CN111733113A - COD (chemical oxygen demand) degrading strain and application thereof - Google Patents
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Abstract
The invention relates to the technical field of environmental microbiology, in particular to a COD degrading strain and application thereof, wherein the COD degrading strain is identified as Chryseobacterium indogenes (Chryseobacterium indicum), and the preservation number is CCTCC NO: m2018651. The chrysobacillus indolens chrysogenum indole CI-B4 is salt-resistant, low-temperature-resistant and wide in temperature application range (5-40 ℃); the capability of efficiently degrading organic pollutants in the wastewater is realized, and the effluent quality of the organic wastewater can be improved; the expanded culture can be applied to biochemical treatment of livestock and poultry breeding wastewater, industrial organic wastewater, river black and odorous water, kitchen waste anaerobic digestion biogas slurry and the like, and has high application value; the method is used for treating organic wastewater, and has the advantages of high treatment efficiency, good economic benefit, convenient operation and no pollution.
Description
Technical Field
The invention relates to the technical field of environmental microorganisms, in particular to a COD degrading strain and application thereof.
Background
With the acceleration of global industrialization process, organic pollution of water environment has become one of global environmental protection issues. The organic pollutants mainly come from the discharge of large-scale high-concentration organic wastewater, and mainly come from the fields of industry, agriculture and the like. The high concentration organic wastewater mainly refers to wastewater with Chemical Oxygen Demand (COD) reaching or exceeding thousands or even tens of thousands of milligrams per liter. The pollutant components in the high-concentration organic wastewater are complex, and the high-concentration organic wastewater poses serious threats to human health and ecological environment after being discharged into a water body. Therefore, the research on the technology for treating high-concentration organic wastewater is a research hotspot in the fields of environmental science and engineering at present.
The reason that the high-concentration organic wastewater is difficult to treat is determined by the characteristics of the high-concentration organic wastewater, and the high-concentration organic wastewater mainly has the following characteristics: high concentration of organic matter, complex composition, high salt content, unstable effluent quality of wastewater and the like. If the wastewater is improperly treated, the wastewater is directly discharged, and due to biodegradation, a receiving water body is anoxic, most aquatic organisms die, water quality is increasingly deteriorated, odor is generated, and environmental sanitation is influenced; meanwhile, pathogenic microorganisms discharged into the water body along with the wastewater can also be propagated in a large amount, and a large amount of toxic organic matters can be continuously accumulated and stored in natural environments such as the water body, soil and the like, and finally enter the human body to harm the health of the human body. At present, the treatment technology of high-concentration organic wastewater is divided into four technologies, namely biological treatment technology, physical treatment technology, chemical treatment technology and physical and chemical treatment technology. Compared with chemical treatment and physical treatment, the biological treatment is a method with lower implementation cost, higher efficiency, more thorough purification, environmental protection and no pollution, and is a mainstream method for solving the organic wastewater pollution in the future. The biological treatment technology is a method for degrading pollutants in the wastewater by using microorganisms to serve as self nutrition and energy and purifying the wastewater. In the implementation process of microbial treatment, the most important part is the strain with high-efficiency degradation capability, and the high-efficiency microbial strain can play a role in strengthening the treatment effect in wastewater treatment in various industries. In the research of organic wastewater treatment, COD is an important organic pollution parameter which can be rapidly measured, and can most directly reflect the pollution degree of wastewater. Therefore, in recent years, the research on the degradation of COD in various organic wastewater by microorganisms has been receiving more and more attention, and the screening of COD degrading bacteria of high-concentration organic wastewater is of great significance.
Disclosure of Invention
In view of the above-mentioned disadvantages of the prior art, the present invention aims to provide a COD degrading strain and its application, which are used to solve the problems in the prior art.
In order to achieve the above and other related objects, the present invention provides in a first aspect a COD degrading bacterial species identified as Chryseobacterium indogenes: (Chryseobacterium indologenes) The strain is preserved in China Center for Type Culture Collection (CCTCC) with the strain name ofChryseobacterium indologenesCI-B4, the preservation date is 2018.9.25, the preservation number is CCTCC NO: m2018651, the preservation address is Wuhan university in China.
