CN107090418B - One strain denitrogen paracoccus and its application in livestock and poultry farm wastewater treatment - Google Patents
One strain denitrogen paracoccus and its application in livestock and poultry farm wastewater treatment Download PDFInfo
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Abstract
The present invention relates to biological technical field, specifically discloses a strain denitrogen paracoccus and its application in livestock and poultry farm wastewater treatment, Paracoccus denitrificans provided by the invention（Paracoccus denitrificans）TDFQ 1, is deposited in China typical culture collection center（CCTCC）, deposit number is：CCTCC NO：M2017238.The bacterial strain has the speed of growth fast, and environmental resistance is strong, the well-grown in organic culture medium.Paracoccus denitrificans of the present invention has heterotrophic nitrification, aerobic denitrification ability, can reduce the harmful substances such as ammonia nitrogen in water body, nitrate, the ammonia nitrogen removal in livestock and poultry farm waste water particularly higher to content of organics, improves breeding environment.The bacterial strain can reduce pollution of the antibiotic to soil and water source, protection human body health when handling animal farm wastewater with efficient degradation fluoroquinolone antibiotics therein.
Description
Technical field
The present invention relates to biological technical field, more particularly to the separation identification of a strain denitrogen paracoccus, cultural method and its
Application in livestock and poultry farm wastewater treatment.
Background technology
With society and expanding economy, intensive livestock and poultry cultivation scale is increasing, and production efficiency improves, economic benefit
Increase, but the generation of a large amount of farm animal excrements and waste water simultaneously, cause the contradiction that poultry industry develops between environmental pollution increasingly
Highlight, fecaluria waste water concentrations bring great pressure to ecological environment.Livestock and poultry breeding industry is the leading production of China's agricultural
One of one of industry, and the main reason for Pollution from livestock and poultry then turns into agricultural non-point source pollution.Substantial amounts of livestock breeding wastewater turns into
It pollution sources, such as cannot get timely processing, high risks will be caused to environment, cause the deterioration of the ecological environment, animal products quality
Decline and jeopardize health, the hysteresis of breeding wastewater Treatment process is also by the serious sustainable development for restricting aquaculture.
Intensive culture waste water is mainly derived from the excreta and rinse water of animal, has the ammonia nitrogen of higher concentration and has
Machine thing, COD is high, and stink is big.There are some researches show ammonia nitrogen concentration is up to 800-2000mg/L in piggery wastewater, chemistry
Oxygen demand is up to 5000-20000mg/L.If waste water is not effectively addressed and is directly discharged into water body and easily causes water eutrophication
Change and underground water pollution.At present, the processing to this kind of waste water mainly includes materialization, biochemistry and natural, ecological etc., wherein biology is de-
Nitrogen technique is simple, and cost is low, small to carrying capacity of environment, and denitrogenation is more thorough.Paracoccus denitrificans (Paracoccus
Denitrificans) it is one of heterotrophic nitrification, aerobic denitrifying bacteria strain, ammoniacal nitrogen directly can be converted into final gaseous state and produced
Thing escapes, and makes it possible Simultaneous Nitrification And Denitrification technology.Compared with traditional Autotrophic nitrification bacterium, heterotrophism nitre
Change rate of bacterial growth is fast, the speed of action is high.
Antibiotic is widely used in livestock and poultry breeding industry to prevent and treat Animal diseases, but these antibiotic only have seldom one
Part is utilized by animal body, and the overwhelming majority is excreted by urine and excrement, and breeding wastewater is the important of antibiotic pollution
Source, in environment antibiotic enrichment ecological environment and human health can be threatened.To the processing method master of antibiotic pollution
There are 2 kinds of non-biodegradation and biodegradation, microbial degradation is one of biodegradable mode.Seek new Degradation of Antibiotics
Microbial strains can provide reference for antibiotic contamination in following microbial resources and technical finesse environment.
