CN109455828A - A kind of application method of immobilized microorganism in livestock breeding wastewater processing - Google Patents
A kind of application method of immobilized microorganism in livestock breeding wastewater processing Download PDFInfo
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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Abstract
The invention discloses a kind of application method of immobilized microorganism in livestock breeding wastewater processing, and techniqueflow includes: the preparation of (1) microbial fermentation solution；(2) preparation of immobilized microorganism；(3) immobilized microorganism Fiber differentiation；(4) domestication culture；(5) cultivating wastewater purification.The present invention constructs multiple-microorganism syntaxial system, these microorganisms can act synergistically, organic matter, nitrogen, phosphorus and the heavy metal contaminants in livestock breeding wastewater of effectively degrading；Microorganism is fixed using compound embedded material, it forms unique microenvironment and good shielding action is generated to external world's infringement, immobilized microorganism is further increased to the repellence and tolerance of breeding wastewater by Fiber differentiation, domestication culture, applied to waste water treatment system, service life internal stability is higher and has good degradation effect to livestock breeding wastewater, lays a good foundation for the practical application of immobilized microorganism in the treatment of waste water.
Description
Technical field
The invention belongs to environment-protection wastewater processing technology fields, and in particular to a kind of immobilized microorganism is in livestock breeding wastewater
Application method in processing.
Background technique
In recent years, with the development of economy with the raising of living standards of the people, people are to animal products such as meat, egg, milk etc.
Demand gradually increase so that the scale in China, intensive livestock and poultry breeding industry flourish, the Production of Livestock and Poultry in China is horizontal
Also large increase has been obtained, but has also resulted in serious problem of environmental pollution simultaneously.It shows according to investigations, livestock and poultry breeding industry discharge
COD (COD) reaches 1268.26 ten thousand tons, and the 41.9% of Zhan Quanguo COD total emission volumn, the discharge amount difference of nitrogen and phosphorus
For 102.48 ten thousand tons and 16.04 ten thousand tons, the 21.7% and 37.9% of national nitrogen and phosphorus total emission volumn is accounted for respectively, Pollution from livestock and poultry is
As the third-largest pollution sources after industrial pollution, living garbage pollution, solve the problems, such as that Pollution from livestock and poultry is very urgent.
Livestock and poultry cultivation mainly includes pig, ox, sheep, horse, chicken, duck, goose etc., and livestock breeding wastewater is mainly by feces of livestock and poultry, urine
Liquid, column home flushing water and part sanitary sewage composition, belong to high organic concentration, high the content of nitrogen and phosphorous and high harmful microorganism number
The " three high " waste water of amount.And highdensity breeding environment provides condition for the generation of infectious disease and prevalence, once epidemic disease invades,
Will spread in china, cause huge economic loss.Therefore, the heavy metal elements such as copper, zinc and antibiotic are also widely used for raiseeing
Fowl cultivation, although having been played in terms of preventing and treating disease, promoting growth of animal and raising important
Effect, but also many negative effects are brought to environment.Since utilization rate is lower in livestock and poultry body for heavy metal element and antibiotic,
Major part can be discharged into water environment with original shape or its metabolite form with Animal fecal pollution, be caused in animal farm wastewater except normal
It advises outside pollutant, also containing multiple pollutants such as heavy metal, antibiotic, surrounding water, atmosphere, soil is caused seriously to pollute,
And jeopardize human health.After livestock breeding wastewater enters water body, make water eutrophication, breed a large amount of pathogenic microorganism,
The toxic gases such as ammonia, hydrogen sulfide can also be generated, not only make water body black foul, can also induce the various respiratory diseases of human body and
Inflammation；Into after soil, waste water through decompose can generate nitrites, may cause interstices of soil blocking, cause Soil ventilation,
Water penetration decline and hardened, salinization, seriously affect soil quality and crop growth, the heavy metal element contained in waste water, disease
The meeting contaminated soil crops such as pathogenic microorganism, helminth and worm's ovum, and human body is entered by food chain, cause disease.
