CN102735671A - Method for determining point of dredging site of species sources of over-wintering cyanobacterial in large shallow lake - Google Patents
Method for determining point of dredging site of species sources of over-wintering cyanobacterial in large shallow lake Download PDFInfo
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Abstract
Provided is a method for determining points of dredging sites of species sources of over-wintering cyanobacterial in a large shallow lake. The method includes the following steps of: during an over-wintering recovery period, sampling a surface layer of lake sediment, analyzing pigment contents of the surface layer of the lake sediment to characterize total biomass and distribution of the cyanobacterial, and preliminarily screening regions as candidate regions where the total biomass of the species sources of the cyanobacteria are quite higher according to the pigment contents; adding sampling points in the candidate regions and adopting a microscope observation method and a flow cytometric method to determine target populations and the number of the target populations; adopting cell photosynthesis activity and/or esterase activity to characterize physiological activity of cyanobacterial cells; and finely screening the candidate regions according to the number of the target populations and performing correction with the help of the physiological activity of the cyanobacterial cells to preferably select regions where the number of the target populations are larger and the physiological activities are higher from the candidate regions so as to determine the points of the dredging sites of the species sources of the over-wintering cyanobacterial through a grid division method and a spatial interpolation method. By adopting the method in the invention, the points of the dredging sites are determined through distribution characteristics of the species sources of the over-wintering cyanobacterial, which provides scientific and reasonable basis for launching a project for dredging the species sources of the over-wintering cyanobacterial.
Description
Technical field
The present invention relates to a kind of improvement method of large-scale shallow water lake blue algae wawter bloom, be specifically related to survive the winter definite method in blue-green algae provenance dredging site of a kind of large-scale shallow water lake.
Background technology
Body eutrophication causes the freshwater resources contaminative deficient.The unusual breeding of blue-green algae and the formation of wawter bloom are one of most important signs of lake eutrophication.Present many researchers recognize, the wawter bloom in summer form with the winter lake sediment in the blue-green algae that survives the winter important relationship is arranged.For example blue-green algae sinks to the bed mud surface survive the winter autumn in the Taihu Lake, and spring in next year, the blue-green algae recovery rose to water surface from the bed mud surface, for wawter bloom forms necessary " provenance " was provided.Based on this understanding; To the control of blue-green alga bloom in summer, just can not wait until that it has formed huge biomass, take measures again after the drift diffusion everywhere with the wind in full lake; The required till that time manpower and materials of paying will increase greatly; And should be in blue-green algae survives the winter dormancy or these algae life processes of recovering spring the weakest link, i.e. algal bloom forms before the wawter bloom, takes measure more targetedly that it is subdued.
To large-scale shallow water lake (like Taihu Lake) research report over the years, blue-green alga bloom is always at first forming in some specific lake region between spring and summer, just spreads subsequently to cause full lake according at present.This is because the effect of lake surface wind field and lake current is assembled moving to the specific region in the large tracts of land wawter bloom sinking process in the fall, forms the provenance of mainly surviving the winter.Bed mud to the blue-green algae provenance is dredged the recovery amount of not only greatly subduing blue-green algae spring and summer in the coming year, reduces the intensity that summer, wawter bloom formed, and can make limited engineering processing power maximum effect, reaches the effect of getting twice the result with half the effort.
The large-size lake of carrying out the at present engineering of desilting and dredging mainly is subduing nutritive salt, place to go bed mud pollution of surface thing or to remove gulf district mud as considering, and the blue-green algae provenance that is not suitable for the surviving the winter characteristics of dredging.Therefore, development is a kind of will help to improve large-scale shallow water lake the survive the winter specific aim and the science of the dredging of blue-green algae provenance to the survive the winter position decision method of blue-green algae provenance dredging of lake.
Summary of the invention
The present invention aims to provide survive the winter definite method in blue-green algae provenance dredging site of a kind of large-scale shallow water lake; Integrated use is ecological, the biological chemistry detection method; From quantity, kind and active three aspects of blue-green algae provenance the distribution character of large-scale shallow water bottom mud in lake surface blue-green algae is analyzed; Through preliminary screening and accurately screening, before dredging, bed mud surface blue-green algae Distribution Pattern is carried out omnibearing monitoring as far as possible truly, to confirm the zone of surviving the winter of whole bottom mud in lake top layer blue-green algae; And then, improve the specific aim and the science of carrying out the measure of blue-green algae provenance dredging work to bottom mud in lake top layer ad-hoc location for the definite site of bottom mud in lake dredging.