The content of the invention
The purpose of the present invention is the deficiency for existing cultivating wastewater purification technology, there is provided a strain denitrogen paracoccus, should
Bacterial strain can remove breeding wastewater while ammonia nitrogen is removed in fluoroquinolone antibiotics.Described Paracoccus denitrificans in
On May 5th, 2017 is preserved in China typical culture collection center (CCTCC), Classification And Nomenclature：Paracoccus denitrificans
(Paracoccus denitrificans) TDFQ-1, deposit number are CCTCC NO：M2017238 addresses：Wuhan, China Wuhan
University.
It is another object of the present invention to provide Paracoccus denitrificans TDFQ-1 preparation method, method is simple, fermentation
Effective bacteria concentration is high in liquid, is adapted to large-scale production.
It is also an object of the present invention to provide Paracoccus denitrificans TDFQ-1 application, denitrogenation pair provided by the invention
Coccus can be used for the ammonia nitrogen and fluoroquinolone antibiotics gone simultaneously in water removal, the processing especially suitable for breeding wastewater.
In order to achieve the above object, the present invention takes following technical measures：
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 acquisition：
Applicant screens efficient allotrophic nitrobacteria from Suizhou, hubei pig factory sewage, by largely screening, physiology life
Change and 16S identifications, final screening obtain a strain denitrogen paracoccus bacterial strain, described Paracoccus denitrificans protects on May 5th, 2017
It is hidden in China typical culture collection center (CCTCC), Classification And Nomenclature：Paracoccus denitrificans (Paracoccus
Denitrificans) TDFQ-1, deposit number are CCTCC NO：M2017238 addresses：Wuhan, China Wuhan University.
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 biological characteristics is as described below：
The bacterial strain can be grown in organic and inorganic medium, and flourish is formed on nutrient agar (NA) culture medium
Circular colonies, bacterium colony size 1-2mm, the smooth moistening in surface.The bacterium microscopy is Gram-negative, single, paired or in heaps, logarithm
Growth period is spherical or subsphaeroidal, and mean size is 1.1 × 1.3um, and resting stage thalline is spherical, diameter 1um.Do not hydrolyze bright
Glue, starch；D-R, arginine are not utilized；Given birth to using sucrose, trehalose, maltose, mannitol for sole carbon source
It is long；Ammonium salt and nitrate can act as nitrogen source；By the use of oxygen or nitrate nitrous oxide can be reduced into as electron acceptor, nitrate
And molecular nitrogen.30 DEG C of optimum temperature, 5-37 DEG C of temperature range.By its 16S rRNA gene sequencing, as a result carried out in NCBI
Blastn is compared, and finds its 16SrRNA and Paracoccus denitrificans (NR 114145.1) sequence homology highest, and similitude is
99%.
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 preparation method, including：
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 seed liquors are pressed into 1-10% (V/V) inoculum concentration
It is inoculated in 50L fermentation tanks and expands culture, fermentation tank liquid amount is 30-35L, and control speed of agitator maintains dissolved oxygen in 15%-
18%, 28-32 DEG C of culture 18-22h collect zymotic fluid.
Described fermentation tank, the formula of its filling fermentation medium include：Peptone 5g, yeast extract 5g, NaCl 2g,
MgSO40.03g, Na2HPO4 0.25g, KH2PO40.75g, FeSO40.03g, MnSO4 0.01g, CaCl20.2g, glycerine
6.0g, add water to be settled to 1L, adjust PH7.5.
In schemes described above, it is preferred that inoculum concentration 3%, the liquid amount of fermentation tank is 35L, dissolved oxygen 16%, is trained
It is 30 DEG C to support temperature.
Paracoccus denitrificans TDFQ-1 application, given up using the Paracoccus denitrificans TDFQ-1 of the present invention for handling livestock and poultry
Water, or for handling the sewage containing high ammonia nitrogen and antibiotic simultaneously.
The preferred Norfloxacin of described antibiotic, Ciprofloxacin or Enrofloxacin.
