CN117887593A - Mixed nutrition type denitrifying bacterium Penicillium sp.N8 and application thereof - Google Patents
Mixed nutrition type denitrifying bacterium Penicillium sp.N8 and application thereof Download PDFInfo
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- 241000195546 Penicillium sp. N8 Species 0.000 title claims abstract description 20
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- -1 pplication Species 0.000 title abstract description 6
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- 229910003208 (NH4)6Mo7O24·4H2O Inorganic materials 0.000 description 1
- IICCLYANAQEHCI-UHFFFAOYSA-N 4,5,6,7-tetrachloro-3',6'-dihydroxy-2',4',5',7'-tetraiodospiro[2-benzofuran-3,9'-xanthene]-1-one Chemical compound O1C(=O)C(C(=C(Cl)C(Cl)=C2Cl)Cl)=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 IICCLYANAQEHCI-UHFFFAOYSA-N 0.000 description 1
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- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 1
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- 239000010413 mother solution Substances 0.000 description 1
- 150000004767 nitrides Chemical class 0.000 description 1
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- 238000006213 oxygenation reaction Methods 0.000 description 1
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Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the technical field of water pollution treatment, and particularly relates to a mixed nutrition type denitrifying bacterium Penicillium sp.N8 and application thereof, wherein the strain is deposited in China center for type culture Collection, the deposited address is the university of Wuhan with the number of 299 of Wuhan, wu Changou, the number of China center for type culture collection, the deposited number is CCTCC M20232691, and the deposited date is 2023, 12 months and 27 days, and the strain is identified as Penicillium.
Description
Technical Field
The invention relates to the technical field of water pollution treatment, in particular to mixed nutrition type denitrifying bacteria Penicillium sp.N8 and application thereof.
Background
The current research shows that most surface water bodies are in a micro-pollution state, the water bodies with the concentration of nitrate, organic matters and other pollutants lower than 10mg/L are called micro-pollution water bodies, and the control of endogenous pollutants of micro-pollution lakes and reservoirs, especially the control of nitrogen pollution, is a key way for inhibiting the eutrophication of the water bodies. Physical, chemical, biological removal methods have achieved a certain effect against the hazards of nitrate accumulation, with biological removal methods being widely used for their efficiency and economy. The biological denitrification technology relates to an aerobic denitrification reaction, which has strong dissolved oxygen resistance, moderate cost, high treatment efficiency and clean byproducts and is harmless to the environment, so that the biological denitrification technology becomes a hot spot for researching the current denitrification method.
Fungi play an important role in the nitrogen cycle, they not only mineralize organic nitrogen and assimilate inorganic nitrogen to grow cells, some fungi also reduce nitrate or nitrite to gaseous nitrides by denitrification or co-denitrification. The denitrification advantages of fungi compared with other microorganisms are mainly as follows: firstly, the denitrification rate is higher and the capability of decomposing organic matters is stronger; second, the fungal spores have complex cell walls, which increase the resistance of fungi to toxic compounds and harsh environments (e.g., lower pH and higher temperature).
Organic pollutants are reduced, the aerobic denitrification nitrogen removal efficiency is lower due to lower concentration of the organic matters, and secondary pollution can be introduced by adding additional carbon sources; oxygenation of water cannot improve the quality of water at all, so that a way to improve denitrification efficiency is needed.
Disclosure of Invention
In order to solve the technical problems, the invention provides a mixed nutrition denitrifying bacterium Penicillium sp.N8 and application thereof, and solves the technical problems that biological denitrification treatment efficiency of micro-polluted water bodies needs to be further improved in the prior art.
The mixed nutrition type denitrifying bacterium Penicillium sp.N 8 has a preservation unit of China center for type culture Collection, a preservation address of Wu Changou of the university of Wuhan No. 299 of Wuhan in Wuhan, a preservation number of CCTCC M20232691, and a preservation date of 2023, 12 months and 27 days.
The application of the mixed nutritional denitrifying bacteria Penicillium sp.N8 in the restoration of nitrogenous water bodies.
Preferably, the nitrogenous water body is remediated to remove nitrogen from the water body.
Preferably, the nitrogen-containing water body is repaired in the presence of an electron donor.
Preferably, when the nitrogenous water body is repaired, the usage amount of the mixed nutritional denitrifying bacteria Penicillium sp.N8 is 1-5% based on the mass fraction of the nitrogenous water body.
Preferably, the carbon-nitrogen ratio in the nitrogen-containing water body is 1-2.5.
