WO2013166611A1 - Method for simultaneous biological removal of nitrogen compounds and xenobiotics of wastewaters - Google Patents
Method for simultaneous biological removal of nitrogen compounds and xenobiotics of wastewaters Download PDFInfo
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- WO2013166611A1 WO2013166611A1 PCT/CH2012/000099 CH2012000099W WO2013166611A1 WO 2013166611 A1 WO2013166611 A1 WO 2013166611A1 CH 2012000099 W CH2012000099 W CH 2012000099W WO 2013166611 A1 WO2013166611 A1 WO 2013166611A1
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- biodegradation
- xenobiotics
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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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- 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
- C02F3/30—Aerobic and anaerobic processes
- C02F3/302—Nitrification and denitrification treatment
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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
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/301—Detergents, surfactants
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/305—Endocrine disruptive agents
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/306—Pesticides
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/308—Dyes; Colorants; Fluorescent agents
Definitions
- the present invention concerns a method for simultaneous biological removal of nitrogen compounds and xenobiotics of wastewaters using aerobic granular biomass.
- Microbial granulation was noticed in anaerobic wastewater treatment and was first described by G. Lettinga, K.C. Pette, R. de Vletter and E. Wind, Anaerobic treatment of beet sugar wastewater on semi-technical scale. CSM-report. Amsterdam, The Netherlands (1977). Whereas, the formation and application of aerobic granules has been reported by the late 1990s (Morgenroth E, Sherden T, van Loosdrecht MCM, Heijnen JJ, Wilderer PA. Aerobic granular sludge in a sequencing batch reactor. Water Res 1997;31 :3191-4.).
- granular sludge formation can be initiated by adding artificial compounds or aggregation with protozoa (Japanese patent JP4354596) or using mycelium yeasts as carriers of bacteria (international application published under Nr. WO9837027). Natural granulation process can also occur without the addition of supporting material. Different theories explain the cause of granular sludge formation.
- PCT application WO 03/070649 "Aerobic biomass granules for waste water treatment" describes the aerobic granular sludge formation in column reactor, with different process cycles i.e. filling, aeration, mixing and settling.
- microbial biomass undergoes starvation process, which increases hydrophobicity and facilitate microbial adhesion and aggregation. Additionally, controlled hydrodynamic shear and flow pattern in bioreactor contribute to granular sludge formation.
- Aerobic granules characteristics depend on the seed sludge, substrate and process conditions. In most studies, aerobic granules were produced by activated sludge as seed. Generally, the greater the number of hydrophobic bacteria in the seed sludge, the faster the aerobic granulation process occurs (Wilen BM, Onuki M, Hermansson M, Lumley D, Mino T. Microbial community structure in activated sludge flock analyzed by fluorescence in situ hybridization and its relation to flock stability. Water Res 2007. doi:10.1016/j.watres.2007.12.013).
- the invention relates only to one specific bacterial formulation, which degrades one specific compound, i.e. phenol, and
- phenol is completely biodegraded during aerobic process i.e. used for microbial growth.
- the object of the present invention is to provide a method to overcome the previously described drawbacks of prior art especially referring WO 2008/020818.
- Simultaneous nitrigen and organic compounds removal is described by Yang et.al. ( Yang SF, Tay JH, Liu Y. Inhibition of free ammonia to the formation of aerobic granules. Biochem Eng J 2004b;17:41-8.), whereas the organic compound was acetate.
- the primary goal of invention is improvement of microbiological quality of nitrogen removing aerobic granules with selected strains of microorganisms for biodegradation of xenobiotics in industrial and mixed industrial and municipal wastewater.
- Industrial wastewaters are considered all wastewaters, which contain artificial organic compounds with complex chemical structure, xenobiotics and/or high nitrogen compounds concentration. These wastewaters include wastewaters from e.g. pharmaceutical and chemical production, such as production of active pharmaceutical ingredients, pharmaceutical intermediaries, antibiotics, dyestuff, pesticides, herbicides etc. Additionally, the following wastewaters can also contain complex organic compounds and nitrogen compounds: coke industry, coal gasification industry, detergent industry, synthetic resins industry, textile industry etc. and need to be treated alone or mixed with municipal wastewater.
- Secondary goal of invention is simultaneous nitrogen removal and biodegradation of complex organic compounds present in the wastewater.
