CN105969659A - Culture device and culture method for determining denitrification of attached organisms of water plants - Google Patents
Culture device and culture method for determining denitrification of attached organisms of water plants Download PDFInfo
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- CN105969659A CN105969659A CN201610574369.2A CN201610574369A CN105969659A CN 105969659 A CN105969659 A CN 105969659A CN 201610574369 A CN201610574369 A CN 201610574369A CN 105969659 A CN105969659 A CN 105969659A
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- denitrification
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- aufwuch
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 80
- 238000012136 culture method Methods 0.000 title abstract description 5
- 239000000523 sample Substances 0.000 claims description 18
- 239000012531 culture fluid Substances 0.000 claims description 17
- 238000000034 method Methods 0.000 claims description 14
- 239000007789 gas Substances 0.000 claims description 9
- 239000011521 glass Substances 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 9
- 239000004033 plastic Substances 0.000 claims description 6
- 229920003023 plastic Polymers 0.000 claims description 6
- 229920005479 Lucite® Polymers 0.000 claims description 4
- 239000004926 polymethyl methacrylate Substances 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 230000017531 blood circulation Effects 0.000 claims description 3
- 125000004122 cyclic group Chemical group 0.000 claims description 3
- 238000007865 diluting Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 3
- 238000011534 incubation Methods 0.000 claims description 3
- 230000000155 isotopic effect Effects 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 230000002085 persistent effect Effects 0.000 claims description 3
- 239000012488 sample solution Substances 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 239000011592 zinc chloride Substances 0.000 claims description 3
- 229920005549 butyl rubber Polymers 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 claims description 2
- 238000004321 preservation Methods 0.000 claims description 2
- 238000006243 chemical reaction Methods 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 3
- 238000011065 in-situ storage Methods 0.000 abstract description 3
- 238000005259 measurement Methods 0.000 abstract description 3
- 238000012258 culturing Methods 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 21
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 229910002651 NO3 Inorganic materials 0.000 description 6
- 239000012790 adhesive layer Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 230000004087 circulation Effects 0.000 description 3
- 238000012851 eutrophication Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000004172 nitrogen cycle Methods 0.000 description 3
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- HSFWRNGVRCDJHI-UHFFFAOYSA-N alpha-acetylene Natural products C#C HSFWRNGVRCDJHI-UHFFFAOYSA-N 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 125000002534 ethynyl group Chemical group [H]C#C* 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 240000004590 Potamogeton malaianus Species 0.000 description 1
- 241000589651 Zoogloea Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012364 cultivation method Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000027734 detection of oxygen Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000007954 hypoxia Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000865 membrane-inlet mass spectrometry Methods 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
- C02F3/327—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae characterised by animals and plants
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
Abstract
The invention discloses a culture device and culture method for determining denitrification of attached organisms of water plants. The culture device is simple and reasonable in structure; a denitrification sample of the attached organisms of the water plants is cultured through a circulating device, so that water which flows and is used for culturing is circulated to form mixed water flow and the water flow is lasted to realize ion concentration and gas distribution in a solution; wild in-situ conditions can be simulated to the greatest extent and a measurement is accurate and reliable; and the tightness is good and the operation is simple and convenient. With the adoption of the culture method, the working efficiency can be improved, operation errors are reduced and a reaction progress is accelerated; and the denitrification potential of the attached organisms can also be fully played, so that an experiment result is closer to an actual value, and the culture method has important meanings on accurate estimation of the denitrification of the attached organisms of the water plants.
Description
Technical field
The present invention relates to a kind of device and method measuring Denitrification, specifically a kind of mensuration water plant is attached
Culture apparatus and the cultural method of biological denitrification.
Background technology
In eutrophication water, water plant stem and leaf surface is often enriched algae, animal, microorganism, zoogloea, organic broken
The many kinds of substances such as bits, inorganic sand grain, form the adhesive layer that thickness does not waits.In adhesive layer heterogeneous microenvironment to water body with plant
Material transport between thing converts has material impact, and then the material circulation of regulation and control water body.The water such as Dan Shi China Hu Ku, river
One of major pollutants of body, regulate and control and cut down the key issue that nitrogen load is water environment management and reparation.Denitrification therein
Effect is the important channel that nitrogen departs from that water body converts to aerial migration, has research whole world nitrogen cycle and body eutrophication
Significance.
At present, research is concentrated mainly in the Denitrification of deposit, and makees water plant aufwuch denitrification
Research relatively fewer.Preliminary study shows: the adhesive layer on water plant stem and leaf surface exists oxygen-enriched micro-environmental hypoxia, and
Containing abundant nitrate, organic matter and microorganism, have detected that Denitrification.Abundant at nutrient, phytomass is bigger
Waters, adhesive layer has become the hot spot region of Denitrification, close to even more than deposit, therefore, accurately measures attached
Biological denitrification and offset pollution-reducing water body nitrogen load and to improve nitrogen cycle theoretical system significant.