Preferably, the Chryseobacterium indolerum isChryseobacterium indologenesCI-B4, which contains a gene sequence shown as SEQ ID NO. 1.
Preferably, the Chryseobacterium indolerum isChryseobacterium indologenesThe highest salt tolerance concentration of CI-B4 was 100 g/L sodium chloride.
Preferably, the Chryseobacterium indolerum isChryseobacterium indologenesThe COD tolerance concentration range of the CI-B4 is 0-30000 mg/L.
Preferably, the indole is producedChryseobacterium aureumChryseobacterium indologenesThe COD degradation rate of CI-B4 in the wastewater liquid culture medium for 48 hours reaches more than 80 percent.
In a second aspect, the invention provides a liquid microbial inoculum, which comprises Chryseobacterium indolerumChryseobacterium indologenesCI-B4, wherein the Chryseobacterium indolerum is produced in the liquid microbial inoculumChryseobacterium indologenesCI-B4 at a concentration of at least 1 × 108cfu/mL。
In a third aspect of the invention, there is provided a Chryseobacterium indolerum strainChryseobacterium indologenesCI-B4 or the application of the liquid microbial inoculum in the degradation of organic wastewater COD.
The organic wastewater comprises livestock and poultry breeding wastewater, industrial organic wastewater, kitchen waste anaerobic digestion biogas slurry, river black and odorous water and domestic wastewater.
The fourth aspect of the present invention provides a COD degradation method, comprising at least the steps of:
(1) golden yellow bacillus indolens is producedChryseobacterium indologenesInoculating the seed solution or liquid microbial inoculum of CI-B4 into the organic wastewater;
(2) aerobic culture is carried out to degrade COD.
Specifically, the inoculation amount of the seed liquid or the liquid microbial inoculum in the step (1) is 1-10% of the volume of the organic wastewater.
Specifically, the temperature of aerobic culture in the step (2) is 5-40 ℃.
Specifically, the pH value of the aerobic culture in the step (2) is 5-9.
Specifically, the DO value of the aerobic culture in the step (2) is 3-7 mg/L.
As mentioned above, the COD degrading strain and the application thereof of the invention have the following beneficial effects:
(1) COD degrading bacterium chrysobacillus indolensChryseobacterium indologenesCI-B4 is salt-resistant, low-temperature-resistant and wide in temperature application range (5-40 ℃);
(2) COD degrading bacterium chrysobacillus indolensChryseobacterium indologenesCI-B4 has the capability of efficiently degrading organic pollutants in wastewater and can improve the effluent quality of organic wastewater；
(3) COD degrading bacterium chrysobacillus indolensChryseobacterium indologenesThe CI-B4 can be applied to biochemical treatment of livestock and poultry breeding wastewater, industrial organic wastewater, river black and odorous water body, kitchen waste anaerobic digestion biogas slurry and the like after expanded culture, and has higher application value;
(4) utilization of Chryseobacterium indolensChryseobacterium indologenesThe CI-B4 has high treatment efficiency, good economic benefit, convenient operation and no pollution when used for treating organic wastewater.
Drawings
FIG. 1 shows a phylogenetic tree map of the COD degrading strain CI-B4 of the present invention.
FIG. 2 shows the COD degradation of cattle wastewater by the COD degrading strain CI-B4 of the present invention.
FIG. 3 shows the degradation of COD in swine wastewater by the COD degrading strain CI-B4 of the present invention.
FIG. 4 shows the COD degrading strain CI-B4 of the invention degrading COD in black and odorous water in river.
FIG. 5 shows the degradation of COD in the anaerobic digestion biogas slurry of kitchen waste by the COD degrading strain CI-B4 of the invention.