- Simple organic compounds further on can be readily used as additional carbon source for denitrification.
- Stabile denitrification favors the aggregation of selected nitrogen removal microorganisms with selected strains of biodegradation bacteria into granular structure.
- said selected strains of heterotrophic bacteria which carry out biodegradation, also detoxify xenobiotics, which ensure stabile nitrification process.
- the xenobiotic biodegradation products such as obtained are simple organic compounds, which are additional carbon source for denitrification and to facilitate a granulation of preselected mixed microbial of nitrogen removing bacteria and selected strain for biodegradation of target xenobiotic.
- the granulation is carried out according to the following steps:
- decanting and filling propagator with a new quantity of liquid Said granulation is advantageously carried out in a separate bioreactor- propagator on the liquid cultivation media under conventional environmental conditions.
- the aerobic granules are produced without addition artificial carriers, or mycelium, filamentous or other type of microorganisms.
- Cultivated aerobic granules from propagator are preferably added into main wastewater treatment bioreactor.
- Other benefits of this invention are better biomass settling characteristics due to dense and compact structure of aerobic granules, lower sludge production and consequently lower sludge treatment costs.
- the present invention is applicable to all biological wastewater treatment systems, which remove nitrogen compounds.
- the present invention and its advantages will be better understood by the following description of embodiments given as non-limiting examples.
- aerobic granules are formed using nitrifying microorganisms (Glancer- Soljan, M., Ban, S., Dragicevic, T., Soljan, V., Matic, V., "Granulated mixed microbial culture suggesting successful employment of bioaugmentation in the treatment of process wastewaters".
- CABEQ 15 (3), 87-94, 2001 from internal microbial collection and isolated and selected heterotrophic strains for biodegradation of target xenobiotics.
- Isolation of heterotrophic strains for biodegradation of target xenobiotics is done from activated sludge on solid microbial media in Petry dishes.
- Solid microbial media is prepared using synthetic substrate, as presented in examples of this invention or the influent or effluent wastewater, which contains target xenobiotics.
- heterotrophic strains are selected based on their growth activity.
- the said media is used for selection of nitrifying strains, based on their tolerance to grow in the presence of xenobiotics.
- Selected microbial strains cultivated on solid media are transferred into bioreactor in order to prepare improved granular biomass and continue cultivation of granular biomass.
- propagators are usually 2-5% of the main bioreactor treatment volume. Cultivation is carried out on synthetic substrate containing the following mineral salts: (NH4)2SO4 2g/L, K2HPO4 1g/L, MgSO4 0.5g/L, FeSO4 0.4g/L, NaCI 0.4g/L, CaCO3 1g/L and MgCO3 1g/L with addition of target xenobiotics.
- the synthetic substrate is diluted up the concentration that are expected to be in the original wastewater.
- synthetic substrate can be mixed with real wastewater in ratio 10-50% of real wastewater. Formation of aerobic granules in propagators is carried out into several steps:
- Steps 3 to 7 are repeated until sufficient quantity of granular biomass is cultivated for harvesting. Once the microbial biomass is cultivated in propagators, it is added into main bioreactor on batch, semi continuous or continuous manner.
- Example 1 Microbial composition and process parameters for biodegradation of wastewater with LAS (linear alkylbenzene sulfonate).
- Mixed microbial culture is prepared using preselected nitrifying strains from our collection, such as: Nitrosomonas europa, Nitrosovibrio tenuis and Nitrobacter agilis and isolated and selected heterotrophic bacteria from activated sludge for LAS biodegradation and denitrification, such as: Pseudomonas putida and Pseudomonas aeruginosa.
- the synthetic substrate for preparation and cultivation of improved granular biomass consists of following mineral salts: (NH4)2S04 2g/L, K2HPO4 1g/L, MgS04 0.5g/L, FeS04 0.4g/L, NaCI 0.4g/L, CaC03 1g/L and MgC03 1g/L
- Preparation and cultivation of improved granular biomass is done in laboratory bioreactor with 5L volume.
- Bioreactor is filled with diluted synthetic media up to the ammonia concentration of 30-50 mg/L and LAS was added in concentration of 50-200 mg/L.
