CN109626596A - A kind of preparation method and applications of effective microorganisms - Google Patents
A kind of preparation method and applications of effective microorganisms Download PDFInfo
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- CN109626596A CN109626596A CN201910039150.6A CN201910039150A CN109626596A CN 109626596 A CN109626596 A CN 109626596A CN 201910039150 A CN201910039150 A CN 201910039150A CN 109626596 A CN109626596 A CN 109626596A
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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Abstract
The invention discloses a kind of preparation method and applications of effective microorganisms, it is the bacterium solution mixing obtained after individually cultivating complex microorganism, it is seeded to fermentation medium and carries out fermented and cultured, supernatant is collected after the zymocyte liquid centrifugal filtration of acquisition, and be uniformly mixed with absorption carrier, the effective microorganisms of high flocculating rate can be obtained.Symbiosis coordination, reasonable compatibility between complex microorganism of the present invention, thallus proliferative speed is fast, and production cost is low, it is environment friendly and pollution-free, the flocculant being prepared effectively flocculates, and component content is high, and flocculating rate is preferable, has positive engineering practicability and wide application value.
Description
Technical field
The present invention relates to a kind of preparation method and applications of effective microorganisms, belong to flocculant technology of preparing neck
Domain.
Background technique
Flocculant, also known as sedimentation agent are mainly used in the processes such as sewage treatment and sludge dewatering, are that one kind can make in water body
The substance of not easily settled solid aerosol cohesion, precipitating is a kind of not only economic but also easy place for improving water treatment efficiency
Reason method.However how flocculating effect often determines the operation conditions and effluent quality and handling of follow-up processing flow
With.Therefore, during flocculation treatment, type, property, the kind of flocculant directly affect treatment effect and flocculation treatment
The core of technology.
Flocculant used at present from its source and is broadly divided into inorganic flocculating agent in nature and (mainly has iron series and aluminium
System), artificial synthesized high polymer coagulant and native biopolymer flocculant.But usually it will appear in its actual application
It is expensive, secondary pollution is also easy to produce, the problems such as insecurity, to limit its extensive exploitation application.In contrast, naturally
Boiomacromolecule flocculant can overcome the defect of inorganic flocculating agent and artificial synthesized high polymer coagulant inherently, final to realize
Non-pollution discharge, thus the research of native biopolymer flocculant especially microbial flocculant have become study instantly it is important
One of project.
Summary of the invention
It is multiple for the production process of current common microbiological flocculant in place of avoiding above-mentioned the shortcomings of the prior art
Miscellaneous, product effectively flocculates the problems such as ingredient is less, and flocculation efficiency is low, the present invention is intended to provide a kind of effective microorganisms
Preparation method and applications.Effective microorganisms production cost of the present invention is low, and safety and environmental protection is pollution-free, and function admirable is high
Effect, flocculation activity is high, can effectively reduce water pollutant ingredient, water body colloid and suspended particulate is removed, to reach preferable
Purifying water effect.
The preparation method of effective microorganisms of the present invention is the bacterium obtained after individually cultivating complex microorganism
Liquid mixing is seeded to fermentation medium and carries out fermented and cultured, collects supernatant after the zymocyte liquid centrifugal filtration of acquisition, and with suction
Appendix body is uniformly mixed, and the effective microorganisms of high flocculating rate can be obtained.
The complex microorganism includes that Klebsiella terrigena (compile by Klebsiella terrigena, CICC preservation, preservation
Numbers 20724, preservation date: on December 23rd, 2006), (Paenibacillus polymyxa, CICC are protected Paenibacillus polymyxa
Hiding, deposit number 23460, preservation date: on October 31st, 2008), Bacillus foecalis alkaligenes (Alcaligenes faecalis,
CICC preservation, deposit number 20602, preservation date: on April 25th, 2006), Rhodococcus ruber (Rhodococcus ruber,
CICC preservation, deposit number 23622, preservation date: on January 18th, 2002) and aspergillus niger (Aspergillus niger, CICC
Preservation, deposit number 2238, preservation date: on June 27th, 1958).
