CN103484523B - The somatic method of counting of a kind of Microcystis aeruginosa group - Google Patents
The somatic method of counting of a kind of Microcystis aeruginosa group Download PDFInfo
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- CN103484523B CN103484523B CN201310425334.9A CN201310425334A CN103484523B CN 103484523 B CN103484523 B CN 103484523B CN 201310425334 A CN201310425334 A CN 201310425334A CN 103484523 B CN103484523 B CN 103484523B
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- microcystis aeruginosa
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Abstract
The invention discloses the somatic method of counting of a kind of Microcystis aeruginosa group, first microcystis waterbloom is quantitatively taked to break out the water sample 1L of the natural water of Sheng phase, again the water sample via hole diameter collected is about the filtering with microporous membrane of 0.45 μm, then the Microcystis aeruginosa population sample of filtration postadhesion on millipore filtration is suspended in containing 10mL10
-3mol/L? in the 100mL beaker of EDTA solution, then 1600rm/min magnetic agitation 100min after 100 little granulated glass spherees is added wherein, obtain fully decentralized unicellular Microcystis aeruginosa sample and without breakage, finally adopt blood counting chamber counting microcystis number.The present invention embodies simplification experimental procedure, consuming time shorter, can the somatic number of accurate counting Microcystis aeruginosa group.
Description
Technical field
The invention belongs to water pollution control field, particularly relate to the somatic method of counting of a kind of Microcystis aeruginosa group.
Background technology
Microcystis aeruginosa (Microcystissp.) is the bloom blue algae kind of blazoning property of the whole world, modal blue-green alga bloom main composition kind in Ye Shi China eutrophication water.Microcystis aeruginosa usually exists with unicellular or two cells form under laboratory culture condition, and is exist with a large amount of macroscopic group form in natural water in the wild.The relevant somatic method of counting of field Microcystis aeruginosa group has whirlpool to shake method, boiling method, titanium dioxide or ultraviolet treatment now, but cannot be separated into unicellular completely at whirlpool concussion method effect Xia Nang algae colony cell, and Microcystis aeruginosa colony cell all has breakage in various degree under boiling method, titanium dioxide or ultraviolet treatment effect, thus above-mentioned method of counting reduces the Cytometric accuracy of Microcystis aeruginosa colony all to some extent.
Summary of the invention
Object of the present invention is just to provide a kind of Microcystis aeruginosa group somatic method of counting, the monitoring of Microcystis aeruginosa density in the water body of blue-green alga bloom is often broken out, for the routine monitoring of microcystis number in natural water and the early warning of microcystis waterbloom and control for various poisons in freshwaters such as artificial water application's system, lake, reservoir and ponds.
In order to solve the problems of the technologies described above, the technical solution used in the present invention is: the somatic method of counting of a kind of Microcystis aeruginosa group, comprises the steps:
(1) microcystis waterbloom is quantitatively taked to break out the water sample of the natural water of Sheng phase;
(2) the water sample via hole diameter collected is not more than the screen filtration of 2 μm, Microcystis aeruginosa sample is attached on screen cloth;
(3) the Microcystis aeruginosa population sample of filtration postadhesion on screen cloth being suspended in concentration is again 10
-3in the beaker of mol/LEDTA solution;
(4) after adding little granulated glass sphere to above-mentioned beaker, magnetic agitation;
(5) draw the unicellular sample of Microcystis aeruginosa that basis of microscopic observation spreads out completely, adopt blood counting chamber counting microcystis number.
In described step (2), described screen cloth is the bolting silk that the millipore filtration in 0.45 μm, aperture or aperture are not more than 2 μm.
In described step (3), utilize liquid-transfering gun to draw EDTA solution and repeatedly rinse screen cloth, until Microcystis aeruginosa colony elutes from screen cloth completely.
In described step (4), in magnetic agitation process, often cross under the microscope and within 5 minutes, check Microcystis aeruginosa colony cell dispersal process, until Microcystis aeruginosa colony cell is separated into unicellular completely.
In described step (4), magnetic agitation speed 1000-2000 turns/min, magnetic agitation time 90-110min.