- Prepared microbial biomass of nitrifying strains and heterotrophic bacteria is added into bioreactor in order to have total biomass concentration of 2-6 g/L.
- Process in done with intermittent aeration and mixing in order to provide dissolved oxygen concentration in range 0.8-2.5 mg/L. Due to drop in pH value, it is necessary to maintained in the range 7.4-8.2 with 2 mol/L NaOH. Reaction time for complete process is 48-72 hours.
- Process is controlled via gas chromatography and UV spectrophotometry, in order to analyze the presence of low molecular weight fatty acids, aldehides and alcohols and nitrate or nitrite under aerobic conditions. After complete conversion of ammonia to nitrite or nitrate within 24-48 hours, the process is switched on denitrification in order to remove nitrite or nitrate and simple organic compounds. Parallel, during denitrification the granulation of biomass occurs.
- Example 2 Microbial composition and process parameters for biodegradation of wastewater with DMF (dimethylformamide).
- Mixed microbial culture is prepared using preselected nitrifying strains from our collection, such as: Nitrosomonas europa, Nitrosovibrio tenuis and Nitrobacter agilis and isolated and selected heterotrophic bacteria from activated sludge for DMF biodegradation and denitrification, such as: Pseudomonas luteola and Chromobacterium violaceum.
- the synthetic substrate for preparation and cultivation of improved granular biomass consists of following mineral salts: (NH4)2S04 2g/L, K2HP04 1 g/L, MgS04 0.5g/L, FeS04 0.4g/L, NaCI 0.4g/L, CaC03 1g/L and MgCO3 1g/L.
- Preparation and cultivation of improved granular biomass is done in laboratory bioreactor with 5L volume. Bioreactor is filled with diluted synthetic media up to the ammonia concentration of 15-20 mg/L and DMF was added in concentration of 300-500 mg/L. Prepared microbial biomass of nitrifying strains and heterotrophic bacteria is added into bioreactor in order to have total biomass concentration of 2-4 g/L. Process in done with intermittent aeration and mixing in order to provide dissolved oxygen concentration in range 0.5-3.5 mg/L. pH value is maintained in the range 7.4-8.5 with 2 mol/L NaOH. Reaction time for complete process is 24-48 hours.
- Process is controlled via gas chromatography and UV spectrophotometry, in order to analyze the presence of methanol, formic acid and nitrate or nitrite under aerobic conditions. After complete conversion of ammonia to nitrite or nitrate within 12-24 hours, the process is switched on denitrification in order to remove nitrite or nitrate and simple organic compounds. Parallel, during denitrification the granulation of biomass occurs.
- Example 3 Microbial composition and process parameters for biodegradation of wastewater with NSA (naphthalenesulfonic acids).
- Mixed microbial culture is prepared using preselected nitrifying strains from our collection, such as: Nitrosomonas europa, Nitrosovibrio tenuis and Nitrobacter agilis and isolated and selected heterotrophic bacteria from activated sludge for NSA biodegradation and denitrification, such as: Pseudomonas desmolityca, Pseudomonas cepacia and Pseudomonas acidovorans.
- the synthetic substrate for preparation and cultivation of improved granular biomass consists of following mineral salts: (NH4)2S04 2g/L, K2HP04 1g/L, MgS04 0.5g/L, FeS04 0.4g/L, NaCI 0.4g/L, CaC03 1g/L and MgC03 1g/L.
- Preparation and cultivation of improved granular biomass is done in laboratory bioreactor with 5L volume.
- Bioreactor is filled with diluted synthetic media up to the ammonia concentration of 10-20 mg/L and NSA (e.g. 6A2NSK - 6-amino-2- naphthalen sulfonic acid) was added in concentration of 300-500 mg/L.
- NSA e.g. 6A2NSK - 6-amino-2- naphthalen sulfonic acid
- Prepared microbial biomass of nitrifying strains and heterotrophic bacteria is added into bioreactor in order to have total biomass concentration of 3-5 g/L.
- Process in done with intermittent aeration and mixing in order to provide dissolved oxygen concentration in range 0.5-4.5 mg/L. Due to drop in pH value, it is necessary to maintained in the range 6.8-8.2 with 2 mol/L NaOH.
- Reaction time for complete process is 48-96 hours.