Detailed Description
In a first aspect, the invention provides a COD degrading strain identified as Chryseobacterium indolerum (A)Chryseobacterium indologenes) The strain is preserved in China Center for Type Culture Collection (CCTCC) with the strain name ofChryseobacterium indologenesCI-B4, the preservation date is 2018.9.25, the preservation number is CCTCC NO: m2018651, wherein the preservation address is Wuhan, Wuhan university in China.
Further, the Chryseobacterium indolerum is producedChryseobacterium indologenesCI-B4, which comprises a gene sequence shown in SEQ ID NO. 1.
Further, the Chryseobacterium indolerum is producedChryseobacterium indologenesCI-B4 is gram-negative bacteria, rod-shaped, and the colony morphology is golden yellow circle, the edge is neat, and the surface is smooth, moist and glossy.
Further, the Chryseobacterium indolerum is producedChryseobacterium indologenesThe highest salt tolerance concentration of CI-B4 was 100 g/L sodium chloride.
The Chryseobacterium indolerum is producedChryseobacterium indologenesThe highest salt tolerance concentration of CI-B4 was tested by the following method:
1) preparing LB solid culture media with different NaCl concentrations;
2) subjecting said Chryseobacterium indolens to a reactionChryseobacterium indologenesInoculating CI-B4 to the culture medium in the step 1) by a streak method;
3) culturing in 30 deg.C constant temperature incubator for 48 hr;
4) after 48 hours, the colony size is larger than or equal to 1mm, and the colony grows and breeds normally, namely the colony is regarded as being tolerant to the sodium chloride with the concentration.
The components of the LB solid culture medium containing different NaCl concentrations are 10 g/L of peptone, 5g/L of yeast extract and 15g/L, NaCl of agar with different concentrations (respectively 10 g/L, 20 g/L, 40 g/L, 60 g/L, 80g/L, 100 g/L and 120 g/L).
The Chryseobacterium indolerum is producedChryseobacterium indologenesThe COD tolerance concentration range of the CI-B4 is 0-30000 mg/L.
The Chryseobacterium indolerum is producedChryseobacterium indologenesThe COD-tolerant concentration range of CI-B4 was determined by the following method:
1) preparing wastewater liquid culture media with different COD concentrations;
2) golden yellow bacillus indolens is producedChryseobacterium indologenesInoculating the CI-B4 strain into the wastewater liquid culture medium in the step 1);
3) culturing in 30 deg.C shaking table;
4) periodically sampling every 24h, observing the growth condition of the strain CI-B4 by using a dilution coating and colony counting method, and after coated solid plates with different gradients are cultured for 48 h, counting colonies, wherein the number of colonies counted twice is increased by 1 × 10 compared with the number counted in the last time２cfu/mL is considered to be strain tolerant to the COD concentration, and a constant or decreasing number of colonies is considered to be intolerant to the COD concentration.
Specifically, the concentrations of COD in the wastewater liquid culture medium with different COD concentrations are respectively as follows: 0mg/L, 100mg/L, 1000 mg/L, 5000mg/L, 10000 mg/L, 20000 mg/L, 30000 mg/L, 40000 mg/L.
Specifically, the preparation method of the wastewater liquid culture medium with different COD concentrations comprises the following steps:
taking organic wastewater (namely liquid obtained after solid-liquid separation in a livestock and poultry breeding wastewater treatment plant) with different volumes, 5g/L sodium chloride, 5g/L tryptone and 2.5 g/L yeast extract, adding water to prepare wastewater liquid culture media with different COD concentrations, and adjusting the pH value to 7.0-7.5.
The Chryseobacterium indolerum is producedChryseobacterium indologenesThe COD degradation rate of CI-B4 in the wastewater liquid culture medium for 48 hours reaches more than 80 percent.