Currently, the processing mode of livestock breeding wastewater mainly includes ecology processing and industrial treatment.Ecology processing
Mode refers to using the microorganism in natural water body and soil the method for handling waste water, mainly there is oxidation pond and artificial swamp
Deng.Oxidation pond is mainly used to reduce the pollutant load of water body, while improving the content of dissolved oxygen, and suitably remove the nitrogen in water
And phosphorus, mitigate the degree of water eutrophication；Artificial swamp can by precipitating, absorption, barrier, microbial assimilation decompose, nitrification,
The approach such as denitrification and plant absorption remove the impurity such as suspended matter, organic matter, the heavy metal in waste water.Ecology processing side
The advantages of formula, is that up-front investment is small, easy to operate, operating cost is low, the disadvantage is that take up a large area, treatment effect by season,
Managing environment influences significantly, and the residence time is long, and if dealt with improperly, will result in the release of pollutant and form new dirt
Dye.Industrial treatment mode refers to the growing environment for remaining suitable by manually adjusting operating parameter, so that the micro- life of sewage treatment
The breeding of object raised growth reduces hydraulic detention time to improve treatment effect, is increasingly becoming the master of livestock breeding wastewater processing
It selects.The industrialized treatment system of livestock breeding wastewater includes system for anaerobic treatment, intermission aerobic processing system, Aerobic Process for Treatment
System is these three types of, but is individually often unable to reach preferable treatment effect using a certain processing system, and uses anaerobism-good
Oxygen combined treatment process is most economical, efficient method generally acknowledged at present.
Immobilized microorganism technique is an emerging technology in bioengineering field, which is by chemically or physically
Free microorganism is fixed on carrier, keeps the activity of microorganism, make microorganism under optimum conditions can be fast by means
Speed, a large amount of proliferation.In the early 1980s, having started to handle industrial wastewater about using immobilized microorganism technique both at home and abroad
With the research of decomposing organic pollutant, and make remarkable progress.Compared with free microorganism handles waste water, immobilized microorganism
Processing has better advantage, and immobilization technology first not easily runs off microorganism, and reaction is easier to control, additionally it is possible to significantly
Improve the microorganism concn for participating in reaction；Secondly immobilized microorganism and reactant long action time, while fixation support pair
External world's infringement generates good shielding action, can improve the tolerance of microorganism；Third, which is separated by solid-liquid separation, to be easy, and secondary pollution is few,
It can repeatedly be utilized.
Livestock breeding wastewater is a sufficiently complex mixed system, and people are main in large-scale cultivation wastewater treatment at present
Consider cutting to conventional pollution index such as COD (COD), biochemical oxygen demand (BOD) (BOD), total nitrogen (TN) and total phosphorus (TP) etc.
Subtract and control, and seldom pays close attention to the removal of other pollutants such as heavy metal, antibiotic in breeding wastewater.Existing about work
Industryization is handled in the report of breeding wastewater, and most of organic matter and suspended matter can be removed after anaerobic system is handled, but good
There is total nitrogen and total tp removal rate are not high enough in oxygen treatment process, the undesirable problem for the treatment of effect.In addition, to realize fixation
The scale for changing microbiological treatment livestock breeding wastewater also needs to consider that the stability of fixation support, mass-transfer performance, machinery are strong
The factors such as degree, service life, cost.
Summary of the invention
The problem of for existing livestock breeding wastewater processing technique, the present invention provides a kind of immobilized microorganisms
Application method in livestock breeding wastewater processing, can not only improve useless Organic substance in water, total nitrogen, total phosphorus and heavy metal pollution
The degradation rate of object, moreover it is possible to keep the efficient degradation effect of microorganism in a long time, there is very high application value.
Technical solution of the present invention is summarized as follows:
A kind of application method of immobilized microorganism in livestock breeding wastewater processing, which is characterized in that the technical flow of this method
Journey is as follows:
(1) preparation of microbial fermentation solution: by Bacillus foecalis alkaligenes, Pseudomonas stutzeri, bacillus coagulans, skin spy's acinetobacter calcoaceticus
It is mixed with the seed liquor of penicillium chrysogenum according to the ratio of 3:3:2:2:5, then seed mixture liquid is accessed in fermentation medium, is connect
Kind amount is 10-15%, and fermentor parameter is set as coefficient 50-60%, ventilatory capacity 0.8-1.2VVM, revolving speed 160-200r/
Min, 28-32 DEG C of temperature, fermented and cultured 60-72h；
(2) preparation of immobilized microorganism: the centrifuge centrifugation 15min that microbial fermentation solution is 7500r/min with revolving speed is collected
Thallus is resuspended in distilled water again, is then sufficiently mixed with compound embedded material with the volume ratio of 1:20, is slowly dropped into saturation boron
It is crosslinked in acid and 3% calcium chloride solution, forms immobilized microorganism gel ball, be put into 4 DEG C of refrigerators and fix for 24 hours, take out gel ball
After being cleaned with 9%NaCl solution, then be washed with distilled water 2-3 times it is spare；
(3) immobilized microorganism Fiber differentiation: the immobilized microorganism gel ball prepared is fitted into three-neck flask, addition accounts for
The induced medium of flask volume 40%, be passed through purification air cultivate 48h under conditions of 28-32 DEG C of temperature, in the training period between
Having a rest property is passed through high-purity hydrogen into flask；
(4) domestication culture: being added breeding wastewater into domestication reactor, and is added solidifying by the immobilized microorganism of Fiber differentiation
Glueballs, domestication reactor aeration quantity are set as keeping content of oxygen dissolved in water in 2.5-3.5mg/L, and the domestication time is 36-48h,
Acclimation temperature is 28-32 DEG C；
(5) cultivating wastewater purification: by the reaction of the immobilized microorganism gel ball tamed investment SBR intermission aerobic processing system
In device, input amount is the 20% of reactor effective volume, and temperature setting is 25-35 DEG C when operation, and the cycle of operation is set as 12h, fortune
Line program is set as standing 3h into water 30min, aeration reaction 8h, is discharged 30min, and to treated, breeding wastewater carries out correlation
Index determining.