Water plant aufwuch Progress of studies on denitrification main cause slowly is a lack of effective culture apparatus
And cultural method.At present, effective apparatus and method are not the most fixed in the cultivation to adhesive layer sample.And to denitrification product
The acetylene inhibition method measuring main employing traditional (surveys N2O), the method is typically by using simple sealing after gathering sample
Serum bottle or lucite tube are collected sample and are taken back laboratory treatment, not only sample loaded down with trivial details, workload big, and easily due to gas
Bulk diffusion uneven cause volumetric errors and NO3 -Cause inhibitory action incomplete when concentration is low, affect the precision of experimental result, shadow
Ring research progress.
Summary of the invention
It is an object of the invention to provide a kind of culture apparatus measuring water plant aufwuch Denitrification and training
Breeding method.This apparatus structure advantages of simple, utilizes circulating device to cultivate water plant aufwuch Denitrification sample, not only
The water circulation that flowing can be made to cultivate forms mixed flow, and makes effects of ion concentration and gas be distributed by constant stream,
And field in-situ condition can also be simulated to greatest extent, it is ensured that measurement result accurately and reliably, and good leak tightness, easy and simple to handle.
This culture fluid can make water plant continue the growth conditions that holding is good, makes aufwuch persistently keep biological activity.This training
Breeding method not only can improve work efficiency, reduce personal error, quickening reaction process, but also aufwuch denitrification can be made to make
Obtain maximum performance by potentiality, and then make experimental result closer to actual value.The present invention is used for instructing and accurately measures water plant
The Denitrification of aufwuch, for abatement polluted-water nitrogen load with improve nitrogen cycle theoretical system and have important function.
The present invention is by the following technical solutions:
A kind of culture apparatus measuring water plant aufwuch Denitrification, it is by culture vessel, Seperating box and makes
The centrifugal pump composition that water circulates；Described culture vessel one end water inlet is sealed even by water pipe with the outlet of Seperating box
Connect；The outlet of described centrifugal pump is connected with the inlet seal of Seperating box；The outlet of described culture vessel and centrifugal pump
Inlet seal connects；Described Seperating box is provided with water inlet, sample tap and thermowell.
Preferably, described culture vessel is transparent airproof lucite tube, glass tubing or plastic tube, manages a diameter of 40-
70mm, pipe range 400-450mm.
Preferably, described connection culture vessel, between Seperating box and centrifugal pump, interconnective water pipe is organic plastics pipe
Or butyl rubber pipe, a diameter of 15-30mm.
Preferably, the two ends of described culture vessel and water pipe use sealing-plug to be tightly connected.
Preferably, described culture vessel places the water plant with aufwuch.
A kind of above-mentioned culture apparatus is utilized to cultivate the method for measuring water plant aufwuch Denitrification,
It comprises the following steps:
1), about raw axis section 20g of water intaking, load in transparent airproof culture vessel, with Seperating box, centrifugal pump group
Dress up Cyclic culture system, after sealing in dark in position at a temperature of preculture 12h, oxygen microelectrode detection make O2Concentration is reduced to
93μmol·L-1Below；
2) after diluting 100 times according to the culture fluid of recipe configuration, Na is added15NO3, make in culture fluid15NO3 -Concentration is
100μmol·L-1；
3) by step 2) the isotopic culture fluid of interpolation after dilution injected in blood circulation by water inlet, and culture fluid leads to
Cross centrifugal pump persistent loop in transparent tube；
4) 4-6 incubation time sequence is set, each time point set 3 parallel, from sample tap draw solution to airproof
In elongated grournd glass pipe, adding 2mL concentration is 0.5g mL-1ZnCl2Solution terminates microbial activity, covers rapidly stopper
And ensure without head space, after covering grinding port plug, then use ParafilmTM；Acquired water sample is less than the 10% of recirculated water；
5) the grournd glass pipe placing sample solution is inverted in the preservation of constant temperature under water less than cultivation temperature 1-4 DEG C, with
Prevent gas from discharging；
6) soluble gas is measured with film interface mass spectrograph in 24h28N2、29N2、30N2Content；
7) finally Denitrification speed is calculated with isotope matching method.