The invention has the advantages that：
Paracoccus denitrificans TDFQ-1 provided by the invention has heterotrophic nitrification-aerobic denitrification effect, organic and inorganic
It can be grown in culture medium, heterotrophic growth speed is much higher than autophyting growth speed.Paracoccus denitrificans TDFQ-1 can contain height
Concentration organic and NH3Grown in-N breeding wastewater, there is high efficiency nitrification and denitrifying capacity, pass through heterotrophic nitrification point
Organic substance is solved, ammonium nitrogen is converted into nitrate nitrogen and nitrite nitrogen, is reduced to nitrite nitrogen by aerobic denitrification
Nitrogen or nitrous oxide.Bacterial strain of the present invention, can be with quinolones thing in degrading cultivation waste water while nitrification is carried out
Matter, the pollution for excessively using and discharging to soil and groundwater of livestock and poultry antibiotic is reduced, avoids antibiotic from passing through food chain
Accumulation effect has an impact to health.
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 of the present invention, which has, is easy to fermented and cultured, raw
Produce cost it is low, can in the waste water containing high-enriched organics growth and breeding, be new bacterium the mineralized nitrogen in breeding wastewater
Body cell and harmless N2, ammonia nitrogen therein, total nitrogen are efficiently removed, is compared to physics and method of chemical treatment, it has investment
The advantages of energy consumption is low less.
18h is cultivated in NB, viable bacteria content reaches 5.84 × 109CFU/ml, cultivate in fermentation medium after optimization
18h, viable bacteria content can reach 8.92 × 109CFU/ml, viable count improve 52.74%.TDFQ-1 preparations are used to handle
Wastewater from Pig Farm, pig farm breeding wastewater is handled with said preparation, the content of ammonia nitrogen and total nitrogen in the breeding wastewater of pig farm is by TDFQ-1
562.08mg/L, 645.66mg/L of before processing are reduced to 54.21mg/L, 85.84mg/L, reduce by 90.35% He respectively
86.71%.Chicken farm cultivation waste water is handled, the content of ammonia nitrogen and total nitrogen in chicken farm cultivation waste water is by TDFQ-1 before processings
662.37mg/L, 695.83mg/L are reduced to 54.36mg/L, 69.11mg/L, reduce 91.79% and 90.07% respectively.
While the ammonia nitrogen and total nitrogen in efficiently removing animal farm wastewater, TDFQ-1 can be with the quinoline promise in degrading waste water
Ketone antibiotic.In the breeding wastewater of pig farm, the test group using TDFQ-1 preparations is compared with the control group without using said preparation
Compared with, Norfloxacin degradation rate is 93.41%, improves 69.36%；Ciprofloxacin degradation rate is 96.93%, is improved
68.59%；Enrofloxacin degradation rate is 93.82%, improves 51.35%.Use 3 kinds of quinolones in TDFQ-1 test group
The degradation rate of antibiotic has reached more than 90%.
Brief description of the drawings
Fig. 1 is griess reagent color developing method primary dcreening operation deammoniation azotobacter schematic diagrames；
Wherein：E holes are TDFQ-1, and H holes are control.
Fig. 2 is Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 16SrDNA phylogenetic evolutions
Tree.
Fig. 3 is Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 light micrographs (400 times).
Fig. 4 is Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 growth curve charts.
Fig. 5 is determination of total nitrogen content standard curve.
Fig. 6 is ammonia nitrogen determination standard curve.
Fig. 7 is Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 to quinolone in the breeding wastewater of pig farm
Class Degradation of Antibiotics effect diagram.
Embodiment
Technical scheme of the present invention, it is the ordinary skill in the art if not otherwise specified；The reagent or material,
If not otherwise specified, commercial channel is derived from.
Used medium of the embodiment of the present invention is as follows：
The formula (1L) of liquid seeds culture collection (NB)：Beef extract 5g, peptone 10g, sodium chloride 5g, water is added to be settled to
1L；
The formula (1L) of solid seed culture medium (NA)：Beef extract 5g, peptone 10g, sodium chloride 5g, agar 15g, add water
It is settled to 1L.
