CN112410259B - Method for quickly forming anaerobic granular sludge - Google Patents

Method for quickly forming anaerobic granular sludge Download PDF

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CN112410259B
CN112410259B CN202011379999.7A CN202011379999A CN112410259B CN 112410259 B CN112410259 B CN 112410259B CN 202011379999 A CN202011379999 A CN 202011379999A CN 112410259 B CN112410259 B CN 112410259B
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郝安琪
邱沪生
袁鹏
杜宛霖
李萌
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Zhongzhi Jiangsu Environmental Construction Co ltd
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Abstract

The invention discloses a method for quickly forming anaerobic granular sludge, which can quickly promote sludge granulation by adding sphingosine-deficient bacterium (NZHB-L1) with rich carbohydrate on the surface. The surface sugar of sphingomonas paucimobilis (NZHB-L1) is favorable for other thalli to be attached to the surface of the microorganism to form large particles, so that the large particles are rapidly settled. The anaerobic granular sludge mainly comprises methane sarcina and methane hyphomycete, and when the components of the methane sarcina are more, the load of treatment is higher, and the degradation speed is higher. However, the methanosarcina sarcina has small particle size and low density and is easy to lose. The methane sarcina is easy to load on the surface of the microbial bacteria added in the invention, thereby forming large-particle thalli, accelerating the sedimentation rate of the thalli, effectively preventing the loss of the methane sarcina and improving the organic load rate of the system.

Description

Method for quickly forming anaerobic granular sludge
Technical Field
The invention belongs to the technical field of biological high technology, and particularly relates to a technology for quickly forming granular sludge by adding easy-to-agglomerate microorganisms.
Technical Field
The biological treatment of waste water is an important technology in environmental engineering and energy engineering, and is one of the important treatment methods for organic waste water. In the past, most of structures used for treating urban domestic sewage, organic waste and high-concentration organic wastewater are common digestion tanks, so that the defects of long hydraulic retention time and low organic load exist, and the application of anaerobic biological treatment in wastewater treatment is greatly limited. In recent decades, the development of various new industries and equipment has greatly improved the application of anaerobic biological treatment in the field of water treatment. Compared with aerobic biological treatment, the method mainly has the advantages of low energy consumption, high treatment load, less excess sludge, less nutrient requirement and the like.
The anaerobic granular sludge is the most central factor of the method, but the generation time of anaerobic microorganisms is long, the growth rate is low, and the sludge growth is slow, so the starting time of an anaerobic generator is generally long, and is usually 6-8 months. At present, various flocculating agents are generally added into an anaerobic reactor in engineering to promote the rapid granulation of the sludge.
Disclosure of Invention
The invention aims to develop a culture method for quickly forming anaerobic granular sludge aiming at actual problems and requirements in production practice, so that the culture time of the sludge is shortened, and the treatment load of the sludge is increased.
The invention achieves the purpose of rapidly promoting the sludge granulation by adding the bacteria with rich sugar coatings on the surfaces, and the bacteria with rich sugar coatings on the surfaces are beneficial to other thalli to be attached to the surfaces of the microorganisms to form large granules, thereby rapidly settling.
A culture method for quickly forming anaerobic granular sludge features that the bacterial strain rich in saccharide on its surface is added to reactor to make other bacterial bodies attached to the surface of said microbe to form large granules, resulting in quick settling.
The bacterial strain with rich glycocalyx on the surface is Sphingomonas paucimobilis (Sphingomonas paucimobilis) NZHB-L1, which is preserved in China general microbiological culture Collection center at 22 months 10 and 2020, address: no. 3 Xilu No. 1 Beijing, Chaoyang, and the preservation number is CGMCC No. 20939.
The method specifically comprises the following steps:
Figure DEST_PATH_IMAGE002
and adding inoculated sludge into the reactor and conveying sewage from a bottom device, wherein the COD of the sewage is 4000-5000 mg/L, the temperature in the device is 30-32 ℃, and the pH is controlled to be 7.0-7.3.
Figure DEST_PATH_IMAGE004
Setting the retention time to be 24h, and adding nutrient elements and trace elements.