- Process is controlled via gas chromatography and UV spectrophotometry, in order to analyze the presence of gentisic acid, pyruvate and nitrate or nitrite under aerobic conditions. After complete conversion of ammonia to nitrite or nitrate within 24-60 hours, the process is switched on denitrification in order to remove nitrite or nitrate and simple organic compounds. Parallel, during denitrification the granulation of biomass occurs.
- Invention can be applied on treatment plants for biological wastewater treatment of industrial and mixed (industrial and municipal) wastewater, using prepared aerobic granules of mixed microbial cultures for nitrogen removal together with selected strains for biodegradation of xenobiotics. No specific working conditions, materials and equipment are necessary.
- the application of this invention can be carried out by persons with general education in wastewater treatment.
Abstract
The present invention concerns a method for simultaneous biological removal of nitrogen compounds and xenobiotics of wastewaters using aerobic granular biomass. Aerobic granules of the aerobic granular biomass, which consist of mixed microbial cultures for nitrogen removal, are improved by selected strains of heterotrophic bacteria for biodegradation of target xenobiotics. These selected strains of heterotrophic bacteria, which carry out biodegradation, also detoxify xenobiotics, which ensure stabile nitrification process. The xenobiotic biodegradation products such as obtained are simple organic compounds, which are additional carbon source for denitrification and to facilitate a granulation of preselected mixed microbial of nitrogen removing bacteria and selected strain for biodegradation of target xenobiotic.
Description
METHOD FOR SIMULTANEOUS BIOLOGICAL REMOVAL OF NITROGEN COMPOUNDS AND XENOBIOTICS OF WASTEWATERS
Technical Field
The present invention concerns a method for simultaneous biological removal of nitrogen compounds and xenobiotics of wastewaters using aerobic granular biomass.
Background Art
Microbial granulation was noticed in anaerobic wastewater treatment and was first described by G. Lettinga, K.C. Pette, R. de Vletter and E. Wind, Anaerobic treatment of beet sugar wastewater on semi-technical scale. CSM-report. Amsterdam, The Netherlands (1977). Whereas, the formation and application of aerobic granules has been reported by the late 1990s (Morgenroth E, Sherden T, van Loosdrecht MCM, Heijnen JJ, Wilderer PA. Aerobic granular sludge in a sequencing batch reactor. Water Res 1997;31 :3191-4.).
Such process is already known and described in PCT application: WO 98/37027, "Method for acquiring grain-shaped growth of a microorganism in a reactor" which describes the aerobic granular sludge preparation under aeration and mixing conditions, using mycelium yeasts as carriers of bacteria in order to facilitate granulation. Later on, PCT application WO 02/18563 "Method for the preparation of stabile and reusable biosensing granules", described granular sludge formation using natural polymers, e.g. sodium alginate, to form biosensing granules.
Based on available literature data and patents, granular sludge formation can be initiated by adding artificial compounds or aggregation with protozoa (Japanese patent JP4354596) or using mycelium yeasts as carriers of bacteria (international application published under Nr. WO9837027).
Natural granulation process can also occur without the addition of supporting material. Different theories explain the cause of granular sludge formation. PCT application WO 03/070649 "Aerobic biomass granules for waste water treatment" describes the aerobic granular sludge formation in column reactor, with different process cycles i.e. filling, aeration, mixing and settling. During the aeration period microbial biomass undergoes starvation process, which increases hydrophobicity and facilitate microbial adhesion and aggregation. Additionally, controlled hydrodynamic shear and flow pattern in bioreactor contribute to granular sludge formation.
