CN101303337B - Method for evaluating water nutrition state - Google Patents
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Abstract
The invention provides an assessment method of water body nutrition status, which comprises the following steps: (1) water bodies samples to be tested are collected; (2) concentrations of algae chlorophylls a, b and c of the water body samples collected by step (1) are measured; (3) the ratio b/a between algae chlorophylls a and b measured by step (2) and the ratio c/a between algae chlorophylls a and c measured by step (2) are calculated; (4) the assessment of water body ecological restoration degree is carried out according to the characteristics of corresponding chlorophylls contained in common algae and the formation rule of algae populations in water bodies under different nutrition status. The assessment method of the invention can be widely applied to various water bodies, and has beneficial effects such as precision, convenience and being capable of making quantitative assessment of water body nutrition status and the like, compared with the prior art.
Description
Technical field
The present invention relates to a kind of water ecology evaluation method, relate in particular to a kind of evaluation method of water nutrition state.
Background technology
Body eutrophication be because supernutrition salt enters the water bodys such as lake, river, under suitable temperature, illumination and flow condition, algal bloom, cause that water transparency descends, dissolved oxygen DO reduces, and fish and other aquatic organisms are dead, has a strong impact on the use value of water body in lake.Under the acting in conjunction of natural cause and artificial interference, the super nutritional status aggravation of urban water-body, the monitoring of water nutrition state and mensuration have become hot issue, although, a lot of mensuration and evaluation methods about the water nutrition degree are arranged now both at home and abroad, but lack a kind of relative simple and the method for effective Accurate Determining.
In the prior art, the evaluation of water nutrition state is chosen usually the transparency, water temperature, Chlorophyll-a Content, total phosphorus, total nitrogen, nitrite nitrogen, chemical oxygen demand (COD), biochemical oxygen demand, permanganate index, dissolved oxygen content etc. of water body as main evaluation index; Evaluation method commonly used has: expert assessment method, comprehensive assessment, principal component analysis evaluation assessment, analytical hierarchy process, fuzzy set theory method and artificial neural network method etc.
Above method weak point is:
(1) number of parameters of measuring is many, and affected by environment large, link is many in sampling, preservation and mensuration process, and uncertainty and error are larger.
(2) comprehensive assessment only has Chlorophyll-a Content to belong to ecological index, only expresses the chlorophyllous summations of Freshwater Phytoplankton such as Cyanophyta in the water body, Chlorophyta and Chrysophyta.Lack the index that quantitative description planktonic algae different population and Community Ecology system change, can't set up the quantitative model of nutritional status level and ecological recovery and phytoplankton population variation and from inquiring in essence the mechanism of Evolution of lake trophic status.
(3) biodiversity index is the international index system of Ecosystem structure and function, and nutritional status water body ubiquity, the crucial biotic population with comparability and group namely are the biodiversity index of planktonic algae.The quantitative method of traditional algae Population and community, the discriminating by phytoplankton in the water body and adopt microscopic examination and counting method based on the algae formalness.This method specialty requires high, waste time and energy, and the personal error of evaluation and count results is larger.
Based on above-mentioned technical background, be necessary to propose a kind of new evaluation method, overcome the deficiency of above-mentioned existing evaluation method, more effective, more objective, the while can be made evaluation to water nutrition state in relative simple ground again.
Summary of the invention
The object of the invention is to: a kind of method for evaluating water nutrition state is provided, can be accurately objective, realize quantitative evaluation and monitoring to water nutrition state quickly and easily.
The objective of the invention is to be achieved through the following technical solutions:
A kind of method for evaluating water nutrition state is provided, may further comprise the steps:
1) gathers water body sample to be monitored;
2) determination step 1) concentration of algae chlorophyll a,b and c in the water body sample that gathers;
3) calculation procedure 2) the concentration ratio c/a of the algae chlorophyll a that records and the concentration ratio b/a of b and algae chlorophyll a and c;
4) according to Measures of Algae in Water Body population component law under the contained corresponding chlorophyllous characteristics of conventional algae and the Different Nutrition state, water nutrition state is estimated.
Wherein, step 1) described collection water body sample to be monitored can gather the equivalent sample simultaneously at a plurality of diverse locations of identical water body.
