CN103937695B - A kind of composite bacteria agent and manufacture method thereof of processing livestock and poultry cultivation sewage - Google Patents
A kind of composite bacteria agent and manufacture method thereof of processing livestock and poultry cultivation sewage Download PDFInfo
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Abstract
The present invention relates to a kind of composite bacteria agent and manufacture method thereof of processing livestock and poultry cultivation sewage, belong to Environmental Biotechnology field. Microbial strains of the present invention is bacillus amyloliquefaciens IAE(<i>Bacillus? amyloliquefaciens?</i>IAE), trichoderma asperellum 12(<i>Trichoderma? asperellum</i>12) and bacillus thuringiensis (<i>Bacillus? thuringiensis</i>). The present invention is applicable to the sewage disposal of the livestock and poultry breeding industry that quantity of wastewater effluent is larger, can decompose by microorganism, the bulky grain solid matter that flocculates (comprising the harmful organisms such as roundworm egg), kill mosquitos and flies harmful organism, reduce the modes such as foul smell discharge and process aquaculture sewage. Sewage solid suspended particle after treatment, COD significantly reduce, after sewage disposal, can effectively reduce the load of using soil as liquid organic fertilizer or clear water mixing irrigation water, reduce the pollution to soil, effectively alleviate intensive culture and produce at certain space-time the difficult problem that high-concentration sewage causes surrounding soil to be difficult to dissolve.
Description
Technical field
The composite bacteria agent and the manufacture method thereof that the present invention relates to process livestock and poultry cultivation sewage, belong to Environmental BiotechnologyField.
Background technology
China's livestock and poultry breeding industry development rapidly, is ensureing town and country animal products supply, is promoting increasing peasant income, is enlivening rural area warpImportant function has been brought into play in Ji aspect. Along with livestock and poultry breeding industry development, breeding pollution thing discharge capacity also significantly increases, to regionEnvironmental security constitutes a threat to. Dynamically update survey data according to Pollutant source investigation, the COD of livestock and poultry breeding industry in 2010,Ammonia nitrogen discharge capacity reaches respectively 1,148 ten thousand tons, 650,000 tons, and the ratio that accounts for national total emission volumn is respectively 45%, 25%, and livestock and poultry are supportedGrow owner want COD in Sewage Water Emissions amount, ammonia nitrogen discharge capacity be respectively 3.23 times of industrial source discharge capacity then,2.30 doubly.
Livestock and poultry large-scale cultivation level progressively improves, and 2010, national live pig, laying hen and milk cow breeding scale ratio were respectivelyReach 65%, 79%, 47%, pig, ox, poultry breeding stock at the year end reach respectively approximately 4.6 hundred million, 1.1 hundred million, 53.5 hundred million, meatClass output ranks first in the world for continuous 22 years, and China live pig, laying hen and milk cow advantage provinces and regions pork, birds, beasts and eggs and milk crop account for respectively92%, 68%, 88% of whole nation total amount, it is raw that (district, city) economized in Hainan, Tianjin, Beijing, Xinjiang, Shanghai, Qinghai, Ningxia, Tibet etc.Pig breeding stock is lower, all less than 1% of the total breeding stock in the whole nation.
So mainly rely on soil utilization to dissolve to livestock and poultry breeding industry offal treatment both at home and abroad. But large-scale scaleCultivation causes producing a large amount of discarded objects at special time and space, and discarded object comprises a large amount of sewage, cannot grow distance fortuneDefeated, waste discharge amount seriously exceedes the soil ability of dissolving, and causes Farmland Soil Pollution, the elements such as the nitrogen phosphorus in feces of livestock and poultry,The materials such as antibiotic in feed addictive, hormone, copper, iron, chromium, zinc, excessive buildup in soil for a long time, causes soil and groundLower water environment pollution. Research shows, the pig farm that year amount of delivering for sale is 100,000, and pollution radius can reach 5 kilometers, and dust and aerosol are takenBand comprises the pathogenic microorganisms such as aftosa, swine plague, ETEC, anthrax, Brucella, fungal spore in a large number, stench,Dust and microorganism discharge capacity far exceed the self-purification capacity of atmosphere, have a strong impact on air quality, pollute human settlement and human body strongHealth.