Fermentation medium (1L)：Peptone 5g, yeast extract 5g, NaCl 2g, MgSO4 0.03g, Na2HPO4 0.25g,
KH2PO4 0.75g, FeSO4 0.03g, MnSO4 0.01g, CaCl2 0.2g, glycerine 6.0g, add water to be settled to 1L, regulation
PH7.5。
Liquid nitrification culture medium (YW) formula (1L)：(NH4) 2SO4 2g, MnSO4 0.01g, MgSO4 0.03g,
Na2HPO4 0.25g, KH2PO4 0.75g, CaCl2 0.2g, FeSO4 0.03g, glycerine 6g, add water to be settled to 1L, regulation
PH7.8。
Embodiment 1：
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 acquisition：
From South Lake, Suizhou, hubei pig farm, Anlu Hubei's chicken house collection lake water and animal farm wastewater, with (NH4)2SO4For
Only nitrogen source, enrichment culture can utilize the bacterium that ammonia nitrogen is nitrogen source growth in YW fluid nutrient mediums, and 30 DEG C, 200rpm is cultivated
72h, it is continuous in YW culture mediums to be enriched with passage 3 times, flat board is applied after nutrient solution dilution, picking single bacterium colony is inoculated in YW Liquid Cultures
Base Shaking culture, the strong bacterial strain of nitrification ability is screened with griess reagent color developing methods (see Fig. 1).
Liquid nitrification culture medium (YW) formula：(NH4)2SO42g, MnSO40.01g, MgSO40.03g, Na2H PO4
0.25g, KH2PO40.75g, CaCl20.2g, FeSO40.03g, glycerine 6g, PH7.8.
Strain 16S is identified
The 7 plants of bacterium obtained to separation are identified, utilize 16S universal primers (27F and 1492R, the primer sequence delivered
For 27F：5’-AGAGTTTGATCCTGGCTCAG-3’；1492R：5 '-GGTTACCTTGTTACGACTT-3 ') to 7 plants of bacterium
16SrRNA gene orders are expanded and are sequenced, and pcr amplification product send Sangon Biotech (Shanghai) Co., Ltd. to survey
Sequence.
Sequencing sequence carries out blastn comparisons in NCBI, and is evolved and sent out using the clusterW constructing systems in MEGA softwares
Tree (chadogram sees Fig. 2) is educated, sequence analysis result shows that this 7 plants of bacterium are respectively 2 bacillus, 2 strain denitrogen paracoccus, 1
Pseudomonas, 1 plant of Rhodococcus erythropolis, 1 plant of particle protozoa, it is respectively designated as bacillus (Bacillus sp) XHYB-
1st, bacillus (Bacillus sp) XHYB-2, Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1, denitrogenation
Secondary coccus (Paracoccus denitrificans) TDFQ-2, Rhodococcus erythropolis (Rhodococcus erythropolis)
HCHQ, pseudomonad (Pseudomonas sp) JDB, particle protozoa (Shinella granuli) KLSS, wherein denitrogenation
Secondary coccus (Paracoccus denitrificans) TDFQ-1 changes colour most substantially during development process separation screening, illustrates it
Heterotrophic nitrification ability is most strong.
Physiology and biochemistry identifies TDFQ-1：
TDFQ-1 flourish on nutrient agar (NA) culture medium forms circular colonies, bacterium colony size 1-2mm, surface light
Sliding moistening.The bacterium microscopy is Gram-negative, single, paired or in heaps, and exponential phase is spherical or subsphaeroidal, mean size
For 1.1 × 1.3um, resting stage thalline is spherical, diameter 1um.Not gelatin hydrolysate, starch；D-R, smart ammonia are not utilized
Acid；Grown using sucrose, trehalose, maltose, mannitol for sole carbon source；Ammonium salt and nitrate can act as nitrogen source；Energy
Electron acceptor is used as by the use of oxygen or nitrate.With《The outstanding Bacteria Identification handbook of uncle》Middle Paracoccus denitrificans physio-biochemical characteristics are consistent, really
It is Paracoccus denitrificans (Paracoccus denitrificans) to recognize the bacterium.TDFQ-1 light micrographs are shown in Fig. 3.Described is de-
Nitrogen pair coccus is preserved in China typical culture collection center (CCTCC), Classification And Nomenclature on May 5th, 2017：Denitrogenation pair ball
Bacterium (Paracoccus denitrificans) TDFQ-1, deposit number are CCTCC NO：M2017238 addresses：Wuhan, China is military
Chinese university.