The method of described collecting sample preferably gathers 1L top layer water sample with the organic glass sampling thief at each sampled point, then takes back immediately and detects the place.
Step 2) method for measurement of concentration of described algae chlorophyll a,b and c can be behind the smudge cells with hot ethanol derect seething chlorophyll, use again the spectrophotometry chlorophyll concentration.
The method for measurement of concentration of described algae chlorophyll a,b and c specifically can be to measure 500～1 000ml water samples, adds 3～5 1%MgCO
3Suspending liquid adopts aperture 0.45 μ m, and the acetate fiber filter membrane of diameter 47mm carries out suction filtration; With qualitative filter paper filter membrane moisture is blotted, filter membrane is shredded put into the 5ml centrifuge tube, add 5ml 90% ethanol, carry out clasmatosis with conventional breaking method, after fragmentation is finished liquid collection is entered centrifuge tube, in 80 ℃ of hot baths, heat 2min, then place the 4 ℃ of lower 4～6h of extraction in dark place; Then use chlorophyll a,b in the spectrophotometry extract and the concentration of c.
Described clasmatosis can preferably use high pressure cell cracker to carry out.
Described preferred scheme with spectrophotometry algae chlorophyll a,b and c concentration is: after extracting algae chlorophyll, with the centrifugal 30min of the speed of 3000r/min, make reference with 90% ethanol, under 750nm, 664nm, 647nm, 630nm, measure its absorbance with spectrophotometer, then use formula 1～3 (being recorded among the disclosed Chinese patent literature CN 1631117A on June 29th, 2005) to calculate the concentration of described chlorophyll a,b, c:
Formula 1:chl a=[12.12 (D
664-D
750)-1.58 (D
647-D
750)-0.08 (D
630-D
750)] V
E/ V
SD
Formula 2:chl b=[-5.55 (D
664-D
750)+21.5 (D
647-D
750)-2.72 (D
630-D
750)] V
E/ V
SD
Formula 3:chl c=[-1.71 (D
664-D
750)-7.77 (D
647-D
750)+25.08 (D
630-D
750)] V
E/ V
SD
Wherein, chl a, chl b, chl c are respectively the concentration of water sample Determination of Chlorophyll a, b, c, and unit is μ g/L; D
664, D
647, D
630, D
750Be respectively extract 630,647,664, the absorbance at 750nm place; V
EBe the constant volume of extract in the centrifuge tube, unit is mL; V
SBe volume of water sample, unit is L; D is the cuvette light path, and unit is cm.
Step 4) described evaluation can be carried out according to two schemes, and scheme (1) can substitution be with following formula A and B respectively with the b/a value that calculates and c/a value, and preferred scheme (2) is that b/a value and the c/a value that will calculate are distinguished substitution formula C and D.Two schemes all can calculate respectively two nutritional status grade x of water body sample
1And x
2, and ask both mean value x as the final nutritional status grade of monitoring water body, the real number representation by 1～8:
Formula A:b/a=0.0012x
1 4-0.0185x
1 3+ 0.0823x
1 2-0.0718x
1+ 0.0024;
Formula B:c/a=0.0029x
2 2-0.0037x
2+ 0.1229;
Formula C:b/a=-0.0306x
1 2+ 0.2481x1-0.334;
Formula D:c/a=0.0054x
2 2-0.0162x
2+ 0.1311;
Wherein 1～8 water nutrition state grade x represents respectively the nutritional status of water body from the severe eutrophy to dystrophic continuous gradation, and be specially: x=1～3 represent the severe eutrophy; X=3～4 represent the moderate eutrophy; X=4～slight the eutrophy of 5 representatives; Nutrition in x=5～7 representatives; X=7～poor the nutrition of 8 representatives.
Evaluation method of the present invention can be applicable to comprise in the Evaluation of Nutritional Status and monitoring of the various water bodys such as lake, river.
It is the process of a gradual change that the nutritional status of water body changes.When water body changed from eutrophic state to poor nutritional status, the preponderant algae population in the water body changed to diatom, dinoflagellate, chrysophyceae from blue-green algae, green alga successively.Specifically, when water body presented nutritional status, blue-green algae and green alga were dominant populations; During middle nutritional status, diatom and green alga are dominant populations; And during poor nutritional status, dinoflagellate and chrysophyceae are preponderated.The inventor has determined that according to list of references of the prior art the population of algae and water under the Different Nutrition state forms, and sees for details shown in the accompanying drawing 1.