Above-mentioned purpose of the present invention is achieved in that
Survive the winter definite method in blue-green algae provenance dredging site of a kind of large-scale shallow water lake is characterized in that: survive the winter the recovery phase blue-green algae, be distributed in the lake region and set the monitoring point to bottom mud in lake top layer sample analysis; Through analyzing total biomass and the space distribution thereof that pigment content in the bed mud top layer characterizes the blue-green algae provenance of surviving the winter, be alternative area according to the total biomass of the pigment content preliminary screening blue-green algae provenance higher zone that distributes; Set up sampled point in alternative area, bed mud top layer blue-green algae population is carried out qualitative and quantitative analysis, confirm to remove the target population and the quantity of the blue-green algae provenance of surviving the winter through microscopic and fluidic cell method; It is active to utilize cell photosynthesis activity and/or esterase active to characterize bed mud top layer blue-green algae cells physiological; Alternative area is further screened according to the target population quantity; Being aided with blue-green algae cells physiological activity proofreaies and correct; In alternative area, preferentially choose target population quantity and the higher zone of physiologically active, carry out the intensive sampling analysis, divide and space interpolation through space lattice; Dredged area is assessed the blue-green algae provenance of confirming to survive the winter dredging site.
Definite method in the blue-green algae provenance dredging site of surviving the winter of the present invention specifically may further comprise the steps:
1) survive the winter the recovery phase blue-green algae, based on lake region coverage of water and feature-set monitoring point, inherent each bed mud top layer, the monitoring point sampling of monitoring phase.
Select quiet for three days on end wind weather,, the sampling of full waters is carried out on full lakebed mud top layer to reduce the influence of wind field to the algae vertical movement.Mud appearance cuts the top layer bed mud of the 2cm of the superiors left and right thickness, and measures each item water quality parameter immediately.Gather 3 parts of parallel sample at each sampled point.
2) utilize pigment analysis to measure the algocyan content in the bed mud, characterize the total biomass of the recovery phase bed mud top layer blue-green algae provenance of surviving the winter and, tentatively confirm as the alternative area of dredging with the higher zone of algocyan content in the Distribution Pattern of lake region.
Algocyan can adopt AAS to measure.The disposal route of bed mud sample is: take by weighing a certain amount of bed mud sample, and air-dry to constant weight, calculate water percentage; Take by weighing a certain amount of sample and put into mortar, add 0.05 M Tris damping fluid (pH 7.0) and grind, under 4 ℃ of dark conditions, leave standstill 8-10h, centrifugal and with supernatant with 90% acetone constant volume.This liquid to be measured is used to measure algocyan content.
Through fluorescent spectrophotometer assay fluorescent intensity; Excitation wavelength is 620nm, wavelength of transmitted light 647nm, typical curve standard measure; Promptly algocyan concentration is made working curve, calculate algocyan content according to bed mud example weight or volume of water sample again with fluorescence intensity.
According to algocyan content in the bed mud; Can confirm the biomass of blue-green algae in the bed mud top layer in different waters; The reflection blue-green algae provenance Spatial Distribution Pattern of recovery phase of surviving the winter in the lake region; And then tentatively confirm the zone of surviving the winter of blue-green algae, and choose the highest region, some monitoring points of algocyan content and further analyze as alternative area.
3) set up sampled point in alternative area; Through Flow Cytometry and microscopic; Further the target population and the quantity of bed mud blue-green algae provenance are carried out qualitative and quantitative analysis, confirm to remove the target population and the quantity of the blue-green algae provenance of surviving the winter, with alternative area according to target the quantity of population sort.
Through to dilution of top layer mud appearance and filtration, adopt Flow Cytometry to be aided with the microscope lens mirror qualitative and quantitative analysis is carried out on bed mud top layer blue-green algae specific population (like microcystic aeruginosa).
