1% inoculation bacterial classification with volume fraction, places 28 ℃, carries out fermentation culture in the shaking table of 200 r/min; Behind the 92h gemma content in the fermented liquid is measured.
Table 1: the technical indicator of complex micro organism fungicide
Table 2: the innoxious index of complex micro organism fungicide
Experimental example 1Indoor rubbish deodorizing test
In the room of a sealing, put the highly rubbish of rotten generation stench of a pile, the complex micro organism fungicide that configures is diluted 20 times with tap water before use, be sprayed directly on the dumping zone.Around rubbish, different collecting locations is set and samples, measure preceding the reaching of sprinkling and spray back 15 minutes hydrogen sulfide and the ammonia concentration in the foul gas, 1 meter away from edge, smelly source be of sampling point, height of sampling is 1.4 meters, 6 point samplings are averaged.Odor concentration adopts 3 comparison expression odor bag methods (GB/T14675); Ammonia adopts clorox-Whitfield's ointment spectrophotometry (GB/T14679); Hydrogen sulfide adopts ammonium alcohol polyvinyl phosphate absorption-methylene-blue colorimetric method (GB/T16489); Total plate count adopts bump method (GB/T17093).Test result sees Table 3
Table 3: the indoor rubbish deodorizing effect experiment of composite microbial bacteria reodorant
Annotate: odor concentration unit: zero dimension; Bacterium sum unit: cfu/m3.
Test-results shows that composite fungus agent has good deodorizing effect.
Experimental example 2
1. preparation is tested and is used microbial inoculum.
Obtain bacterium liquid by aforementioned spawn culture method, various bacterium liquid are mixed in proportion: 5 parts of EM bacterium liquid, 3 parts of subtilis liquid, 1 part of muscardine liquid, 1 part of thuringiensis bacillus liquid is in the test tube of packing into.
2. the insect of test usefulness has: locust, pine moth, the big aphid of cypress, cutworm, looper, fly, longicorn worm, aphid.
3. test method.
Get test with mixed bacteria liquid 200 mL, respectively 80 insects are sealed up gauze with the bottle packing, it is indoor below 25 ℃ to put normal temperature, is sprayed in the bottle of dress worm with different diluents again.48 hours observation and inspections also calculate the insect mortality ratio.Identify that dead method is: in the test bottle, add 1-5 5%NaCl solution, observe stiff, the then curling or twisting of worm of living of dead worm behind 2 clocks.
Mortality ratio=(Chong that lives Shuo – test back live borer population) ÷ borer population * 100% of always living
Table 4: 2 times of diluent desinsections of composite fungus agent stoste kind synopsis
Table 5: the composite fungus agent of different concns is to the effect of deinsectization
| Composite fungus agent concentration | 24h mortality ratio % | 48h mortality ratio % | 72 mortality ratio % |
| 5 times of diluents of composite fungus agent | 40 | 80 | 100 |
| 10 times of diluents of composite fungus agent | 40 | 75 | 100 |
| 20 times of diluents of composite fungus agent | 30 | 70 | 100 |
| 30 times of diluents of composite fungus agent | 30 | 60 | 95 |
| 50 times of diluents of composite fungus agent | 20 | 60 | 90 |
The result shows: the insect of listed plant has good killing effect in the composite microorganism viable bacteria preparation his-and-hers watches of the present invention, and very wide insecticidal spectrum is arranged.
Claims (9)
1. complex micro organism fungicide, it is characterized in that: it comprises EM bacterium, subtilis, muscardine, Bacillus thuringiensis.
2. a kind of complex micro organism fungicide according to claim 1, it is characterized in that: described complex micro organism fungicide is mixed by the fermented liquid of EM bacterium, subtilis, muscardine, Bacillus thuringiensis.
3. a kind of complex micro organism fungicide according to claim 2, it is characterized in that: the total viable count of effective bacterium in the described complex micro organism fungicide is no less than 20 * 10
8Cfu/ml, the quantity of each effective bacterium is no less than 0.01 hundred million/ml.
4. the preparation method of a complex micro organism fungicide: it is characterized in that, prepare the fermented liquid of EM bacterium, subtilis, muscardine, Bacillus thuringiensis at first respectively; Then above-mentioned fermented liquid is mixed by following weight proportion and get product: 4～8 parts of EM fermented liquids, 3 parts of fermentation of bacillus subtilis liquid, 1 part of fermentation of white muscardine fungi liquid, 1 part of Bacillus thuringiensis.
5. the preparation method of a kind of complex micro organism fungicide according to claim 4, it is characterized in that: the preparation method of described EM fermented liquid is:
Earlier 0.1 kilogram in brown sugar is added in 1 kilogram the water, add EM bacterial classification 10 grams, sealed fermenting after being mixed with solution: 35～37 ℃ of leavening temperatures, fermentation time 3～5 days namely gets the activated liquid bacterial classification; The liquid spawn that activation is good adds second order fermentation jar sealed fermenting, 35～37 ℃ of leavening temperatures, fermentation time 3～5 days, inoculum size 10%; The proportioning of second order fermentation liquid: brown sugar 10%, VITAMIN 0.3%, amino acid 0.1%; The finished product standard: smell is sour-sweet, pH value=4.0～6.0, viable bacteria content 〉=10,000,000,000/ml.
6. the preparation method of a kind of complex micro organism fungicide according to claim 4, it is characterized in that: the preparation method of described fermentation of bacillus subtilis liquid is:
Subtilis is seeded in the substratum, and culture medium prescription is dextrose anhydrous 3.50 g/L, peptone 0.83 g/L, yeast extract paste 0.50 g/L, potassium primary phosphate 0.35 g/L and calcium carbonate 0.25 g/L, and 34 ± 2 ℃ of temperature are cultivated 16 h.
7. the preparation method of a kind of complex micro organism fungicide according to claim 4, it is characterized in that: the preparation method of described fermentation of white muscardine fungi liquid is:
Muscardine is seeded in the substratum, and culture medium prescription is (g/L): corn steep liquor: 2, and sucrose: 3, KH
2PO
4: 0.226%, MgSO
47H
2O:0.0123%, FeSO
47H
20:0.0023%, ZnSO
4: 0.002%, CaCO
3: 0.4, potato juice: 20, glucose: 20, potassium primary phosphate: 3, agar: 15, peptone: 5, yeast extract paste: 5, pH6; Fermentation condition is: at constant temperature 25 degree, vibration 140r/min cultivates 96h, inoculum size 5%.
8. the preparation method of a kind of complex micro organism fungicide according to claim 4, it is characterized in that: described Bacillus thuringiensis preparation of fermentation liquid method is:
Bacillus thuringiensis is seeded in the substratum, and culture medium prescription is (g/L): peptone: 5, and extractum carnis: 30, NaCl: 5, agar: 15, W-Gum, glucose, sucrose are carbon source: 10, soybean cake powder: 1.5, yeast powder: 0.4, K
2HPO
4: 0.03, MgSO
47H
2O:0.02, CaCO
3: 0.04, ZnSO
4: 0.02, pH 7.0-7.2;
Fermentation condition is: initial pH7.2, and autoclaving 20min. is 1% inoculation bacterial classification with volume fraction, places 28 ℃, carries out fermentation culture in the shaking table of 200 r/min; Behind the 92h gemma content in the fermented liquid is measured.
9. according to each described a kind of complex micro organism fungicide application on biological deodorizing or botanical insect-proof of claim 1～4.
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