Preferably, the temperature of the nitrogen-containing water body is 25-30 ℃.
Preferably, the moving speed of the nitrogenous water body is 80-160rpm.
Preferably, the addition amount of the iron is 10-15 g/L.
A method for treating nitrate in polluted water body comprises inoculating mixed nutritional denitrifying bacteria Penicilliumsp.N8 into nitrogen-containing water body, and adding iron as electron donor.
The invention also has the following technical characteristics:
The culture medium of the aerobic denitrifying fungus is light yellow and divergent; the similarity of the gene sequence of Penicillium goetzii and Penicillium sp.N 8 is found to be more than 99% by adopting the ITS sequencing technology and the phylogenetic tree established based on the neighbor cell connection method, which indicates that N8 belongs to the Penicillium genus.
The application method comprises the following steps: setting the iron addition amount of different inorganic electron donors to be 0, 5, 10, 15 and 20g/L; different C/N,1, 1.5, 2.5; different rotation speeds, rpm=40, 80, 160 and different temperatures, t=5, 10, 15, 20 ℃, explored denitrification performance under different environmental conditions; and then exploring the joint influence of the C/N, rpm and the temperature change condition on the denitrification effect through a dynamic model.
Compared with the prior art, the invention has the beneficial effects that:
When the C/N of the aerobic denitrifying fungus is reduced to 1.5 and 1 from 2, the removal rate of TN by the strain N8 is reduced to 90% and 95% from 100%; along with the continuous increase of the oscillation speed, the denitrification rate of the strain N8 is continuously increased, and the nitrate removal rate can reach 100% at 160 rpm; with the gradual decrease of the temperature, the removal rate of TN by the strain N8 is reduced from 93.36% to 75.34%, 50.80% and 29.18%; in addition, the denitrification rate under the conditions of C/N, rpm and temperature change more accords with a half-order rate equation, which shows that the reaction rate is influenced by external environment conditions.
Drawings
FIG. 1 shows phylogenetic tree (a) and strain morphology (b) of the aerobic denitrifying fungus Penicillium sp.N8;
FIG. 2 is a graph of the denitrification performance of Penicillium sp.N8 at various iron additions, wherein (a) nitrate nitrogen concentration varies; (b) a change in nitrite nitrogen concentration; (c) ammonia nitrogen concentration variation; (d) total nitrogen concentration variation;
FIG. 3 shows denitrification performance of Penicillium sp.N8 at different carbon to nitrogen ratios, wherein (a) carbon to nitrogen ratio is 1, (d) carbon to nitrogen ratio is 1.5, and (c) carbon to nitrogen ratio is 2.5;
Fig. 4 is denitrification performance of Penicillium sp.n8 at different rotational speeds, wherein (a) rpm=40, (b) rpm=80, (c) rpm=160;
Fig. 5 shows denitrification performance of Penicillium sp.n8 at different temperatures, wherein (a) t=5, (b) t=10, (c) t=15, (d) t=20;
FIG. 6 shows the zero-order (a), half-order (b) and first-order (C) kinetic models of aerobic denitrifying fungi at different C/N; zero-order (d), half-order (e) and first-order (f) kinetic models of strain N8 at different rpm; the strain N8 has a kinetic model of zero order (g), half order (h) and first order (i) at different temperatures;
FIG. 7 shows denitrification of aerobic denitrifying fungi and cell growth and DOC removal under optimal conditions, wherein (a) shows denitrification performance and (b) shows results of cell growth and DOC removal;
FIG. 8 shows the release of Fe 3+ and Fe 2+ in an aerobic denitrifying fungal system;
FIG. 9 is a model of kinetics of nitrate removal by aerobic denitrifying fungi under different parameters.
Detailed Description
The following detailed description of specific embodiments of the invention is, but it should be understood that the invention is not limited to specific embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The experimental methods described in the examples of the present invention are conventional methods unless otherwise specified.
The fungus culture medium (DRBC) adopts a Bengalese rose bengal agar culture medium known in the prior art, and has the formula: 5.6-5.8 g/L peptone 、0.025g/L C20H2Cl4I4Na2O5、10g/L C6H12O6、0.002g/L C6H4Cl2N2O2、1 g/L KH2PO4、0.1g/L C11H12Cl2N2O5、0.5g/L MgSO4,pH. The fungus solid culture medium is prepared by adding 20g/L agar powder based on the formula.