The fluidic cell method is: adopt flow cytometer, optical maser wavelength is respectively 488nm and 600nm, and the autofluorescence of pigment is collected through FL3 (675/20nm, chlorophyll a fluorescence) and two sense channels of FL4 (660/20nm, algocyan fluorescence).Non-algae particle or gathering cell can pass through the difference of autofluorescence (FL3/FL4) and morphosis parameter (FSC/SSC) and remove.Bi-component absolute counting microballoon is used in cell count in the microscope lens mirror.
4) measure the photosynthesis activity and/or the esterase active of frustule, the blue-green algae cellular physiological activity of alternative area is sorted.
The sample of each collection is measured the photosynthesis ability and the esterase active of frustule respectively; Wherein carry out the analysis of algae photosynthetic activity through the pigment fluorescence analyser; When temperature was reduced to 1～4 ℃, algocyan photosynthesis might disappear, and was aided with the esterase detection method this moment and analyzed algae cell activity.
The concrete operations of algae photosynthesis activity analysis are following: get mud appearance dilute filtration; The measuring cup of packing in right amount; Dark adatpation 15min opens measuring light (ML) earlier, treats that initial fluorescence obtains Fo after stable, opens saturated light intensity (ST); Obtain the maximum photosynthetic efficiency of the Fm and the photosystem II of sample, and measure the quick optical response curve.
The concrete grammar of esterase active detection method analysis algae cell activity is following: get mud appearance dilute filtration, in algae liquid, add an amount of fluorescent dye, after the dyeing of room temperature lucifuge, detect the intensity that enzyme is lived through flow cytometer.
The Fv/Fm, the maximum photosynthetic efficiency of photosystem II and the photosynthesis ability that the quick optical response curve can characterize frustule comprehensively that obtain through the photosynthetic activity analysis, and esterase active detects through the power of the back FL1 passage fluorescence signal that dyes and representes the esterase active size.The both characterizes the activated state of frustule from different perspectives, thereby alternative area is further screened, and preferentially chooses the active higher zone of specific objective population as the dredging site.
5) according to the target stock biomass of alternative area and the ordering of blue-green algae cellular physiological activity; In alternative area, preferentially choose target population quantity and the higher zone of physiologically active; Carry out the intensive sampling analysis,, divide and space interpolation through space lattice according to target stock biomass and blue-green algae cellular physiological activity; Calculate and assess dredging the zone, thereby obtain to survive the winter blue-green algae provenance dredging site.
Provenance to the bed mud surface is dredged the blue-green algae kind source cell that is primarily aimed at the surperficial 2-3cm of bed mud, on mode, should take low-intensity dredging mode, and this not only reduces the destruction to ecologic environment, also can avoid discharging the secondary pollution that causes because of the endogenous nutritive salt of bed mud.In addition, also can take lake substrate remodeling method according to the site that this patent is confirmed: like modified clay technology, the water-bed ploughing and weeding technology of improving the soil, water-bed ploughshare is ploughed the technology etc. of improving the soil; Or adopt biological algal control method such as rice straw that the blue-green algae provenance is removed.
Advantage of the present invention and effect: definite method in blue-green algae provenance dredging site is survived the winter tightly around eliminating bed mud this target of blue-green algae provenance amount of surviving the winter in large-scale shallow water of the present invention lake; Select and set up a series of analysis indexes and method; Gather mud appearance through lay the monitoring point, many places in the lake region; Carry out multinomial physics and chemistry, cell biology analysis; Distribute from the total biomass of blue-green algae and to carry out primary dcreening operation, alternative site target stock biomass dusting cover and cell physiological ecologic active and proofread and correct this three aspect, the blue-green algae provenance dredging site of surviving the winter, bottom mud in lake top layer is judged to have stronger accuracy and operability progressively.The inventive method is analyzed blue-green algae provenance amount and activity from different perspectives, through mutual checking (as: the blue-green algae total biomass and the specific population biomass of correlation analysis; Photosynthesis and esterase active or the like), thus the accuracy of judging improved.The inventive method can be confirmed the blue-green algae provenance dredging site of surviving the winter, large-scale shallow water bottom mud in lake top layer; For the feasibility and the specific aim of lake treatment engineering measure (like winter dredging) provides scientific and reasonable foundation; Make limited engineering processing power maximum effect, reach the effect of getting twice the result with half the effort.
