CN109706237A - A method of evaluation polluted-water recovery - Google Patents
A method of evaluation polluted-water recovery Download PDFInfo
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- CN109706237A CN109706237A CN201910144415.9A CN201910144415A CN109706237A CN 109706237 A CN109706237 A CN 109706237A CN 201910144415 A CN201910144415 A CN 201910144415A CN 109706237 A CN109706237 A CN 109706237A
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Abstract
The invention belongs to technical field of sewage, and in particular to a method of polluted-water recovery is evaluated using biomembrane fungi composition and community diversity.This method judges the recovery of water body entirety with this using the composition and community diversity of fungi in high-flux sequence detection biomembrane, achievees the effect that judge that water body restores.This method is that fungi variation is closely related in novel water body recovery judgment method, with water body, and then can be changed by fungi in water body come comprehensive descision water body recovery situation.It is proved according to existing test data and embodiment, this method is that effective water body restores judgment method.
Description
Technical field:
The invention belongs to technical field of sewage, and in particular to a kind of to utilize biomembrane fungi composition and community diversity
The method for evaluating polluted-water recovery.
Background technique:
Traditional water body restoration evaluation mainly based on the physical and chemical index including each pollutant concentration, passes through
Pollutional condition locating for river system is evaluated in the qualitative or quantitative analysis of these indexs, this kind of index can be measured
Certain in environment is some or the levels of certain a class of pollutant matter, and monitoring result is exactly accurate, but can not be accurate
The summation polluting effect for distinguishing heterogeneity pollutant can not directly reflect all kinds of external source pollution inputs to the ecosystem
The influence of middle biology.
Most of microbe is so that existing for this attachment state of biomembrane, surface coatings in natural passes through in natural water
Microorganism extracellular polymeric in the surface of solids, constantly grows numerous the adherency of cell, cohesion using the nutriment in water as nutrient
It grows, plays an important role to water body substance circulation and energy flow, while the variation of biomembrane feature also can be by water body
The influence of middle environmental factor and nutrient, the xenobiotic pollutants matter in water environment can obtain conversion degradation on biomembrane.Mesh
The research of preceding structure composition and function to microorganism in biomembrane is concentrated mainly in bioreactor, the life to practical river
The research of object membrane sample is also less.In the natural environment, micropopulation has oneself defense mechanism in biomembrane, can be for outer
Boundary's environmental pressure is made a response.The microbiologic population of microorganism specific ionization in biomembrane can more resist severe extraneous ring
Border, such as the toxic action of poor nutritional and pollutant.
The present invention provides a kind of method using biomembrane fungi composition and community diversity evaluation water body recovery situation.It is logical
High-flux sequence detection biomembrane fungal organism diversity and its dominant bacteria relative abundance are crossed, above data comprehensive study is passed through
Restored with achieving the effect that evaluate water body.
Summary of the invention:
To achieve the goals above, the present invention will provide it is a kind of using biomembrane fungi composition and community diversity evaluation water
The method of body recovery situation.This method detects the composition and community diversity of fungi in aqueous bio film using high-flux sequence,
The recovery that water body entirety is judged with this achievees the effect that judge that water body restores.This method is that novel water body restores judgement
Fungi variation is closely related in method, with water body, and then can be by fungi variation in water body come comprehensive descision water body recovery situation.
It is proved according to existing test data and embodiment, this method is that effective water body restores judgment method.
The method includes the following steps:
(1) fungal organism film composition analysis
Cultivate respectively and acquire before water pollution and water body restore after biomembrane, after extracting DNA, carry out 18SrDNA high
Flux sequencing, determine before water pollution and water body restore after fungal species in biomembrane, specific to kind；
(2) fungal organism film community diversity is analyzed
It is similar to the fungal species progress in biomembrane after restoring before water pollution with water body using ANOSIM analysis method
Property analysis, obtain R value (β-diversity) and P value；
Compared with water body fungal species before pollution, value > 0.05 P can then be sentenced water body fungal species after recovery
Break the level that the water body is restored to before pollution, otherwise then prove the water body do not return to it is contaminated before it is horizontal；P value simultaneously
When > 0.05, R value is smaller, and it is higher to represent water body recovery extent.