Aerobic granules characteristics depend on the seed sludge, substrate and process conditions. In most studies, aerobic granules were produced by activated sludge as seed. Generally, the greater the number of hydrophobic bacteria in the seed sludge, the faster the aerobic granulation process occurs (Wilen BM, Onuki M, Hermansson M, Lumley D, Mino T. Microbial community structure in activated sludge flock analyzed by fluorescence in situ hybridization and its relation to flock stability. Water Res 2007. doi:10.1016/j.watres.2007.12.013). Referring to microbial diversity in aerobic granules, their application in wastewater treatment varies from simple chemical compounds removal, such as nitrogen and phosphorous (Yang SF, Tay JH, Liu Y. Inhibition of free ammonia to the formation of aerobic granules. Biochem Eng J 2004b; 17:41 -8.), up to the treatment of high strength organic wastewater (Moy BYP, Tay JH, Toh SK, Liu Y, Tay STL. High organic loading influences the physical characteristics of aerobic sludge granules. Lett Appl Microbiol 2002;34:407-12.). A.M. Maszenen et. al. in "Bioremediation of wastewater with recalcitrant organic compounds and metals by aerobic granules. Biotechnology Advances, 29 (201 1), 1 1 1-123", describe potential application of aerob:c granular sludge in treatment of different organic compounds. PCT application WO 2008/020818 "Granular microbial formulation capable of self aggregation used in treatment of wastewater" discloses a method for preparing specific microbial formulation in granular form for successful phenol removal. Said microbial formulation is prepared after isolation of the microbial strains growing
on specific substrate, i.e. phenol, and granulated as described in the PCT application WO 98/37027. >
The drawbacks of this invention are the following:
- the invention relates only to one specific bacterial formulation, which degrades one specific compound, i.e. phenol, and
most of industrial wastewaters contain complex organic compounds, xenobiotics and nitrogen,
phenol is completely biodegraded during aerobic process i.e. used for microbial growth.
Disclosure of Invention
The object of the present invention is to provide a method to overcome the previously described drawbacks of prior art especially referring WO 2008/020818. In particular, to provide method for producing improved aerobic granules for nitrogen removal with selected strains of microorganisms for biodegradation of target xeniobiotics and using biodegradation products as a carbon source for simultaneous nitrogen compounds and xenobiotics removal. Simultaneous nitrigen and organic compounds removal is described by Yang et.al. ( Yang SF, Tay JH, Liu Y. Inhibition of free ammonia to the formation of aerobic granules. Biochem Eng J 2004b;17:41-8.), whereas the organic compound was acetate. According to our best knowledge, there is no published results of simultaneous nitrogen compounds removal and xenobiotics. The primary goal of invention is improvement of microbiological quality of nitrogen removing aerobic granules with selected strains of microorganisms for biodegradation of xenobiotics in industrial and mixed industrial and municipal wastewater. Industrial wastewaters are considered all wastewaters, which contain artificial organic compounds with complex chemical structure, xenobiotics and/or high nitrogen compounds concentration. These wastewaters include wastewaters from e.g. pharmaceutical and chemical production, such as production of active pharmaceutical ingredients, pharmaceutical
intermediaries, antibiotics, dyestuff, pesticides, herbicides etc. Additionally, the following wastewaters can also contain complex organic compounds and nitrogen compounds: coke industry, coal gasification industry, detergent industry, synthetic resins industry, textile industry etc. and need to be treated alone or mixed with municipal wastewater.
Secondary goal of invention is simultaneous nitrogen removal and biodegradation of complex organic compounds present in the wastewater. Simple organic compounds further on can be readily used as additional carbon source for denitrification. Stabile denitrification favors the aggregation of selected nitrogen removal microorganisms with selected strains of biodegradation bacteria into granular structure.
These goals are reached by the method of the present invention such as defined in the preamble and characterized in that aerobic granules of the aerobic granular biomass, which consist of mixed microbial cultures for nitrogen removal, are improved by selected strains of heterotrophic bacteria for biodegradation of target xenobiotics. Additionally, parallel with biodegradation of complex organic compounds, xenobiotics, wastewater is detoxificated, which ensure stabile nitrification process.
Advantageously, said selected strains of heterotrophic bacteria, which carry out biodegradation, also detoxify xenobiotics, which ensure stabile nitrification process.
According to a preferred embodiment of the method of the invention, the xenobiotic biodegradation products such as obtained are simple organic compounds, which are additional carbon source for denitrification and to facilitate a granulation of preselected mixed microbial of nitrogen removing bacteria and selected strain for biodegradation of target xenobiotic.
Preferably, the granulation is carried out according to the following steps:
regeneration of preselected mixed culture of microorganisms containing autotrophic strains of bacteria on above said chemical substrate, addition of selected bacterial strain for biodegradation of target xenobiotic into a propagator,
addition of a liquid cultivation media into the propagator,
aeration of the propagator in order to ensure nitrification of ammonia and biodegradation target xenobiotics,
mixing in order to provide aggregation of preselected mixed culture of microorganisms and selected bacterial strain for biodegradation and use of biodegradation products as a carbon source for denitrification, settling of said aggregated biomass, and
decanting and filling propagator with a new quantity of liquid. Said granulation is advantageously carried out in a separate bioreactor- propagator on the liquid cultivation media under conventional environmental conditions.