The sewage that the reduces breeding enterprise surrounding soil load of dissolving, must reduce organic concentration and cause of disease in sewageMicro organism quantity reduces the discharge of foul smell simultaneously in processing procedure. The method of existing processing aquaculture wastewater mostly is compost or natural pondGas fermentation. Composting mode processing need to be added the absorbent materials such as stalk, plant ash, sawdust, peat in large quantities, increase treating capacity andProcessing cost. After biogas is processed, solid matter is by the material of anaerobic digestion Cheng Geng little, and sewage total amount changes little, but natural pond slagSeparation of Solid and Liquid difficulty, natural pond slag subsequent treatment is also a great problem. Therefore, develop a kind of method, make sewage disposal process compared toConventional method, foul smell, dust and aerosol amount are less, and the harmful organism amount of multiplying mosquitos and flies is less, and sewage after treatment is organicSubstrate concentration is low, and pathogenic microorganism quantity is few, can be directly as liquid organic fertilizer or the mixed water of filling with of clear water, very urgent and mustWant.
Three, summary of the invention
The object of invention is to provide a kind of composite bacteria agent of processing livestock and poultry breeding industry sewage, supports for intensive livestock and poultryGrow sewage disposal application.
A kind of livestock and poultry cultivation sewage disposal composite bacteria agent, is characterized by composite bacteria agent and contains solution starch gemma barBacterium IAE(BacillusamyloliquefaciensIAE)
), trichoderma asperellum 12(Trichoderma.asperellum12) and bacillus thuringiensis (B.thuringiensis）；
A wherein said bacillus amyloliquefaciens IAE(BacillusamyloliquefaciensIAE), inOn March 15th, 2013 is preserved in Chinese Typical Representative culture collection center, and preservation address is Luo Jia Shan, wuchang, wuhan, deposit numberCCTCCNO:M2013086, referred to as bacillus amyloliquefaciens CCTCCNO:M2013086;
A described strain trichoderma asperellum 12(Trichoderma.asperellum12), in preservation on May 16 in 2013In Chinese Typical Representative culture collection center, preservation address is Luo Jia Shan, wuchang, wuhan, deposit number CCTCCNO:M2013213, referred to as trichoderma asperellum CCTCCNO:M2013213.
Above-mentioned bacillus amyloliquefaciens CCTCCNO:M2013086, is characterized in that: from culture dish, choose with oeseThis bacterium of getting growth 24h is inoculated in the 250ml triangular flask that 100ml flocculant synthetic media is housed, inoculum concentration 104cfuml-1, 37 DEG C, 150rpm are cultivated 60h, bacterial strain synthesized micro-organism flocculation dosage 21.8gL-1。
The above-mentioned bacillus amyloliquefaciens CCTCCNO:M2013086 nutrient solution compound method used of fermenting is, to prepare 1LNutrient solution is example: glucose 80g, sodium glutamate 50g, (NH4)2SO48g，NaCl5g，K2HPO4·3H2O2g，MnSO4·7H2O0.25g，MnSO4?H2O0.03g, distilled water 1000ml, pH value nature, 115 DEG C of sterilizing 30min.
Above-mentioned trichoderma asperellum CCTCCNO:M2013213 fermentation method for producing is: trichoderma asperellum CCTCCNO:M2013213 are inoculated in nutrient solution and cultivate, and its condition of culture is 25 DEG C of cultivation temperature, shaking speed 120rpm, incubation time 72Hour, this bacterial strain fermentation liquor cellulase activity 25.2Uml-1。
Above-mentioned nutrient solution compound method used is (to prepare 1L culture medium as example): after peeling with 100g potato, be cut into littlePiece is put in water and boils, and boils 30min after boiling, adds after filtering 5g common sucrose and 2g rice straw powder in filtrate, is settled to1000ml, pH value nature, 115 DEG C of sterilizing 30min.
Be used for bacillus thuringiensis (B.thuringiensis) bacterial strain of biological control mosquitos and flies purchased from the raw agriculture of a specified duration in JiangsuChange Co., Ltd, also can be to other commercial companies or Microbe Inst., Chinese Academy of Sciences, Agricultural University Of Nanjing, Hohai University etc.R&D institution buys. Bacterial strain has biological control effect to fly and mosquito, for fly killing rate higher than 60%, for mosquitoKilling rate reaches 80%, can effectively suppress multiplying of mosquitos and flies in sewage.
The assay method that above-mentioned bacillus thuringiensis zymotic fluid is killed mosquitos and flies is: bacillus thuringiensis is inoculated into bacterial classificationCulture medium, cultivates, and its condition is: shaking table concussion 170rpm, 37 DEG C of cultivation temperature, cultivate CFU 24 hours≥1×109cfuml-1, bacterium liquid sparges in the wide-mouth bottle of cultivation fly and mosquito, measures compared to the processing of spray sterilized water, bacteriumThe death rate of mosquitos and flies is processed in liquid spraying.
