CN104845905B - The high efficiency composition microbial inoculum of removal COD containing salad oil wastewater a kind of and its application - Google Patents
The high efficiency composition microbial inoculum of removal COD containing salad oil wastewater a kind of and its application Download PDFInfo
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Abstract
The invention discloses the high efficiency composition microbial inoculum of removal COD containing salad oil wastewater a kind of and its application.The composite bacteria agent is made up of four kinds of high-efficiency grease degradation bacterias, respectively candida tropicalis HY A 3, Serratieae SH AB 1, candida tropicalis SZ2 A 3 and serratia marcescens LB1 AB 2.The present invention gathers bacterium source sample from sludge sewage, dining room effluent sewerage etc., and the single bacterium of the COD containing salad oil wastewater is efficiently removed through separating；Then high efficiency composition microbial inoculum is built by combine experiment, as a result the microbial inoculum to be built by HY A 3, SH AB 1, SZ2 A 3 and LB1 AB 2 is combined to COD clearance highests.When the concentration of composite bacteria agent is 1 5%, more than 75% is up to salad oil wastewater COD clearance.The microbial inoculum has good application prospect and value in salad oil wastewater handles engineering.
Description
Technical field
The present invention relates to a kind of removal COD containing salad oil wastewater high efficiency composition microbial inoculum, further relates to remove salad oil wastewater
COD composite bacteria agent application process.
Background technology
As salad oil demand steeply rises, the highly concentrated oil-bearing wastewater in salad oil processing influences to caused by environment
Concern is caused.Salad oil manufacturing wastewater refer to soybean etc. squeeze or leach gained crude oil carry out degumming, depickling, decolouring,
The waste water caused by processing such as deodorization, dewaxing, de- ester.Substantial amounts of organic wastewater, the spy of this waste water are produced in this refining process
Point is high COD (ρCODUp to more than 10000mg/L), the big (ρ of oil contentoilUp to more than 3000mg/L), B/C 0.5-0.6.Mesh
The preceding processing for salad oil manufacturing wastewater is more using physico-chemical process pretreatment oil removing and activated sludge process or anaerobic technique processing.
After amount of grease enters sewage disposal system, it will usually cause to produce bubble, and the growth of der Pilz can be promoted, influence at sewage
Manage the operation of biochemical system.Therefore to strengthen the deoiling function of the biochemical system in salad oil wastewater processing procedure, accelerate to oil
The degraded of fat, the exceeded possibility of reduction system water outlet, foam caused by reduction, salad oil wastewater processing provided by the invention
Core --- enhancement microbiological, the functional flora formed using the saccharomycete for screening to obtain from nature and bacterium, aerobic
Efficient-decomposition is carried out to the aliphatic acid of high concentration, glyceride and other organic matters in biological treatment, to realize raising sewage
The effluent quality of processing system.
The content of the invention
For in the prior art the defects of, efficient remove salad oil wastewater COD's it is an object of the invention to provide a kind of
Composite bacteria agent and preparation method thereof.
It is a further object of the present invention to provide the application process of above-mentioned composite bacteria agent.
In order to realize foregoing invention purpose, the technical solution adopted by the present invention is as follows：
A kind of removal COD containing salad oil wastewater composite bacteria agent, it is characterised in that the composite bacteria agent includes bacterial strain：Heat
Band Candida HY-A-3, CGMCC No.10424；Serratieae SH-AB-1, CGMCC No.10427；Candida tropicalis
SZ2-A-3, CGMCC No.10425；Serratia marcescens LB1-AB-2, CGMCC No.10426.
Further, the candida tropicalis HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and viscous
Matter Serratieae LB1-AB-2 bacterium amount accounts for 25-45%, 25-45%, 8-25% and 2-25% of total bacterium amount volume respectively.
Further, the candida tropicalis HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and viscous
Matter Serratieae LB1-AB-2 bacterium amount accounts for 30-40%, 30-40%, 15-20%, 3-20% of total bacterium amount volume respectively.
Further, the candida tropicalis HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and viscous
Matter Serratieae LB1-AB-2 bacterium amount accounts for 33%, 33%, 17%, the 17% of total bacterium amount volume respectively.