Described step 2) in the formula of culture fluid consist of:
The invention has the beneficial effects as follows: the present invention utilizes circulating device to cultivate water plant aufwuch Denitrification sample
Product, the water circulation being possible not only to make flowing cultivate forms mixed flow, and makes effects of ion concentration stable by constant stream,
Gas is evenly distributed, and can also limits simulation field in-situ condition.Apparatus of the present invention are simple, reasonable in design, it is ensured that survey
Measure result accurately and reliably, good leak tightness, easy and simple to handle, with low cost, there is good application prospect.This water plant cultural method
Culture fluid used by can make water plant continue the growth conditions that holding is good, makes aufwuch persistently keep biological activity.
This cultural method not only can improve work efficiency, reduce operating error, quickening reaction process, but also can make the anti-nitre of aufwuch
Change effect potentiality obtain maximum performance, and then make experimental result closer to actual value, make water plant aufwuch denitrification
Accurate budget significant.
Accompanying drawing explanation
Fig. 1 is apparatus of the present invention structural representations.
In figure, 1 is culture vessel, and 2 is centrifugal pump, and 3 is Seperating box, and 4 is thermowell, and 5 is sample tap, and 6 is into water
Mouthful.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described in further detail.
Embodiment 1
A kind of culture apparatus measuring water plant aufwuch Denitrification, it is by culture vessel 1, Seperating box 3 and
The centrifugal pump 2 making water circulate forms；Described culture vessel 1 one end water inlet and the outlet of Seperating box 3 are close by water pipe
Envelope connects；The outlet of described centrifugal pump 2 is connected with the inlet seal of Seperating box 3；The outlet of described culture vessel 1 with from
The inlet seal of heart pump 2 connects；Described Seperating box 3 is provided with water inlet 6, sample tap 5 and thermowell 4.
Described culture vessel 1 is transparent airproof lucite tube, glass tubing or plastic tube, manages a diameter of 40-70mm, pipe
Long 400-450mm.
Between described connection culture vessel 1, Seperating box 3 and centrifugal pump 2, interconnective water pipe is organic plastics pipe or fourth
Base rubber tube, a diameter of 15-30mm.
The two ends of described culture vessel 1 and water pipe use sealing-plug to be tightly connected.
Described culture vessel 1 places the water plant with aufwuch.
The device of the present embodiment is used for submerged plant p.malaianus aufwuch denitrification in eutrophication water
The enforcement of effect experiment.Concrete grammar is as follows:
The cultivation of water plant adhesive layer sample:
1), about raw axis section 20g of water intaking, (pipe range 450mm, interior diameter in transparent airproof culture vessel 1 is loaded
50mm), and connect and compose Cyclic culture system by water pipe between Seperating box 3, centrifugal pump 2, after being sealed by culture vessel 1 in position
At a temperature of preculture 12h in dark, the detection of oxygen microelectrode makes O2Concentration is reduced to 93 μm ol L-1Below.
2) after diluting 100 times according to the culture fluid of table 1 recipe configuration, Na is added15NO3(99.3 15N atom%,
Sigma-Aldrich), make in culture fluid15NO3 -Concentration is 100 μm ol L-1。
3) by step 2) the isotopic culture fluid of interpolation after dilution injects in blood circulation by water inlet 6, culture fluid
By centrifugal pump 2 persistent loop in transparent tube.
4) 4-6 incubation time sequence (0,0.5,1.0,2.0,4.0,6.0h) is set, each time point set 3 parallel.
From sample tap 5 draw solution to airproof elongated grournd glass pipe, add ZnCl2Solution (2mL, 0.5g mL-1) terminate micro-
Biological activity, covers rapidly stopper and ensures without head space, after covering grinding port plug, then using ParafilmTM.Acquired water sample does not surpasses
Cross the 10% of recirculated water.
5) the grournd glass pipe placing sample solution is inverted in constant temperature (less than cultivation temperature 1-4 DEG C) under water to preserve, with
Prevent gas from discharging.
6) sample is measured with film interface mass spectrograph (Membrane Inlet Mass Spectrometer, MIMS) in 24h
Soluble gas28N2、29N2、30N2Content.
7) finally Denitrification speed is calculated with isotope matching method.
The culture fluid composition used in table 1 experiment
Additionally, the present embodiment is also provided with original position Aquaponic+isotope groups simultaneously, Solution culture method is not added with isotope groups,
Acetylene inhibition group, operating procedure is implemented according to appropriate technical solution, be the results are shown in Table 2.
Table 2 relevant experimental data
Above-mentioned test result indicate that, this device is scientific and reasonable, good leak tightness, and measurement result is the most reliable, it is possible to exactly
Carry out cultivation and the research of aufwuch Denitrification, there is well application and promotion prospect.