In described step (5), before drawing the unicellular sample of Microcystis aeruginosa, draw again after mixing, and blood counting chamber is counted the microcystis quantity that microcystis number is converted into every premium on currency sample.
The invention has the beneficial effects as follows: whirlpool concussion method effect Xia Nang algae colony cell can be overcome and cannot be separated into the defect that Microcystis aeruginosa colony cell under unicellular and boiling method, titanium dioxide or ultraviolet treatment effect all has breakage in various degree completely, can the somatic number of Microcystis aeruginosa group in accurate counting natural water.Experimental procedure is simple, consuming time shorter, and accurately can improve the somatic number of counting Microcystis aeruginosa group.Be widely used in the prevention and control of the various poisons in freshwater microcystis waterblooms such as artificial water application's system, lake, reservoir and pond.
Accompanying drawing explanation
The change of Tu1Shi Microcystis aeruginosa colony form of the first embodiment under the inventive method effect.
The change of Tu2Shi Microcystis aeruginosa colony form of the second embodiment under the inventive method effect.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail:
The somatic method of counting of Microcystis aeruginosa group of the present invention, comprises the steps:
(1) microcystis waterbloom is quantitatively taked to break out the water sample of the natural water of Sheng phase;
(2) the water sample via hole diameter collected is not more than the screen filtration of 2 μm, Microcystis aeruginosa sample is attached on screen cloth;
(3) the Microcystis aeruginosa population sample of filtration postadhesion on screen cloth being suspended in concentration is again 10
-3in the beaker of mol/LEDTA solution;
(4) after adding little granulated glass sphere to above-mentioned beaker, magnetic agitation;
(5) draw the unicellular sample of Microcystis aeruginosa that basis of microscopic observation spreads out completely, adopt blood counting chamber counting microcystis number.
In described step (2), described screen cloth is the bolting silk that the millipore filtration in 0.45 μm, aperture or aperture are not more than 2 μm.
In described step (3), utilize liquid-transfering gun to draw EDTA solution and repeatedly rinse screen cloth, until Microcystis aeruginosa colony elutes from screen cloth completely.
In described step (4), in magnetic agitation process, often cross under the microscope and within 5 minutes, check Microcystis aeruginosa colony cell dispersal process, until Microcystis aeruginosa colony cell is separated into unicellular completely.
In described step (4), magnetic agitation speed 1000-2000 turns/min, magnetic agitation time 90-110min.
In described step (5), before drawing the unicellular sample of Microcystis aeruginosa, draw again after mixing, and blood counting chamber is counted the microcystis quantity that microcystis number is converted into every premium on currency sample.
The exocellular polysaccharide of bacterium and fungal cell, in conjunction with the effect of granulated glass sphere, can elute and not destroy the integrity of bacterium or fungal cell by high speed magnetic stirring under the cooperation of EDTA solution.
Embodiment 1 the inventive method is applied to the example of natural water microcystis measuring density
Use the present invention counts microcystis number during Tianjin, Tianjin river valley eight li of platform sections outburst microcystis waterblooms on July 3rd, 2013:
(1) water sampler of 1L is utilized to take the water sample 1L of the natural water of river, Tianjin microcystis waterbloom outburst Sheng phase; (2) Tianjin river sample via hole diameter collected is about the filtering with microporous membrane of 0.45 μm, Microcystis aeruginosa sample is then attached on millipore filtration;
(3) the Microcystis aeruginosa population sample of filtration postadhesion on millipore filtration is suspended in again containing 10mL10
-3in the 100mL beaker of mol/LEDTA solution;
(4) after adding 100 little granulated glass spherees to above-mentioned 100mL beaker, 1600rm/min magnetic agitation 90-110min; (see figure 1)
(5) draw the unicellular sample of Microcystis aeruginosa that basis of microscopic observation spreads out completely, adopt hemocytometer
Number plate counting microcystis number.
In described step (3), utilize liquid-transfering gun to draw EDTA solution and repeatedly rinse screen cloth, until Microcystis aeruginosa colony elutes from screen cloth completely.