The denitrification liquid culture medium (DM) adopts a conventional denitrification liquid culture medium known in the prior art, and the formula and the preparation method are as follows: KNO 3 is 0.108 g/L, KH 2PO4 is 1.5 g/L, glucose is 0.413 g/L, mgSO 4·7H2 O is 0.1 g/L, na 2HPO4·12H2 O is 5.0 g/L, and trace element mother liquor is 2mL; adding the components into ultrapure water to a constant volume of 1L, stirring until the components are completely dissolved, adjusting the pH value to 7.0-7.2, and sterilizing at the high temperature of 121 ℃ for 30 minutes for later use.
The formula and the preparation method of the trace element mother solution are as follows: 4.4 mg of ZnSO 4, ethylenediamine tetraacetic acid of 100 mg, mnCl 2·4H2 O of 10.2 mg, caCl 2 of 11 mg, feSO 4·7H2 O of 10mg, cuSO 4·5H2 O of 3.2 mg and CoCl 2·6H2 O of 2.2 mg (NH 4)6Mo7O24·4H2 O,3.2 mg), adding the above components into ultrapure water to a constant volume of 1L, stirring until the components are completely dissolved, then adjusting the pH value to 7.0-7.2, and sterilizing at 121 ℃ for 30 minutes for later use.
The iron in the invention is zero-valent iron, and can be zero-valent iron powder or iron rod.
Example 1
The screening and identifying method of the aerobic denitrifying fungus specifically comprises the following steps:
step one, enrichment of aerobic denitrifying fungi:
collecting water samples at 0.5m part of a reservoir, taking 100 mL water samples, enriching fungi by using a 0.22 mu m filter membrane, attaching the filter membrane to a DRBC solid culture medium plate, inverting a culture dish in a biochemical incubator, and culturing for 3-4 days at 30 ℃ until colony formation occurs on the culture medium.
Screening aerobic denitrifying fungi:
(1) After colony formation on the petri dish, the fungus colony on DRBC was scraped into a 150 mL conical flask (n=3) containing 100 mL low C/N (C/n=2) denitrification liquid medium (DM) with an inoculating loop, which is the case without inorganic electron donor addition;
(2) 1.0 g/L of inorganic electron donor was added to the shaker for 1h, and the fungal colonies on the same DRBC were scraped with the inoculating loop into another 150mL conical flask (n=3) containing 100mL low C/N (C/N=2) DM medium to eliminate stress of the inorganic electron donor on the fungus.
Culturing the two conical flasks at 30 ℃ under the conditions of 125+/-5 rpm, sampling and measuring every 24: 24h, measuring the concentration of nitrate nitrogen and nitrite nitrogen, and comparing the two phases, wherein the two conical flasks have a certain nitrate removal rate under the condition of no inorganic electron donor and have a nitrate removal rate of more than 80% under the condition of no inorganic electron donor, so that fungi are further researched.
Step three, strain identification:
and (3) carrying out Mulberry sequencing on the selected high-efficiency aerobic denitrifying fungi, carrying out naming numbering on the fungi after determining the corresponding species of the fungi by sequencing, storing the fungi in 50% glycerol (1:1), and storing the fungi at the temperature of-20 ℃ for further research.
PCR was performed using the primers.
The primer is as follows:
ITS1:5'-TCCGTAGGTGAACCTGCGG-3', denoted SEQ ID NO.2,
ITS4：5’-TCCTCCGCTTATTGATATGC-3’，SEQ ID NO.3。
The PCR reaction system (25 mu L) is as follows: the genome DNA template is 20-50 ng/mu L, the PCR Premix is 12.5 mu L, the primer ITS1 (10 mu M) is 1 mu L, the primer ITS4 (10 mu M) is 1 mu L, and the ddH 2 O is 9.5 mu L.
The PCR procedure was: 95 ℃ for 5min;94 ℃ for 30s;57 ℃,30s;72 ℃,90s; amplifying for 30 cycles, then extending for 10min at 72 ℃, obtaining a 16S rDNA fragment after PCR is finished, and obtaining a 16S rDNA nucleotide sequence of the 16S rDNA fragment after sequencing, wherein the nucleotide sequence is specifically as follows:
CTGCGGAAGGATCATTACCGAGTGAGGGCCCTCTGGGTCCAACCTCCCACCCGTGTTTATTTTACCTTGTTGCTTCGGCGGGCCCGCCTTAACTGGCCGCCGGGGGGCTTACGCCCCCGGGCCCGCGCCCGCCGAAGACACCCTCGAACTCTGTCTGAAGATTGTAGTCTGAGTGAAAATATAAATTATTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATACGTAATGTGAATTGCAAATTCAGTGAATCATCGAGTCTTTGAACGCACATTGCGCCCCCTGGTATTCCGGGGGGCATGCCTGTCCGAGCGTCATTTCTGCCCTCAAGCACGGCTTGTGTGTTGGGCCCCGTCCTCCGATCCCGGGGGACGGGCCCGAAAGGCAGCGGCGGCACCGCGTCCGGTCCTCGAGCGTATGGGGCTTTGTCACCCGCTCTGTAGGCCCGGCCGGCGCTTGCCGATCAACCCAAATTTTTATCCAGGTTGACCTCGGATCAGGTAGGGATACCCGCTGA, The sequence is shown as SEQ ID NO.1, beginning at the 5 'end and ending at the 3' end.