The formula of above-mentioned fluid nutrient medium used is to prepare according to 1L culture medium: beef extract 3g, peptone 10g, NaCl10g, running water 1000ml, pH scope 7.0-7.5,121 DEG C of sterilizing 20min.
The preparation of the composite bacteria agent of described livestock and poultry cultivation sewage disposal, comprising: 1) by bacillus amyloliquefaciens CCTCCNO:M2013086 inoculation is to liquid seed culture medium, and the condition of cultivation is: shaking table concussion 170rpm, cultivation temperature 37DEG C, cultivate CFU >=1 × 10 24 hours9cfuml-1; Liquid seed culture medium formula used is to train according to 1LThe basigamy system of supporting: beef extract 3g, peptone 10g, NaCl10g, running water 1000ml, pH scope 7.0-7.5,121 DEG C of sterilizings20min; Bacillus amyloliquefaciens CCTCCNO:M2013086 seed is inoculated into liquid fermentation training according to 5% percent by volumeSupport base, carry out liquid fermentation production, the condition of its fermenting and producing is: 28 ~ 35 DEG C of cultivation temperature, and dissolved oxygen throughput scope is 30～100%, 150 ~ 220rpm, CFU >=1 × 10 of this bacterial strain of fermentation later stage fermentation liquid9cfuml-1, fermentation Low Temperature LiquidStore for future use; The formula of liquid fermentation medium used is to prepare according to 1L culture medium: glucose 80g, and sodium glutamate 50g,(NH4)2SO48g，NaCl5g，K2HPO4·3H2O2g，MnSO4·7H2O0.25g，MnSO4?H2O0.03g, distilled water1000ml, pH value nature, 115 DEG C of sterilizing 30min.
2) trichoderma asperellum CCTCCNO:M2013213 fermenting and producing, trichoderma asperellum CCTCCNO:M2013213 is inoculated intoIn PDA nutrient solution, carry out liquid seeds cultivation, condition of culture is: shaking table concussion 120rpm, and 25 DEG C of cultivation temperature, cultivation 72 is littleTime, CFU >=0.5 × 108cfuml-1; The formula of the seed PDA culture medium of liquid used is to cultivate according to 1LBasigamy system: be cut into small pieces to be put in water after peeling with 200g potato and boil, boil 30min after boiling, with after filtered through gauze, filtrate adds20g common sucrose, is settled to 1000ml, pH value nature, 115 DEG C of sterilizing 30min; By trichoderma asperellum CCTCCNO:M2013213 bacterial classification are inoculated into liquid fermentation medium according to 10% percent by volume, carry out liquid fermentation production, and it ferments rawThe condition of producing is: initial pH nature, cultivate 19 ~ 22 DEG C of temperature, and dissolved oxygen throughput scope is 30～100%, 90 ~ 170rpm, fermentationMiddle and later periods forms mycelium pellet rotating speed 250rpm stirring and is broken into mycelia fragment, zymotic fluid trichoderma asperellum CCTCCNO:MCFU >=0.5 × 10 of 2013213 bacterial strains8cfuml-1, the storage of zymotic fluid low temperature is for subsequent use; Liquid fermentation training usedNutrient solution compound method is (to prepare 1L culture medium as example): after peeling with 100g potato, be cut into small pieces to be put in water and boil, after boilingBoil 30min, in filtrate, add after filtering 5g common sucrose and 2g rice straw powder, be settled to 1000ml, pH value nature, 115DEG C sterilizing 30min.