The nutrient elements comprise the following components in percentage by weight: 0-1% of glucose, NH4Cl 0~1%，KH2PO4 0~1%，MgSO4 0~1%，NaCl 0~1%，CaCO30-1% of yeast extract and 0-0.1% of yeast extract.
Preferably, the nutrient elements comprise the following components in percentage by weight: glucose 0.6%, NH4Cl 0.2%，KH2PO4 0.1％，MgSO40.05％，NaCl 0.02％，CaCO30.3 percent of yeast extract and 0.01 percent of yeast extract. The pH value is maintained between 7.0 and 7.3.
The trace elements are as follows: fe. Co and Ni.
The mass ratio of the trace elements is as follows: fe 0-20 mg/L, Co 0-1 mg/L, Ni 0-1 mg/L.
Preferably, the dosage of the trace elements is as follows: fe 10mg/L, Co 0.1mg/L, Ni 0.2 mg/L.
Figure DEST_PATH_IMAGE006
And adding sphingosine paucimobilis NZHB-L1 into the reactor in a one-time adding mode. The mass ratio of the addition amount of the strain to the sludge inoculation is 0.1-10%.
The sphingomonas paucimobilis is obtained by screening sludge from certain lake bottom of Nanjing by the inventor, and is cultured by using an LB culture medium, wherein the culture medium comprises the following components: 5g/L yeast extract and 10g/L peptone, and the pH value is kept between 7.0 and 7.3.
The morphological characteristics of the strain of the invention are as follows:
thick and long rod-shaped with blunt ends and thick capsule around the thallus. Spore formation: thick wall, oval, round colony, colorless, raised, sticky, transparent or translucent, and neat edge.
The culture method of the strain comprises the following steps:
(1) and (3) activation: and (3) selecting a single colony of sphingomonas paucimobilis from the solid plate, transferring the single colony into an LB liquid culture medium, and carrying out shake culture at 37 ℃ for 3-4 h until the logarithmic phase at the rotation speed of 150 rpm on a shaking table. The LB culture medium comprises the following components: 5g/L yeast extract and 10g/L peptone, and the pH value is kept between 7.0 and 7.3.
(2) Transferring: transferring the sphingomonas paucimobilis liquid activated in the logarithmic phase in the process 1 to a 50L seeding tank according to the inoculation amount of 5% for culturing, controlling the temperature of the seeding tank to be maintained at 30-37 ℃, the rotating speed to be maintained at 220-250 rpm, controlling the dissolved oxygen DO to be 3-7 mg/L, and culturing for 24-48 h. The culture medium of the seeding tank comprises the following components in percentage by mass: glucose 0.6%, NH4Cl0.2%，KH2PO4 0.1％，MgSO4 0.05％，NaCl 0.02％，CaCO30.3 percent of yeast extract, 0.01 percent of yeast extract and the pH value maintained between 7.0 and 7.3.
(3) Expanding culture: transferring the sphingomonas paucimobilis solution cultured in the seeding tank of the process 2 into a 1000L fermentation tank according to the inoculum size of 2-5% for amplification culture, wherein the components of a culture medium of the fermentation tank are the same as those of the seeding tank, and the indexes of various physical and chemical parameters are as follows: the temperature is 37 ℃, the rotating speed is 250rpm, the dissolved oxygen is 8 mg/L, and the fermentation time is 48-60 h. After the fermentation is finished, the effective viable count of the tank body bacterial liquid can reach more than 300/ml, and the sphingomonas paucimobilis bacterial agent can be obtained after the fermentation culture liquid is taken out of the tank and packaged by a plastic barrel.
The present invention preferably employs the following conditions:
I. the mixing mode of water inlet at the bottom and water outlet at the top of the internal circulation is adopted.
II. The microelements such as Fe, Co, Ni and the like are directly supplemented into the reactor.
III, adopting a reactor produced by Wuhan Henghaxing experimental equipment Limited company, wherein the ratio H/D of the height to the diameter is 4-8.