Further, when the bacterium solution mixing that complex microorganism obtains after individually cultivating, Klebsiella terrigena bacterium solution,
Paenibacillus polymyxa bacterium solution, Bacillus foecalis alkaligenes bacterium solution, the volume ratio of Rhodococcus ruber bacterium solution and aspergillus niger bacterium solution are 2.8~3.2:
0.8~1.2:2.8~3.2:1.8~2.2:0.8~1.2, preferred volume ratio 3:1:3:2:1.
Further, the inoculation volume ratio for fermentation medium being seeded to after bacterium solution mixing is 5~15%.
Further, when fermented and cultured, fermentation time is 3~4d, and mixing speed is 160~200r/min, and ventilatory capacity is
1:1.5~2.2, stirring interval time are 2h, stir 2min, and cultivation temperature is 30~37 DEG C.Fermented and cultured viable bacteria into bacterium solution
Number reaches 5 × 107CFU/mL or more can collect supernatant by centrifugal filtration.
The fermentation medium is mixed by the raw material of following weight proportion: 18~25g of brown sugar, 7~9g of peptone,
3~5g of beef extract, 2~5g of NaCl, K2HPO42.8~3.1g, MgSO4.7H21.2~1.7g of O, vitamin B150mg is mended
Cooling uses after filling 5 ° of B é brewer's worts to 1L, pH=7,121 DEG C of sterilizing 30min.
The condition of the centrifugal filtration is 4 DEG C~6 DEG C, 6000~8000r/min, is centrifuged 25~30min.
The absorption carrier mainly includes turfy soil, montmorillonite, diatomite and chitosan, is constituted by mass fraction as follows:
8 parts of turfy soil, 5 parts of montmorillonite, 5 parts of diatomite, 2 parts of chitosan.
The turfy soil, montmorillonite, diatomite and chitosan are 100 mesh.
Mass ratio when supernatant is mixed with absorption carrier is 4:6.
Further, in preparation method of the present invention, the process that complex microorganism is individually cultivated includes the following steps:
The Klebsiella terrigena, Paenibacillus polymyxa and Bacillus foecalis alkaligenes bacterium solution the preparation method comprises the following steps: choosing respectively
It takes above-mentioned 3 kinds of strains to be seeded in solid slant culture, is placed in incubator and cultivates, cultivation temperature is 28 DEG C, and incubation time is
For 24 hours, after its slant medium grows bacterium colony, progress shaking table shaken cultivation, shaking speed in fluid nutrient medium are seeded to
For 150~180r/min, cultivation temperature is 30~37 DEG C, and incubation time is 24~36h, when viable count reaches 1 × 108CFU/mL
When bacterium solution preparation complete；The solid slope culture medium is mixed by the raw material of following weight proportion: peptone 5g, beef
It is cooling after medicinal extract 3g, NaCl 5g, agar 15g, supplement distilled water to 1L, pH=7,121 DEG C of sterilizing 30min to use；The liquid
Culture medium is mixed by the raw material of following weight proportion: peptone 5g, beef extract 3g, NaCl 5g, and supplement distilled water is extremely
It is cooling after 1L, pH=7,121 DEG C of sterilizing 30min to use.
The Rhodococcus ruber bacterium solution the preparation method comprises the following steps: picking Rhodococcus ruber strain is seeded in solid slant culture, set
It is cultivated in incubator, cultivation temperature is 37 DEG C, and incubation time 48h is inoculated with after its slant medium grows bacterium colony
Shaking table shaken cultivation is carried out into fluid nutrient medium, shaking speed is 150~180r/min, and cultivation temperature is 35~37 DEG C, training
Supporting the time is 48~72h, when viable count reaches 1 × 108Bacterium solution preparation is completed when CFU/mL；The solid slope culture medium be by
The raw material of following weight proportion is mixed: glucose 20g, peptone 9g, K2HPO4 3g、MgSO4.7H2O 1.5g, vitamin
B1It is cooling after 50mg, agar 15g, supplement potato extracting solution to 1L, pH=6,121 DEG C of sterilizing 30min to use；The liquid
Culture medium is mixed by the raw material of following weight proportion: glucose 20g, peptone 9g, K2HPO4 3g、MgSO4.7H2O
1.5g, vitamin B1Cooling makes after 50mg, agar 15g, supplement potato extracting solution to 1L, pH=6,121 DEG C of sterilizing 30min
With.