Describe the present invention below in conjunction with specific embodiment.Protection scope of the present invention is not exceeded with embodiment, but is limited claim.
Description of drawings
Lake region, Fig. 1 Taihu Lake monitoring site distribution schematic diagram.
Bed mud surface, Fig. 2 Taihu Lake algocyan content spatial distribution map.
Fig. 3 north Taihu Lake dredging alternative area monitoring site distribution schematic diagram.
The fluidic cell figure that bed mud surface, Fig. 4 Taihu Lake Microcystis aeruginosa is distinguished.
Fig. 5 dredges alternative area microcystic aeruginosa biomass relatively.
Fig. 6 dredges alternative area microcystic aeruginosa esterase active relatively.
Bed mud surface, Fig. 7 Mei Liang gulf blue-green algae provenance dredging site synoptic diagram of surviving the winter.
Embodiment
According to survive the winter definite method in blue-green algae provenance dredging site of large-scale shallow water of the present invention lake, be applied to confirm in the provenance dredging work of Taihu Lake the dredging site, taken a sample in the bottom mud in lake top layer in the monitoring point of setting in blue-green algae recovery phase of surviving the winter; Confirm to remove target population (like the microcystic aeruginosa) quantity of the blue-green algae that survives the winter through pigment analysis, mirror mirror and Flow Cytometry; In conjunction with the physiologically active analysis of blue-green algae, confirm the blue-green algae provenance dredging site of surviving the winter, bed mud top layer, Taihu Lake.
Described method specifically may further comprise the steps:
1. survive the winter the recovery phase blue-green algae, based on lake region coverage of water and feature-set monitoring point, inherent each bed mud top layer, the monitoring point sampling of monitoring phase.
Based on TAIHU LAKE bulk area and feature-set 14 place monitoring point (as shown in Figure 1), select quiet for three days on end wind weather, the sampling of full waters is carried out on full lakebed mud top layer.Use internal diameter to gather the bed mud sample as the column bottom sampler of 62mm; The top layer bed mud that cuts the 2cm of the superiors left and right thickness moves in the sealing bag; And (Yellow Spring Instruments USA) measures water quality parameter to use multi-functional water quality appearance (YSI6600-V2) immediately.Gather 3 parts of parallel sample at each sampled point.
2. utilize pigment analysis to make up to survive the winter before the dredging space distribution on blue-green algae bed mud top layer in Taihu Lake.
Adopt algocyan content in each monitoring point bed mud sample of spectrophotometry.Take by weighing the bed mud sample after a certain amount of the thawing, to constant weight, and take by weighing dry weight, calculate water percentage in room-dry.Take by weighing the ground sample in the 5g left and right sides and put into mortar, add 0.05 M Tris damping fluid (pH 7.0), carefully grind 2-5min, under 4 ℃ of dark conditions, leave standstill 8-10h.With the centrifugal 5min of the rotating speed of 4000g, supernatant is transferred in the 10mL volumetric flask, is settled to 10mL with 90% acetone at last then, obtains the bed mud sample extracting solution and is used for measuring bed mud algocyan content.
Measure algocyan content (Yan Rong, 2004) through Tianjin, island fluorospectrophotometer RF-5301, excitation wavelength is 620nm, wavelength of transmitted light 647nm, and sweep velocity 60nm/min makes curve with fluorescence intensity to algocyan concentration workmanship.Under the condition identical, measure the fluorescence intensity of bed mud sample extracting solution, thereby on working curve, try to achieve algocyan concentration in the extract, calculate algocyan content according to the bed mud example weight again with standard model series.