Has the advantages that: the rapid culture method of the anaerobic granular sludge has the advantages that:
the added sphingosine paucimobilis (NZHB-L1) is easy to combine with microorganism bacteria in the sludge to form large-particle bacteria, thereby effectively preventing the loss of the bacteria and greatly shortening the culture time of the granular sludge.
The invention achieves the technology of rapidly promoting sludge granulation by adding sphingomonas paucimobilis (NZHB-L1) with rich carbohydrate on the surface. The microbial surface sugar is a layer of transparent colloidal substance outside a cell wall, the components of the transparent colloidal substance are xanthan gum and polypeptide, and the specific characteristics of the sugar-coated components are more favorable for other thalli to be attached to the surface of the microbe to form large particles. The anaerobic granular sludge mainly comprises methane sarcina and methane hyphomycete, and when the components of the methane sarcina are more, the load of treatment is higher, and the degradation speed is higher. However, the methanosarcina sarcina has small particle size and low density and is easy to lose. The methane sarcina is easy to load on the surface of the microbial bacteria added in the invention, thereby forming large-particle thalli. The culture time of the sludge granules is shortened from 120 days to 70 days.
Description of the drawings:
FIG. 1 is a diagram showing a colony of Sphingomonas paucimobilis (NZHB-L1).
FIG. 2 shows the anaerobic granular sludge formed after 10% Sphingomonas paucimobilis (NZHB-L1) was added to the inoculated sludge.
Detailed Description
The digested sludge of the industrial wastewater treatment plant described in the examples originates from: a secondary sedimentation tank of a pesticide chemical plant in Jiangsu salt city.
The sphingomonas paucimobilis is obtained by screening sludge at the bottom of a certain lake in Nanjing by the inventor, and the strain has the morphological characteristics that: thick and long rod-shaped with blunt ends and thick capsule around the thallus. Spore formation: wall thickness, oval, mesogenic or subtenogenic, no or micro-swelling of the blastocysts. The colony is round, colorless, raised, sticky, transparent or semitransparent, and has regular edges.
The culture method of sphingomonas paucimobilis NZHB-L1 comprises the following steps:
(1) and (3) activation: and (3) selecting a single colony of sphingomonas paucimobilis from the solid plate, transferring the single colony into an LB liquid culture medium, and carrying out shake culture at 37 ℃ for 3-4 h until the logarithmic phase at the rotation speed of 150 rpm on a shaking table. The LB culture medium comprises the following components: 5g/L yeast extract and 10g/L peptone, and the pH value is kept between 7.0 and 7.3.
(2) Transferring: transferring the sphingomonas paucimobilis liquid activated in the logarithmic phase in the process 1 to a 50L seed tank for culture according to the inoculation amount of 5 percent, controlling the temperature of the seed tank to be maintained at 30-37 ℃, and keeping the rotating speed at 220-25 DEG CControlling the dissolved oxygen DO at 3-7 mg/L at 0rpm, and culturing for 24-48 h. The culture medium of the seeding tank comprises the following components in percentage by mass: glucose 0.6%, NH4Cl0.2%，KH2PO4 0.1％，MgSO4 0.05％，NaCl 0.02％，CaCO30.3 percent of yeast extract, 0.01 percent of yeast extract and the pH value is maintained between 7.0 and 7.3.
(3) Expanding culture: transferring the sphingomonas paucimobilis solution cultured in the seeding tank of the process 2 into a 1000L fermentation tank according to the inoculum size of 2-5% for amplification culture, wherein the components of a culture medium of the fermentation tank are the same as those of the seeding tank, and the indexes of various physical and chemical parameters are as follows: the temperature is 37 ℃, the rotating speed is 250rpm, the dissolved oxygen is 8 mg/L, and the fermentation time is 48-60 h. After the fermentation is finished, the effective viable count of the tank body bacterial liquid can reach more than 300/ml, and the sphingosine-less monad microbial inoculum can be obtained after the fermentation culture liquid is taken out of the tank and packaged by a plastic barrel.