The aspergillus niger bacterium solution the preparation method comprises the following steps: picking aspergillus niger strain is seeded in solid slant culture, be placed in training
It supports and is cultivated in case, cultivation temperature is 37 DEG C, and incubation time 48h is seeded to liquid after its slant medium grows bacterium colony
Shaking table shaken cultivation is carried out in body culture medium, shaking speed is 150~180r/min, and cultivation temperature is 35~37 DEG C, when culture
Between be 48~72h, when viable count reaches 1 × 108Bacterium solution preparation is completed when CFU/mL；The solid slope culture medium is by following
The raw material of weight proportion is mixed: 5 ° of B é brewer's wort 1L, agar 15g, natural pH, cooling after 121 DEG C of sterilizing 30min to use；
The fluid nutrient medium is mixed by the raw material of following weight proportion: 5 ° of B é brewer's wort 1L, agar 15g, natural pH, 121
It is cooling after DEG C sterilizing 30min to use.
The application for the effective microorganisms that the present invention prepares is that the effective microorganisms are added to give birth to
In sewage living, flocculation sedimentation, to improve purifying water effect.
The best input amount of the effective microorganisms is 50g/m3。
Compared with prior art, the invention has the advantages that:
1, effectively flocculate component content height in the microbial flocculant that the present invention is prepared, and action effective is fast, at low cost,
It working well, safety and environmental protection is pollution-free, and it is easy to operate, there is positive engineering practicability and wide application value；
2, selected microorganism has stronger combination and cooperative ability, does not repel mutually, can produce each other more
Flocculation effective substance, wherein Bacillus foecalis alkaligenes can improve the alkalinity of compound microorganism ferments product, and wadding can be improved under alkaline environment
The crushing effect of solidifying complex microorganism, facilitates the release of effective flocculate；
3, selected absorption carrier contains a certain number of sticking grains, it is made to have different degrees of electricity negative in water body
Property, the processes such as charge neutrality, absorption can occur with metastable suspended particulate is presented in raw sewage, the current potential for destroying raw sewage is flat
Weighing apparatus aggravates the collision between suspended particulate, enhances its flocculation sedimentation effect.The sun that turfy soil, montmorillonite and diatomite have
Ion exchange capacity plays booster action in flocculation process；Chitosan is cation polymeric flocculant, in sewage treatment
It plays its gantry and net catches function, greatly improve flocculation adsorption sedimentation effect.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention,
Technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is in the present invention
A part of the embodiment, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art are not having
Every other embodiment obtained under the premise of creative work is made, the scope of the present invention is belonged to.
Embodiment 1: the preparation of effective microorganisms
Effective microorganisms of the present invention the preparation method is as follows:
1, complex microorganism is individually cultivated, detailed process is as follows:
The Klebsiella terrigena, Paenibacillus polymyxa and Bacillus foecalis alkaligenes bacterium solution the preparation method comprises the following steps: choosing respectively
It takes above-mentioned 3 kinds of strains to be seeded in solid slant culture, is placed in incubator and cultivates, cultivation temperature is 28 DEG C, and incubation time is
For 24 hours, after its slant medium grows bacterium colony, progress shaking table shaken cultivation, shaking speed in fluid nutrient medium are seeded to
For 150~180r/min, cultivation temperature is 30~37 DEG C, and incubation time is 24~36h, when viable count reaches 1 × 108CFU/mL
When bacterium solution preparation complete；The solid slope culture medium is mixed by the raw material of following weight proportion: peptone 5g, beef
It is cooling after medicinal extract 3g, NaCl 5g, agar 15g, supplement distilled water to 1L, pH=7,121 DEG C of sterilizing 30min to use；The liquid
Culture medium is mixed by the raw material of following weight proportion: peptone 5g, beef extract 3g, NaCl 5g, and supplement distilled water is extremely
It is cooling after 1L, pH=7,121 DEG C of sterilizing 30min to use.