The result of the alignment of the 16S rDNA nucleotide sequences is shown in FIG. 1. As can be seen from FIG. 1, the nucleotide sequence of the 16S rDNA has more than 99% similarity with Penicillium goetzii, belongs to Penicillium sp (Penicillium), is named Penicillium sp.N8, has a preservation unit of China center for type culture Collection, has a preservation address of Wu Changou eight-channel No. 299 of Wuhan university in Wuhan, has a preservation number of CCTCC M20232691, and has a preservation date of 2023, 12 months and 27 days.
Example 2
In this example, aerobic denitrification capability of the aerobic denitrification fungus Penicillium sp.N8 (hereinafter abbreviated as N8) was tested, and fungus N8 with a mass concentration of 1% was inoculated into sterilized DM medium with N=5mg/L, and different iron addition amounts, carbon nitrogen ratios, rotational speeds, and temperatures were set. After samples were filtered through a pre-burning 0.45 μm GF/F glass fiber filter every 2 days, the concentrations of the nitrate nitrogen (NO 3 - -N), the nitrite nitrogen (NO 2 - -N), the ammonia nitrogen (NH 4 + -N) and the Total Nitrogen (TN) of each system were determined as follows:
(1) The effect of different iron additions (0, 5, 10, 15, 20g/L, i.e. 0, 5, 10, 15, 20g of zero-valent iron powder added to each liter of treated water) on the N8 denitrification efficiency was examined at C/n=2, 30 ℃, and 120 rpm.
As can be seen from fig. 2, in the case where no exogenous inorganic electron donor was added, the removal rate of nitrate by N8 in the medium with C/n=2 was about 55%, and no significant nitrous acid accumulation was observed. The nitrate removal rate of N8 is gradually accelerated along with the increase of the iron addition amount, and the complete removal can be realized for 6 days under the maximum addition amount; meanwhile, the accumulation amount of ammonia nitrogen is continuously increased, which shows that the proportion of the iron which is mainly used for chemical action is gradually increased, and the conditions are synthesized, so that the nitrate removal rate is faster, and the 10g/L addition amount of ammonia nitrogen which is accumulated in a small amount and can be finally removed through nitrification is selected as the optimal addition amount.
(2) Under the condition that the iron addition amount is 10g/L, the temperature is 30 ℃, and the rotating speed is 120rpm, the influence of different C/N (1, 1.5 and 2.5) on N8 nitrogen reduction is examined.
As shown in FIG. 3, when the C/N is reduced from 2.5 to 1.5 and 1, the TN removal rate of the strain N8 is reduced from 100% to 90% and 95%, nitrate nitrogen can be completely removed, but ammonia nitrogen re-accumulation occurs in the later stage of cultivation, which may be caused by ammonia nitrogen release due to death of cells after the decay period, and it is also demonstrated that the organic carbon available to the strain at low C/N is lower, which may limit the denitrification activity and cell activity. Along with the increase of C/N to 2.5, the removal rate of TN by the strain N8 reaches the 10 th day to completely remove, and meanwhile, no ammonia nitrogen accumulation occurs in the later period, which indicates that the TN removal efficiency can be promoted by improving the C/N. The choice of lower nutrient conditions that enable complete removal of C/n=2 was further investigated taking into account the nutrient-poor conditions of the reservoir.
(3) The oscillation speed reflects the concentration of DO (dissolved oxygen), and the change of DO concentration is critical to aerobic denitrifying bacteria, and the influence of different rotation speeds on N8 nitrogen reduction is examined under the conditions that the iron addition amount is 10g/L and the temperature is 30 ℃ and C/N=2.