Under above-mentioned Different Nutrition state, in the corresponding algae population, blue-green algae is with microcystic aeruginosa (Microcystis aeruginosa, BG11 medium) and anabena (Anabaena sp., SE medium) is representative, only contains chlorophyll a, do not contain b, c; Green alga is representative with chlorella pyrenoidosa (Chlorella pyrenoidosa, SE medium), contains chlorophyll a and b; Diatom is representative with Han Shi rhombus algae (Nitzschia hantzschiana, D1 medium), contains chlorophyll a and c; Dinoflagellate is representative with thin dinoflagellate (Glenodinium pulvisculus, HB 119 medium), also contains chlorophyll a and c; Chrysophyceae is decided whip chrysophyceae (Prymnesiumparvum, AF-6 medium) for representative with little, also contains chlorophyll a and c.The inventor obtains the concentration of above-mentioned algae chlorophyll a,b, c and the ratio of chlorophyll b/a, c/a through the research of great many of experiments chamber, sees for details shown in the accompanying drawing 2.
In conjunction with the research conclusion of above-mentioned two aspects, can calculate chlorophyll b/a, the c/a ratio of algal community under the Different Nutrition state (available x represents).And then obtain chlorophyll b/a, c/a ratio change curve (seeing shown in the accompanying drawing 3) and the corresponding following functional expression A of curve, the B of algal community under the Different Nutrition state:
A：b/a＝0.0012x
4-0.0185x
3+0.0823x
2-0.0718x+0.0024
B：c/a＝0.0029x
2-0.0037x+0.1229
Also can further accurately obtain in addition the chlorophyll b/a of algal community under the Different Nutrition state, Optimal Curve (seeing shown in the accompanying drawing 4) and following preferred functional expression C, the D that c/a ratio changes:
C：b/a＝-0.0306x
2+0.2481x-0.334
D：c/a＝0.0054x
2-0.0162x+0.1311
According to above-mentioned curve obtained as can be known, when water body changes from eutrophic state to poor nutritional status, show as chlorophyll b/a ratio and increase first, rear minimizing, the value of Chlorofucsin/a then increases gradually.Fig. 3 (a), (b) and Fig. 4 (a), (b) have reflected respectively above-mentioned variation tendency, especially Fig. 4 reflect therefrom, the severe eutrophic state to the variation between the middle nutritional status more near real conditions, more practical in nearly reality.
Therefore, water body for a unknown nutritional status, as long as measure its chlorophyll b/a and c/a value, can judge qualitatively its nutritional status by the curve in Fig. 3 or 4, or estimating quantitatively its nutritional status by the result of calculation average of above-mentioned functional expression A, B or C, D, the result who wherein especially calculates with functional expression C, D is more accurate practical.
Compare with the comprehensive nutrition state index of prior art, chlorophyll ratioing technigue of the present invention need not to measure a plurality of parameters, under the condition that does not increase instrument and operation easier, include unexistent chlorophyll b and c in the routine monitoring when only needing to measure chlorophyll-a concentration in, thereby not only guaranteed result's accuracy more to have greatly reduced workload.Its main beneficial effect may be summarized to be:
(1) quantification
Utilize algae chlorophyll ratioing technigue of the present invention, can be on the Population and community aspect qualitative assessment water nutrition state.
(2) simplicity
Compared with prior art, simplified original 5-10 monitoring parameter, the concentration of single-time measurement chlorophyll a,b and c, and take the West Beijing sea as case (embodiment of the invention 1) contrast comprehensive nutrition state index, verified the reliability of the method for the invention.
(3) precision
Indoor sample pre-treatment and field monitoring sampling and preserve simplely, systematic error is little, can monitor simultaneously a large amount of samples, and disposable sample preparation can obtain 3 data simultaneously, and has good repeatability and comparability.