The preparation of the Rhodococcus ruber bacterium solution are as follows: picking Rhodococcus ruber strain is seeded in solid slant culture, is placed in training
It supports and is cultivated in case, cultivation temperature is 37 DEG C, and incubation time 48h is seeded to liquid after its slant medium grows bacterium colony
Shaking table shaken cultivation is carried out in body culture medium, shaking speed is 150~180r/min, and cultivation temperature is 35~37 DEG C, when culture
Between be 48~72h, when viable count reaches 1 × 108Bacterium solution preparation is completed when CFU/mL；The solid slope culture medium is by following
The raw material of weight proportion is mixed: glucose 20g, peptone 9g, K2HPO4 3g、MgSO4.7H2O 1.5g, vitamin B1
It is cooling after 50mg, agar 15g, supplement potato extracting solution to 1L, pH=6,121 DEG C of sterilizing 30min to use；The Liquid Culture
Base is mixed by the raw material of following weight proportion: glucose 20g, peptone 9g, K2HPO4 3g、MgSO4.7H2O 1.5g,
Vitamin B1It is cooling after 50mg, agar 15g, supplement potato extracting solution to 1L, pH=6,121 DEG C of sterilizing 30min to use.
The preparation of the aspergillus niger bacterium solution are as follows: picking aspergillus niger strain is seeded in solid slant culture, is placed in incubator
Middle culture, cultivation temperature is 37 DEG C, incubation time 48h, after its slant medium grows bacterium colony, is seeded to liquid training
It supports and carries out shaking table shaken cultivation in base, shaking speed is 150~180r/min, and cultivation temperature is 35~37 DEG C, and incubation time is
48~72h, when viable count reaches 1 × 108Bacterium solution preparation is completed when CFU/mL；The solid slope culture medium is by following weight
The raw material of proportion is mixed: 5 ° of B é brewer's wort 1L, agar 15g, natural pH, cooling after 121 DEG C of sterilizing 30min to use；It is described
Fluid nutrient medium is mixed by the raw material of following weight proportion: 5 ° of B é brewer's wort 1L, agar 15g, natural pH, 121 DEG C go out
It is cooling after bacterium 30min to use.
2, the bacterium solution for obtaining step 1 mixes, and Klebsiella terrigena bacterium solution, Paenibacillus polymyxa bacterium solution, excrement produce alkali
The volume ratio of bacillus bacterium solution, Rhodococcus ruber bacterium solution and aspergillus niger bacterium solution be 2.8~3.2:0.8~1.2:2.8~3.2:1.8~
2.2:0.8~1.2, preferred volume ratio 3:1:3:2:1 are seeded to fermentation medium and carry out fermented and cultured, and inoculation volume ratio is 5
~15%, fermentation time is 3~4d, and mixing speed is 160~200r/min, and ventilatory capacity is 1:1.5~2.2, when stirring is spaced
Between be 2h, stir 2min, cultivation temperature be 30~37 DEG C, fermented and cultured viable count into bacterium solution reaches 5 × 107CFU/mL with
On；
3, supernatant will be collected after zymocyte liquid centrifugal filtration that step 2 obtains, and is uniformly mixed with absorption carrier, supernatant
Mass ratio when liquid is mixed with absorption carrier is 4:6, and the effective microorganisms of high flocculating rate can be obtained.
The absorption carrier is made of turfy soil, montmorillonite, diatomite and chitosan, is constituted by mass fraction as follows: grass
8 parts, 5 parts of montmorillonite, 5 parts of diatomite, 2 parts of chitosan of charcoal soil；The turfy soil, montmorillonite, diatomite and chitosan are 100
Mesh.
Embodiment 2: application of the effective microorganisms to sewage treatment
Effective microorganisms prepared by embodiment 1 are applied to the processing of sanitary sewage.Concrete operations are as follows: will give birth to
Sewage raw water (COD 765mg/L, total phosphorus 21.8mg/L, ammonia nitrogen 40.7mg/L) living is placed in the flocculation with agitating function
Effective microorganisms 50g/m prepared by embodiment 1 is added in device and to it3, quickly it is stirred with 150r/min
60s, then 90s is slowly stirred with 50r/min, after standing 30min, its supernatant is taken to be detected.
Not use sanitary sewage of the invention as blank sample, with Berthelot spectrophotometry (HJ 535-2009) point
Not Ce Ding ammonia-nitrogen content in sewage, measure the total phosphorus in sewage respectively with ammonium molybdate spectrophotometric method (GB 11893-89) and contain
Amount, the COD content in sewage is measured with rapid-digestion spectrophotometry (HJ/T 399-2007), on the basis of blank sample respectively
Degradation rate is calculated, using the removal rate of sanitary sewage ammonia nitrogen of the invention up to 89.7%, COD removal rate up to 92.9%, total phosphorus drops
Solution rate is up to 88.7%, and flocculating rate is up to 96.2%.
