CN105002127A - Composite bacterium and application thereof to treatment of garlic processing wastewater - Google Patents

Composite bacterium and application thereof to treatment of garlic processing wastewater Download PDF

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CN105002127A
CN105002127A CN201510524020.3A CN201510524020A CN105002127A CN 105002127 A CN105002127 A CN 105002127A CN 201510524020 A CN201510524020 A CN 201510524020A CN 105002127 A CN105002127 A CN 105002127A
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substratum
bacterium
content
fungus agent
composite fungus
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CN105002127B (en
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周俊冉
王曙光
韩新玉
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周俊冉
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14Fungi; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/30Organic compounds
    • C02F2101/40Organic compounds containing sulfur
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/32Nature of the water, waste water, sewage or sludge to be treated from the food or foodstuff industry, e.g. brewery waste waters

Abstract

The invention discloses a composite bacterium, which can be used for the degradation of COD (Chemical Oxygen Demand) and/or organic sulfides and pretreatment for the degradation of garlic processing wastewater. The composite bacterium is obtained by mixing fermentation broth of bacillus megaterium, pseudomonas stutzeri, aspergillus oryzae, klebsiella oxytoca and desulfovibrio desulfuricans, of which bacterial cell concentrations are 20 to 30 percent, 10 to 15 percent, 15 to 35 percent, 10 to 35 percent and 5 to 15 percent respectively. The composite bacterium can be applied to the degradation of COD and/or organic sulfides, and can also be applied to the pretreatment for the degradation of the garlic processing wastewater. By such a biological pretreatment method for the degradation of the garlic processing wastewater, the influence of changes in the conditions of COD and organic sulfide concentrations, water volumes and the like on a degradation effect can be reduced, and use of a plurality of chemical reagents in a subsequent process can be reduced and even avoided; the pretreated wastewater can be discharged up to the standard in an activated sludge treatment system.

Description

Composite fungus agent and the application in the process of garlic processing waste water thereof
Technical field
The present invention relates to a kind of composite fungus agent, particularly relate to that one can be applied to COD degradation, organic sulfide degraded, garlic processing waste water are degraded pretreated composite fungus agent, belong to industrial microorganism field.
Background technology
Garlic processing waste water is the waste water discharged in Natural Garlic processing.Garlic processing waste water is based on natural organic matter (as polysaccharide, lipid material etc.), and not containing toxic substance, biodegradable composition is many.Although garlic processing waste water is not containing toxic substance, containing a large amount of biodegradable organic substance, and COD content is about about 20000mg/L.If directly enter water body without process, need to consume dissolved oxygens a large amount of in water, cause water hypoxia, make hydrobiont dead.Suspended substance in waste water sinks under water, and worsens water quality.Under anaerobic decompose, the organic sulfide be rich in garlic processing waste water produces special foul smell, serious environment pollution.
Garlic processing waste water generally adopts conventional biochemical method to administer, but there is inherent defect: one, garlic processing waste water is a kind of very special waste water, garlicin is wherein a kind of wide-spectrum bactericide, adopt common activated sludge aeration method process, garlicin has strong restraining effect to active bacteria, reduce sludge activity, the degradation capability of water body COD is very poor.Two, garlic processing waste water COD is about about 20000mg/L, far above the dosis tolerata (about <3000mg/L) of active sludge, the restraining effect of garlicin in addition, as adopted the process of activated sludge aeration method, can use after raw wastewater must being diluted 10 ~ 20 times, garlic pieces per ton consumes about 20 ~ 40 cubic metres, tap water, causes the serious waste of water resources.