Example 1
Sludge inoculation: the method is characterized in that digested sludge of an industrial wastewater treatment plant is used as inoculated sludge, the initial COD of the wastewater is 4500mg/L, the temperature of a reactor is 31 ℃, the pH is controlled at 7.3, the VSS of the inoculated sludge is 15g/L, the SS is 35g/L, and the retention time is 24 h.
And (3) nutrient elements: glucose 0.6%, NH4Cl 0.2%，KH2PO4 0.1％，MgSO4 0.05％，NaCl 0.02％，CaCO30.3 percent of yeast extract and 0.01 percent of yeast extract.
Trace elements: the added trace elements Fe is 10mg/L, Co is 0.1mg/L, and Ni is 0.2 mg/L.
The reactor configuration: a reactor manufactured by Wuhan Hengmega xing laboratory facilities Ltd was used, and the ratio H/D of the height to the diameter of the reactor was 6.
Time to form granular sludge: and (4) 120 days.
Example 2
Sludge inoculation: the method is characterized in that digested sludge of an industrial wastewater treatment plant is used as inoculated sludge, the initial COD of the wastewater is 4500mg/L, the temperature of a reactor is 31 ℃, the pH is controlled at 7.3, the VSS of the inoculated sludge is 15g/L, the SS is 35g/L, and the retention time is 24 h.
Adding sphingosine paucimobilis (NZHB-L1) into a reactor filled with inoculated sludge, and adding sphingosine paucimobilis (NZHB-L1) into the reactor, wherein the adding amount of the sphingosine paucimobilis is estimated by the sludge inoculation amount, and the mass ratio of the sphingosine paucimobilis to the sludge inoculation amount is as follows: 1 percent.
And (3) nutrient elements: glucose 0.6%, NH4Cl 0.2%，KH2PO4 0.1％，MgSO4 0.05％，NaCl 0.02％，CaCO30.3 percent of yeast extract and 0.01 percent of yeast extract.
Trace elements: the added trace elements Fe is 10mg/L, Co is 0.1mg/L, and Ni is 0.2 mg/L.
The reactor configuration: a reactor manufactured by Wuhan Hengmega xing laboratory facilities Ltd was used, and the ratio H/D of the height to the diameter of the reactor was 6.
Time to form granular sludge: for 100 days.
Example 3
The inoculated sludge takes the digested sludge of an industrial wastewater treatment plant as inoculated sludge, the initial COD of the wastewater is 4500mg/L, the temperature of the reactor is 31 ℃, the pH is controlled at 7.3, the VSS of the inoculated sludge is 15g/L, the SS is 35g/L, and the retention time is 24 h.
Adding sphingosine paucimobilis (NZHB-L1) into a reactor filled with inoculated sludge, and adding sphingosine paucimobilis (NZHB-L1) into the reactor, wherein the adding amount of the sphingosine paucimobilis is estimated by the sludge inoculation amount, and the mass ratio of the sphingosine paucimobilis to the sludge inoculation amount is as follows: 5 percent.
And (3) nutrient elements: glucose 0.6%, NH4Cl 0.2%，KH2PO4 0.1％，MgSO4 0.05％，NaCl 0.02％，CaCO30.3 percent of yeast extract and 0.01 percent of yeast extract.
Trace elements: the added trace elements Fe is 10mg/L, Co is 0.1mg/L, and Ni is 0.2 mg/L.
The configuration of the reactor is as follows: a reactor manufactured by Wuhan Hengmega Happy laboratory facilities Ltd is used, and the ratio H/D of the height of the reactor to the diameter of the reactor is 6.
Time to form granular sludge: for 90 days.
Example 4
Sludge inoculation: the method is characterized in that digested sludge of an industrial wastewater treatment plant is used as inoculated sludge, the initial COD of the wastewater is 4500mg/L, the temperature of a reactor is 31 ℃, the pH is controlled at 7.3, the VSS of the inoculated sludge is 15g/L, the SS is 35g/L, and the retention time is 24 h.
Adding sphingosine paucimobilis (NZHB-L1) into a reactor filled with inoculated sludge, and adding sphingosine paucimobilis (NZHB-L1) into the reactor, wherein the adding amount of the sphingosine paucimobilis is estimated by the sludge inoculation amount, and the mass ratio of the sphingosine paucimobilis to the sludge inoculation amount is as follows: 8 percent.