The 48 h COD degradation rate is detected by the following method:
inoculating the bacterial liquid into a wastewater liquid culture medium, culturing for 48 h, sampling, determining the COD concentration by adopting a national standard HJ 828-2017 dichromate method, and calculating the COD degradation rate by utilizing a formula (I):
COD degradation rate (%) = [ COD(0)-COD(48)]/ COD(0)×100% ①
Wherein, COD(0)Inoculating a bacterial liquid into a wastewater liquid culture medium, and then obtaining the initial COD concentration of a mixed system; COD(48)The COD concentration of the wastewater liquid culture medium is measured after the inoculation of the bacterial liquid and the culture for 48 hours.
Wherein the components of the wastewater liquid culture medium are as follows: the content of organic wastewater (liquid obtained after solid-liquid separation in a livestock and poultry breeding wastewater treatment plant) is 20% (volume ratio, namely, 200 mL of organic wastewater is contained in 1L of liquid culture medium), 5g/L of sodium chloride, 5g/L of tryptone and 2.5 g/L of yeast extract, and the pH value is 7.0-7.5.
In a second aspect, the invention provides a liquid microbial inoculum, which comprises Chryseobacterium indolerumChryseobacterium indologenesCI-B4, wherein the Chryseobacterium indolerum is produced in the liquid microbial inoculumChryseobacterium indologenesCI-B4 at a concentration of at least 1 × 108cfu/mL。
The preparation method of the liquid microbial inoculum comprises the following steps: golden yellow bacillus indolens is producedChryseobacterium indologenesCI-B4 pure bacteriumThe seeds are inoculated into an LB liquid culture medium for culture for 16-24 h.
Specifically, the culture temperature is 30 ℃.
Specifically, the rotating speed of the shaking table during the cultivation is 160-200 r/min.
In a third aspect of the invention, there is provided a Chryseobacterium indolerum strainChryseobacterium indologenesCI-B4 or the application of the liquid microbial inoculum in the degradation of organic wastewater COD.
The Chryseobacterium indolerum is produced in the liquid microbial inoculumChryseobacterium indologenesCI-B4 at a concentration of at least 1 × 108cfu/mL。
The organic wastewater comprises livestock and poultry breeding wastewater, industrial organic wastewater, kitchen waste anaerobic digestion biogas slurry, river black and odorous water, domestic wastewater and the like.
The fourth aspect of the present invention provides a COD degradation method, comprising at least the steps of:
(1) golden yellow bacillus indolens is producedChryseobacterium indologenesInoculating the seed solution or liquid microbial inoculum of CI-B4 into the organic wastewater;
(2) aerobic culture is carried out under the conditions of proper temperature, pH value and DO (dissolved oxygen) value to degrade COD.
Specifically, the seed solution or the liquid microbial inoculum in the step (1) contains chrysobacillus indoleniniChryseobacterium indologenesCI-B4 at a concentration of at least 1 × 108cfu/mL。
Specifically, the inoculation volume of the seed liquid or the liquid microbial inoculum in the step (1) is 1-10% of the volume of the organic wastewater, and preferably 2-5%.
Specifically, the temperature in the step (2) is 5-40 ℃, and preferably 20-30 ℃.
Specifically, the pH value in the step (2) is 5-9, preferably 7-7.5.
Specifically, the DO value in the step (2) is 3-7 mg/L, preferably 3-5 mg/L.
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments, and is not intended to limit the scope of the present invention; in the description and claims of the present application, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.
The media and components used in the examples were as follows:
wastewater liquid culture medium: the content of organic wastewater (liquid obtained after solid-liquid separation in a livestock and poultry breeding wastewater treatment plant, the COD concentration is 21680 mg/L, and the pH value is 7.5) is 20% (volume ratio, namely, 200 mL of organic wastewater is contained in 1L of liquid culture medium), 5g/L of sodium chloride, 5g/L of tryptone, 2.5 g/L of yeast extract, and the pH value is 7.0-7.5.
Wastewater solid culture medium: the content of organic wastewater (liquid obtained after solid-liquid separation in a livestock and poultry breeding wastewater treatment plant, the COD concentration is 21680 mg/L and the pH value is 7.5) is 20% (volume ratio, namely, 1L of liquid culture medium contains 200 mL of organic wastewater), 5g/L of sodium chloride, 5g/L of tryptone, 2.5 g/L of yeast extract, 15g/L of agar and the pH value is 7.0-7.5.