The key that biochemical process improvement process garlic processing waste water exists inherent defect is, garlic processing waste water contains antibacterial, the natural organic sulfides of sterilization such as garlicin, to bacterium, there is very strong lethality, thus T suppression cell division, the eubolism of destroy microorganisms, cause severe jamming to active sludge treatment below, active sludge biological system is very easily subject to the paralysis impacting, even cause system.Have research that garlic wastewater is introduced in municipal sewage plant at present and carry out aeration biochemical process, the germicidal action due to the garlic sulfide of high density makes sludge decomposition and counts out; Traditional anaerobic biological treatment method ends in failure equally, and its reason is also that the garlic wet goods of high density has kill activity to virus, bacterium, makes its efficiency being difficult to reach industrial application and level.In addition quantity discharged changes greatly, and concentration is high, and COD value can reach the high concentrated organic wastewater of 10000mg/L ~ 20000mg/L, further increases the difficulty of garlic wastewater process.Therefore pre-treatment must be carried out to high concentrated organosulfur compound garlic wastewater, to alleviate the impact on subsequent biological treatment unit.
The removal of heat pre-treatment method to organic sulfides such as garlicins has certain effect, but its action effect can not meet needs of production.Fe-C Micro Electrolysis Method is widely used in waste water treatment engineering, and domestic have research to adopt Fe-C Micro Electrolysis Method to carry out pre-treatment to garlic section waste water, has certain effect to Wastewater Pretreatment.But this technique is unstable in process waste water process, and a cycle of operation of light electrolysis post is about one month, and after this need to rinse with diluted acid, reproducibility is poor.A large amount of iron carbon can be consumed simultaneously, adjust the soda acid of pH and the expenditure on power of aeration, in man power and material, all can not reach good effect.Run through some scene, water treatment plant domestic at present and show, acid-base pretreatment-Fe-C micro electrolysis-flocculation-Fenton combined pretreatment technique has effect to removal garlic wastewater organic sulfide.But employ the harmful influence such as a large amount of acid, alkali, PAC, PAM, fenton reagent in this technological process; huge threat is caused to environment; also bring pressure for follow-up biochemical treatment, the consumption of annual a large amount of material such as chemical reagent and iron carbon also substantially increases cost of water treatment.
Summary of the invention
Object of the present invention is exactly for the deficiencies in the prior art, provides one can be applied to the pretreated mixed bacterial of garlic processing waste water, composite fungus agent.
For achieving the above object, first the present invention provides a kind of mixed bacterial, and its technical scheme is as follows:
A kind of composite fungus agent, is characterized in that: mixed by the fermented liquid of bacillus megaterium (Bacillus megaterium), pseudomonas stanieri (Pseudomonas stutzeri), aspergillus oryzae (Aspergillus oryzae), acid-producing Klebsiella bacterium (Klebsiella oxytoca), desulfovibrio desulfurican (Desulfovibrio desulfuricans).
With optimal conditions, in above-mentioned composite fungus agent, mycetocyte percentage is bacillus megaterium content 20% ~ 30%, pseudomonas stanieri content 10% ~ 15%, aspergillus oryzae content 15% ~ 35%, acid-producing Klebsiella bacterium content 10% ~ 35%, desulfovibrio desulfurican content 5% ~ 15%.
Further, in above-mentioned composite fungus agent, mycetocyte percentage is bacillus megaterium content 25%, pseudomonas stanieri content 15%, aspergillus oryzae content 25%, acid-producing Klebsiella bacterium content 25%, desulfovibrio desulfurican content 10%.
Through verification experimental verification, above-mentioned arbitrary composite fungus agent can be applied to COD degradation and/or organic sulfide degraded.In garlic processing field, above-mentioned arbitrary composite fungus agent can be applied to the pre-treatment of garlic processing waste water, for removing COD degradation and/or organic sulfide.
Adopting above-mentioned arbitrary composite fungus agent can be applied to the degradation method of COD and/or organic sulfide, is that composite fungus agent is inoculated into degraded substrate, and degraded substrate is containing COD and/or organic sulfide; Adopt fermentative degradation, culture condition is 5 DEG C ~ 35 DEG C, shaking table or micro-aeration, control dissolved oxygen≤3mg/L.