In the step (1), Bacillus foecalis alkaligenes, Pseudomonas stutzeri and Pi Te acinetobacter calcoaceticus seed liquor cultural method
Are as follows: the strain of these three microorganisms is inoculated on nutrient agar using spread plate, 30 DEG C of constant temperature incubation 48-
72h, picking well-grown single colonie on plate are inoculated into nutrient broth medium under aseptic technique, in 30 DEG C,
160r/min shaking table shaken cultivation 48h.
In the step (1), the cultural method of bacillus coagulans seed liquor are as follows: gemma will be condensed using spread plate
The strain of bacillus is inoculated on LB solid medium, 35 DEG C of constant temperature incubation 48-72h, picking well-grown single bacterium on plate
Fall, be inoculated into LB liquid medium under aseptic technique, in 35 DEG C, 180r/min shaking table shaken cultivation for 24 hours.
In the step (1), the cultural method of penicillium chrysogenum seed liquor are as follows: using spread plate by the bacterium of penicillium chrysogenum
Kind is inoculated in PDA culture medium, 28 DEG C of constant temperature incubation 4-6d, washes lower spore with sterile saline, shaken well forms spore
Suspension accesses in PDB culture medium according to 5% inoculum concentration, in 28 DEG C, 150r/min shaking table shaken cultivation 72h.
In the step (1), the ingredient of fermentation medium are as follows: wheat bran 24-30g/L, molasses 11-15g/L, yeast powder 4-
6g/L, (NH4)2SO43-5g/L, NaCl 8.5-9.3g/L, KNO34.3-4.7g/L, K2HPO4 0.4-0.6g/L, KH2PO4
1.5-1.9g/L, MgSO4·7H2O1.2-1.4g/L, CaCl20.4-0.5g/L, CoCl2·6H2O 3.5-4.0mg/L,
FeSO4·7H2O 5.2-5.6mg/L, MnSO4·H2O 1.6-2.2mg/L, surplus are water, and pH value is natural.
In the step (2), the ingredient of embedded material is polyvinyl alcohol 4%, sodium alginate 3%, diatomite 0.2%, Fe3O4
Nano particle 0.3%, active carbon 0.2%.
In the step (3), the ingredient of induced medium are as follows: sucrose 12-16g/L, bean cake powder 5.1-5.5g/L, urea
3.3-3.8g/L NaCl 3.6-4.0g/L, KH2PO4 1.3-1.7g/L, MgSO4·7H2O0.7-0.8g/L, glutathione 14-
18mg/L, sodium citrate 6.5-7.5mg/L, sodium nitroprussiate 48-52 μm of ol/L, glycine betaine 1.6-2.0mg/L, fulvic acid 0.24-
0.28g/L, health polyphenol 0.1-0.2mg/L, pH value are natural.
In the step (3), interval ventilation refer to culture 12h, for 24 hours, 36h be passed through high-purity hydrogen, ventilatory capacity is
Full of entire three-neck flask.
In the step (4), domestication reactor should set up feed system, aerating system and temperature control system, the fixation of addition
The volume ratio for changing microorganism gel ball and breeding wastewater is 1:3-1:5.
The beneficial effect that the present invention generates mainly includes the following aspects:
1) present invention constructs multiple-microorganism syntaxial system according to the difference of different microorganisms degradation efficiency and substrate of degrading,
Bacillus foecalis alkaligenes, Pseudomonas stutzeri, bacillus coagulans can effectively in degrading waste water organic matter and nitrogen-containing pollutant, skin
Special acinetobacter calcoaceticus can contain phosphor contaminant effectively in degrading waste water, and penicillium chrysogenum inherently has tolerance to heavy metal ion
And repellence, and the heavy metal in dissolution waste water can be adsorbed, these microorganisms can share out the work and help one another, mutualism, thus
Realize effective degradation to livestock breeding wastewater.