LB liquid medium: 10 g/L sodium chloride, 10 g/L tryptone, 5g/L yeast extract and a pH value of 7.0-7.5.
LB solid medium: 10 g/L sodium chloride, 10 g/L tryptone, 5g/L yeast extract, 15g/L agar and a pH value of 7.0-7.5.
The prepared culture medium is sterilized by high pressure steam at 121 deg.C for 20 min.
In the examples, COD concentration was determined by the national standard HJ 828-2017 dichromate method.
Example 1: separation screening and performance determination of COD degrading bacteria
Enrichment culture: the experimental sample organic wastewater is collected from sewage treatment pools from different sources, 5 mL of organic wastewater is taken to be put into 45 mL of sterilized wastewater liquid culture medium, and the organic wastewater liquid culture medium is uniformly mixed and then placed into a shaking table with the temperature of 30 ℃ and the speed of 180 r/min for enrichment culture for 96 hours.
Separation and purification: and (3) after the cultured enrichment solution is subjected to gradient dilution, uniformly coating the enrichment solution on a wastewater solid culture medium, culturing the enrichment solution in a constant-temperature incubator at 30 ℃ for 24-48 hours, selecting monoclones with different colony morphologies, and marking, purifying and numbering for preservation. A total of 16 strains are obtained by separation and purification.
Preliminary verification: respectively inoculating 16 strains obtained by separation into an LB liquid culture medium for enrichment culture, centrifuging 5% (volume ratio) bacterial liquid, washing with sterile normal saline, resuspending, inoculating into a sterilized wastewater liquid culture medium, culturing in a shaking table at 30 ℃ and 180 r/min, periodically sampling every 12 h, determining the concentration of COD, and detecting by adopting a national standard HJ 828-2017 dichromate method, thereby evaluating the capacity of each strain for degrading COD.
Calculating the COD degradation rate according to the measured data and the following calculation formula:
COD degradation rate = [ COD(0)-COD(x)]/ COD(0)×100%
COD(0)Inoculating the waste water liquid culture medium with the bacterial liquid, and then obtaining the initial COD concentration of the mixed system.
COD(x)COD concentration of the samples taken at time x.
The result shows that the strain CI-B4 has the highest COD degradation rate in the mixed solution after 48 hours, and can reach 80% in 48 hours.
And (3) performance measurement:
(1) the strain CI-B4 is inoculated to LB solid culture media with different NaCl concentrations (10 g/L, 20 g/L, 40 g/L, 60 g/L, 80g/L, 100 g/L and 120 g/L) by a scribing method, the strain is cultured for 48 hours in a 30 ℃ constant temperature incubator, and the growth condition of the strain is regularly observed, so that the strain CI-B4 is found to grow well on the LB solid culture media containing 100 g/L NaCl, the colony size is 2-3 mm, and the strain does not grow on the LB solid culture media containing 120 g/L NaCl, and the highest salt tolerance concentration of the strain CI-B4 which grows well is 100 g/L.
(2) Inoculating CI-B4 strain to LB liquid culture medium, culturing at different temperature (5 deg.C, 10 deg.C, 20 deg.C, 30 deg.C, 40 deg.C, 50 deg.C, 60 deg.C) in 180 r/min shaking table, sampling periodically every 24h, and measuring OD by spectrophotometer600The result shows that the strain CI-B4 can grow at 5-40 ℃ and grows well.
(3) The strain CI-B4 is respectively inoculated into wastewater liquid culture media with different COD concentrations (0 mg/L, 100mg/L, 1000 mg/L, 5000mg/L, 10000 mg/L, 20000 mg/L, 30000 mg/L and 40000 mg/L), samples are periodically taken every 24h, the growth condition of the strain CI-B4 is observed by using a dilution coating and colony counting method, and the result shows that the strain CI-B4 can grow in organic wastewater with the COD concentration range of 0-30000 mg/L.