Embodiment 2：
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 preparation：
1) Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 growth curve
Inoculation by the Paracoccus denitrificans activated (Paracoccus denitrificans) TDFQ-1 by 1% (V/V)
Amount is inoculated in 200mlNB and 200ml fermentation mediums respectively, 30 DEG C, 200rpm shaking table cultures, OD600 is measured by sampling every 2h
Value, using the time as abscissa, OD600 values are that ordinate draws growth curve, see Fig. 4.
The Paracoccus denitrificans of table 1 (Paracoccus denitrificans) TDFQ-1 growth curve data
Growth curve represents the growth characteristics of microorganism, and the growth of microorganism can be specified according to the growth curve of microorganism
Rule, the time for determining its arrival stationary phase can determine suitable fermented incubation time, has certain finger to production practices
Lead meaning.
As a result show, in NB culture mediums, TDFQ-1 enters exponential phase from 4h, and 18h reaches stationary phase, 4-18h
Between be in fast-growth exponential phase；TDFQ-1 enters exponential phase from 4h in the fermentation medium, and 18h reaches steady
Periodically, the exponential phase of fast-growth is between 4-18h.From TDFQ-1 growth curve, bacterial strain of the present invention is in 18h
Stationary phase is can reach, growth cycle is short, under large scale fermentation condition of culture, can reduce production cost.
2) Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 preparation：
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 seed liquors are pressed to 1% (V/V) inoculum concentration
200ml fermentation mediums are inoculated in, 30 DEG C, 200rpm shaking table cultures 18h (reaches time point stationary phase), and viable count reaches 8.92
×109CFU/ml.And TDFQ-1 be inoculated under similarity condition in NB culture mediums cultivate 18h when, viable count be 5.84 ×
109CFU/ml, the viable count that compares improve 52.74%.
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 seed liquors are connect by 3% (V/V) inoculum concentration
Kind expands culture in 50L fermentation tanks, and fermentation tank liquid amount is to maintain dissolved oxygen to be trained at 16%, 30 DEG C for 35L, control speed of agitator
Support 18h and collect zymotic fluid, zymotic fluid bacterium amount reaches 9.87 × 109CFU/ml。
The formula (g/L) of described fermentation medium：Peptone 5g, yeast extract 5g, NaCl 2g, MgSO40.03g,
Na2HPO40.25g, KH2PO40.75g, FeSO40.03g, MnSO40.01g, CaCl20.2g, glycerine 6.0, regulation
PH7.5；
Embodiment 3：
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 removes to ammonia nitrogen in YW culture mediums and total nitrogen
Effect
Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 is inoculated in 200ml NB fluid nutrient mediums
In, 30 DEG C, 200rpm is cultivated 18 hours, and supernatant is removed in the centrifugation of zymotic fluid liquid, adds sterilizing is single to steam water washing 2 times, adds sterilizing is single to steam water
It is resuspended, regulation cell concentration OD600 values are 1.5；Seed liquor is resuspended and is inoculated in 200ml YW culture mediums by 5% inoculum concentration, 30
DEG C, 200rpm culture 54h, 2ml nutrient solutions are taken within every 6 hours, for determining the content of total nitrogen and ammonia nitrogen in zymotic fluid.Total nitrogen uses
Potassium persulfate oxidation scene photometry, ammonia nitrogen use Nessler's reagent photometer.
Total nitrogen and ammonia nitrogen determination standard curve：Fig. 5 and Fig. 6, as a result as shown in table 2 and table 3.
In Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 difference incubation time zymotic fluids total nitrogen and
Ammonia-nitrogen content determines.
The Paracoccus denitrificans of table 2 (Paracoccus denitrificans) TDFQ-1 is to nitrogen removal rate in culture medium
The Paracoccus denitrificans of table 3 (Paracoccus denitrificans) TDFQ-1 is to ammonia nitrogen removal frank in culture medium
From table 2 and table 3, Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 is to total in culture medium
The final clearance of nitrogen is 92.41%, and ammonia nitrogen, the decrement of total nitrogen are all higher than when the clearance of ammonia nitrogen is 94.72%, 18h
85%.Increase over time afterwards, the change of the decrement of ammonia nitrogen and total nitrogen be not very greatly, therefore, in actual applications, denitrogenation
Secondary coccus TDFQ-1 is defined as 18h to the processing time of breeding wastewater.