2) present invention is fixed microorganism using compound embedded material, polyvinyl alcohol and sodium alginate as carrier,
Diatomite, bean cake powder, Fe3O4Nano particle, active carbon, as crosslinking agent, are suitably being matched as additive, boric acid and calcium chloride
Immobilized microorganism gel ball is prepared than under, is had the advantages that performance is stable, intensity is high, the service life is long, resistance to mass tranfer is small, is formed
Microenvironment to the external world infringement have good shielding action, microorganism can be improved to the tolerance of environment.
3) of the invention that immobilized microorganism is further increased to feeding by immobilized microorganism Fiber differentiation and domestication culture
The repellence and tolerance of waste water are grown, the Fiber differentiation stage is passed through hydrogen and combines specific induced medium, can be enhanced
Microorganism oxidation resistance, improves reverse resistance of microorganisms, and domestication cultivation stage makes microorganism gradually adapt to wastewater environment, micro- life
Object growth reaches preferable state, so that microorganism be made to have better degradation property.
4) present invention puts into immobilized microorganism in the reactor of SBR intermission aerobic processing system, substantially increases poultry
Organic matter in fowl breeding wastewater, total nitrogen, total phosphorus and heavy metal contaminants degradation rate, reached good wastewater degradation effect
Fruit.Immobilized microorganism is higher in service life internal stability, can also repeatedly utilize, so that treatment effeciency is improved, at reduction
Manage cost.
Detailed description of the invention
Influence of Fig. 1 immobilized microorganism input amount to wastewater degradation effect；
Influence of Fig. 2 temperature to wastewater degradation effect；
Influence of Fig. 3 cycle of operation to wastewater degradation effect.
Specific embodiment
Application method of the immobilized microorganism of the present invention in livestock breeding wastewater processing is described with reference to embodiments, is answered
Should it is realized that, these description merely to explain the present invention, its content is not defined.
Strain used in following embodiment: Bacillus foecalis alkaligenes (Alcaligenes faecalis), deposit number
BNCC189345, Pseudomonas stutzeri (Pseudomonas stutzeri), deposit number BNCC209988 condenses gemma bar
Bacterium (Bacillus coagulans), deposit number BNCC220722 is purchased from Su Zhoubei Na Chuanlian Bioisystech Co., Ltd；
Skin spy acinetobacter calcoaceticus (Acinetobacter pittii), deposit number CICC10526, penicillium chrysogenum (Penicillium chrysogenum), deposit number CICC40209 is purchased from Chinese industrial Microbiological Culture Collection administrative center.
Embodiment 1
(1) preparation of seed liquor: using spread plate by Bacillus foecalis alkaligenes, the bacterium of Pseudomonas stutzeri and Pi Te acinetobacter calcoaceticus
Kind is inoculated on nutrient agar, 30 DEG C of constant temperature incubation 48-72h, and picking well-grown single colonie on plate is sterile
It is inoculated into nutrient broth medium under operating condition, in 30 DEG C, 160r/min shaking table shaken cultivation 48h；Bud will be condensed with method
The strain of spore bacillus is inoculated on LB solid medium, 35 DEG C of constant temperature incubation 48-72h, picking well-grown list on plate
Bacterium colony is inoculated into LB liquid medium under aseptic technique, in 35 DEG C, 180r/min shaking table shaken cultivation for 24 hours；
The strain of penicillium chrysogenum is inoculated in PDA culture medium with method, 28 DEG C of constant temperature incubation 4-6d are washed down with sterile saline
Spore, shaken well form spore suspension, according in 5% inoculum concentration access PDB culture medium, shake in 28 DEG C, 150r/min
Bed shaken cultivation 72h.