3) bacillus thuringiensis zymotic fluid is produced, and bacillus thuringiensis is inoculated into liquid seed culture medium, cultivates barPart is: shaking table concussion 170rpm, 37 DEG C of cultivation temperature, cultivate CFU >=1 × 10 24 hours8cfuml-1; UsedThe formula of seed culture medium of liquid be to prepare according to 1L culture medium: beef extract 3g, peptone 10g, NaCl10g, from the beginningWater 1000ml, pH scope 7.0-7.5,121 DEG C of sterilizing 20min; Bacillus thuringiensis seed is connect according to 5% percent by volumePlant liquid fermentation medium, carry out liquid fermentation production, the condition of its fermenting and producing is: initial pH scope is 7.0 ~ 7.5, trainingSupport 25 ~ 33 DEG C of temperature, dissolved oxygen throughput scope is 30～100%, 150 ~ 220rpm, the bacterium colony of this bacterial strain of fermentation later stage fermentation liquidForm unit >=1 × 109cfuml-1, zymotic fluid stores for future use. The formula of liquid fermentation medium used is to train according to 1LThe basigamy system of supporting: beef extract 3g, peptone 10g, NaCl10g, running water 1000ml, pH scope 7.0-7.5,121 DEG C of sterilizings20min。
4) by microbial fermentation solution bacterial classification according to bacillus amyloliquefaciens CCTCCNO:M2013086: trichoderma asperellumCCTCCNO:M2013213: bacillus thuringiensis=10:8:1 volume ratio is compound, with calcium hydroxide regulate pH value be 8.0 ~8.5, stir and within 0.5 ~ 1 hour, allow microbial cells stable and enter resting state with mixer. Compound leaves standstill 2 little at normal temperatureTime, filling, be aquaculture sewage purification flora.
Described processing aquaculture sewage flora can be used for aquaculture sewage storeroom directly to be sprayed, mixes or in conjunction with workJourney measure is added, the various microorganisms that effectively in degradation of sewage, macromolecular substances, precipitation suspend and solid particulate matter,Effectively suppress to multiply mosquitos and flies class harmful organism in sewage, reduce the generation of foul smell, aerosol, dust in processing procedure, make placeOrganic matter of sewage concentration after reason significantly reduces.
The invention provides a kind of composite bacteria agent and the production method thereof that can process aquaculture sewage, by high yieldBiological flocculant, High Cellulase Production and have high-performance bio and kill the microorganism high density fermentation of mosquitos and flies, utilize zymotic fluidProduce and stablize composite bacteria agent capable, this microbial inoculum process aquaculture sewage fast, low cost, low environment pollute, and processes rear organic matter of sewageConcentration significantly reduces, and is conducive to the application in the sewage disposal of modern intensive livestock and poultry cultivation.
Brief description of the drawings
Fig. 1 treatment tank profile, wherein 1 is sludge outlet; 2 is treatment pond 1; 3 is grid; 4 is treatment pond 2; 5For delivery port.
Four, detailed description of the invention
Embodiment 1
Bacterial strain obtains
Produce the acquisition of biological flocculant bacterial strain
Gather the plant rhizosphere surrounding soil low temperature of in the exposed beach of tidal flat of Jiangsu Province, sporadicly growing and preserve, adoptGlutamic acid culture medium separating bio flocculant is γ-polyglutamic acid synthesized micro-organism bacterial strain. The standard of screening is that bacterium colony is in trainingSupport on base and form high protuberance, bulge has very strong viscosity and absorptive bacterial strain. Synthetic γ-the poly of screening liquid fermentationThe strongest bacterial strain of ability of glutamic acid, obtains bacterial strain through 16srDNA sequential evolution analysis, and qualification belongs to bacillus amyloliquefaciens(BacillusamyloliquefaciensIAE). Its synthesising biological flocculation dosage is 21.8gL-1, its biological property is:It is shaft-like that thalline is, even dyeing, tool motility, amphimicrobian, gemma ovalize, Gram-positive, bacterium on culture mediumThe opaque colony that falls to being white in color, edge is irregular, has protuberance, surface folding. Described bacillus amyloliquefaciens IAE(B.amyloliquefaciensIAE) be preserved in Chinese Typical Representative culture collection center, preservation address on March 15th, 2013For Luo Jia Shan, wuchang, wuhan, deposit number CCTCCNO:M2013086, referred to as bacillus amyloliquefaciens CCTCCNO:M2013086；
The formula of above-mentioned culture medium is to prepare according to preparation 1L amount: glucose 80g, sodium glutamate 50g, (NH4)2SO48g，NaCl5g，K2HPO4·3H2O2g，MnSO4·7H2O0.25g，MnSO4?H2O0.03g, distilled water 1000ml, pH valueNature, 115 DEG C of sterilizing 30min.
Generation cellulase strain obtains
Gather aeration poor soil, beach aleuritic texture soil, diving mud, the sample such as the fungal bacterial strain that has, lowTemperature is preserved, and adopts Congo red culture medium to separate fungi, and the larger bacterial strain of hydrolysis circle saves backup.