The present invention provides with above-mentioned arbitrary composite fungus agent as activeconstituents simultaneously, simultaneously prepare routine techniques according to composite bacteria and be equipped with the necessary Organic nutrient such as glucose, sucrose, peptone, and the composite fungus agent of the component of the organic slat solution such as a small amount of ammonium salt, phosphoric acid salt, sylvite.Gained can be applied to COD degradation and/or organic sulfide degraded.In garlic processing field, the pre-treatment of garlic processing waste water can be applied to.
Adopting above-mentioned arbitrary composite fungus agent can be applied to the degradation method of COD and/or organic sulfide, is that composite fungus agent is inoculated into degraded substrate, and degraded substrate is containing COD and/or organic sulfide; Adopt fermentative degradation, culture condition is 5 DEG C ~ 35 DEG C, shaking table or micro-aeration, control dissolved oxygen≤3mg/L.
Compared with prior art, the invention has the beneficial effects as follows: (1) provides a kind of composite fungus agent be made up of 5 kinds of bacterium, COD degradation and/or organic sulfide degraded can be applied to, also can be applied to the pre-treatment of garlic processing waste water degradation treatment.(2) application method of composite fungus agent in COD degradation and/or organic sulfide degraded is provided.Composite fungus agent of the present invention utilizes 5 kinds of bacterial classification component populations mutually to work in coordination with, and strengthens the degraded degradation efficiency of substrate and shock-resistant ability, degradation method is with strong points, the time of response is fast, simple to operate.(3) biologic pretreatment method of the garlic processing waste water degraded realized by composite fungus agent is provided, by the advantage of composite fungus agent, the condition such as COD and organic sulfide substrate concentration, the water yield in garlic processing waste water that can greatly reduce changes the impact produced degradation effect, ensures that pretreating effect is stablized.And effectively can reduce the use to the chemical reagent such as acid, alkali, PAC, PAM, Fenton in the garlic processing waste water degradation technique even avoided after pre-processing; Pretreated waste water enters active sludge processing system, can realize qualified discharge.
Accompanying drawing explanation
Fig. 1 is that embodiment seven garlic processing waste water COD is containing spirogram.
Fig. 2 is that embodiment seven garlic processing waste water organic sulfide is containing spirogram.
Fig. 3 is that embodiment eight garlic processing waste water COD is containing spirogram.
Fig. 4 is that embodiment nine garlic processing waste water organic sulfide is containing spirogram.
Fig. 5 is that embodiment eight garlic processing waste water COD is containing spirogram.
Fig. 6 is that embodiment nine garlic processing waste water organic sulfide is containing spirogram.
Embodiment
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are further described.
Embodiment one: the preparation of composite fungus agent
1, bacterial classification
A group bacterial classification:
Bacillus megaterium (Bacillus megaterium): bacillus megaterium SWSB401
Pseudomonas stanieri (Pseudomonas stutzeri): pseudomonas stanieri SWSP001
Aspergillus oryzae (Aspergillus oryzae): aspergillus oryzae SWSA002
Acid-producing Klebsiella bacterium (Klebsiella oxytoca): Klebsiella oxytoca SWSK003
Desulfovibrio desulfurican (Desulfovibrio desulfuricans): desulfovibrio desulfurican SWSD001
B group bacterial classification:
Bacillus megaterium (Bacillus megaterium): bacillus megaterium ATCC12872
Pseudomonas stanieri (Pseudomonas stutzeri): pseudomonas stanieri ATCC17588
Aspergillus oryzae (Aspergillus oryzae): aspergillus oryzae ATCC 42149
Acid-producing Klebsiella bacterium (Klebsiella oxytoca): Klebsiella oxytoca ATCC43086
Desulfovibrio desulfurican (Desulfovibrio desulfuricans): desulfovibrio desulfurican ATCC27774
2, substratum
No. I substratum: gsh 1.5g, extractum carnis 7g, halfcystine 1g, glucose 15g, KCl 2g, (NH 4) 3pO 43H 2o 3g, K 2sO 42g, MgCl 23g, NaCl 4g, distilled water 1000mL, autoclaving, obtained substratum; Substratum mixes with degerming, autoclaved garlicin 2g after filtration again, obtained No. I substratum.