(2) preparation of microbial fermentation solution: Bacillus foecalis alkaligenes, Pseudomonas stutzeri, bacillus coagulans, Pi Te is motionless
The seed liquor of bacillus and penicillium chrysogenum is mixed according to the ratio of 3:3:2:2:5, and seed mixture liquid is then accessed fermentation medium
In (the ingredient of fermentation medium are as follows: wheat bran 26g/L, molasses 14g/L, yeast powder 5g/L, (NH4)2SO44g/L, NaCl 8.9g/
L, KNO34.5g/L, K2HPO4 0.5g/L, KH2PO4 1.7g/L, MgSO4·7H2O1.3g/L, CaCl20.45g/L, CoCl2·
6H2O 3.75mg/L, FeSO4·7H2O 5.4mg/L, MnSO4·H2O 1.9mg/L, surplus are water, and pH value is natural), inoculum concentration
It is 12%, fermentor parameter is set as coefficient 55%, ventilatory capacity 1.0VVM, revolving speed 180r/min, and 30 DEG C of temperature, fermented and cultured
72h。
Influence of 2 embedded material of embodiment to immobilized microorganism gel ball performance
The preparation of immobilized microorganism: microbial fermentation solution is centrifuged with the centrifuge that revolving speed is 7500r/min in embodiment 1
15min collects thallus and is resuspended in distilled water again, is then sufficiently mixed with compound embedded material with the volume ratio of 1:20, slowly drips
Enter to be saturated and be crosslinked in boric acid and 3% calcium chloride solution, form immobilized microorganism gel ball, is put into 4 DEG C of refrigerators and fixes for 24 hours, take
After gel ball is cleaned with 9%NaCl solution out, then it is washed with distilled water 2-3 times.
Mass-transfer performance measurement: the basal salt media of 100mL is added into 250mL triangular flask, immobilized microorganism is coagulated
Glueballs is added thereto, and adds 4-5 drop red ink, is taken out, is rinsed well with clear water, and observe gel ball section every a period of time
Color change interval.The range that mass-transfer performance is changed colour by gel ball reflects that, closer to center, mass-transfer performance is better indirectly.
Mechanical strength measurement: the basal salt media of 20mL is added into 50mL triangular flask, 100 gel balls are put into base
In plinth salt culture medium, and 20 beades are added, take out gel ball after 30 DEG C, 180r/min shaking table shaken cultivation, 7d, observes
Breakage counts breakage rate, to judge the mechanical strength of gel ball.
1, polyvinyl alcohol and sodium alginate with the influence for comparing gel ball mass-transfer performance and mechanical strength
The total concentration of polyvinyl alcohol and sodium alginate is 7% constant in compound embedded material, sets 2:5,3 for the proportion of the two:
4,4:3,5:2,6:1, and diatomite 0.2%, Fe are added respectively3O4Nano particle 0.3%, active carbon 0.2% are observed and measure gel
Ball mass-transfer performance and mechanical strength, the results are shown in Table 1.
As known from Table 1, when the proportion of polyvinyl alcohol and sodium alginate is 4:3, i.e., polyvinyl alcohol concentration is 4%, alginic acid
When na concn is 3%, the mass-transfer performance of gel ball is preferable, and breakage rate is relatively low, i.e., mechanical strength is preferable.Polyvinyl alcohol and sea
Mosanom is the main embedded material of immobilization technology, and polyvinyl alcohol is organic synthesis high molecular material, is had preferable mechanical
Intensity, but mass-transfer performance is weaker；Sodium alginate is natural macromolecular material, has preferable mass-transfer performance, but mechanical strength is not
It is good, it is easy to be degraded by microorganisms, therefore suitable proportion is selected to mix polyvinyl alcohol and sodium alginate, as the micro- life of immobilization
The occlusion vehicle of object has more preferable effect.
2, influence of the additive to gel ball mass-transfer performance and mechanical strength in compound embedded material
One group of compound embedded material is polyvinyl alcohol 4%, and sodium alginate 3% is free of additive；Another group is polyvinyl alcohol 4%, sea
Mosanom 3%, diatomite 0.2%, Fe3O4Nano particle 0.3%, active carbon 0.2% are observed and measure gel ball mass-transfer performance and machine
Tool intensity, the results are shown in Table 2.
As known from Table 2, the gel ball containing additive is more preferable than the mass-transfer performance without additive, and the breakage rate of the two
It is not significantly different.Diatomite, Fe3O4Nano particle and active carbon are all porous materials, have biggish specific surface area, and have
Very strong adsorption capacity, adding suitable amount can be improved mass-transfer performance, while not influence mechanical strength again.
Embodiment 3
Immobilized microorganism gel ball, technical solution according to the invention are prepared using best complex embedded material in embodiment 2
Lab scale processing is carried out to the waste water that Jiangsu Province Lianyungang livestock breeding wastewater processing center is collected, which has been subjected to early period
Precipitating and Anaerobic Treatment, test the following steps are included:
(1) immobilized microorganism Fiber differentiation: the immobilized microorganism gel ball prepared is fitted into three-neck flask, addition accounts for
Induced medium (the ingredient are as follows: sucrose 14g/L, peptone 5.3g/L, urea 3.6g/L, NaCl 3.8g/ of flask volume 40%
L, KH2PO4 1.5g/L, MgSO4·7H2O0.75g/L, glutathione 16mg/L, sodium citrate 7.0mg/L, 50 μ of sodium nitroprussiate
Mol/L, glycine betaine 1.8mg/L, fulvic acid 0.26g/L, health polyphenol 0.15mg/L, pH value are natural), purification air is passed through in temperature
Cultivate 48h under conditions of 30 DEG C, culture 12h, for 24 hours, 36h be passed through high-purity hydrogen, ventilatory capacity is to burn full of entire three neck
Bottle.