The main biological characteristics of the strain CI-B4 are gram-negative bacteria and rod-shaped, the size of a bacterial colony cultured on an LB solid plate for 24 hours is 3-4 mm, and the bacterial colony is golden yellow and round, has neat edges and smooth, moist and glossy surface. The 16S rDNA sequence of the strain CI-B4 is shown as SEQ ID NO.1, the sequence length is 1394 bp, the 16S rRNA sequence is submitted at NCBI, the homologous sequence comparison analysis is carried out by software and GenBank, and MEGA 7 software is used for constructing a phylogenetic tree of the strain (figure 1). With the above information in mind, the strain CI-B4 was identified as Chryseobacterium indolerum (A)Chryseobacterium indologenes）。
Example 2: COD degrading bacteria CI-B4 degrading COD in livestock and poultry breeding wastewater
The livestock and poultry breeding wastewater is taken from liquid obtained after solid-liquid separation of excrement and sewage of a certain cattle farm and a certain pig farm, the COD concentration of a stock solution of the cattle breeding wastewater is 22000 mg/L, the pH value is 7.5, the COD concentration of a stock solution of the pig breeding wastewater is 18000 mg/L, and the pH value is 7.3.
Inoculating CI-B4 strain into LB liquid medium, culturing at 30 deg.C in 180 r/min shaking table for 24 hr, and culturing at 5% volume ratio in logarithmic growth phase (density of bacteria is at least 1 × 10)8cfu/mL), centrifuging and washing the bacterial liquid, adding the bacterial liquid into a reactor, wherein 2L of wastewater is filled in the reactor, and the control conditions are as follows: the rotating speed of the electric stirrer is 180 r/min, the DO value is 5 mg/L, the temperature is 30 ℃, and the pH value is adjusted to 7-7.5. Samples were taken every 12 hours during the reaction, and the COD degradation rate in the wastewater was measured and calculated according to the COD measurement method in example 1.
FIG. 2 shows the COD degradation in the wastewater of the cattle farm in 96h by the strain CI-B4, and it can be seen that the highest value of the COD degradation rate by the strain CI-B4 reaches 80.2%, which is 47.7% higher than the COD degradation rate of the control group of which the strain is not inoculated by 54.3%. The bacterial strain CI-B4 is proved to have remarkable degradation effect on organic pollutants in the wastewater of the cattle farm.
FIG. 3 shows the degradation of COD in pig farm wastewater by the strain CI-B4 in 96h, and it can be seen that the highest value of COD degradation rate by the strain CI-B4 reaches 84.6%, which is 28.6% higher than the 65.8% degradation rate of COD in the control group without inoculation. The strain CI-B4 is proved to have remarkable degradation effect on organic pollutants in the wastewater of the pig farm.
Example 3: COD degrading effect of COD degrading bacteria CI-B4 on COD in black and odorous water body of river channel
The river water is taken from black and odorous water body of a river, the COD concentration of the river water is 260 mg/L, and the pH value of the river water is 6.9.
Inoculating the strain CI-B4 into LB liquid medium, culturing at 20 deg.C in 180 r/min shaking table for 24h, and culturing at 5% volume ratio in logarithmic growth phase (density of bacteria is at least 1 × 10)8cfu/mL), and adding the bacteria liquid into a reactor after centrifugation and washing, wherein 2L of wastewater liquid is filled in the reactor, and the control conditions are as follows: the rotating speed of the electric stirrer is 180 r/min, the DO value is 3 mg/L and the temperature is highThe temperature is 20 ℃, and the pH value is adjusted to 7-7.5. Samples were taken every 12 hours during the reaction, and the degradation rate of COD in the wastewater was measured and calculated according to the method for measuring COD in example 1.