Pass through said method; The survive the winter Spatial Distribution Pattern of blue-green algae of the Taihu Lake bed mud that can obtain to survive the winter the phase (for example Dec) blue-green algae is seen Fig. 2; Can be known by Fig. 2: the blue-green algae Taihu Lake northern bed mud algocyan content of surviving the winter in the phase is higher; Particularly Zhu Shan gulf, Mei Liang gulf, this three gulfs district of tribute arm of lake, the band of lakeside among a small circle in lake, South Pacific is also slightly high in addition, and the algocyan content on lake region, middle part bed mud surface (p < 0.05) obviously on the low side.Therefore; Can judge this moment because the influence of wind field and lake current; The blue-green algae that sinks autumn and winter concentrates on 3 arm of lakes of northern lake region, Taihu Lake and the zone among a small circle in lake, South Pacific gradually, and this can be used as alternative area and for next step accurate judgement the basis is provided in the zone everywhere.
3. set up sampled point in alternative area, carry out qualitative and quantitative analysis through microscopic and Flow Cytometry, with alternative area according to target the quantity of population sort.
Respectively get 5 points (Fig. 3) respectively in above-mentioned 4 alternative area and carry out further qualitative and quantitative analysis.Each monitoring point is got 10 g mud appearance respectively, with 10 times of dilution methods with mud appearance towards rare to 10
-2With 10
-3PBS suspension (can suitably adjust) according to mud appearance character, it is centrifugal behind 300 order membrane filtrations that (2500 rpm 7min) to can surveying concentration, process cell suspension three times repeatedly.Adopt Flow Cytometry to be aided with the mirror mirror quantitative test is carried out on bed mud top layer blue-green algae specific population (like microcystic aeruginosa).
The fluidic cell method adopts FACSVantage SE flow cytometer (Becton Dickinson; USA); First laser is Coherent water-cooled Argon ion laser (excitation wavelength is 488nm); The fluorescence signal that produces is collected (FL1:530/30nm, FDA fluorescence through 2 sense channels; FL3:675/20nm, chlorophyll a fluorescence); Second laser is Coherent dye laser (excitation wavelength is 600nm), sends out with Coherent water-cooled Argon ion laser pump, and the sense channel wavelength is 660/20nm (FL4, an algocyan fluorescence).Non-algae particle or gathering cell can pass through the difference of autofluorescence (FL3/FL4) and morphosis (FSC/SSC) and remove.The analysis flow velocity of sample is 1 μ L/s, and single kind consuming time is 20s.The microscope lens mirror is assisted qualitative judgement, and cell count use bi-component absolute counting microballoon (Caltag Laboraties Inc, CA).
As shown in Figure 4; Fluorescence analysis (FL3 or FL4) and morphosis analysis (FSC or SSC) through Flow Cytometry can be distinguished microcystic aeruginosa in the bed mud; And (Caltag Laboraties Inc CA) counts cell in the suspension can to pass through bi-component absolute counting microballoon.As shown in Figure 5, according to quantitative test, can judge that the biomass ordering of microcystic aeruginosa on the bed mud surface of surviving the winter this moment is followed successively by the Mei Liang gulf to microcystic aeruginosa cell number in the suspension>the tribute arm of lake>the Zhu Shan gulf>lake, South Pacific.Wherein with the monitoring point numerical value of west bank, Mei Liang gulf for the highest.
4. photosynthesis activity through frustule and/or esterase active sort to the blue-green algae cellular physiological activity of alternative area.
Each collected specimens of above-mentioned four alternative area is measured the photosynthesis ability and the esterase active of frustule respectively.The concrete operations of algae photosynthesis activity analysis are following: get the mud appearance dilute filtration of just adopting back; The measuring cup of packing in right amount; Dark adatpation 15min opens measuring light (ML) earlier, treats that initial fluorescence obtains Fo after stable, opens saturated light intensity (ST); Obtain the maximum photosynthetic efficiency of the Fm and the photosystem II of sample, and measure the quick optical response curve.Because temperature has been reduced to 4 °, the active basic disappearance of the photosynthesis of bed mud blue-green algae or extremely faint, so data are unlisted.