No. II substratum: gsh 1g, peptone 6g, glucose 5g, sucrose 5g, (NH 4) 3pO 43H 2o 3g, KCl 2g, NaCl 4g, MgCl 23g, K 2sO 41g, distilled water 1000mL, autoclaving, obtained substratum; Substratum mixes with degerming, autoclaved garlicin 5g after filtration again, obtained No. II substratum.
No. III substratum: peptone 3g, sucrose 10g, (NH 4) 3pO 43H 2o 2g, KCl 2g, K 2sO 41g, NaCl 4g, distilled water 1000mL, autoclaving, obtained substratum; Substratum mixes with degerming, autoclaved garlicin 20g after filtration again, obtained No. III substratum.
No. IV substratum: peptone 3g/L, sucrose 10g/L, wheat stalk (pulverizing) 3g/L, (NH 4) 3pO 43H 2o 2g/L, (NH 4) 2sO 45g/L, KCl 2g/L, NaCl 3g/L, distilled water 1000mL, garlic (crushing and beating) 15g/L, mixing obtained aqueous solution, makes No. IV substratum.
Nutrient agar plate medium: tryptone 10g, yeast extract 5g, sodium-chlor 5g, agar 15g, distilled water 1000mL; Pour flat board into and make nutrient agar plate medium.
LB substratum: peptone 10g, extractum carnis 3g, sodium-chlor 5g, agar 20g, distilled water 1000mL.
3, microbial inoculum preparation
Prepared by composite fungus agent A:
Step S1,5 kinds of bacterium of A group bacterial classification are inoculated into No. I substratum respectively, shaking table is cultivated, 20 DEG C, 150rpm cultivates 30h ~ 60h; After each reaches exponential phase, switching, until detected result often plants mycetocyte concentration>=10 in substratum 9cFU/mL;
Step S2,5 kinds of bacterium are forwarded to No. II substratum from No. I substratum respectively, 20 DEG C, 150rpm cultivates 30h ~ 60h; After each reaches exponential phase, switching, until mycetocyte concentration>=10 in often kind of substratum 9cFU/mL;
Step S3,5 kinds of bacterium are forwarded to No. III substratum from No. II substratum respectively, 20 DEG C, 150rpm cultivates 30h ~ 60h; After each reaches exponential phase, switching, until mycetocyte concentration>=10 in often kind of substratum 9cFU/mL;
Step S4, by 5 kinds of bacterium respectively from No. III culture medium inoculated to nutrient agar plate medium; Cultivate 36h ~ 64h for 25 DEG C ~ 30 DEG C, line, separation, switching, until grow more homogeneous, regular bacterium colony;
Step S5,5 kinds of bacterium are seeded to LB medium slant from nutrient agar plate medium respectively; Cultivate 48h, then respectively join in sterilized water for 25 DEG C, vibration 30min, until mycetocyte concentration>=10 in often kind of substratum 9cFU/mL;
Step S6, according to the mycetocyte percentage concentration ratio of bacillus megaterium 25%, pseudomonas stanieri 15%, aspergillus oryzae 25%, acid-producing Klebsiella bacterium 25%, desulfovibrio desulfurican 10%, 5 kinds of bacterium to be mixed, obtained composite fungus agent A of the present invention.
Prepared by composite fungus agent B:
According to above-mentioned steps S1 ~ step S6 same procedure and operation, 5 kinds of bacterium of B group bacterial classification are cultivated, obtains composite fungus agent B of the present invention.
Embodiment two: the preparation of composite fungus agent
Prepare composite fungus agent, itself and embodiment one something in common no longer repeat, and its difference is that in step S6, each bacterial classification mixture ratio example is different, specifically:
Prepared by composite fungus agent A:
Step S6, the mycetocyte percentage concentration ratio of 5 kinds of bacterial classifications of A group bacterial classification according to bacillus megaterium 20%, pseudomonas stanieri content 10%, aspergillus oryzae content 15%, acid-producing Klebsiella bacterium content 10%, desulfovibrio desulfurican content 5% to be mixed, obtained composite fungus agent A of the present invention.