Claims (7)
1. the culture apparatus measuring water plant aufwuch Denitrification, it is characterised in that it is by culture vessel
(1), Seperating box (3) and the centrifugal pump (2) making water circulate form；Described culture vessel (1) one end water inlet and Seperating box
(3) outlet is tightly connected by water pipe；The outlet of described centrifugal pump (2) is connected with the inlet seal of Seperating box (3)；
The outlet of described culture vessel (1) is connected with the inlet seal of centrifugal pump (2)；Described Seperating box (3) is provided with water inlet
(6), sample tap (5) and thermowell (4).
The culture apparatus measuring water plant aufwuch Denitrification the most according to claim 1, it is characterised in that
Described culture vessel (1) is transparent airproof lucite tube, glass tubing or plastic tube, manages a diameter of 40-70mm, pipe range 400-
450mm。
The culture apparatus measuring water plant aufwuch Denitrification the most according to claim 1, it is characterised in that
Between described connection culture vessel (1), Seperating box (3) and centrifugal pump (2), interconnective water pipe is organic plastics pipe or butyl
Rubber tube, a diameter of 15-30mm.
The culture apparatus measuring water plant aufwuch Denitrification the most according to claim 1, it is characterised in that
The two ends of described culture vessel (1) and water pipe use sealing-plug to be tightly connected.
The culture apparatus measuring water plant aufwuch Denitrification the most according to claim 1, it is characterised in that
Described culture vessel (1) is placed the water plant with aufwuch.
6. one kind utilizes the culture apparatus according to any one of claim 1-5 to cultivate for measuring water plant aufwuch
The method of Denitrification, it is characterised in that it comprises the following steps:
1), about raw axis section 20g of water intaking, load in transparent airproof culture vessel, be assembled into Seperating box, centrifugal pump
Cyclic culture system, after sealing in dark in position at a temperature of preculture 12h, oxygen microelectrode detection make O2Concentration is reduced to 93 μ
mol·L-1Below；
2) after diluting 100 times according to the culture fluid of recipe configuration, Na is added15NO3, make in culture fluid15NO3 -Concentration is 100 μ
mol·L-1；
3) by step 2) the isotopic culture fluid of interpolation after dilution injected in blood circulation by water inlet, culture fluid by from
Heart pump persistent loop in transparent tube；
4) 4-6 incubation time sequence is set, each time point set 3 parallel, elongated to airproof from sample tap draw solution
In grournd glass pipe, adding 2mL concentration is 0.5g mL-1ZnCl2Solution terminates microbial activity, covers rapidly stopper and protects
Card, without head space, after covering grinding port plug, then uses ParafilmTM；Acquired water sample is less than the 10% of recirculated water；
5) the grournd glass pipe placing sample solution is inverted in the preservation of constant temperature under water less than cultivation temperature 1-4 DEG C, to prevent
Gas is discharged；
6) soluble gas is measured with film interface mass spectrograph in 24h28N2、29N2、30N2Content；
7) finally Denitrification speed is calculated with isotope matching method.
Cultivation the most according to claim 6 is for the method measuring water plant aufwuch Denitrification, its feature
Be, described step 2) in the formula of culture fluid consist of:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610574369.2A CN105969659A (en) | 2016-07-20 | 2016-07-20 | Culture device and culture method for determining denitrification of attached organisms of water plants |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610574369.2A CN105969659A (en) | 2016-07-20 | 2016-07-20 | Culture device and culture method for determining denitrification of attached organisms of water plants |
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| Publication Number | Publication Date |
|---|---|
| CN105969659A true CN105969659A (en) | 2016-09-28 |
Family
ID=56951996
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610574369.2A Pending CN105969659A (en) | 2016-07-20 | 2016-07-20 | Culture device and culture method for determining denitrification of attached organisms of water plants |
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| CN (1) | CN105969659A (en) |
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| CN111964993A (en) * | 2020-08-21 | 2020-11-20 | 水利部交通运输部国家能源局南京水利科学研究院 | Deep water sediment flow culture system for simulating in-situ water pressure |
| CN111977621A (en) * | 2020-08-12 | 2020-11-24 | 福建农林大学 | Production method of light-driven high-purity nitrous oxide |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109142549A (en) * | 2018-08-27 | 2019-01-04 | 睿藻生物科技（苏州）有限公司 | A kind of microalgae metabolic rate measurement chamber for mass spectrometer |
| CN111977621A (en) * | 2020-08-12 | 2020-11-24 | 福建农林大学 | Production method of light-driven high-purity nitrous oxide |
| CN111977621B (en) * | 2020-08-12 | 2022-10-14 | 福建农林大学 | Production method of light-driven high-purity nitrous oxide |
| CN111964993A (en) * | 2020-08-21 | 2020-11-20 | 水利部交通运输部国家能源局南京水利科学研究院 | Deep water sediment flow culture system for simulating in-situ water pressure |
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Application publication date: 20160928 |