It is following that the esterase active detection method is analyzed the concrete grammar of algae cell activity: get 10 g mud appearance, with 10 times of dilution methods with mud appearance towards rare to 10
-2With 10
-3PBS suspension (can suitably adjust) according to mud appearance character, it is centrifugal behind 300 order membrane filtrations that (2500 rpm 7min) to can surveying concentration, process cell suspension three times repeatedly.Get adding an amount of fluorescent dye (Fluorescence dieastrate) in the 1ml algae liquid, behind the room temperature lucifuge dyeing 8min, detect the intensity that enzyme is lived through flow cytometer FL1 passage.Can know that like Fig. 6 the cytoactive difference of Zhu Shan gulf, tribute arm of lake, Mei Liang gulf microcystic aeruginosa is little, the lake, South Pacific is obviously on the low side (p 0.05).The littoral pasture and water that distribute in lake, South Pacific are more in addition, recovery and the activity of algae had certain inhibiting effect, so can from alternative area, remove the lake, South Pacific.
5. three aspect results of comprehensive above algocyan content, target stock biomass and blue-green algae cellular physiological activity; If carry out the survive the winter blue-green algae provenance dredging of Taihu Lake bed mud this moment; Can preferentially select the Mei Liang gulf to carry out intensive sampling; And combine the difference of bottom mud in lake physicochemical property and the attachment characteristic of frond, confirm the distribution of this lake region bed mud surfactivity blue-green algae provenance.Can use Arcgis software and divide and space interpolation, and dredged area is assessed, thereby obtain to survive the winter blue-green algae provenance dredging site through space lattice.
Space lattice is divided and interpolation: according to coverage of water size setting space number of grid, the laying of grid adopts GIS-Geographic Information System (GIS) to lay.Adopt the grid of 200 * 200m to this waters, Mei Liang gulf.Adopt the anti-distance weighting method of Arcgis software to carry out space interpolation and data storing.
Bed mud blue-green algae dredged area assessment: adopt analytical hierarchy process and natural break method that target stock biomass and cellular physiological activity are carried out COMPREHENSIVE CALCULATING and assessment; The waters of same classification is merged; Be divided into 4 grades from high to low according to the comprehensive grading value, and, see Fig. 7 through the mapping of Arcgis software.The preferred higher grid (grey is darker among Fig. 7) of score value of selecting is as the blue-green algae provenance dredging site of surviving the winter.
Further improve the accuracy in dredging site like need, can increase detection site or detect index, adopt use the same method comparison microcystic aeruginosa biomass and activated state, and then make the more judgement of science.From the time; Also can take the method for periodic monitoring; When the microcystic aeruginosa biomass accumulation of bed mud surface is the highest, carry out dredging when promptly algae at utmost sinks down into the bed mud surface in the water body, this will further improve the specific aim and the science of directed dredging work.In addition; On applicability, this patent be except instructing bed mud blue-green algae provenance dredging, also can be applicable to some and directly survives the winter in the removal technology of blue-green algae to the bottom mud in lake surface; For it handles the reference that the position provides science; The physics remodeling method that comprises the lake substrate, like modified clay technology, the water-bed ploughing and weeding technology of improving the soil, water-bed ploughshare is ploughed the technology etc. of improving the soil; The chemical method of chemicals treatment such as copper sulphate, hydrogen peroxide; Biological methods such as rice bar processing.
Claims (6)
1. a large-scale shallow water lake definite method in blue-green algae provenance dredging site of surviving the winter is characterized in that: survive the winter the recovery phase blue-green algae, be distributed in the lake region and set the monitoring point to bottom mud in lake top layer sample analysis; Through analyzing total biomass and the space distribution thereof that pigment content in the bed mud top layer characterizes the blue-green algae provenance of surviving the winter, be alternative area according to the total biomass of the pigment content preliminary screening blue-green algae provenance higher zone that distributes; Set up sampled point in alternative area, bed mud top layer blue-green algae population is carried out qualitative and quantitative analysis, confirm to remove the target population and the quantity of the blue-green algae provenance of surviving the winter through microscopic and fluidic cell method; It is active to utilize cell photosynthesis activity and/or esterase active to characterize bed mud top layer blue-green algae cells physiological; Alternative area is further screened according to the target population quantity; Being aided with blue-green algae cells physiological activity proofreaies and correct; In alternative area, preferentially choose target population quantity and the higher zone of physiologically active, carry out the intensive sampling analysis, through the method for gridding and space interpolation; Dredged area is assessed, confirmed the blue-green algae provenance dredging site of surviving the winter, large-scale shallow water bottom mud in lake top layer.