Present invention also offers a kind of preparation method of removal COD containing salad oil wastewater composite bacteria agent, including following step
Suddenly：Candida HY-A-3 and SZ2-A-3 single bacterium colony is taken to be inoculated in 100mL yeast extract powder peptone dextrose culture-mediums respectively,
Serratieae SH-AB-1 and serratia marcescens LB1-AB-2 single bacterium colonies is taken to be inoculated in 100mL beef extract-peptone Liquid Cultures
In base, the shaken cultivation 18h at 180rpm, 28 DEG C；Four plants of bacterium for cultivating 18h are carried out compound.
Present invention also offers a kind of removal COD containing salad oil wastewater method, comprise the following steps：It is false the torrid zone will to be included
Silk yeast HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and serratia marcescens LB1-AB-2 it is compound
Microbial inoculum is inoculated in salad oil wastewater by 25-45%, 25-45%, 8-25% and 2-25% for accounting for total bacterium amount volume proportioning respectively
In, and cultivate 2~3 days.
Further, the candida tropicalis HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and viscous
Matter Serratieae LB1-AB-2 bacterium amount accounts for 30-40%, 30-40%, 15-20% and 3-20% of total bacterium amount volume respectively.
Further, the candida tropicalis HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and
Serratia marcescens LB1-AB-2 bacterium amount accounts for 33%, 33%, 17% and the 17% of total bacterium amount volume respectively.
Further, the composite bacteria agent is inoculated with the form of bacterium solution, and its inoculum concentration is the 1-5% (v/v) of processing wastewater volume.
Further, described inoculum concentration is the 1.5-2.5% (v/v) of processing wastewater volume.
Effect of the invention is that：Using composite bacteria agent of the present invention and its application, to the COD's containing salad oil wastewater
Clearance is up to more than 75%.
Brief description of the drawings
Fig. 1 candida tropicalis Candida tropicalis HY-A-3 systematic evolution tree；
Fig. 2 Serratieae Serratia sp.SH-AB-1 systematic evolution tree；
Fig. 3 candida tropicalis Candida tropicalis SZ2-A-3 systematic evolution tree；
Fig. 4 serratia marcescens Serratia marcescens LB1-AB-2 systematic evolution tree.
Classification And Nomenclature:Candida tropicalis, Latin name：Candida tropicalis HY-A-3
Preservation date：On January 22nd, 2015；
Depositary institution's full name and abbreviation：China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC)
Deposit number:CGMCC No.10424；
Classification And Nomenclature:Serratieae, Latin name：Serratia sp.SH-AB-1
Preservation date：On January 22nd, 2015；
Depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC)
Deposit number:CGMCC No.10427；
Classification And Nomenclature:Candida tropicalis, Latin name：Candida tropicalis SZ2-A-3
Preservation date：On January 22nd, 2015；
Depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC)
Deposit number:CGMCC No.10425；
Classification And Nomenclature:Serratia marcescens, Latin name：Serratia marcescens LB1-AB-2
Preservation date：On January 22nd, 2015；
Depositary institution：China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC)
Deposit number:CGMCC No.10426；
Embodiment
The present invention illustrated below, but the protection domain being not intended to limit the present invention.
In order to solve the above problems, the present invention is gathered under certain shopping center dining room effluent sewerage, market of farm produce meat stand side
The sample such as body refuse by ditch sewage, sewage treatment plant's biochemical reaction tank sludge and certain vegetable oil processing factory treatment tank, through richness
Collection, domestication, separation, purifying, are filtered out with the pure bacterial strain for removing salad oil wastewater COD, then through primary dcreening operation, secondary screening, are selected efficiently
Function bacterium, structure remove salad oil wastewater COD composite bacteria agent.