As shown in FIG. 4, when the shaking speed was 40rpm, the nitrate removal rate of the strain N8 was reduced to 85%, which was caused by insufficient DO available to the aerobic denitrification strain, which was unfavorable for denitrification; meanwhile, insufficient DO may result in a decrease in the oxidation rate of iron, a decrease in the electron donor supply amount, and a decrease in the denitrification rate. In addition, the ammonia nitrogen accumulation amount in the later culture period reaches 0.45mg/L at the highest, which is probably caused by the cell death autolysis of the strain. Along with the continuous increase of the oscillation speed, the denitrification rate of the strain N8 is continuously increased, and the nitrate removal rate is increased from 100% on the 14 th day to 10 th day; finally, no ammonia nitrogen is accumulated, which is probably because the increase of DO concentration can accelerate the mass transfer rate of oxygen and nitrate, and the oxidation rate of iron can be accelerated to a certain extent, thereby improving the activity of nitrogen metabolizing enzyme; furthermore, no accumulation of nitrite nitrogen was observed throughout the experiment. Thus, the optimal rotation speed for strain N8 was 160rpm.
(4) The effect of different temperatures (5, 10, 15 and 20 ℃) on the reduction of N8 nitrogen was examined at an iron dosage of 10g/L, C/N=2, rpm=120.
As shown in FIG. 5, with the gradual decrease of the temperature, the removal rate of TN by the strain N8 is decreased from 93.36% to 75.34%, 50.80% and 29.18%, and ammonia nitrogen re-accumulation occurs in the later stage of culture under the low temperature condition, probably because the ammonia nitrogen is released due to death of cells after entering the decay period, and the lower activity of cells with the strain degree under the low temperature condition is also indicated. When the iron addition amount is examined, when the iron addition amount is 10g/L, C/N=2, the rotating speed is 120rpm, and the temperature is 30 ℃, the TN removal rate is 100%, and the overall water level and nitrogen removal condition after reservoir mixing are considered, so that the 30 ℃ with higher removal efficiency is selected as the optimal temperature.
Example 3
This example shows the effect of the aerobic denitrifying fungus of example 1 on denitrification C/N, rpm (DO) and temperature by using a kinetic model.
Three different parameters including C/N, rpm and temperature are set according to the actual seasonal change of reservoirs in spring, summer, autumn and winter and the thermal stratification phenomenon of reservoirs, and in order to specifically analyze the influence on denitrification performance under different conditions, continuously measured data are fitted into different models in the study.
Wherein C is the nitrate concentration (mg/L) corresponding to the reaction time (T, h). K 0V, R (mg/L),K1/2V, R (mg1 /2 (L1/2 h)-1) and K 1V, R (h-1) are the reaction rate constants of the zero order, half order and first order kinetic models. Zero order kinetic models are often used to represent that the reaction rate is not limited by the contaminant concentration; semi-order kinetic models are commonly used to represent environmental factors that are limiting steps in contaminant removal; a first order kinetic model is often used to represent that environmental factors are limiting steps in contaminant removal, nitrate concentration will become a limiting factor in reaction rate.
As can be seen from fig. 6 and 9, as the C/N ratio increases, R 2 of the half-order model (R 2 =0.8445 at C/N ratio=1.0, R 2 = 0.9593 at C/N ratio=2.0) and the first-order model (R 2 =0.7733 at C/N ratio=1.0, R 2 = 0.9449 at C/N ratio=2.0) increases gradually, and the corresponding maximum coefficients at C/N ratio of 2.0 are 0.1326 mg 1/2 (L1/2h)-1 and 0.1650 h -1, respectively. It is shown that the denitrification rate is continuously accelerated with the increase of C/N.
As can be seen from fig. 6 and 9, strain N8 more conforms to the half-order kinetic model with a change in rpm, and as rpm increases, the half-order model (R 2 =0.9418 at rpm=40, R 2 = 0.9712 at rpm=160) increases gradually at R 2, with a corresponding maximum coefficient of 0.1628 mg 1/2 (L1/2h)-1 at an rpm ratio of 160. Indicating that the denitrification rate is continuously accelerated with the increase of rpm. In the first order kinetic model, when the rpm was increased to 160, R 2 and 0.9449 were decreased to 0.9213, the denitrification rate was decreased to some extent, indicating that too high dissolved oxygen would rather hinder the mass transfer rate and thus decrease the denitrification rate.
As can be seen from fig. 6 and 9, strain N8 more conforms to the half-order kinetic model under the temperature change condition, and as the temperature increases, the half-order model (R 2 =0.8646 at t=5 and R 2 = 0.9565 at t=20) gradually increases R 2, and the corresponding maximum coefficient at T is 0.0836 mg 1/2 (L1/2h)-1 at 20 ℃. It is shown that as T increases, the denitrification rate increases.