The aerobic granules are produced without addition artificial carriers, or mycelium, filamentous or other type of microorganisms.
Cultivated aerobic granules from propagator are preferably added into main wastewater treatment bioreactor. Other benefits of this invention are better biomass settling characteristics due to dense and compact structure of aerobic granules, lower sludge production and consequently lower sludge treatment costs.
In broad embodiment, the present invention is applicable to all biological wastewater treatment systems, which remove nitrogen compounds.
The present invention and its advantages will be better understood by the following description of embodiments given as non-limiting examples.
Modes for carrying out the Invention
According to a scheme of preparation and cultivation of improved aerobic granules, aerobic granules are formed using nitrifying microorganisms (Glancer- Soljan, M., Ban, S., Dragicevic, T., Soljan, V., Matic, V., "Granulated mixed microbial culture suggesting successful employment of bioaugmentation in the treatment of process wastewaters". CABEQ 15 (3), 87-94, 2001), from internal microbial collection and isolated and selected heterotrophic strains for biodegradation of target xenobiotics. Isolation of heterotrophic strains for biodegradation of target xenobiotics is done from activated sludge on solid microbial media in Petry dishes. Solid microbial media is prepared using synthetic substrate, as presented in examples of this invention or the influent or effluent wastewater, which contains target xenobiotics. On the solid microbial media heterotrophic strains are selected based on their growth activity. In addition, the said media is used for selection of nitrifying strains, based on their tolerance to grow in the presence of xenobiotics. Selected microbial strains cultivated on solid media are transferred into bioreactor in order to prepare improved granular biomass and continue cultivation of granular biomass.
According to a on-site cultivation of granular biomass in propagators, propagators are usually 2-5% of the main bioreactor treatment volume. Cultivation is carried out on synthetic substrate containing the following mineral salts: (NH4)2SO4 2g/L, K2HPO4 1g/L, MgSO4 0.5g/L, FeSO4 0.4g/L, NaCI 0.4g/L, CaCO3 1g/L and MgCO3 1g/L with addition of target xenobiotics. In order to avoid toxic effects of ammonia and target xenobiotics, the synthetic substrate is diluted up the concentration that are expected to be in the original wastewater. In order to pre-adapt granular biomass on real wastewater, synthetic substrate can be mixed with real wastewater in ratio 10-50% of real wastewater.
Formation of aerobic granules in propagators is carried out into several steps:
1. regeneration of preselected mixed culture of microorganisms containing autotrophic strains of bacteria on above said chemical substrate,
2. addition of selected bacterial strain for biodegradation of target xenobiotic into the propagator,
3. addition of said liquid cultivation media into the propagator,
4. aeration of propagator in order to ensure nitrification of ammonia and biodegradation target xenobiotics,
5. mixing in order to provide aggregation of preselected mixed culture of microorganisms and selected bacterial strain for biodegradation and use of biodegradation products as a carbon source for denitrification,
6. settling of said aggregated biomass, and
7. decanting and filling propagator with new quantity of liquid Steps 3 to 7 are repeated until sufficient quantity of granular biomass is cultivated for harvesting. Once the microbial biomass is cultivated in propagators, it is added into main bioreactor on batch, semi continuous or continuous manner. The prepared aerobic granules according to the procedure described above in which aerobic granules have size between 0.5-2 mm, depending on xenobiotics added
The following three examples will explain in detail the process of the present invention.
Example 1 : Microbial composition and process parameters for biodegradation of wastewater with LAS (linear alkylbenzene sulfonate).
Mixed microbial culture is prepared using preselected nitrifying strains from our collection, such as: Nitrosomonas europa, Nitrosovibrio tenuis and Nitrobacter agilis and isolated and selected heterotrophic bacteria from activated sludge for
LAS biodegradation and denitrification, such as: Pseudomonas putida and Pseudomonas aeruginosa.