The utility model has the advantages that
General water body recovery situation can only be carried out according to the levels of certain class or certain polluters in environment
Judgement, but the summation polluting effect of heterogeneity pollutant can not be accurately distinguished, can not directly it reflect all kinds of outer
Source pollution input is to influence biological in the ecosystem.It is proposed by the invention according to biomembrane fungi composition and community diversity
Method to judge water body recovery, fungi composition and group by biomembrane after restoring before detection water pollution with water body
Structure diversity judges that the ecological functions of water body are horizontal before whether being restored to, simple and effective.
Detailed description of the invention:
The fungal community of 1 background group of Fig. 1 embodiment, pollution group and recovery group is analyzed
Wherein, (a) is three different group fungi principal component analysis figures, is (b) three different group fungi superiority bacteria spps
Abundance situation；
2 bacterial community principal coordinate analysis of Fig. 2 embodiment
Wherein, M represents spring, and S represents autumn, and C represents background group, and P represents pollution group, and R represents recovery group.
Specific embodiment:
The present invention is further described referring to specific embodiment, it will be appreciated by those skilled in the art that this hair
It is bright to be not limited to these specific embodiments.
Method in following embodiments is unless otherwise instructed conventional method, used in reagent, such as without especially
Illustrate, is conventional commercial reagent.
Embodiment 1, sample used of the present invention acquisition and quality measuring method
The acquisition of 1.1 samples
It is uncontaminated water sample that we, which choose a kind of sugar, produced in chao'an county river valley alpine region basin water sample, chooses a kind of sugar, produced in chao'an county river valley urban area section
Water sample is pollution water sample.Experimental setup is three experimental groups: background group (control), pollution group (polluted) and recovery group
And every group of three Duplicate Samples (recover),.Biomembrane incubation time is 60 days, and specific incubation is as follows: first with multi-pass
Water sample in 5L plastic barrel is continuously injected into culturing room, culturing room's size by exhaust valve by road peristaltic pump are as follows: and 24mm ×
40mm × 8mm, volume are 7.7 cubic centimetres, and there are inlet and outlet in culturing room, and culturing room is discharged the plastics for being directly discharged into 5L
Bucket.Culturing room top and bottom end are transparent glass layer, can provide the adsorbable layer surface of biofilm formation.Wherein, background group
Water inlet be the uncontaminated water sample in alpine region, pollution group and recovery group, will be at the 21st day in the preceding water sample for being passed through alpine region for 20 days
Water is changed to the water sample of metropolitan district pollution, changes the water inlet of recovery group into clean alpine region river water again at the 41st day.Continuous training
After supporting 60 days, peristaltic pump is closed, the careful all biomembranes for scraping culturing room and being formed are added in 200ml sterile water, magnetic force stirs
Mixing device is uniformly mixed suspension.
1.2 biomembrane DNA are extracted and high-flux sequence
Biomembrane suspension is crossed to the filter membrane of 0.45 micron pore size, and illustrates to require to extract according to DNA extraction kit
The DNA sample extracted is sent to Hua Da gene and carries out 18SrDNA ITS high-flux sequence by DNA, and to the alkaloid after sequencing
Motif column data is analyzed, and is filtered particular by QIIME software to lower machine data, removal redundant sequence, and according to
97% similarity carries out cluster OTU analysis to sequence, each OTU is considered a kind of species, by data analysis we have found that
The superiority bacteria spp of background group is Rhodotorula, Acremonium and Cladosporium；The superiority bacteria spp of pollution group be Cladosporium,
Gloeotinia belongs to and Tuber；The superiority bacteria spp of recovery group is Acremonium, rhodotorula Pseudomonas and Cladosporium.We send out simultaneously
It include 9577 kinds of OTU in existing fungal organism film, and background group is more identical as the OTU relative abundance of recovery group, and pollution group
OTU relative abundance and background group and recovery group difference are larger.Since this experimental result data is huge, it is related to nearly ten thousand kinds of species,
Result can not be completely presented in the description, therefore setting table 1 does example results displaying, as shown in table 1.