In described step (4), in magnetic agitation process, often cross under the microscope and within 5 minutes, check Microcystis aeruginosa colony cell dispersal process, until Microcystis aeruginosa colony cell is separated into unicellular completely.
In described step (5), before drawing the unicellular sample of Microcystis aeruginosa, draw again after mixing.
Microcystis (containing microcystic aeruginosa M.aeruginosa and fish evil Microcystis aeruginosa M.ichthyoblabe) density when enforcement through method of the present invention obtains the eight li of platform section outburst microcystis waterblooms in Jin in July 3 river in 2013 is 2.42 × 10
8ind/L.
Embodiment 2 the inventive method is applied to the example of aquaculture water microcystis measuring density
On July 5th, 2013, microcystis (containing microcystic aeruginosa M.aeruginosa and the fish evil Microcystis aeruginosa M.ichthyoblabe) density recording carp seed multiplication farm No. 3 pool cultivated water bodys according to the inventive method was 6.42 × 10 at TanJin Agricultural College carp seed multiplication farm
7ind/L.
Method, with embodiment 1, just changes the millipore filtration in 0.45 μm, aperture into bolting silk that aperture is not more than 2 μm, the results are shown in Figure 2.
Above-described embodiment is only for illustration of technological thought of the present invention and feature, its object is to enable those skilled in the art understand content of the present invention and implement according to this, only can not limit the scope of the claims of the present invention with the present embodiment, namely the equal change done of all disclosed spirit or modification, still drop in the scope of the claims of the present invention.
Claims (3)
1. the somatic method of counting of Microcystis aeruginosa group, is characterized in that, comprise the steps:
(1) microcystis waterbloom is quantitatively taked to break out the water sample of the natural water of Sheng phase;
(2) by the water sample that collects through screen filtration, Microcystis aeruginosa sample is attached on screen cloth, and described screen cloth is the bolting silk that the millipore filtration in 0.45 μm, aperture or aperture are not more than 2 μm;
(3) the Microcystis aeruginosa population sample of filtration postadhesion on screen cloth being suspended in concentration is again 10
-3in the beaker of mol/LEDTA solution;
(4) after adding little granulated glass sphere to above-mentioned beaker, magnetic agitation, magnetic agitation speed 1000-2000 turns/min, magnetic agitation time 90-110min, in magnetic agitation process, often cross under the microscope and within 5 minutes, check Microcystis aeruginosa colony cell dispersal process, until Microcystis aeruginosa colony cell is separated into unicellular completely;
(5) draw the unicellular sample of Microcystis aeruginosa that basis of microscopic observation spreads out completely, adopt blood counting chamber counting microcystis number.
2. the Cytometric method of Microcystis aeruginosa colony according to claim 1, is characterized in that, in described step (3), utilizes liquid-transfering gun to draw EDTA solution and repeatedly rinses screen cloth, until Microcystis aeruginosa colony elutes from screen cloth completely.
3. the Cytometric method of Microcystis aeruginosa colony according to claim 1, it is characterized in that, in described step (5), before drawing the unicellular sample of Microcystis aeruginosa, draw again after mixing, and blood counting chamber is counted the microcystis quantity that microcystis number is converted into every premium on currency sample.
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| CN106018245A (en) * | 2016-05-20 | 2016-10-12 | 河海大学 | Method for enriching and detecting microcystis single cell and colony biomass |
| CN111562210B (en) * | 2020-06-16 | 2023-01-03 | 北京挑战农业科技有限公司 | Method for detecting number of viable bacteria in pre-coated feed microecological preparation product |
| CN112881632A (en) * | 2021-01-20 | 2021-06-01 | 深圳市水文水质中心 | Method and device for counting algae in water sample |
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Non-Patent Citations (2)
| Title |
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| 低功率超声波去除铜绿微囊藻技术;舒天阁等;《华侨大学学报(自然科学版)》;20080131;第29卷(第1期);摘要、第72页最后一段 * |
| 蓝藻伪空胞的测定及其前处理方法研究;杨波等;《安徽农业科学》;20111231;第39卷(第23期);第1.2.3.1节 * |
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