In addition, the denitrification rate under the conditions of C/N, rpm and temperature change more accords with a half-order rate equation, which shows that the reaction rate is influenced by external environment conditions. In addition, the release of iron ions and thus the denitrification rate may be affected by changes in the external environment. Thus, changes in these several environmental conditions under natural conditions would collectively affect nitrate removal by strain N8, rather than a single effect.
Example 4
In this example, N8 was tested for denitrification ability and cell growth, as shown in FIG. 7.
Under the condition of no electron donor addition, the cell number is slightly reduced from 1.54 multiplied by 10 6 to 1.07 multiplied by 10 6 in the adaptation period of 0-1 day, and the nitrate nitrogen removal rate is also slower from 5mg/L to 4.71mg/L; during the logarithmic phase of cell growth (2-5 d), the nitrate concentration of strain N8 decreased from 4.51mg/L to 2.91mg/L, and the fluctuation was changed after the nitrate removal rate reached a maximum of 64.6% by day 8. Under the condition of electron donor addition, the cell number is slightly reduced from 1.54×10 6 to 9.45×10 5 in the adaptation period of 0-2 days, and the adaptation time is slightly longer than that of the case without inorganic electron donor, probably due to the selection of iron; meanwhile, the nitrate nitrogen removal rate is also slower, and is reduced from 5mg/L to 4.51mg/L; during the logarithmic phase of cell growth (3-6 d), the nitrate concentration of strain N8 was reduced from 4.51mg/L to 1.82mg/L, and the nitrate removal rate reached 100% by day 11. At the same time, the DOC content gradually decreases with the continuous removal of nitrate, which may be due to the self-propagation of DOC consumed by the cells, while providing the necessary electron donor for denitrification.
As shown in fig. 8, the concentration of Fe 3+ and the concentration of Fe 2+ show a trend of increasing and then decreasing, the concentration of Fe 3+ varies widely, and the content of Fe 3+ reaches 1.483mg/L at the highest on the 4 th day, and then rapidly decreases to about 0.5mg/L to fluctuate up and down, which means that the oxidation speed of iron gradually slows down from rapid to slow, and the surface-generated iron oxide blocks the oxidation of subsequent iron, but still proceeds slowly, resulting in content fluctuation. The ferrous content was lower, after reaching the peak value of 0.12mg/L on day 2, followed by a fluctuation in the range of 0.067-0.042mg/L, probably due to the faster rate of conversion of ferrous to ferric due to aerobic conditions, and lower concentrations.
It should be noted that, when the claims refer to numerical ranges, it should be understood that two endpoints of each numerical range and any numerical value between the two endpoints are optional, and the present invention describes the preferred embodiments for preventing redundancy.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. It is therefore intended that the following claims be interpreted as including the preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.
Claims (10)
1. The mixed nutrition type denitrifying bacterium Penicillium sp.N 8 has a preservation unit of China center for type culture Collection, a preservation address of Wu Changou of the university of Wuhan No. 299 of Wuhan in Wuhan, a preservation number of CCTCC M20232691, and a preservation date of 2023, 12 months and 27 days.
2. Use of the mixed nutritional denitrifying bacteria Penicillium sp.N8 according to claim 1 in the remediation of nitrogen-containing water bodies.
3. The use according to claim 2, wherein the nitrogen-containing water body is remediated to remove nitrogen from the water body.
4. Use according to claim 3, characterized in that the remediation of the nitrogen-containing water is carried out in the presence of an electron donor.
5. The use according to claim 4, wherein the amount of the mixed nutrient denitrifying bacteria Penicillium sp.N8 used in the remediation of the nitrogen-containing water body is 1% -5% by mass of the nitrogen-containing water body.
6. The use according to claim 4, wherein the carbon to nitrogen ratio in the nitrogen containing water is 1-2.5.
7. The use according to claim 4, wherein the temperature of the nitrogen-containing water body is 25-30 ℃.
8. The use according to claim 4, wherein the rotation speed of the nitrogen-containing water body is 80-160rpm.
9. The use according to claim 4, wherein the electron donor is iron and the amount of iron added is 10-15 g/L.
10. A method for treating nitrate in a polluted water body, which is characterized in that mixed nutritional denitrifying bacteria Penicillium sp.N8 according to claim 1 are inoculated into a nitrogen-containing water body, and iron is added as an electron donor.
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