(4) be widely used
Can be widely used in Chinese Different Waters, seriously polluted, the super nutritional status of water environment water and demand carrying out the ecological recovery water body urgently particularly, and the dynamic process that water ecology recovers carried out the quantification monitoring and estimate.
Description of drawings
Fig. 1 is the population composition diagram of Measures of Algae in Water Body under the Different Nutrition state;
The concentration of chlorophyll a when Fig. 2 (a) is conventional algae Exponential growth stage;
The concentration of chlorophyll b, c and chlorophyll b/a, c/a ratio when Fig. 2 (b) is conventional algae Exponential growth stage;
Fig. 3 (a) uses in the evaluation method of aforementioned schemes (1) chlorophyll b/a ratio variation diagram under the Different Nutrition state;
Fig. 3 (b) uses in the evaluation method of aforementioned schemes (1) chlorophyll c/a ratio variation diagram under the Different Nutrition state;
Fig. 4 (a) uses in the evaluation method of aforementioned schemes (2) chlorophyll b/a ratio variation diagram under the Different Nutrition state
Fig. 4 (b) uses in the evaluation method of aforementioned schemes (2) chlorophyll c/a ratio variation diagram under the Different Nutrition state;
Fig. 5 is the Xihai sea, Beijing sampling point position figure.
Embodiment
Mode by the following examples further elaborates technical scheme of the present invention and effect, but protection scope of the present invention is not limited to described embodiment.
1. field sampling
Choosing the Xihai sea, Beijing is sample ground, respectively in May, 2007 and October gathering water sample at its water inlet, three sampling points of water delivering orifice and hydrostatic place, sampled point setting is seen Fig. 5, gathers 1L top layer water sample with the organic glass sampling thief at each sampled point, measures the transparency (SD) of water body with the Sa Shi dish.Take back immediately the laboratory after the sampling;
2. measure the content of water sample chlorophyll a,b, c
Measure the 500ml water sample, add 3～5 1%MgCO
3Suspending liquid adopts aperture 0.45 μ m, and the acetate fiber filter membrane of diameter 47mm carries out suction filtration; With qualitative filter paper filter membrane moisture is blotted, filter membrane shredded put into the 5ml centrifuge tube, add 5ml 90% ethanol, carry out after the fragmentation liquid collection being entered centrifuge tube with conventional method of cell disruption, in 80 ℃ of hot baths, heat 2min, then place the 4 ℃ of lower 4～6h of extraction in dark place; Then with the speed CEF 30min of 3000r/min, make reference with 90% ethanol, under 750nm, 664nm, 647nm, 630nm, measure its absorbance with spectrophotometer, then use following formula 1～3 (being recorded among disclosed Chinese patent literature CN 1631117 A on June 29th, 2005) to calculate the concentration of described chlorophyll a,b, c:
Formula 1:chl a=[12.12 (D
664-D
750)-1.58 (D
647-D
750)-0.08 (D
630-D
750)] V
E/ V
SD
Formula 2:chl b=[-5.55 (D
664-D
750)+21.5 (D
647-D
750)-2.72 (D
630-D
750)] V
E/ V
SD
Formula 3:chl c=[-1.71 (D
664-D
750)-7.77 (D
647-D
750)+25.08 (D
630-D
750)] V
E/ V
SD
Wherein, chl a, chl b, chl c are respectively the concentration of water sample Determination of Chlorophyll a, b, c, and unit is μ g/L; D
664, D
647, D
630, D
750Be respectively extract 630,647,664, the absorbance at 750nm place; V
EBe the constant volume of extract in the centrifuge tube, unit is mL; V
SBe volume of water sample, unit is L; D is the cuvette light path, and unit is cm.
Then calculate the value of chl b/chl a and chl c/chl a in the above-mentioned water sample.
3. judge Xihai sea nutritional status level with evaluation method of the present invention
The chl b/chl a that step 2 is calculated and chl c/chl a value be two relational expressions below the substitution respectively:
b/a＝0.0012x
1 4-0.0185x
1 3+0.0823x
1 2-0.0718x
1+0.0024
c/a＝0.0029x
2 2-0.0037x
2+0.1229
Calculate x
1And x
2Mean value x as the water nutrition state grade point, this result compares with the result of integrated nutritional index method of the prior art evaluation, obtains table 1:
As seen, two kinds of methods all draw Mays three place's water bodys and are in severe nutritional status state; October three, place's water quality situation all took a turn for the better to some extent, and all was in slight eutrophy-middle nutritional status.Thereby can get, method of the present invention and comprehensive nutrition state index acquired results are substantially identical, but method of the present invention mensuration process is simple, and favorable repeatability is more suitable for widespread use.