FIG. 4 shows that the strain CI-B4 degrades COD in black and odorous water in a river within 96h, and it can be seen that the highest degradation rate of the strain CI-B4 to COD is 53.1%, which is 28.0% higher than the degradation rate of 41.5% of the strain of the control group without inoculation. The bacterial strain CI-B4 has obvious degradation effect on organic pollutants in black and odorous water in riverways.
Example 4: COD degrading effect of COD degrading bacteria CI-B4 on COD in anaerobic digestion biogas slurry of kitchen waste
The biogas slurry obtained by anaerobic digestion of the kitchen waste is taken from biogas slurry discharged from a biogas fermentation tank of a kitchen waste treatment plant, the COD concentration of the stock solution is 6900 mg/L, and the pH value is 7.
Inoculating CI-B4 strain into LB liquid medium, culturing at 40 deg.C in 180 r/min shaking table for 24 hr, and culturing at 5% volume ratio in logarithmic growth phase (density of bacteria is at least 1 × 10)8cfu/mL), centrifuging and washing the bacterial liquid, adding the bacterial liquid into a reactor, wherein 2L of wastewater is filled in the reactor, and the control conditions are as follows: the rotation speed of the electric stirrer is 180 r/min, the DO value is 3 mg/L, the temperature is 40 ℃, and the pH value is adjusted to 7-7.5. Samples were taken every 12 hours during the reaction, and the COD degradation rate in the wastewater was measured and calculated according to the COD measurement method in example 1.
FIG. 5 shows that the strain CI-B4 degrades COD in the anaerobic digestion biogas slurry of kitchen waste within 96h, and it can be seen that the highest value of the degradation rate of the strain CI-B4 to COD is 76.9%, which is 39.1% higher than the degradation rate of 55.3% of the COD of the control group without inoculation. The bacterial strain CI-B4 is proved to have obvious degradation effect on organic pollutants in the anaerobic digestion biogas slurry of the kitchen waste.
The above examples are intended to illustrate the disclosed embodiments of the invention and are not to be construed as limiting the invention. In addition, various modifications of the invention set forth herein, as well as variations of the methods of the invention, will be apparent to persons skilled in the art without departing from the scope and spirit of the invention. While the invention has been specifically described in connection with various specific preferred embodiments thereof, it should be understood that the invention should not be unduly limited to such specific embodiments. Indeed, various modifications of the above-described embodiments which are obvious to those skilled in the art to which the invention pertains are intended to be covered by the scope of the present invention.
Sequence listing
<110> Shanghai higher research institute of Chinese academy of sciences
<120> COD degrading strain and application thereof
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>1394
<212>DNA
<213> Chryseobacterium indogenes (Chryseobacterium indolium)
<400>1
gcagctcctg ttacggtcac cgacttcagg taccccagac ttccatggct tgacgggcgg 60
tgtgtacaag gcccgggaac gtattcaccg cgccatggct gatgcgcgat tactagcgat 120
tccagcttca tagagtcgag ttgcagactc caatccgaac tgagaccggc tttcgagatt 180
tgcatcacat cgctgtgtag ctgccctctg taccggccat tgtattacgt gtgtggccca 240
aggcgtaagg gccgtgatga tttgacgtca tccccacctt cctctctact tgcgtaggca 300
gtctcactag agtccccaac ttaatgatgg caactagtga caggggttgc gctcgttgca 360
ggacttaacc taacacctca cggcacgagc tgacgacaac catgcagcac cttgaaaaat 420
gtccgaagaa aagtctattt ctaaacctgt catttcccat ttaagccttg gtaaggttcc 480
tcgcgtatca tcgaattaaa ccacataatc caccgcttgt gcgggccccc gtcaattcct 540