HY-A-3 in the present invention is located away from certain shopping center dining room effluent sewerage；SH-AB-1 is located away from Beijing
In the other lower ditch sewage in market of farm produce meat stand；SZ2-A-3 is located away from Hebei sewage treatment plant biochemical reaction tank sludge；LB1-
AB-2 is located away from body refuse by Beijing vegetable oil processing factory treatment tank.4 plants of bacterium in composite bacteria agent are carried out with form spy respectively
Sign observation and the identification of part Physiology and biochemistry.HY-A-3 thalli morphologies are oval, and size is about 5~6 × 7~9 μm, soaked in yeast
Go out rounded bacterium colony on powder peptone dextrose culture-medium (YPD culture mediums), milky, smooth, raised, neat in edge, 10%
Remain to preferably grow in NaCl solution；SH-AB-1 thalline are in rod-short, and size is about 0.5~0.8 × 0.9~1.3 μm, no bud
Spore, bacterium colony is rounded, raised, smooth, milky, Gram-negative, contacts enzyme positive, and Starch Hydrolysis is negative, it is impossible to
Grown in NaCl solution more than 7% concentration；SZ2-A-3 thalli morphologies are oval, and size is about 5~7 × 8~9 μm,
Bacterium colony is circular on YPD culture mediums, smooth, raised, milky, contacts enzyme positive, is remained to more quite in 10% NaCl solution
It is long；LB1-AB-2 thalline are in rod-short, and size is about 0.5~0.8 × 0.6~1.0 μm, no gemma, and bacterium colony is circular, smooth, convex
Rise, be red, Gram-negative contacts enzyme positive, and Starch Hydrolysis is negative, it is impossible in the NaCl solution more than 7% concentration
Growth.4 plants of bacterium are carried out with molecular biology identification respectively, carries out 18S rDNA's after HY-A-3 and SZ2-A-3 extraction gene DNAs
PCR is expanded, and SH-AB-1 and LB1-AB-2 progress 16S rDNA PCR amplifications, the fragment of acquisition is used to be sequenced.Postorder is sequenced
Row result carries out Blast comparisons, and constructing system chadogram in Genbank, as Figure 1-Figure 4.
By sequencing result and the systematic evolution tree of structure, with reference to 4 plants of bacterium morphological features, growth conditions, Physiology and biochemistry identification
As a result, can be using Preliminary Identification bacterial strain HY-A-3 and SZ2-A-3 as candida tropicalis (Candida tropicalis), LB1-
AB-2 is serratia marcescens (Serratia marcescens), and SH-AB-1 belongs to Serratia (Serratia sp.).
Above-mentioned four kinds of bacterial strains are deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (address：Court of Beijing
The positive institute 3 of area's North Star West Road 1).
In composite bacteria agent of the present invention, candida tropicalis HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-
A-3, serratia marcescens LB1-AB-2 bacterium amount account for 25-45%, 25-45%, 8-25% and 2- of total bacterium amount volume respectively
25%, 30-40%, 30-40%, 15-20% and 3-20% of total bacterium amount volume are preferably accounted for respectively.
The preparation method of above-mentioned composite bacteria agent is to take Candida HY-A-3 and SZ2-A-3 single bacterium colony to be inoculated in respectively
In 100mL yeast extract powder peptones dextrose culture-medium (YPD fluid nutrient mediums), Serratieae SH-AB-1 and Serratia are taken
Bacterium LB1-AB-2 single bacterium colonies are inoculated in 100mL beef extract-peptone fluid nutrient mediums, the shaken cultivation at 180rpm, 28 DEG C
18h.18h candida tropicalis HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3, cement sand will be cultivated
Thunder Salmonella LB1-AB-2 is compound by 30-40%, 30-40%, 15-20% and 3-20% progress of the bacterium amount than accounting for total bacterium amount volume.
The application of above-mentioned composite bacteria agent, it is 1000- that composite bacteria agent is inoculated in into oil mass containing salad by 1-5% concentration
2000mg/L oil wastewater, to the clearance of waste water COD up to more than 75%.
Embodiment 1：The acclimating and separation screening of grease degrading strain.
The present invention gathers the rich grease-contained bacterium source sample in many places first, then carries out acclimating training to bacterium source sample respectively
Support, then therefrom isolate pure bacterial strain, then high-efficiency grease degradation bacteria is obtained through primary dcreening operation, secondary screening.
Specific implementation step is as follows：
(1) the acclimating culture and initial gross separation of grease degrading strain
3g mud samples or 3mL water samples are taken, it is inoculated in the triangular flask of the liquid of enrichment culture containing 100mL, and in 28 DEG C, 180rpm
3d is cultivated in shaking table.The bacterium solution after 3mL enrichments is taken, is inoculated in the triangular flask of the domestication nutrient solution containing 100mL, 28 DEG C, 180rpm
Shaking table culture, 3d are 1 cycle, and bacterium solution is moved on new domestication culture medium and cultivated by each cycle after terminating.
The bacterium solution after 1mL domestications is taken, is inoculated in bacterium containing 100mL or saccharomycete liquid basal medium, is placed in respectively
28 DEG C, cultivate on 180rpm shaking table.After 3d, after the bacterium solution gradient dilution of culture, corresponding total amount of binder is respectively coated on
In culture medium flat plate, in 30 DEG C of incubator culture 48-72h.The microbial strains of picking colony morphological feature significant difference, respectively
Streak inoculation is in corresponding basal medium, 30 DEG C of incubated 48-72h, the repeated isolated single bacterium colony pure culture of line
Thing, inclined-plane preserve.