Above-mentioned Congo red culture medium compound method is (to prepare 1L culture medium as example): sodium carboxymethylcellulose 15g, albumenPeptone 5g, KH2PO41.0g，MgSO4·7H2O0.2g, NaCl5g, Congo red 0.2g, pH7.0, agar 20g, running water1000ml。
Produce cellulase strain and belong to trichoderma asperellum (Trichoderma.asperellum12), main biological characteristicsProperty be: on PDA culture medium 28 DEG C cultivate 3 days, mycelia can be covered with whole culture dish, mycelial growth initial stage white, felted, byCenter to edge forms ring-type gradually, and the ring-type that keeps to the side mycelia color is more shallow, produces dense on close centres circleDirty-green conidium, the culture dish of growth mycelia gives out very heavy Coconut Juice aromatic odor, back side buff; ConidiophoreTo growth, be arborization, dirty-green, bottle stalk is short, and base portion attenuates, and expand centre; Conidium is spherical in shape, rough surface,Size is 3.5～5.0 μ m × 2.0～5.0 μ m; This bacterial strain fermentation liquor cellulase activity is 25.2Uml-1; ITS sequence is evolvedAnalyze and show that this bacterial strain is trichoderma asperellum (T.asperellum). This bacterial strain is preserved in Chinese Typical Representative training on May 16th, 2013Support thing preservation center, preservation address is Luo Jia Shan, wuchang, wuhan, and deposit number CCTCCM2013213, referred to as trichoderma asperellumCCTCCNO：M2013213。
The bacillus thuringiensis bacterial strain of killing mosquitos and flies obtains
Have on the bacillus thuringiensis market of killing mosquitos and flies effect and had very ripe commercial product, bacterial strain is purchasedFrom Jiangsu Shengjiu Agrochemicals Co., Ltd., kill mosquitos and flies effect by checking,, kill for mosquito higher than 60% for fly killing rateThe rate of going out reaches 80%, can effectively suppress multiplying of mosquitos and flies in sewage.
The assay method that above-mentioned bacillus thuringiensis zymotic fluid is killed mosquitos and flies is: bacillus thuringiensis is inoculated into bacterial classificationCulture medium, cultivates, and its condition is: shaking table concussion 170rpm, 37 DEG C of cultivation temperature, cultivate CFU 24 hours≥1×109cfuml-1, bacterium liquid sparges in the wide-mouth bottle of cultivation fly and mosquito, measures compared to the processing of spray sterilized water, bacteriumThe death rate of mosquitos and flies is processed in liquid spraying.
The formula of above-mentioned fluid nutrient medium used is to prepare according to 1L culture medium: beef extract 3g, peptone 10g, NaCl10g, running water 1000ml, pH scope 7.0-7.5,121 DEG C of sterilizing 20min.
Microbial inoculum is produced
Bacillus amyloliquefaciens CCTCCNO:M2013086 fermenting and producing, by bacillus amyloliquefaciens CCTCCNO:M2013086 inoculation are to liquid seed culture medium, and the condition of cultivation is: shaking table concussion 170rpm, 37 DEG C of cultivation temperature, cultivate24 hours, CFU >=1 × 109cfuml-1; Liquid seed culture medium formula used is to cultivate basigamy according to 1LSystem: beef extract 3g, peptone 10g, NaCl10g, running water 1000ml, pH scope 7.0-7.5,121 DEG C of sterilizing 20min; To separateBacillus amyloliquefaciens CCTCCNO:M2013086 seed is inoculated into liquid fermentation medium according to 5% percent by volume, carries out liquidBody fermenting and producing, the condition of its fermenting and producing is: 28 ~ 35 DEG C of cultivation temperature, dissolved oxygen throughput scope is 30～100%, 150 ~220rpm, CFU >=1 × 10 of this bacterial strain of fermentation later stage fermentation liquid9cfuml-1, zymotic fluid stores for future use; UsedThe formula of liquid fermentation medium be to prepare according to 1L culture medium: glucose 80g, sodium glutamate 50g, (NH4)2SO48g，NaCl5g，K2HPO4·3H2O2g，MnSO4·7H2O0.25g，MnSO4?H2O0.03g, distilled water 1000ml, pH value is certainlySo, 115 DEG C of sterilizing 30min.