(2) domestication culture: being added a certain amount of waste water into domestication reactor, domestication reactor should set up feed system,
Aerating system and temperature control system, and the immobilized microorganism gel ball by Fiber differentiation, the immobilized microorganism of addition is added
The volume ratio of gel ball and breeding wastewater is 1:4, and domestication reactor aeration quantity is set as that content of oxygen dissolved in water is kept to exist
3.0mg/L, domestication time are 42h, and acclimation temperature is 30 DEG C.
(3) cultivating wastewater purification: by the immobilized microorganism gel ball tamed investment SBR intermission aerobic processing system
In reactor, reactor effective volume is 300L, and input amount is 20% i.e. 60L, and hydraulic detention time is 3 days, and temperature is set when operation
It is set to 25-35 DEG C, the cycle of operation is set as 12h, and operation program is set as standing 3h, water outlet into water 30min, aeration reaction 8h
30min；Index of correlation measurement is carried out to water inlet waste water before operation, measures COD 1579mg/L, TN 512mg/L, TP
136mg/L, Cu2+9.78mg/L Zn2+ 5.30mg/L carries out same index determining to water outlet after operation.
COD (COD) measurement: rapid-digestion spectrophotometry；Total nitrogen (TN) concentration mensuration: salicylic acid is divided light
Degree method；Total phosphorus (TP) concentration mensuration: ammonium molybdate spectrophotometric method；Cu2+Concentration mensuration: atomic absorption spectrophotometry.
Influence of 4 Fiber differentiation of embodiment to wastewater degradation effect
The present embodiment carries out three groups of tests, and first group is immobilized microorganism without Fiber differentiation, i.e. cancellation 3 step of embodiment
(1), other same as Example 3；Second group is not passed through high-purity hydrogen when being Fiber differentiation, other same as Example 3；Third
Group only adds sucrose 14g/L, peptone 5.3g/L, urea 3.6g/L, NaCl 3.8g/L for induced medium to be substituted for,
KH2PO4 1.5g/L, MgSO4·7H2The basal medium of O0.75g/L, it is other same as Example 3；Respectively to going out at every group
Water carries out index of correlation measurement, calculates COD, TN, TP, Cu2+Degradation rate, the results are shown in Table 3.
The data of analytical table 3, embodiment 3 is to use immobilized microorganism method for inducing and cultivating of the invention, each in waste water
The degradation rate of item index reaches highest, illustrates best to the degradation effect of waste water.Fiber differentiation of the invention is using intermittent
The method for being passed through high-purity hydrogen, hydrogen are a kind of gaseous signal molecules, have various biological effect, can participate in micro- life
The growth metabolism process of object improves microorganism to the tolerance of biotic and abiotic stress；Induced medium of the invention
Sodium nitroprussiate is added to as nitric oxide donors, nitric oxide is also a kind of gaseous signal molecule, there is various biological effect,
The nitric oxide of low concentration, which resists extraneous adverse circumstances, promotion microorganism growth metabolism etc. to microorganism, important work
With, and glutathione, sodium citrate, glycine betaine, fulvic acid, health polyphenol all have enhancing microorganism inoxidizability, improve micro- life
The effect of object resistance.Therefore immobilized microorganism significantly improves the repellence and tolerance of waste water after Fiber differentiation,
Better degradation property can be played.
Influence of the domestication culture of embodiment 5 to wastewater degradation effect
The present embodiment cancels 3 step of embodiment (2), will direct plunge into SBR by the immobilized microorganism gel ball of Fiber differentiation
It is other same as Example 3 in the reactor of intermission aerobic processing system, index of correlation measurement is carried out to water outlet, is calculated
COD、TN、TP、Cu2+Degradation rate, the results are shown in Table 4.
The data of analytical table 4, compared with without domestication culture, immobilized microorganism refers to items in waste water after domestication culture
Target degradation rate is obviously improved, and illustrates that domestication culture can generate beneficial effect to the degradation of waste water.Domestication culture can make
Microorganism gradually adapts to wastewater environment, and microorganism growth is made to reach preferable state, so that immobilized microorganism be made to play
Better degradation property out.