Prepared by composite fungus agent B:
Step S6, the mycetocyte percentage concentration ratio of 5 kinds of bacterial classifications of B group bacterial classification according to bacillus megaterium 20%, pseudomonas stanieri content 10%, aspergillus oryzae content 15%, acid-producing Klebsiella bacterium content 10%, desulfovibrio desulfurican content 5% to be mixed, obtained composite fungus agent B of the present invention.
Embodiment three: the preparation of composite fungus agent
Prepare composite fungus agent, itself and embodiment one something in common no longer repeat, and its difference is that in step S6, each bacterial classification mixture ratio example is different, specifically:
Prepared by composite fungus agent A:
Step S6, the mycetocyte percentage concentration ratio of 5 kinds of bacterial classifications of A group bacterial classification according to bacillus megaterium 30%, pseudomonas stanieri content 15%, aspergillus oryzae content 35%, acid-producing Klebsiella bacterium content 35%, desulfovibrio desulfurican content 15% to be mixed, obtained composite fungus agent A of the present invention.
Prepared by composite fungus agent B:
Step S6, the mycetocyte percentage concentration ratio of 5 kinds of bacterial classifications of B group bacterial classification according to bacillus megaterium 30%, pseudomonas stanieri content 15%, aspergillus oryzae content 35%, acid-producing Klebsiella bacterium content 35%, desulfovibrio desulfurican content 15% to be mixed, obtained composite fungus agent B of the present invention.
Embodiment four: the amplification culture of composite fungus agent
Composite fungus agent A, B obtained by Example one, be inoculated in 2L No. II substratum, 25 DEG C, 150rpm shaking table cultivation>=48h respectively, to effective bacterium total concn 10 9more than CFU/mL, obtains one-level microbial inoculum seed liquor; Being inoculated in 20L No. IV substratum by cultivating the one-level microbial inoculum seed liquor well, cultivating in upper retort, control temperature 20 DEG C ~ 25 DEG C, dissolved oxygen 8mg/L, cultured continuously>=40h, to effective bacterium total concn 3 × 10 9cFU/mL, obtains secondary microbial inoculum seed liquor; Secondary microbial inoculum seed liquor is inoculated in 300L No. IV substratum, cultivates in the retort of upper 1m3, control temperature 20 DEG C ~ 25 DEG C, dissolved oxygen 8mg/L, cultured continuously>=42h, to effective bacterium total concn 3.5 × 10 9cFU/mL, obtains A, B two kinds of composite fungus agent seed liquor.
Embodiment five: the amplification culture of composite fungus agent
Composite fungus agent A, B obtained by Example two, according to embodiment four same operation, obtain A, B two kinds of composite fungus agent seed liquor.
Embodiment six: the amplification culture of composite fungus agent
Composite fungus agent A, B obtained by Example three, according to embodiment four same operation, obtain A, B two kinds of composite fungus agent seed liquor.
Embodiment seven: composite fungus agent is applied to garlic processing waste water Biological Pretreatment
1, garlic processing waste water
Garlic processing waste water takes from the garlic processing waste water of Jining of Shandong Province food garden discharge, and COD concentration 12000, organic sulfide 7000mg/L are determined in test.Garlic processing waste water distributes the distinctive irritating smell of organic sulfide.
2, test process and result
Getting triangular flask composite fungus agent seed liquor obtained for 50mL embodiment four and 300ml garlic processing waste water are mixed as adding bacterium group, separately getting triangular flask and 300ml garlic processing waste water is mixed as blank group with 50mL deionized water.
To add bacterium group and blank group be placed in simultaneously shaking table cultivate 25 DEG C, cultivate under 30rpm condition.During cultured continuously 38h, add the obfuscation of bacterium group waste water, and occur obvious aerogenesis phenomenon; Blank group is without considerable change.During cultured continuously 60h, adding bacterium group waste water is furvous, and irritating smell disappears; Blank group slightly in faint yellow, has obvious irritation smell.During cultured continuously 100h, add bacterium group and blank group difference is strengthened further.