Trichoderma asperellum CCTCCNO:M2013213 fermenting and producing, trichoderma asperellum CCTCCNO:M2013213 is inoculated intoIn PDA nutrient solution, carry out liquid seeds cultivation, condition of culture is: shaking table concussion 120rpm, and 25 DEG C of cultivation temperature, cultivation 72 is littleTime, CFU >=0.5 × 108cfuml-1; The formula of the seed PDA culture medium of liquid used is to cultivate according to 1LBasigamy system: be cut into small pieces to be put in water after peeling with 200g potato and boil, boil 30min after boiling, with after filtered through gauze, filtrate adds20g common sucrose, is settled to 1000ml, pH value nature, 115 DEG C of sterilizing 30min; By trichoderma asperellum CCTCCNO:M2013213 bacterial classification are inoculated into liquid fermentation medium according to 10% percent by volume, carry out liquid fermentation production, and it ferments rawThe condition of producing is: initial pH nature, cultivate 19 ~ 22 DEG C of temperature, and dissolved oxygen throughput scope is 30～100%, 90 ~ 170rpm, fermentationMiddle and later periods forms mycelium pellet rotating speed 250rpm stirring and is broken into mycelia fragment, zymotic fluid trichoderma asperellum CCTCCNO:MCFU >=0.5 × 10 of 2013213 bacterial strains8cfuml-1, the storage of zymotic fluid low temperature is for subsequent use; Liquid fermentation training usedNutrient solution compound method is (to prepare 1L culture medium as example): after peeling with 100g potato, be cut into small pieces to be put in water and boil, after boilingBoil 30min, in filtrate, add after filtering 5g common sucrose and 2g rice straw powder, be settled to 1000ml, pH value nature, 115DEG C sterilizing 30min.
Bacillus thuringiensis zymotic fluid is produced, and bacillus thuringiensis is inoculated into liquid seed culture medium, condition of cultureFor: shaking table concussion 170rpm, 37 DEG C of cultivation temperature, cultivate CFU >=1 × 10 24 hours9cfuml-1; UsedThe formula of the seed culture medium of liquid is to prepare according to 1L culture medium: beef extract 3g, peptone 10g, NaCl10g, running water1000ml, pH scope 7.0-7.5,121 DEG C of sterilizing 20min; Bacillus thuringiensis seed is inoculated according to 5% percent by volumeTo liquid fermentation medium, carry out liquid fermentation production, the condition of its fermenting and producing is: initial pH scope is 7.0 ~ 7.5, cultivates25 ~ 33 DEG C of temperature, dissolved oxygen throughput scope is 30～100%, 150-220rpm, the bacterium colony shape of this bacterial strain of fermentation later stage fermentation liquidBecome unit >=1 × 109cfuml-1, zymotic fluid stores for future use. The formula of liquid fermentation medium used is to cultivate according to 1LBasigamy system: beef extract 3g, peptone 10g, NaCl10g, running water 1000ml, pH scope 7.0-7.5,121 DEG C of sterilizing 20min.
Processing aquaculture wastewater composite flora produces
By bacillus amyloliquefaciens CCTCCNO:M2013086, trichoderma asperellum CCTCCNO:M2013213 and Su YunjinFermentation of bacillus liquid is compound according to the volume ratio of 10:8:1, adopts calcium hydroxide to regulate between pH value to 8.0 ~ 8.5, with stirringMachine stirs and within 0.5 ~ 1 hour, allows microbial cells stable and enter resting state. Compound leaves standstill 2 hours at normal temperature, filling and sealing,Product is aquaculture wastewater and processes flora.
Process the application of aquaculture wastewater flora and compliance test result thereof
For trying aquaculture wastewater: 2000 cultivation scale pig farm sewage, daily blowdown water amount 50 ~ 60m3, external environment: temperature22~35℃。
Sewage disposal adopts the cement structures treatment tank of two series connection, and centre has grid to separate; Previous sewage placeReason pond is long × wide is 500cm × 100cm, bottom angled, and water inlet depth of water 200cm, delivery port depth of water 30cm, uncovered, gridThickness 15cm, the middle rice husk of filling, second treatment tank be long × wide × and be 500cm × 150cm × 200cm deeply, two stringsConnection treatment pond section is illustrated as Fig. 1.
In the sewage of first treatment tank, spray or spread fertilizer over the fields composite bacteria agent, dosage is that every day is sooner or later eachSpray 3L microbial inoculum, control treatment is not used composite bacteria agent, after 12 hours measuring 10cm deep water body and second before gridIndividual treatment tank delivery port water body COD, roundworm egg quantity and Escherichia coli quantity, adopt mosquitos and flies to paste paper and measure first dirtThe mosquitos and flies quantity that water treating pond unit are produces.