Influence of the 6 immobilized microorganism input amount of embodiment to wastewater degradation effect
The present embodiment sets 10% for the input amount of immobilized microorganism gel ball in 3 step of embodiment (3), 15%, 20%,
25%, 30%, it is other same as Example 3, index of correlation measurement is carried out to water outlet respectively, calculates COD, TN, TP, Cu2+Drop
Solution rate, as a result as shown in Figure 1.
From fig. 1, it can be seen that when immobilized microorganism input amount is gradually increased, COD, TN, TP, Cu in waste water2+And Zn2+'s
Degradation rate also increases accordingly, and has reached higher level when input amount is 20%, continues to increase input amount, items refer in waste water
Target degradation rate does not significantly improve, and from cost consideration, selects 20% immobilized microorganism input amount.
Influence of 7 temperature of embodiment to wastewater degradation effect
The present embodiment is by temperature setting is 20 DEG C, 25 DEG C, 30 DEG C, 35 DEG C, 40 DEG C, Qi Tayu when operation in 3 step of embodiment (3)
Embodiment 3 is identical, carries out index of correlation measurement to water outlet respectively, calculates COD, TN, TP, Cu2+Degradation rate, as a result such as Fig. 2
It is shown.
As can be seen from Figure 2, within the scope of 25-35 DEG C, COD, TN, TP, Cu in waste water2+Degradation rate all in higher level,
In the degradation rate highest of 30 DEG C of indices, wastewater degradation effect is best.Temperature is the important Survival Factor of microorganism, temperature
Variation will affect growth and the metabolic rate of microorganism, and fixation support can provide metastable environment to microorganism,
Influence it by temperature change smaller, so that the ability for changing immobilized microorganism adaptive temperature is stronger.
Influence of 8 cycle of operation of embodiment to wastewater degradation effect
Immobilized microorganism gel ball is put into the anti-of SBR intermission aerobic processing system according to the method for embodiment 3 by the present embodiment
It answers in device, 60 periods of continuous operation, index of correlation is carried out to water outlet respectively in the 12nd, 24,36,48,60 period of operation
Measurement, calculates COD, TN, TP, Cu2+Degradation rate, as a result as shown in Figure 3.
As can be seen from Figure 3, in 60 periods of continuous operation, COD, TN, TP, Cu in waste water2+Degradation rate variation do not show
It writes, all in higher level, wastewater degradation effect is preferable, illustrates that immobilized microorganism is preferable in service life internal stability, energy
Keep preferable degradation property.
The above embodiment of the present invention scheme is only the description of the invention and cannot limit the present invention, is pointed out in claim
Product of the present invention constituent, component ratio, the range of preparation method parameter, and above-mentioned explanation does not point out ginseng of the present invention
Therefore several ranges any change within the meaning and scope equivalent to the claims of the present invention, is all considered to be
It is included within the scope of the claims.
Claims (9)
1. a kind of application method of immobilized microorganism in livestock breeding wastewater processing, which is characterized in that the technology of this method
Process is as follows:
(1) preparation of microbial fermentation solution: by Bacillus foecalis alkaligenes, Pseudomonas stutzeri, bacillus coagulans, skin spy's acinetobacter calcoaceticus
It is mixed with the seed liquor of penicillium chrysogenum according to the ratio of 3:3:2:2:5, then seed mixture liquid is accessed in fermentation medium, is connect
Kind amount is 10-15%, and fermentor parameter is set as coefficient 50-60%, ventilatory capacity 0.8-1.2VVM, revolving speed 160-200r/
Min, 28-32 DEG C of temperature, fermented and cultured 60-72h；
(2) preparation of immobilized microorganism: the centrifuge centrifugation 15min that microbial fermentation solution is 7500r/min with revolving speed is collected
Thallus is resuspended in distilled water again, is then sufficiently mixed with compound embedded material with the volume ratio of 1:20, is slowly dropped into saturation boron
It is crosslinked in acid and 3% calcium chloride solution, forms immobilized microorganism gel ball, be put into 4 DEG C of refrigerators and fix for 24 hours, take out gel ball
After being cleaned with 9%NaCl solution, then be washed with distilled water 2-3 times it is spare；
(3) immobilized microorganism Fiber differentiation: the immobilized microorganism gel ball prepared is fitted into three-neck flask, addition accounts for
The induced medium of flask volume 40%, be passed through purification air cultivate 48h under conditions of 28-32 DEG C of temperature, in the training period between
Having a rest property is passed through high-purity hydrogen into flask；
(4) domestication culture: being added breeding wastewater into domestication reactor, and is added solidifying by the immobilized microorganism of Fiber differentiation
Glueballs, domestication reactor aeration quantity are set as keeping content of oxygen dissolved in water in 2.5-3.5mg/L, and the domestication time is 36-48h,
Acclimation temperature is 28-32 DEG C；
(5) cultivating wastewater purification: by the reaction of the immobilized microorganism gel ball tamed investment SBR intermission aerobic processing system
In device, input amount is the 20% of reactor effective volume, and temperature setting is 25-35 DEG C when operation, and the cycle of operation is set as 12h, fortune
Line program is set as standing 3h into water 30min, aeration reaction 8h, is discharged 30min, and to treated, breeding wastewater carries out correlation
Index determining.