Sample stage by stage in process of the test, adopt extinction turbidimetry for Determination garlic processing waste water organic sulfide, adopt potassium dichromate standard method to measure COD.Add bacterium group and blank group COD, organic sulfide content curve as shown in Figure 1, Figure 2.
Embodiment eight: composite fungus agent is applied to garlic processing waste water Biological Pretreatment
The method identical according to embodiment seven and operation, composite fungus agent seed liquor embodiment five obtained carries out Degrading experiment, adds bacterium group and blank group COD, organic sulfide content curve respectively as Fig. 3 ~ Fig. 4.
Embodiment nine: composite fungus agent is applied to garlic processing waste water Biological Pretreatment
The method identical according to embodiment seven and operation, composite fungus agent seed liquor embodiment six obtained carries out Degrading experiment, adds bacterium group and blank group COD, organic sulfide content curve respectively as Fig. 5 ~ Fig. 6 (embodiment six).
Above embodiment seven, eight, nine testing data display, composite fungus agent of the present invention enhances the degraded of COD, organic sulfide in garlic processing waste water greatly.
Embodiment ten: composite fungus agent is applied to garlic processing waste water Biological Pretreatment
1, waste water
Garlic processing waste water source is with embodiment seven, and COD concentration 11000, organic sulfide 7000mg/L are determined in test.Garlic processing waste water distributes the distinctive irritating smell of organic sulfide.
Sanitary wastewater takes from the sanitary wastewater discharge of food garden, Jining sewage work, and COD concentration 1000 is determined in test.
Waste water mixture: by garlic processing waste water and sanitary wastewater by volume 3:10 be mixedly configured into 1000L waste water mixture.
2, test process and result
1000L waste water mixture is placed in reaction flask, mud (water ratio 70%) 30g that embodiment four obtained composite fungus agent seed liquor 60L, EPP expanded particle 5g, settleability are good is added in reaction flask, in 30 DEG C of micro-aerations, under dissolved oxygen control≤3mg/L condition, cultured continuously 96h.Cultivate and terminate, waste water is furvous, and the distinctive smell of garlic processing waste water disappears.
Inject COD concentration 11000 garlic processing waste water to reaction flask, adopt continuum micromeehanics, dilution control is at 0.2/d.Continuous operation, waste water COD clearance is stabilized in about 73%, and organic sulfide clearance is stabilized in about 84%.Process terminates, and by A/O Sludge System on the waste water after process, COD stable water outlet is within 50mg/L.

Claims (8)

1. a composite fungus agent, is characterized in that: mixed by the fermented liquid of bacillus megaterium (Bacillus megaterium), pseudomonas stanieri (Pseudomonas stutzeri), aspergillus oryzae (Aspergillus oryzae), acid-producing Klebsiella bacterium (Klebsiella oxytoca), desulfovibrio desulfurican (Desulfovibriodesulfuricans).
2. composite fungus agent according to claim 1, is characterized in that: each component mycetocyte percentage is bacillus megaterium content 20% ~ 30%, pseudomonas stanieri content 10% ~ 15%, aspergillus oryzae content 15% ~ 35%, acid-producing Klebsiella bacterium content 10% ~ 35%, desulfovibrio desulfurican content 5% ~ 15%.
3. composite fungus agent according to claim 1, is characterized in that: each component mycetocyte percentage is bacillus megaterium content 25%, pseudomonas stanieri content 15%, aspergillus oryzae content 25%, acid-producing Klebsiella bacterium content 25%, desulfovibrio desulfurican content 10%.
4. the application of the arbitrary described composite fungus agent of claims 1 to 3 in COD degradation and/or organic sulfide degraded, or the application in garlic processing waste water degradation treatment.
5. application method according to claim 4, is characterized in that: mixed bacterial is inoculated into degraded substrate, adopts fermentative degradation, and culture condition is 5 DEG C ~ 35 DEG C, shaking table or micro-aeration, control dissolved oxygen≤3mg/L; Described degraded substrate is containing COD and/or organic sulfide and/or garlic processing waste water.