The synthetic substrate for preparation and cultivation of improved granular biomass consists of following mineral salts: (NH4)2S04 2g/L, K2HPO4 1g/L, MgS04 0.5g/L, FeS04 0.4g/L, NaCI 0.4g/L, CaC03 1g/L and MgC03 1g/L
Preparation and cultivation of improved granular biomass is done in laboratory bioreactor with 5L volume. Bioreactor is filled with diluted synthetic media up to the ammonia concentration of 30-50 mg/L and LAS was added in concentration of 50-200 mg/L. Prepared microbial biomass of nitrifying strains and heterotrophic bacteria is added into bioreactor in order to have total biomass concentration of 2-6 g/L. Process in done with intermittent aeration and mixing in order to provide dissolved oxygen concentration in range 0.8-2.5 mg/L. Due to drop in pH value, it is necessary to maintained in the range 7.4-8.2 with 2 mol/L NaOH. Reaction time for complete process is 48-72 hours. Process is controlled via gas chromatography and UV spectrophotometry, in order to analyze the presence of low molecular weight fatty acids, aldehides and alcohols and nitrate or nitrite under aerobic conditions. After complete conversion of ammonia to nitrite or nitrate within 24-48 hours, the process is switched on denitrification in order to remove nitrite or nitrate and simple organic compounds. Parallel, during denitrification the granulation of biomass occurs.
Example 2: Microbial composition and process parameters for biodegradation of wastewater with DMF (dimethylformamide).
Mixed microbial culture is prepared using preselected nitrifying strains from our collection, such as: Nitrosomonas europa, Nitrosovibrio tenuis and Nitrobacter agilis and isolated and selected heterotrophic bacteria from activated sludge for
DMF biodegradation and denitrification, such as: Pseudomonas luteola and Chromobacterium violaceum.
The synthetic substrate for preparation and cultivation of improved granular biomass consists of following mineral salts: (NH4)2S04 2g/L, K2HP04 1 g/L, MgS04 0.5g/L, FeS04 0.4g/L, NaCI 0.4g/L, CaC03 1g/L and MgCO3 1g/L.
Preparation and cultivation of improved granular biomass is done in laboratory bioreactor with 5L volume. Bioreactor is filled with diluted synthetic media up to the ammonia concentration of 15-20 mg/L and DMF was added in concentration of 300-500 mg/L. Prepared microbial biomass of nitrifying strains and heterotrophic bacteria is added into bioreactor in order to have total biomass concentration of 2-4 g/L. Process in done with intermittent aeration and mixing in order to provide dissolved oxygen concentration in range 0.5-3.5 mg/L. pH value is maintained in the range 7.4-8.5 with 2 mol/L NaOH. Reaction time for complete process is 24-48 hours. Process is controlled via gas chromatography and UV spectrophotometry, in order to analyze the presence of methanol, formic acid and nitrate or nitrite under aerobic conditions. After complete conversion of ammonia to nitrite or nitrate within 12-24 hours, the process is switched on denitrification in order to remove nitrite or nitrate and simple organic compounds. Parallel, during denitrification the granulation of biomass occurs.
Example 3: Microbial composition and process parameters for biodegradation of wastewater with NSA (naphthalenesulfonic acids).
Mixed microbial culture is prepared using preselected nitrifying strains from our collection, such as: Nitrosomonas europa, Nitrosovibrio tenuis and Nitrobacter agilis and isolated and selected heterotrophic bacteria from activated sludge for
NSA biodegradation and denitrification, such as: Pseudomonas desmolityca, Pseudomonas cepacia and Pseudomonas acidovorans.
The synthetic substrate for preparation and cultivation of improved granular biomass consists of following mineral salts: (NH4)2S04 2g/L, K2HP04 1g/L, MgS04 0.5g/L, FeS04 0.4g/L, NaCI 0.4g/L, CaC03 1g/L and MgC03 1g/L.