(2) screening of high-efficiency grease degradation bacteria
The bacterial strain point for ruling isolated is connected on lipase checking culture medium flat plate, 30 DEG C of constant incubator cultures
48-72h, just sifts out the bacterial strain that transparent circle is relatively large in diameter, and foundation is provided for next step secondary screening.
With colony inoculation in oese picking primary dcreening operation flat board into the triangular flask of the basal medium containing 100mL, 28 DEG C,
180rpm shaking table cultures, transferred after cultivating 18h by 2% inoculum concentration in the triangular flask of the salad oil wastewater containing 100mL, 28 DEG C,
180rpm shaker fermentations, COD is measured by sampling respectively at 48h, 72h.Last bound fat enzyme circle checking test, obtain 8 plants of degradeds
The stronger bacterial strain of grease ability.
Culture medium needed for the present invention prepares as follows：
1. enriched medium：K2HPO43g, NaCl 5g, MgSO4·7H2O 0.5g, peptone 10g, yeast extract 5g,
Salad oil 10g, distilled water 1000mL, pH=7.2.
2. tame culture medium：Yeast extract 1g, (NH4)2SO43g, K2HPO41g, NaH2PO41g, NaCl 0.5g,
MgSO4·7H2O 0.2g, salad oil add 5g, 10g, 15g, distilled water 1000mL, pH7.0 successively.
3. lipase verifies culture medium：KH2PO40.3g, MgSO4·7H2O 0.1g, K2HPO41.5g, (NH4)2SO4
1g, NaCl 5g, FeSO4·7H2O 0.01g, Tween 80 20mL, salad oil 6g, agar 18g, distilled water 1000mL,
pH7.0。
4. autogamy oily waste water：KH2PO40.3g, MgSO4·7H2O 0.1g, K2HPO41.5g, (NH4)2SO41g,
NaCl 5g, FeSO4·7H2O 0.01g, salad oil 2g, distilled water 1000mL, pH7.0.
Embodiment 2：The structure of composite bacteria agent in the present invention.
8 plant heights effect grease degrading strain is filtered out, selects wherein 4 plants progress combine experiments, 4 kinds of bacterium bacterium amount ratios in every kind of combination
It is identical, it is inoculated in by total inoculum concentration 2% in autogamy oily waste water, the COD of salad oil wastewater is determined after 72h.COD clearance highests
The combination for being combined as HY-A-3, SH-AB-1, SZ2-A-3 and LB1-AB-2, its COD clearance be 80.18%, it is identified, 4
Strain bacterium is respectively candida tropicalis HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3, serratia marcescens
LB1-AB-2.The horizontal quadrature of four factor three that 4 kinds of bacterium in combinations thereof are carried out with different vaccination ratio is tested.Research shows,
Most preferably inoculation volume proportion HY-A-3 between each bacterial strain:SH-A-1:SZ2-A-3:LB1-AB-2 is 33%, 33%, 17% and 17%,
The total bacterium amount of mix bacterium agent is inoculated in oily waste water by 2% inoculative proportion, and oil wastewater COD clearances are 81.3% after 72h
(raw water COD1426.2mg/L).
Embodiment 3：Part unlike the embodiments above is, by candida tropicalis HY-A-3, Serratieae SH-AB-
1st, tetra- plants of bacterium of candida tropicalis SZ2-A-3 and serratia marcescens LB1-AB-2 account for total bacterium amount volume by bacterium amount respectively
25%th, 25%, 25% and 25% proportioning is carried out compound, and mix bacterium agent is seeded in oily waste water in 1% ratio, after 72h, oil
Fat waste water COD clearance is 75.98% (raw water COD1076.32mg/L).
Embodiment 4：Part unlike the embodiments above is, by candida tropicalis HY-A-3, Serratieae SH-AB-
1st, tetra- plants of bacterium of candida tropicalis SZ2-A-3 and serratia marcescens LB1-AB-2 account for total bacterium amount volume by bacterium amount respectively
45%th, 45%, 8% and 2% proportioning is carried out compound, and mix bacterium agent is seeded in oily waste water in 5% ratio, after 72h, grease
Waste water COD clearance is 80.15% (raw water COD1058.49mg/L).
Embodiment 5：Composite bacteria agent is tested actual oil wastewater COD degradation in the present invention.