6. application method according to claim 5, is characterized in that: described culture condition is 25 DEG C, 30rpm shaking table, or, 30 DEG C, dissolved oxygen≤3mg/L.
7. a composite bacteria system, is characterized in that: with the arbitrary described composite fungus agent of claims 1 to 3 for activeconstituents.
8. the preparation method of the arbitrary described composite fungus agent of claims 1 to 3, is characterized in that implementing according to following steps:
Step S1,5 kinds of bacterium are inoculated into No. I substratum respectively, shaking table is cultivated, 20 DEG C, 150rpm cultivates 30h ~ 60h; After each reaches exponential phase, switching, until detected result often plants mycetocyte concentration>=10 in substratum 9cFU/mL;
Step S2,5 kinds of bacterium are forwarded to No. II substratum from No. I substratum respectively, 20 DEG C, 150rpm cultivates 30h ~ 60h; After each reaches exponential phase, switching, until mycetocyte concentration>=10 in often kind of substratum 9cFU/mL;
Step S3,5 kinds of bacterium are forwarded to No. III substratum from No. II substratum respectively, 20 DEG C, 150rpm cultivates 30h ~ 60h; After each reaches exponential phase, switching, until mycetocyte concentration>=10 in often kind of substratum 9cFU/mL;
Step S4, by 5 kinds of bacterium respectively from No. III culture medium inoculated to nutrient agar plate medium; Cultivate 36h ~ 64h for 25 DEG C ~ 30 DEG C, line, separation, switching, until grow more homogeneous, regular bacterium colony;
Step S5,5 kinds of bacterium are seeded to LB medium slant from nutrient agar plate medium respectively; Cultivate 48h, then respectively join in sterilized water for 25 DEG C, vibration 30min, until mycetocyte concentration>=10 in often kind of substratum 9cFU/mL;
Step S6, by each bacterium component according to the obtained composite fungus agent of mycetocyte percentage concentration ratio mixing;
Described No. I substratum: gsh 1.5g, extractum carnis 7g, halfcystine 1g, glucose 15g, KCl 2g, (NH 4) 3pO 43H 2o 3g, K 2sO 42g, MgCl 23g, NaCl 4g, distilled water 1000mL, autoclaving, obtained substratum; Substratum mixes with degerming, autoclaved garlicin 2g after filtration again, obtained No. I substratum;
Described No. II substratum: gsh 1g, peptone 6g, glucose 5g, sucrose 5g, (NH 4) 3pO 43H 2o 3g, KCl 2g, NaCl 4g, MgCl 23g, K 2sO 41g, distilled water 1000mL, autoclaving, obtained substratum; Substratum mixes with degerming, autoclaved garlicin 5g after filtration again, obtained No. II substratum;
Described No. III substratum: peptone 3g, sucrose 10g, (NH 4) 3pO 43H 2o 2g, KCl 2g, K 2sO 41g, NaCl 4g, distilled water 1000mL, autoclaving, obtained substratum; Substratum mixes with degerming, autoclaved garlicin 20g after filtration again, obtained No. III substratum;
Described No. IV substratum: peptone 3g/L, sucrose 10g/L, wheat stalk (pulverizing) 3g/L, (NH 4) 3pO 43H 2o 2g/L, (NH 4) 2sO 45g/L, KCl 2g/L, NaCl 3g/L, distilled water 1000mL, garlic (crushing and beating) 15g/L, mixing obtained aqueous solution, makes No. IV substratum;
Described nutrient agar plate medium: tryptone 10g, yeast extract 5g, sodium-chlor 5g, agar 15g, distilled water 1000mL; Pour flat board into and make nutrient agar plate medium;
LB substratum: peptone 10g, extractum carnis 3g, sodium-chlor 5g, agar 20g, distilled water 1000mL.
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Cited By (2)

* Cited by examiner, † Cited by third party
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