Embodiment 4：
Processing of Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 to plant's high organic waste water
First, to the processing of pig farm breeding wastewater
1. ammonia nitrogen and total nitrogen removal effect in pair pig farm breeding wastewater
The breeding wastewater on Suizhou pig farm is taken, is filtered off except the particle suspensions in breeding wastewater, the sewage after filtering
Experimental water as Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 denitrifications.By 5% (V/V's)
Measure 12.5ml (viable counts 5 × 109CFU/ml TDFQ-1 liquid preparations) are added in 250ml sewage, at room temperature (25 DEG C) place
18h is managed, determines the changes of contents of ammonia nitrogen in sewage and total nitrogen.
The Paracoccus denitrificans of table 4 (Paracoccus denitrificans) TDFQ-1 is to ammonia nitrogen in the breeding wastewater of pig farm and always
The removal effect of nitrogen
As shown in Table 4, by Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 processing, pig farm is supported
The content for growing the ammonia nitrogen in waste water and total nitrogen is reduced to 54.21mg/ by 562.08mg/L, 645.66mg/L of TDFQ-1 before processings
L, 85.84mg/L, 90.35% and 86.71% is reduced respectively.
2. the degradation of fluoroquinolone antibiotics in pair pig farm breeding wastewater
The breeding wastewater on Suizhou pig farm is taken, is filtered off except the particle suspensions in breeding wastewater, the sewage after filtering
Experimental water as Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 degraded carbostyril antibiotics.Press
5% (V/V) amount is by 12.5ml (viable counts 5 × 109CFU/ml TDFQ-1 liquid preparations) are added in 250ml sewage, room temperature
Under (25 DEG C) processing certain times, determine sewage in quinolones (Norfloxacin, Ciprofloxacin, Enrofloxacin) antibiotic content
Change (table 5, Fig. 7).The measure of antibiotic concentration uses high performance liquid chromatography (HPLC) in breeding wastewater.
The Paracoccus denitrificans of table 5 (Paracoccus denitrificans) TDFQ-1 is to quinolones in the breeding wastewater of pig farm
The degradation (unit ng/L) of antibiotic
As shown in Table 5, it is useless with Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 processing pig farm cultivation
The test group of water and unused TDFQ-1 control group compare, and Norfloxacin degradation rate brings up to 93.41% from 24.05%, drop
Solution rate improves 69.36%；Ciprofloxacin degradation rate brings up to 96.93% from 28.34%, and degradation rate improves 68.59%；Grace
Promise sand star degradation rate brings up to 93.82% from 42.47%, improves 51.35%.Use 3 kinds of quinoline promises in TDFQ-1 test group
The degradation rate of ketone antibiotic has reached more than 90%.
2nd, to ammonia nitrogen in chicken farm cultivation waste water and total nitrogen removal effect
The breeding wastewater of Anlu chicken house is taken, is filtered off except the particle suspensions in breeding wastewater, the sewage after filtering
Experimental water as Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 denitrifications.By 5% (V/V's)
Measure 12.5ml (viable counts 5 × 109CFU/ml TDFQ-1 liquid preparations) are added in 250ml sewage, at room temperature (25 DEG C) place
18h is managed, determines the changes of contents of ammonia nitrogen in sewage and total nitrogen.
The Paracoccus denitrificans of table 6 (Paracoccus denitrificans) TDFQ-1 is to ammonia nitrogen in chicken farm cultivation waste water and always
The removal effect of nitrogen
| Index | Before processing | After processing | Clearance (%) |
| Ammonia nitrogen (mg/L) | 662.37 | 54.36 | 91.79 |
| Total nitrogen (mg/L) | 695.83 | 69.11 | 90.07 |
As shown in Table 6, supported by Paracoccus denitrificans (Paracoccus denitrificans) TDFQ-1 processing, chicken house
The content for growing the ammonia nitrogen in waste water and total nitrogen is reduced to 54.36mg/ by 662.37mg/L, 695.83mg/L of TDFQ-1 before processings
L, 69.11mg/L, 91.79% and 90.07% is reduced respectively.