1 biomembrane fungi OUT table of table
Wherein difference OTU represents different fungal species, and the numerical value in table represents the relative abundance of corresponding species, we
Accordingly it will be seen that the ANOSIM analysis of the composition situation of fungal organism film and progress next step.
1.3 similarity analysis methods
Similarity analysis is carried out using fungal species of the ANOSIM analysis method to background group, pollution group and recovery group to obtain
R value, wherein R value is that 1 to represent two groups of samples entirely different, and 0 is identical.
1.3 biomembrane fungies composition and community diversity
Each experimental group biomembrane fungi composition and structure of community are shown in Fig. 1, from figure 1 it appears that pollution group and background group,
There are notable differences for the fungal community of recovery group.Simultaneously by the biomembrane fungal species composition data of pollution group and background group
R=1, P=0.001 are obtained after carrying out ANOSIM analysis, show that the biomembrane fungal community of background group and pollution group is constituted
It is entirely different, R=is obtained after the biomembrane fungal species composition data of background group and recovery group is carried out ANOSIM analysis
0.374, P=0.07 this show water body restore after biomembrane fungal community tendency of changes before pollution group knot
Structure, recovery are good.
Meanwhile it can be seen that the fungi dominant bacteria phase of two experimental groups of background group and recovery group with pollution group by Fig. 1-b
To abundance, there were significant differences, and the main advantage strain relative abundance of recovery group and the relative abundance of background group reach unanimity.
By carrying out multiple sampling analysis to the sample before different location, different quality pollution and after restoring, we determined that
Water body restores threshold value: water body fungal community after recovery compared with water body fungal community before pollution its P value >
0.05, then it is horizontal before to may determine that the water body is restored to, it is contaminated horizontal before otherwise then to prove that the water body does not return to,
R is smaller simultaneously, and recovery extent is higher.
Embodiment 2
Using method same as Example 1, Caobai River basin water sample is acquired in spring and autumn respectively and background is set
Group (control), pollution group (polluted) and recovery group (recover) extract DNA, and carry out after carrying out biological Membrance cuiture
16SrDNA high-flux sequence, and determine bacterial species, specific to kind, to the bacterial species of background group, pollution group and recovery group,
By R language, to treated, bacterial species information carries out principal component analysis, and bacterial biof iotalm group beta diversity is as shown in Figure 2.
We have found after carrying out ANOSIM analysis to the other biofilm bacteria information of three groups: point between pollution group and background group
Analysis result is R=0.858, P=0.003；Analysis result between recovery group and background group is R=0.791, P=0.001.This
Showing the biofilm bacteria composition of recovery group and background group has a notable difference, and as shown in Fig. 2, autumn in principal coordinate analysis
Recovery group and the bacterial community difference of background group are even bigger than the bacterial community difference of pollution group and background group, therefore, it is considered that carefully
Flora falls the recovery situation that can not react water body.
Claims (1)
1. a kind of method for evaluating polluted-water recovery, which is characterized in that steps are as follows:
(1) fungal organism film composition analysis
Cultivate respectively and acquire before water pollution and water body restore after biomembrane in water body, after extracting DNA, carry out 18SrDNA
High-flux sequence, determine before water pollution and water body restore after fungal species in biomembrane, specific to kind；
(2) fungal organism film community diversity is analyzed
Using ANOSIM analysis method, similitude point is carried out to the fungal species before water pollution and after water body recovery in biomembrane
Analysis, obtains R value and P value；
For water body fungal species after recovery compared with water body fungal species before pollution, value > 0.05 P then may determine that this
Water body is restored to the level before pollution, and it is contaminated horizontal before otherwise then to prove that the water body does not return to；While P value >
When 0.05, R value is smaller, and it is higher to represent water body recovery extent.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111944914A (en) * | 2020-07-16 | 2020-11-17 | 中国科学院生态环境研究中心 | Method for evaluating water health risk based on resistance gene and virulence factor gene |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111944914A (en) * | 2020-07-16 | 2020-11-17 | 中国科学院生态环境研究中心 | Method for evaluating water health risk based on resistance gene and virulence factor gene |
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