First treatment tank grid place COD of sewage concentration, roundworm egg quantity, Escherichia coli quantitative measurement result showAs table 1, application composite bacteria agent process water body COD concentration, roundworm egg quantity, Escherichia coli quantity respectively lower by 68% than contrast,81% and 67%. Processing delivery port sewage water body COD concentration, the Escherichia coli quantity of composite bacteria agent are lower by 72% than contrast respectivelyWith 82%, use composite bacteria agent processing and Escherichia coli (table 2) do not detected. Use composite bacteria agent processing, grid goes outWater is very fast, is not easy to stop up. Compared to control treatment, use composite bacteria agent processing, unit are mosquito and fly quantityDecline respectively 50% and 62%;
Table 1 treatment pond 1 sewage index
Table 2 treatment pond 2 sewage indexs
Claims (1)
1. a livestock and poultry cultivation sewage disposal composite bacteria agent, is characterized by composite bacteria agent and contains bacillus amyloliquefaciens(IAEBacillusamyloliquefaciensIAE), trichoderma asperellum 12 (Trichodermaasperellum12) andBacillus thuringiensis (Bacillusthuringiensis);
Wherein said bacillus amyloliquefaciens IAE (BacillusamyloliquefaciensIAE), in March, 2013Within 15th, be preserved in Chinese Typical Representative microorganism fungus kind preservation center, preservation address is Luo Jia Shan, wuchang, wuhan, deposit number CCTCCNO:M2013086, referred to as bacillus amyloliquefaciens CCTCCNO:M2013086;
Described trichoderma asperellum 12 (Trichodermaasperellum12), is preserved in Chinese allusion quotation on May 16th, 2013Type microorganism fungus kind preservation center, preservation address is Luo Jia Shan, wuchang, wuhan, deposit number CCTCCNO:M2013213, letterBe called trichoderma asperellum CCTCCNO:M2013213;
By described bacillus amyloliquefaciens CCTCCNO:M2013086, trichoderma asperellum CCTCCNO:M2013213 and Su YunGold fermentation of bacillus liquid is compound according to the volume ratio of 10:8:1;
Described zymotic fluid, prepares as follows:
1) bacillus amyloliquefaciens CCTCCNO:M2013086 inoculation is arrived to liquid seed culture medium, the condition of cultivationFor: shaking table concussion 170rpm, 37 DEG C of cultivation temperature, cultivate CFU >=1 × 10 24 hours9cfuml-1; UsedLiquid seed culture medium formula is to prepare according to 1L culture medium: beef extract 3g, peptone 10g, NaCl10g, running water1000ml, pH scope 7.0-7.5,121 DEG C of sterilizing 20min; Bacillus amyloliquefaciens CCTCCNO:M2013086 seed is pressedBe inoculated into liquid fermentation medium according to 5% percent by volume, carry out liquid fermentation production, the condition of its fermenting and producing is: cultivate temperatureSpend 28 ~ 35 DEG C, dissolved oxygen throughput scope is 30～100%, 150 ~ 220rpm, and the bacterium colony of this bacterial strain of fermentation later stage fermentation liquid formsUnit >=1 × 109cfuml-1, zymotic fluid low-temperature storage is for subsequent use; The formula of liquid fermentation medium used is: glucose80g, sodium glutamate 50g, (NH4)2SO48g，NaCl5g，K2HPO4·3H2O2g，MnSO4·7H2O0.25g，MnSO4?H2O0.03g, distilled water 1000ml, pH value nature, 115 DEG C of sterilizing 30min;
2) trichoderma asperellum CCTCCNO:M2013213 is inoculated in PDA nutrient solution, carries out liquid seeds cultivation, condition of cultureFor: shaking table concussion 120rpm, 25 DEG C of cultivation temperature, cultivate CFU >=0.5 × 10 72 hours8cfuml-1; UsedThe formula of seed PDA culture medium of liquid be to prepare according to 1L culture medium: after peeling with 200g potato, be cut into small pieces and be put intoIn water, boil, boil 30min after boiling, with after filtered through gauze, filtrate adds 20g common sucrose, is settled to 1000ml, pH value nature,115 DEG C of sterilizing 30min; Trichoderma asperellum CCTCCNO:M2013213 seed is inoculated into liquid according to 10% percent by volume to be sent outFerment culture medium, carries out liquid fermentation production, and the condition of its fermenting and producing is: initial pH nature, cultivate 19 ~ 22 DEG C of temperature, and dissolved oxygen is logicalTolerance scope is 30～100%, 90 ~ 170rpm, and the fermentation middle and later periods forms mycelium pellet rotating speed 250rpm stirring and is broken into myceliaFragment, CFU >=0.5 × 10 of fermentation trichoderma asperellum CCTCCNO:M2013213 bacterial strain8cfuml-1, fermentationThe storage of liquid low temperature is for subsequent use; Liquid fermentation and culture liquid making method used is: after peeling with 100g potato, be cut into small pieces and be put in waterBoil, after boiling, boil 30min, in filtrate, add after filtering 5g common sucrose and 2g rice straw powder, be settled to 1000ml, pH valueNature, 115 DEG C of sterilizing 30min;