ttgagtttca aacttgcgtt cgtactcccc aggtggctaa cttatcactt tcgcttagtc 600
tctgaacccg aaagcccaaa aacgagttag catcgtttac ggcgtggact accagggtat 660
ctaatcctgt tcgctcccca cgctttcgtc catcagcgtc agttgttgct tagtaacctg 720
ccttcgcaat tggtgttcta agtaatatct atgcatttca ccgctacact acttattcca 780
gctacttcaa caacactcaa gacctgcagt atcaatggca gtttcacagt taagctgtga 840
gatttcacca ctgacttaca gatccgccta cggacccttt aaacccaata aatccggata 900
acgcttgcac cctccgtatt accgcggctg ctggcacgga gttagccggt gcttattcgt 960
atagtacctt cagctactct cacgagagta ggtttatccc tatacaaaag aagtttacaa 1020
cccatagggc cgtcgtcctt cacgcgggat ggctggatca ggctctcacc cattgtccaa 1080
tattcctcac tgctgcctcc cgtaggagtc tggtccgtgt ctcagtacca gtgtggggga 1140
tcaccctctc aggcccccta aagatcgcag acttggtgag ccgttacctc accaactatc 1200
taatcttgcg cgtgcccatc tctatccacc ggagttttca atgtcgaatg atgccatcca 1260
acatattatg gggtattaat cttcctttcg aaaggctatc ccccagataa aggcaggttg 1320
cacacgtgtt ccgcacccgt acgccgctct caagatcccg aaagatctct accgctcggc 1380
ttgcatgtgt tagc 1394
Claims (10)
1. Chryseobacterium indolerum producing strainChryseobacterium indologenesCI-B4, the preservation number is CCTCCNO: m2018651.
2. Chryseobacterium indogenes according to claim 1Chryseobacterium indologenesCI-B4, wherein said Chryseobacterium indolerum is characterized in thatChryseobacterium indologenesCI-B4 contains a gene sequence shown in SEQ ID NO. 1.
3. Chryseobacterium indogenes according to claim 1Chryseobacterium indologenesCI-B4, wherein said Chryseobacterium indolerum is characterized in thatChryseobacterium indologenesThe highest salt tolerance concentration of CI-B4 was 100 g/L sodium chloride.
4. Chryseobacterium indogenes according to claim 1Chryseobacterium indologenesCI-B4, wherein said Chryseobacterium indolerum is characterized in thatChryseobacterium indologenesThe COD tolerance concentration range of the CI-B4 is 0-30000 mg/L.
5. Chryseobacterium indogenes according to claim 1Chryseobacterium indologenesCI-B4, wherein said Chryseobacterium indolerum is characterized in thatChryseobacterium indologenesThe COD degradation rate of CI-B4 in the wastewater liquid culture medium for 48 hours reaches more than 80 percent.
6. A liquid microbial inoculum comprising the Chryseobacterium indolent of any of claims 1-5Chryseobacterium indologenesCI-B4, wherein the Chryseobacterium indolerum is produced in the liquid microbial inoculumChryseobacterium indologenesCI-B4 at a concentration of at least 1 × 108cfu/mL。
7. Chryseobacterium indogenes according to any one of claims 1 to 5Chryseobacterium indologenesUse of CI-B4 or the liquid microbial inoculum according to claim 6 in COD degradation of organic wastewater.
8. The application of claim 7, wherein the organic wastewater comprises livestock and poultry breeding wastewater, industrial organic wastewater, anaerobic digestion biogas slurry of kitchen waste, black and odorous water body in river channels and domestic wastewater.
9. A COD degradation method is characterized by comprising the following steps:
(1) the Chryseobacterium indogenes of any one of claims 1-5Chryseobacterium indologenesInoculating the seed solution of CI-B4 or the liquid microbial inoculum of claim 6 into organic wastewater;
(2) aerobic culture is carried out to degrade COD.
10. The COD degradation method of claim 9, further comprising one or more of the following features:
1) in the step (1), the inoculation volume of the seed liquid or the liquid microbial inoculum is 1-10% of the volume of the organic wastewater;
2) the temperature of aerobic culture in the step (2) is 5-40 ℃;
3) the pH value of aerobic culture in the step (2) is 5-9;
4) the DO value of the aerobic culture in the step (2) is 3-7 mg/L.
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