1. field sampling
Choosing the Baiyang Lake, Hebei province is sample ground, gather water sample in April, 2008 respectively at listed each sampled point in the table 2, gather 1L top layer water sample with the organic glass sampling thief at each sampled point, on-the-spot its water temperature and the pH value surveyed is with the transparency (SD) of Sa Shi dish measurement water body.Take back immediately the laboratory after the sampling.
2. measure the content of each water sample chlorophyll a,b, c
Measure the 1000ml water sample, add 3～5 1%MgCO
3Suspending liquid adopts aperture 0.45 μ m, and the acetate fiber filter membrane of diameter 47mm carries out suction filtration; With qualitative filter paper filter membrane moisture is blotted, filter membrane is shredded put into the 5ml centrifuge tube, add 5ml 90% ethanol, carry out fragmentation with high pressure cell cracker, after fragmentation is finished liquid collection is entered centrifuge tube, in 80 ℃ of hot baths, heat 2min, then place the 4 ℃ of lower 4～6h of extraction in dark place; Then with the speed CEF 30min of 3000r/min, make reference with 90% ethanol, under 750nm, 664nm, 647nm, 630nm, measure its absorbance with spectrophotometer, then use following formula 1～3 (being recorded among disclosed Chinese patent literature CN 1631117 A on June 29th, 2005) to calculate the concentration of described chlorophyll a,b, c:
Formula 1:chl a=[12.12 (D
664-D
750)-1.58 (D
647-D
750)-0.08 (D
630-D
750)] V
E/ V
SD
Formula 2:chl b=[-5.55 (D
664-D
750)+21.5 (D
647-D
750)-2.72 (D
630-D
750)] V
E/ V
SD
Formula 3:chl c=[-1.71 (D
664-D
750)-7.77 (D
647-D
750)+25.08 (D
630-D
750)] V
E/ V
SD
Wherein, chl a, chl b, chl c are respectively the concentration of water sample Determination of Chlorophyll a, b, c, and unit is μ g/L; D
664, D
647, D
630, D
750Be respectively extract 630,647,664, the absorbance at 750nm place; V
EBe the constant volume of extract in the centrifuge tube, unit is mL; V
SBe volume of water sample, unit is L; D is the cuvette light path, and unit is cm.
Then calculate the value of chl b/chl a and chl c/chl a in above-mentioned each water sample.
3. judge Baiyang Lake nutritional status level with evaluation method of the present invention
The chl b/chl a that step 2 is calculated and chl c/chl a value be two relational expressions below the substitution respectively:
b/a＝-0.0306x
1 2+0.2481x
1-0.334
c/a＝0.0054x
2 2-0.0162x
2+0.1311
Calculate x
1And x
2Mean value x, as the water nutrition state grade point, this result compares with the result of integrated nutritional index method of the prior art evaluation, obtains table 2:
By as seen from Table 2, evaluation method acquired results of the present invention and integrated nutritional index method of the prior art are basically identical, and good reproducibility is objective reliable in actual applications, but evaluation procedure is easier, is more suitable for widespread use.
In a word, method for evaluating water nutrition state of the present invention is worthy to be popularized as a kind of fast and accurately method for evaluating water nutrition state.