3) bacillus thuringiensis is inoculated into liquid seed culture medium, and condition of culture is: shaking table concussion 170rpm, cultivation temperature 37DEG C, cultivate CFU >=1 × 10 24 hours9cfuml-1; The formula of the seed culture medium of liquid used is, pressesPrepare according to 1L culture medium: beef extract 3g, peptone 10g, NaCl10g, running water 1000ml, pH scope 7.0-7.5,121 DEG C go outBacterium 20min; Bacillus thuringiensis seed is inoculated into liquid fermentation medium according to 5% percent by volume, carries out liquid fermentationProduce, the condition of its fermenting and producing is: initial pH scope is 7.0 ~ 7.5,25 ~ 33 DEG C of cultivation temperature, and dissolved oxygen throughput scope is30～100%, 150-220rpm, CFU >=1 × 10 of this bacterial strain of fermentation later stage fermentation liquid9cfuml-1, fermentationLiquid stores for future use; The formula of liquid fermentation medium used is to prepare according to 1L culture medium: beef extract 3g, and peptone 10g,NaCl10g, running water 1000ml, pH scope 7.0-7.5,121 DEG C of sterilizing 20min;
4) by microbial fermentation solution bacterial classification according to bacillus amyloliquefaciens CCTCCNO:M2013086: trichoderma asperellum CCTCCNO:M2013213: bacillus thuringiensis=10:8:1 volume ratio is compound, regulating pH value with calcium hydroxide is 8.0 ~ 8.5, afterContinue mixer stirring and within 0.5 ~ 1 hour, allow microbial cells stablize and enter resting state, compound leaves standstill 2 hours at normal temperature,Filling, be a kind of livestock and poultry cultivation sewage disposal composite bacteria agent.
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| CN104450530B (en) * | 2014-10-24 | 2017-06-23 | 浙江省农业科学院 | One plant of Trichoderma asperellum strain, microbial inoculum and its application in organic garbage treatment |
| CN104671432B (en) * | 2015-01-27 | 2016-08-31 | 河海大学 | A kind of water body purification of aquaculture agent and preparation method thereof |
| CN106190906A (en) * | 2016-07-19 | 2016-12-07 | 贵州欧瑞信环保科技有限责任公司 | A kind of microorganism fly eradication complexing agent |
| CN106085923B (en) * | 2016-08-22 | 2019-09-06 | 安徽农业大学 | A kind of preparation method and application of bacillus amyloliquefaciens and its biological flocculant |
| CN106479921A (en) * | 2016-10-19 | 2017-03-08 | 中国科学院海洋研究所 | The bacillus amyloliquefaciens of high-efficient purification aquaculture wastewater and its application |
| CN106800330A (en) * | 2017-02-14 | 2017-06-06 | 厦门大学 | A kind of method for processing refuse leachate |
| CN107964521B (en) * | 2017-12-28 | 2020-10-30 | 江苏世邦生物工程科技有限公司 | Composite bacterium culture composition resistant to heavy metal ion toxicity and preparation method and application thereof |
| CN108517304A (en) * | 2018-03-27 | 2018-09-11 | 丽水学院 | A kind of composite microbial Tetramune, preparation method and its application in sewage disposal |
| CN112273187A (en) * | 2020-10-09 | 2021-01-29 | 河海大学 | Municipal sludge-based sponge urban water storage matrix and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN102433284A (en) * | 2011-12-21 | 2012-05-02 | 浙江理工大学 | Preparation method of dedicated microbial flocculant for dairy cattle breeding waste water and strain used in method |
-
2013
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102433284A (en) * | 2011-12-21 | 2012-05-02 | 浙江理工大学 | Preparation method of dedicated microbial flocculant for dairy cattle breeding waste water and strain used in method |
Non-Patent Citations (1)
| Title |
|---|
| 解淀粉芽孢杆菌对水中丁草胺的降解及影响;谭文捷等;《环境科学研究》;20051231;第18卷(第3期);71-74 * |
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