And (3) nutrient elements: glucose 0.6%, NH4Cl 0.2%，KH2PO4 0.1％，MgSO4 0.05％，NaCl 0.02％，CaCO30.3 percent of yeast extract and 0.01 percent of yeast extract.
Trace elements: the added trace elements Fe is 10mg/L, Co is 0.1mg/L, and Ni is 0.2 mg/L.
The reactor configuration: a reactor manufactured by Wuhan Hengmega Happy laboratory facilities Ltd is used, and the ratio H/D of the height of the reactor to the diameter of the reactor is 6.
Time to form granular sludge: and (5) 85 days.
Example 5
Sludge inoculation: digested sludge of an industrial wastewater treatment plant is used as inoculated sludge, the initial COD of the wastewater is 4500mg/L, the temperature of a reactor is 31 ℃, the pH is controlled at 7.3, the VSS of the inoculated sludge is 15g/L, the SS is 35g/L, and the retention time is 24 h.
Adding sphingosine paucimobilis (NZHB-L1) into a reactor filled with inoculated sludge, and adding sphingosine paucimobilis (NZHB-L1) into the reactor, wherein the adding amount of the sphingosine paucimobilis is estimated by the sludge inoculation amount, and the mass ratio of the sphingosine paucimobilis to the sludge inoculation amount is as follows: 10 percent.
And (3) nutrient elements: glucose 0.6%, NH4Cl 0.2%，KH2PO4 0.1％，MgSO4 0.05％，NaCl 0.02％，CaCO30.3 percent of yeast extract and 0.01 percent of yeast extract.
Trace elements: the added trace elements Fe is 10mg/L, Co is 0.1mg/L, and Ni is 0.2 mg/L.
The reactor configuration: a reactor manufactured by Wuhan Hengmega Happy laboratory facilities Ltd is used, and the ratio H/D of the height of the reactor to the diameter of the reactor is 6.
Time to form granular sludge: for 70 days.

Claims (8)

1. A method for rapidly forming anaerobic granular sludge, characterized by: adding bacterial strains with surfaces rich in saccharide quilt into a reactor, and enabling other thalli to be attached to the surfaces of the bacterial strains with surfaces rich in saccharide quilt to form large particles so as to rapidly settle; the bacterial strain with rich glycocalyx on the surface is Sphingomonas paucimobilis (Sphingomonas paucimobilis) NZHB-L1, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, and has the preservation number of CGMCC No. 20939.
2. The method of claim 1, wherein the anaerobic granular sludge is formed by: the reactor conveys sewage from a bottom device, the COD of the sewage is 4000-5000 mg/L, the temperature in the device is 30-32 ℃, and the pH value is controlled to be 7.0-7.3.
3. The method of claim 1, wherein the anaerobic granular sludge is formed by: the strain is added into the reactor in a one-time adding mode.
4. The method for rapidly forming anaerobic granular sludge as claimed in claim 1, wherein: adding nutrient elements into the reactor, wherein the proportion of the nutrient elements is as follows: 0-1% of glucose, NH4Cl 0~1%，KH2PO4 0~1%，MgSO4 0~1%，NaCl 0~1%，CaCO30-1% of yeast extract, 0-0.1% of yeast extract and pH value maintained between 7.0-7.3.
5. The method for rapidly forming anaerobic granular sludge as claimed in claim 1, wherein: the ratio H/D of the height to the diameter of the reactor is 4-8.
6. The method of claim 4, wherein the anaerobic granular sludge is formed by: 0-0.1% of Fe, 0-0.1% of Co and 0-0.1% of Ni are added into the reactor.
7. A bacterial strain capable of quickly forming anaerobic granular sludge is classified and named as Sphingomonas paucimobilis (Sphingomonas paucimobilis) NZHB-L1, is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, and has a preservation number of CGMCC No. 20939.
8. Use of the strain of claim 7 for the rapid formation of anaerobic granular sludge.
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