Claims (7)
1. method for evaluating water nutrition state may further comprise the steps:
1) gathers water body sample to be monitored;
2) determination step 1) concentration of algae chlorophyll a,b and c in the water body sample that gathers;
3) calculation procedure 2) the concentration ratio c/a of the algae chlorophyll a that records and the concentration ratio b/a of b and algae chlorophyll a and c;
4) according to Measures of Algae in Water Body population component law under the contained corresponding chlorophyllous characteristics of conventional algae and the Different Nutrition state, water nutrition state is made evaluation, concrete grammar is as follows:
B/a value and c/a value that step 3) calculates are distinguished substitution with following formula A and B, calculate respectively two nutritional status grade x of water body sample
1And x
2, and ask both mean value x as the final nutritional status grade of monitoring water body, the real number representation by 1 ~ 8;
Formula A:b/a=0.0012x
1 4-0.0185x
1 3+ 0.0823x
1 2-0.0718x
1+ 0.0024
Formula B:c/a=0.0029x
2 2-0.0037x
2+ 0.1229
Perhaps, with b/a value and c/a value difference substitution following formula C and the D that step 3) calculates, calculate the nutritional status grade x of water body sample, the real number representation by 1 ~ 8;
Formula C:b/a=-0.0306x
2+ 0.2481x-0.334
Formula D:c/a=0.0054x
2-0.0162x+0.1311
Wherein 1 ~ 8 water nutrition state grade represents respectively the nutritional status of water body from the severe eutrophy to dystrophic continuous gradation, is specially: x=1 ~ 3 represent the severe eutrophy; X=3 ~ 4 represent the moderate eutrophy; X=4 ~ slight the eutrophy of 5 representatives; Nutrition in x=5 ~ 7 representatives; X=7 ~ poor the nutrition of 8 representatives.
2. method for evaluating water nutrition state claimed in claim 1 is characterized in that: the described collection of step 1) water body sample to be monitored is that the diverse location at identical water body gathers the equivalent sample simultaneously.
3. method for evaluating water nutrition state claimed in claim 2 is characterized in that: the method for described collecting sample is to gather 1L top layer water sample with the organic glass sampling thief at each sampled point, then takes back immediately and detects the place.
4. method for evaluating water nutrition state claimed in claim 1 is characterized in that step 2) method for measurement of concentration of described algae chlorophyll a,b and c be behind the smudge cells with hot ethanol derect seething chlorophyll, use again the spectrophotometry chlorophyll concentration.
5. method for evaluating water nutrition state claimed in claim 4, it is characterized in that: the method for measurement of concentration of described algae chlorophyll a,b and c is to measure 500 ~ 1000ml water sample, adds 3 ~ 5 1%MgCO
3Suspending liquid adopts aperture 0.45 μ m, and the acetate fiber filter membrane of diameter 47mm carries out suction filtration; With qualitative filter paper filter membrane moisture is blotted, filter membrane is shredded put into the 5ml centrifuge tube, add 5ml 90% ethanol, carry out clasmatosis with conventional breaking method, after fragmentation is finished liquid collection is entered centrifuge tube, in 80 ℃ of hot baths, heat 2min, then place the 4 ℃ of lower 4 ~ 6h of extraction in dark place; Then use chlorophyll a,b in the spectrophotometry extract and the concentration of c.
6. method for evaluating water nutrition state claimed in claim 5 is characterized in that: described clasmatosis uses high pressure cell cracker to carry out.
7. method for evaluating water nutrition state claimed in claim 5, it is characterized in that: the chlorophyll a,b in the described spectrophotometry extract and the concentration of c are after extracting algae chlorophyll, with the centrifugal 30min of the speed of 3000r/min, make reference with 90% ethanol, under 750nm, 664nm, 647nm, 630nm, measure its absorbance with spectrophotometer, then use formula 1 ~ 3 to calculate the concentration of described chlorophyll a,b, c:
Formula 1:chl a=[12.12 (D
664-D
750)-1.58 (D
647-D
750)-0.08 (D
630-D
750)] V
E/ V
SD
Formula 2:chl b=[-5.55 (D
664-D
750)+21.5 (D
647-D
750)-2.72 (D
630-D
750)] V
E/ V
SD
Formula 3:chl c=[-1.71 (D
664-D
750)-7.77 (D
647-D
750)+25.08 (D
630-D
750)] V
E/ V
SD
Wherein, chl a, chl b, chl c are respectively the concentration of water sample Determination of Chlorophyll a, b, c, and unit is μ g/L; D
664, D
647, D
630, D
750Be respectively extract 630,647,664, the absorbance at 750nm place; V
EBe the constant volume of extract in the centrifuge tube, unit is mL; V
SBe volume of water sample, unit is L; D is the cuvette light path, and unit is cm.
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