Claims (5)
1. the Paracoccus denitrificans of one plant of separation, it is characterised in that described Paracoccus denitrificans is Paracoccus denitrificans (Paracoccus
Denitrificans) TDFQ-1, deposit number are CCTCC NO：M2017238.
2. the preparation method of the Paracoccus denitrificans described in claim 1, including：By Paracoccus denitrificans (Paracoccus
Denitri ficans) TDFQ-1 seed liquors by 1-10% (V/V) inoculum concentration be inoculated in 50L fermentation tanks expand cultivate, fermentation
Canned liquid measure is 30-35L, and control speed of agitator maintains dissolved oxygen to collect zymotic fluid in 15%-18%, 28-32 DEG C of culture 18-22h；
Described fermentation tank, the formula of its filling fermentation medium include：Peptone 5g, yeast extract 5g, NaCl 2g, MgSO4
0.03g, Na2HPO40.25g, KH2PO40.75g, FeSO40.03g, MnSO40.01g, CaCl20.2g, glycerine 6.0g, adds
Water is settled to 1L, adjusts PH7.5.
3. application of the Paracoccus denitrificans in livestock and poultry farm wastewater treatment described in claim 1.
4. application of the Paracoccus denitrificans in antibiotic of degrading described in claim 1, described antibiotic is Norfloxacin, ring
Third husky star or Enrofloxacin.
5. application of the Paracoccus denitrificans in sewage of the processing simultaneously containing high ammonia nitrogen and antibiotic described in claim 1, institute
The antibiotic stated is Norfloxacin, Ciprofloxacin or Enrofloxacin.
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| CN110117567B (en) * | 2019-06-04 | 2023-03-03 | 西安工程大学 | Paracoccus denitrificans strain screening and application thereof in deodorization |
| CN110452846B (en) * | 2019-08-16 | 2022-07-12 | 鹭滨环保科技（上海）股份有限公司 | Paracoccus denitrificans and biological denitrification method by combining paracoccus denitrificans with mineralization bed |
| CN110452858B (en) * | 2019-09-12 | 2021-04-27 | 中国科学院微生物研究所 | Paracoccus B54 and application thereof |
| CN110591962B (en) * | 2019-09-28 | 2020-11-06 | 华中农业大学 | Proteus mirabilis BSFLG-CIP5 derived from stratiomyiid intestinal tract and application thereof |
| CN112723558B (en) * | 2020-12-16 | 2021-12-14 | 青岛尚德生物技术有限公司 | Application of paracoccus denitrificans in preparation of microbial agent for degrading ammoniacal nitrogen in landfill leachate |
| CN112522152B (en) * | 2020-12-16 | 2021-10-01 | 青岛尚德生物技术有限公司 | Salt-tolerant paracoccus denitrificans GBW-HB1904 and application thereof |
| CN113830902A (en) * | 2021-09-23 | 2021-12-24 | 青岛尚德生物技术有限公司 | Application of paracoccus denitrificans in removal of nitrate nitrogen in high-salinity wastewater |
| CN114181856B (en) * | 2021-11-30 | 2023-12-15 | 南京农业大学 | Microbial combination capable of effectively inhibiting odor production of biomass fermentation from source and application thereof |
| CN114933990B (en) * | 2022-05-31 | 2023-09-05 | 南京理工大学 | N-methylpyrrolidone degradation synchronous denitrifying bacterium and application thereof |
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| CN102465104B (en) * | 2010-11-04 | 2013-03-20 | 中国石油化工股份有限公司 | Aerobic denitrifying Paracoccus denitrificans and application thereof |
| CN103045520A (en) * | 2013-01-05 | 2013-04-17 | 福建省农业科学院农业生物资源研究所 | Heterotrophic nitrification bacterial strain |
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Denomination of invention: A strain of denitrifying Paracoccus and its application in livestock wastewater treatment Effective date of registration: 20210401 Granted publication date: 20180410 Pledgee: Wuhan Branch of China Everbright Bank Co.,Ltd. Pledgor: WUHAN KEYUAN BIOLOGICAL DEVELOPMENT Co.,Ltd. Registration number: Y2021420000018 |