Actual waste water is derived from certain Peasants Joy food and drink waste water, is crossed two layers gauze using preceding and is removed dish leaf leftovers, at the beginning of waste water
Beginning COD is 1723.6mg/L, pH 6.0.The actual oily waste waters of 100mL are taken in 250mL triangular flasks, after adjusting pH to 7.2, are pressed
2% inoculum concentration inoculating compound bacterium agent, the oily waste water for not being inoculated with microbial inoculum is set as control, it is same to be placed in 180rpm, 28 DEG C of conditions
Lower fermented and cultured, COD is determined after 72h.Composite bacteria agent is 78.95% to actual oily waste water COD clearance.
It should be noted that what above-mentioned specific embodiment was merely exemplary, under the above-mentioned teaching of the present invention, this area
Technical staff can carry out various improvement and deformation on the basis of above-described embodiment, and these are improved or deformation falls in this hair
In bright protection domain.It will be understood by those skilled in the art that specific descriptions above are intended merely to explain the purpose of the present invention,
It is not intended to limit the present invention.Protection scope of the present invention is limited by claim and its equivalent.
Claims (9)
1. a kind of removal COD containing salad oil wastewater composite bacteria agent, it is characterised in that the composite bacteria agent includes bacterial strain：The torrid zone
Candida HY-A-3, CGMCC No.10424；Serratieae SH-AB-1, CGMCCNo.10427；Candida tropicalis SZ2-
A-3, CGMCC No.10425；Serratia marcescens LB1-AB-2, CGMCC No.10426,
The candida tropicalis HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and serratia marcescens
LB1-AB-2 bacterium amount accounts for 25-45%, 25-45%, 8-25% and 2-25% of total bacterium amount volume respectively.
A kind of 2. removal COD containing salad oil wastewater according to claim 1 composite bacteria agent, it is characterised in that：The heat
With Candida HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and serratia marcescens LB1-AB-2
Bacterium amount accounts for 30-40%, 30-40%, 15-20% and 3-20% of total bacterium amount volume respectively.
A kind of 3. removal COD containing salad oil wastewater according to claim 2 composite bacteria agent, it is characterised in that：The heat
With Candida HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and serratia marcescens LB1-AB-2
Bacterium amount accounts for 33%, 33%, 17% and the 17% of total bacterium amount volume respectively.
4. the preparation method of the composite bacteria agent described in claim 1-3 any claims, comprises the following steps：False silk is taken respectively
Yeast HY-A-3 and SZ2-A-3 single bacterium colony is inoculated in 100mL yeast extract powder peptone dextrose culture-mediums, takes Serratieae SH-
AB-1 and serratia marcescens LB1-AB-2 single bacterium colonies are inoculated in 100mL beef extract-peptone fluid nutrient mediums, 180rpm,
Shaken cultivation 18h at 28 DEG C；Four plants of bacterium for cultivating 18h are carried out compound.
5. a kind of removal COD containing salad oil wastewater method, comprises the following steps：Candida tropicalis HY-A-3, sand will be included
Thunder Salmonella SH-AB-1, candida tropicalis SZ2-A-3 and serratia marcescens LB1-AB-2 composite bacteria agent are respectively by accounting for total bacterium
25-45%, 25-45%, 8-25% and 2-25% proportioning of amount volume are inoculated in salad oil wastewater, and are cultivated 2~3 days.
A kind of 6. removal COD containing salad oil wastewater according to claim 5 method, it is characterised in that：The torrid zone is false
Silk yeast HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and serratia marcescens LB1-AB-2 bacterium amount
30-40%, 30-40%, 15-20% and 3-20% of total bacterium amount volume are accounted for respectively.
A kind of 7. removal COD containing salad oil wastewater method as claimed in claim 6, it is characterised in that：The false silk in the torrid zone
Yeast HY-A-3, Serratieae SH-AB-1, candida tropicalis SZ2-A-3 and serratia marcescens LB1-AB-2 bacterium amount point
33%, 33%, 17% and the 17% of total bacterium amount volume is not accounted for.
A kind of 8. method of removal COD containing salad oil wastewater as described in claim any one of 5-7, it is characterised in that：It is described
Composite bacteria agent is inoculated with the form of bacterium solution, and its inoculum concentration is the 1-5% (v/v) of processing wastewater volume.
A kind of 9. removal COD containing salad oil wastewater method as claimed in claim 8, it is characterised in that：Described inoculum concentration
To handle the 1.5-2.5% (v/v) of wastewater volume.
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