The measuring method of flocculating rate: 4g/L Kaolin clay suspension and 0.5g/L CaCl are prepared2Solution is added by volume ratio
In addition stating Kaolin clay suspension 98% and CaCl2Solution 2% mixes, and flocculation system is made.It is added to 500mL flocculation system
0.1mL fermented supernatant fluid is quickly stirred 60s with 150r/min, then is slowly stirred 90s with 50r/min, after standing 30min,
It takes its supernatant to measure the OD value A under wavelength 550nm, aforesaid operations is repeated with distilled water substitution fermented supernatant fluid and measure OD value B.
Flocculating rate={ (B-A)/B } × 100%.
The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although with reference to the foregoing embodiments
Invention is explained in detail, those skilled in the art should understand that: it still can be to aforementioned each implementation
Technical solution documented by example is modified or equivalent replacement of some of the technical features；And these modification or
Replacement, the spirit and scope for technical solution of various embodiments of the present invention that it does not separate the essence of the corresponding technical solution.
Claims (10)
1. a kind of preparation method of effective microorganisms, it is characterised in that: be after individually cultivating complex microorganism
The bacterium solution of acquisition mixes, and is seeded to fermentation medium and carries out fermented and cultured, collects supernatant after the zymocyte liquid centrifugal filtration of acquisition
Liquid, and be uniformly mixed with absorption carrier, the effective microorganisms of high flocculating rate can be obtained.
2. preparation method according to claim 1, it is characterised in that:
The complex microorganism includes Klebsiella terrigena, Paenibacillus polymyxa, Bacillus foecalis alkaligenes, Rhodococcus ruber and black song
It is mould.
3. preparation method according to claim 2, it is characterised in that:
When the bacterium solution mixing that complex microorganism obtains after individually cultivating, Klebsiella terrigena bacterium solution, mostly viscous class brood cell bar
Bacterium bacterium solution, Bacillus foecalis alkaligenes bacterium solution, the volume ratio of Rhodococcus ruber bacterium solution and aspergillus niger bacterium solution are 2.8~3.2:0.8~1.2:2.8
~3.2:1.8~2.2:0.8~1.2.
4. preparation method according to claim 1,2 or 3, it is characterised in that:
The inoculation volume ratio that fermentation medium is seeded to after bacterium solution mixing is 5~15%.
5. the preparation method according to claim 4, it is characterised in that:
When fermented and cultured, fermentation time is 3~4d, and mixing speed is 160~200r/min, and ventilatory capacity is 1:1.5~2.2, is stirred
Mixing interval time is 2h, stirs 2min, and cultivation temperature is 30~37 DEG C.
6. preparation method according to claim 5, it is characterised in that:
Fermented and cultured viable count into bacterium solution reaches 5 × 107CFU/mL or more can collect supernatant by centrifugal filtration.
7. the preparation method according to claim 4, it is characterised in that:
The fermentation medium is mixed by the raw material of following weight proportion: 18~25g of brown sugar, 7~9g of peptone, beef
3~5g of medicinal extract, 2~5g of NaCl, K2HPO42.8~3.1g, MgSO4·7H21.2~1.7g of O, vitamin B150mg, supplement
It is cooling after 5 ° of B é brewer's worts to 1L, pH=7,121 DEG C of sterilizing 30min to use.
8. preparation method according to claim 1, it is characterised in that:
The absorption carrier includes turfy soil, montmorillonite, diatomite and chitosan, is constituted by mass fraction as follows: turfy soil 8
Part, 5 parts of montmorillonite, 5 parts of diatomite, 2 parts of chitosan；Mass ratio when supernatant is mixed with absorption carrier is 4:6.
9. the application for the effective microorganisms that any method prepares in claim 1-8, it is characterised in that: be by
The effective microorganisms are added in sanitary sewage, flocculation sedimentation, to improve purifying water effect.
10. application according to claim 9, it is characterised in that:
The input amount of the effective microorganisms is 50g/m3。
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Application publication date: 20190416 |