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Quick counting method for microcystis aeruginosa

A microcystis aeruginosa, counting method technology, applied in the direction of microorganism-based method, microorganism determination/inspection, biochemical equipment and method, etc. Counting and other issues to achieve the effect of improving detection efficiency and accuracy, simple pretreatment, and reducing sample loss

Inactive Publication Date: 2013-09-04
NANJING INST OF GEOGRAPHY & LIMNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Microscopic examination requires continuous observation of multiple fields of view, which is not only inefficient, but also has low accuracy due to easy omission and repeated counting
Although flow cytometry is accurate and fast, the instrument is expensive and not widely used
At present, the most commonly used method for the determination of phytoplankton chlorophyll-a is the spectrophotometer method, but this method has cumbersome analysis and operation steps, and generally cannot be used for the determination of a large number of samples
Moreover, this method needs to use organic solvents (acetone, methanol, ethanol, etc.) to extract the pigment, so it has a certain poisonous effect on the operator
[0004] In view of the lack of means and low efficiency of algae monitoring and counting methods in my country, it is of great significance to establish a simple and accurate counting method for Microcystis aeruginosa for the monitoring of harmful algae

Method used

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  • Quick counting method for microcystis aeruginosa
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  • Quick counting method for microcystis aeruginosa

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] According to the rapid counting method of Microcystis aeruginosa of the present invention, at first establish a calibration curve or a regression equation between the algae liquid absorbance and the cell density of Microcystis aeruginosa, the specific steps are as follows:

[0029] 1. Preparation of algae solution. Microcystis aeruginosa was cultured with sterilized BG-11 in a light incubator, and the culture conditions were: light intensity 40μE·(m 2 s) -1 , temperature 25°C, light-to-dark ratio 12h: 12h, shake regularly 3 times a day. After culturing for 7-10 days, take 1ml of the algae liquid and dilute it 2, 5, 8, 10, 20, 50, 100, 1000 times with sterilized BG-11 medium respectively.

[0030] 2. Cell count. Accurately count the cells of each concentration with a microscope, repeat 5 times for each sample, and take the average value to obtain the cell density C i .

[0031] 3. Measure the absorbance. Add the algae solution of the blank group and different conce...

Embodiment 2

[0038] According to the calibration curve or regression equation established in Example 1, the rapid counting method of Microcystis aeruginosa of the present invention is used to measure the cell density of the algae liquid, and compared with the flow cytometer counting method, the specific steps are as follows:

[0039] 1. Preparation of algae solution. Microcystis aeruginosa was cultured with sterilized BG-11 in a light incubator, and the culture conditions were: light intensity 40μE·(m 2 s) -1 , temperature 25°C, light-to-dark ratio 12h: 12h, shake regularly 3 times a day. After culturing for 10 days, take 1ml of algae fluid and dilute it with sterilized BG-11 medium to make algae fluid with three concentration gradients of high, medium and low, and add sterilized BG-11 to the blank group.

[0040] 2. Cell count. The cell densities of the algae liquid in the high, medium and low concentration gradients were 9.64×10 by precise counting using a flow cytometer. 4 pcs / mL, 5...

Embodiment 3

[0049] According to the rapid counting method of Microcystis aeruginosa of the present invention, the impact of rice straw methanol crude extract on the growth of Microcystis aeruginosa is studied, and the specific steps are as follows:

[0050] 1. Sample preparation. Take 50g of crushed dry rice straw (passed through a 40-mesh sieve) and place it in a 1L Erlenmeyer flask, add 500ml of analytically pure methanol, ultrasonically treat for 1 hour at room temperature, and repeat twice. After the extraction, the residue was removed, the particle interference was removed through a 0.22 μm filter membrane, and the extract was obtained by rotary evaporation at 65° C. and weighed.

[0051] Dimethyl sulfoxide (DMSO) was used to dilute the methanol extract (experiments have confirmed that when the proportion of DMSO added is less than 1%, it has no effect on the growth of Microcystis aeruginosa. Hong Yu, Hu Hongying, Huang Jingjing, Sakoda A, Sagehashi M. Different Effects of solvent e...

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Abstract

The invention discloses a quick counting method for microcystis aeruginosa. The quick counting method comprises the following steps of: measuring the absorbancy of microcystis aeruginosa with different concentrations at wavelength of 680nm through a microplate reader, accurately counting frustules through a microscope at the same time, creating a good linear relation between the absorbancy and cell density, and quickly counting the microcystis aeruginosa in a sample to be measured through an obtained correction curve or regression equation. The simple, safe, effective and quick quantifying method for microcystis aeruginosa provided by the invention overcomes the defects of lack of algae monitoring and counting measures and low monitoring and counting efficiency in China.

Description

technical field [0001] The invention relates to a method for monitoring and counting algae, in particular to a method for rapidly counting Microcystis aeruginosa. Background technique [0002] The eutrophication of the water body causes algae to multiply and form water blooms, and cyanobacteria blooms are the most common, among which Microcystis aeruginosa is the most important algae to form water blooms, and it often produces microcystins, which seriously threaten other aquatic organisms Even human health, so Microcystis aeruginosa is also the most harmful algae among cyanobacteria. In order to effectively prevent and control cyanobacterial blooms, the monitoring of Microcystis aeruginosa is particularly important. [0003] Usually, methods to reflect algae biomass include cell counting and chlorophyll a content determination. Cell counting methods can be counted by traditional microscopic examination and flow cytometry. Microscopic examination requires continuous observ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/06G01N21/31C12R1/01
Inventor 苏文孔繁翔赵旭辉
Owner NANJING INST OF GEOGRAPHY & LIMNOLOGY
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