2. a kind of application method of the immobilized microorganism according to claim 1 in livestock breeding wastewater processing, special
Sign is, in the step (1), Bacillus foecalis alkaligenes, Pseudomonas stutzeri and Pi Te acinetobacter calcoaceticus seed liquor cultural method are as follows:
The strain of these three microorganisms is inoculated on nutrient agar using spread plate, 30 DEG C of constant temperature incubation 48-72h,
Picking well-grown single colonie on plate is inoculated into nutrient broth medium under aseptic technique, in 30 DEG C,
160r/min shaking table shaken cultivation 48h.
3. a kind of application method of the immobilized microorganism according to claim 1 in livestock breeding wastewater processing, special
Sign is, in the step (1), the cultural method of bacillus coagulans seed liquor are as follows: gemma will be condensed using spread plate
The strain of bacillus is inoculated on LB solid medium, 35 DEG C of constant temperature incubation 48-72h, picking well-grown single bacterium on plate
Fall, be inoculated into LB liquid medium under aseptic technique, in 35 DEG C, 180r/min shaking table shaken cultivation for 24 hours.
4. a kind of application method of the immobilized microorganism according to claim 1 in livestock breeding wastewater processing, special
Sign is, in the step (1), the cultural method of penicillium chrysogenum seed liquor are as follows: using spread plate by the bacterium of penicillium chrysogenum
Kind is inoculated in PDA culture medium, 28 DEG C of constant temperature incubation 4-6d, washes lower spore with sterile saline, shaken well forms spore
Suspension accesses in PDB culture medium according to 5% inoculum concentration, in 28 DEG C, 150r/min shaking table shaken cultivation 72h.
5. a kind of application method of the immobilized microorganism according to claim 1 in livestock breeding wastewater processing, special
Sign is, in the step (1), the ingredient of fermentation medium are as follows: wheat bran 24-30g/L, molasses 11-15g/L, yeast powder 4-6g/
L, (NH4)2SO43-5g/L, NaCl 8.5-9.3g/L, KNO34.3-4.7g/L, K2HPO4 0.4-0.6g/L, KH2PO4 1.5-
1.9g/L, MgSO4·7H2O1.2-1.4g/L, CaCl20.4-0.5g/L, CoCl2·6H2O 3.5-4.0mg/L, FeSO4·
7H2O 5.2-5.6mg/L, MnSO4·H2O 1.6-2.2mg/L, surplus are water, and pH value is natural.
6. a kind of application method of the immobilized microorganism according to claim 1 in livestock breeding wastewater processing, special
Sign is, in the step (2), the ingredient of embedded material is polyvinyl alcohol 4%, sodium alginate 3%, diatomite 0.2%, Fe3O4It receives
Rice grain 0.3%, active carbon 0.2%.
7. a kind of application method of the immobilized microorganism according to claim 1 in livestock breeding wastewater processing, special
Sign is, in the step (3), the ingredient of induced medium are as follows: sucrose 12-16g/L, bean cake powder 5.1-5.5g/L, urea
3.3-3.8g/L NaCl 3.6-4.0g/L, KH2PO4 1.3-1.7g/L, MgSO4·7H2O0.7-0.8g/L, glutathione 14-
18mg/L, sodium citrate 6.5-7.5mg/L, sodium nitroprussiate 48-52 μm of ol/L, glycine betaine 1.6-2.0mg/L, fulvic acid 0.24-
0.28g/L, health polyphenol 0.1-0.2mg/L, pH value are natural.
8. a kind of application method of the immobilized microorganism according to claim 1 in livestock breeding wastewater processing, special
Sign is, in the step (3), interval ventilation refer to culture 12h, for 24 hours, 36h be passed through high-purity hydrogen, ventilatory capacity is to fill
Full entire three-neck flask.
9. a kind of application method of the immobilized microorganism according to claim 1 in livestock breeding wastewater processing, special
Sign is, in the step (4), domestication reactor should set up feed system, aerating system and temperature control system, the immobilization of addition
The volume ratio of microorganism gel ball and breeding wastewater is 1:3-1:5.
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