Preparation and cultivation of improved granular biomass is done in laboratory bioreactor with 5L volume. Bioreactor is filled with diluted synthetic media up to the ammonia concentration of 10-20 mg/L and NSA (e.g. 6A2NSK - 6-amino-2- naphthalen sulfonic acid) was added in concentration of 300-500 mg/L. Prepared microbial biomass of nitrifying strains and heterotrophic bacteria is added into bioreactor in order to have total biomass concentration of 3-5 g/L. Process in done with intermittent aeration and mixing in order to provide dissolved oxygen concentration in range 0.5-4.5 mg/L. Due to drop in pH value, it is necessary to maintained in the range 6.8-8.2 with 2 mol/L NaOH. Reaction time for complete process is 48-96 hours. Process is controlled via gas chromatography and UV spectrophotometry, in order to analyze the presence of gentisic acid, pyruvate and nitrate or nitrite under aerobic conditions. After complete conversion of ammonia to nitrite or nitrate within 24-60 hours, the process is switched on denitrification in order to remove nitrite or nitrate and simple organic compounds. Parallel, during denitrification the granulation of biomass occurs.
In this manner produced aerobic granules have capacity for nitrogen removal and parallel target xenobiotics. Since the granules are much heavier than activated sludge, they cannot be easily washed out of the biological treatment system and they further improve the settling properties of existing activated sludge onto they are added. While the foregoing written description of the invention enables one of ordinary skill to make and use what is considered
presently to be the best mode thereof, those of ordinary skill will understand and appreciate the existence of variations, combinations, and equivalents of the specific embodiment, method, and examples herein. The invention should therefore not be limited by the above described embodiment, method, and examples, but by all embodiments and methods within the scope and spirit of the invention.
Industrial Applicability
Invention can be applied on treatment plants for biological wastewater treatment of industrial and mixed (industrial and municipal) wastewater, using prepared aerobic granules of mixed microbial cultures for nitrogen removal together with selected strains for biodegradation of xenobiotics. No specific working conditions, materials and equipment are necessary. The application of this invention can be carried out by persons with general education in wastewater treatment.
Claims
Method for simultaneous biological removal of nitrogen compounds and xenobiotics of wastewaters using aerobic granular biomass, characterized in that aerobic granules of the aerobic granular biomass, which consist of mixed microbial cultures for nitrogen removal, are improved by selected strains of heterotrophic bacteria for biodegradation of target xenobiotics.
Method according to claim 1 , characterized in that said selected strains of heterotrophic bacteria, which carry out biodegradation, also detoxify xenobiotics, which ensure stabile nitrification process.
Method according to claims 1 and 2, characterized in that the xenobiotic biodegradation products such as obtained are simple organic compounds, which are additional carbon source for denitrification and to facilitate a granulation of preselected mixed microbial of nitrogen removing bacteria and selected strain for biodegradation of target xenobiotic.
Method according to claim 3, characterized in that the granulation is carried out according to the following steps:
- regeneration of preselected mixed culture of microorganisms containing autotrophic strains of bacteria on above said chemical substrate,
- addition of selected bacterial strain for biodegradation of target xenobiotic into a propagator,
- addition of a liquid cultivation media into the propagator,
- aeration of the propagator in order to ensure nitrification of ammonia and biodegradation target xenobiotics,
- mixing in order to provide aggregation of preselected mixed culture of microorganisms and selected bacterial strain for biodegradation and use of biodegradation products as a carbon source for denitrification,
- settling of said aggregated biomass, and
- decanting and filling propagator with a new quantity of liquid.
Method according to claim 4, characterized in that said granulation is carried out in the separate bioreactor-propagator on the liquid cultivation media under conventional environmental conditions.
Method according to claim 1 , characterized in that aerobic granules are produced without addition artificial carriers, or mycelium, filamentous or other type of microorganisms. 7. Method according to anyone of the preceding claims, characterized in that cultivated aerobic granules from propagator are added into main wastewater treatment bioreactor.
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| CN103787510A (en) * | 2014-02-28 | 2014-05-14 | 中国水产科学研究院淡水渔业研究中心 | Nutritional type filler for enriching indigenous microorganisms to purify water and preparation method thereof |
| CN104478091A (en) * | 2014-11-27 | 2015-04-01 | 新疆德蓝股份有限公司 | High-efficiency ammonia nitrogen degradation composite strain culture method |
| CN107585875A (en) * | 2016-07-08 | 2018-01-16 | 江西盖亚环保科技有限公司 | A kind of method of more biofacies zoogloea filler processing high concentrated organic wastewaters |
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| CN111285473A (en) * | 2019-12-28 | 2020-06-16 | 北京翰祺环境技术有限公司 | Method for promoting quick start of denitrification and decarburization system by using high-salt-resistant strain |
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