2. survive the winter definite method in blue-green algae provenance dredging site of large-scale shallow water according to claim 1 lake is characterized in that described method may further comprise the steps:
1) survives the winter the recovery phase blue-green algae,, be distributed in the lake region and set the monitoring point, taken a sample in the bottom mud in lake top layer in each monitoring point based on lake region coverage of water and characteristic;
2) adopt pigment analysis to measure the algocyan content in the bed mud top layer; Characterize the total biomass of bed mud top layer blue-green algae provenance and in the space distribution of lake region; Confirm the zone of surviving the winter of blue-green algae provenance, tentatively confirm as the alternative area of dredging with the higher zone of algocyan content;
3) set up sampled point in alternative area,, confirm in the bed mud top layer to remove the target population and the quantity of the blue-green algae provenance of surviving the winter through fluidic cell method and microscopic, and with alternative area according to target the quantity of population sort;
4) the photosynthesis activity and/or the esterase active of frustule in the mensuration bed mud top layer, it is active to utilize cell photosynthesis activity and/or esterase active to characterize the blue-green algae cells physiological, and alternative area is sorted by the blue-green algae cellular physiological activity;
5) according to the target population quantity of alternative area and the ordering of blue-green algae cellular physiological activity; In alternative area, preferentially choose target population quantity and the higher zone of physiologically active; Carry out the intensive sampling analysis,, divide and space interpolation through space lattice according to target stock biomass and blue-green algae cellular physiological activity; Calculate and assess dredging the zone, confirm the blue-green algae provenance dredging site of surviving the winter, bottom mud in lake top layer.
3. survive the winter definite method in blue-green algae provenance dredging site of large-scale shallow water according to claim 2 lake is characterized in that step 2) in, described algocyan content adopts AAS to measure, and excitation wavelength is 620nm, wavelength of transmitted light 647nm.
4. survive the winter definite method in blue-green algae provenance dredging site of large-scale shallow water according to claim 2 lake; It is characterized in that; In the step 3); Described fluidic cell method is: adopt flow cytometer, optical maser wavelength is respectively 488nm and 600nm, and the autofluorescence of pigment is collected through two sense channels of FL3, FL4 of wavelength 675/20nm and 660/20nm; Non-algae particle or gathering cell are removed through the difference of autofluorescence FL3/FL4 and morphosis parameter F SC/SSC.
5. survive the winter definite method in blue-green algae provenance dredging site of large-scale shallow water according to claim 2 lake is characterized in that, in the step 4); The active assay method of the photosynthesis of described frustule is to get bed mud top layer mud appearance dilute filtration, the measuring cup of packing in right amount; Dark adatpation 15min opens measuring light ML earlier, treats that initial fluorescence obtains Fo after stable, opens saturated light intensity ST; Obtain the maximum photosynthetic efficiency of the Fm and the photosystem II of sample, and measure the quick optical response curve.
6. survive the winter definite method in blue-green algae provenance dredging site of large-scale shallow water according to claim 2 lake; It is characterized in that in the step 4), the detection method of described esterase active does; Get bed mud top layer mud appearance dilute filtration; In algae liquid, add an amount of fluorescent dye, the dyeing of room temperature lucifuge is through the strong and weak esterase active that detects of back flow cytometer FL1 passage fluorescence that dyes.
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| CN103616490A (en) * | 2013-12-03 | 2014-03-05 | 中国科学院南京地理与湖泊研究所 | Method for estimating total stock of water-blooming cyanobacteria in large-size shallow lake |
| WO2023221509A1 (en) * | 2022-05-18 | 2023-11-23 | 长江水利委员会长江科学院 | Method for separating, extracting and quickly classifying and counting living algae cells of soil/sediment |
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| CN103616490A (en) * | 2013-12-03 | 2014-03-05 | 中国科学院南京地理与湖泊研究所 | Method for estimating total stock of water-blooming cyanobacteria in large-size shallow lake |
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| WO2023221509A1 (en) * | 2022-05-18 | 2023-11-23 | 长江水利委员会长江科学院 | Method for separating, extracting and quickly classifying and counting living algae cells of soil/sediment |
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