微囊藻细胞计数法专题研究英

1、Study on Counting Method for Microcystis Cells Yingke- ZHOUCollege of Mining Engineering，Hebei United UniversityTangShan，HeBei ，063009，This paper expounds the ultrasonic short-term effect to treatment water in order to find a method which counts the cells of Microcystis in water more scientifically.

2、 By experiment, it is further study of ultrasonic frequency and reaction time influence on treatment effect.Key words: Microcystis; count; ultrasonicWater environment eutrophication caused by algal blooms has become more and more serious in our country. Fresh water lakes such as Taihu 1, Dianchi2,3

3、and Chaohu2 every summer and autumn explode cyanobacterial bloom, which is capable of generating microcystin microcystis. Toxic algal blooms occurring not only bring serious harm to aquatic ecosystem 3, but also algal toxins to human health having direct or potential harm4, so controling algal bloom

4、 and eliminating toxins harm have been brooking no delay. Microcystis cell count monitoring will provide first hand information for algal warning and control.Algal density counting method is divided into individual counting and cell counting . Because of the biological individual differences, especi

5、ally in clusters of Microcystis, individual of Microcystis varies very largly, the bigger can reach hundreds or even thousands of times than the smaller. So cell counting method is more scientific to determine the algae growth. But the cell counting method is time-consuming and more counting errors

6、uses, so using the short-term effect of ultrasound to break up large individuals makes the individual of Microcystis in water become smaller and be more distributly.RADITIONAL MICROSCOPY AND ULTRASONIC TREATMENT METHOD COMPARISONExperiments select A and B two containing Microcystis water samples, th

7、en each takes the 30mL of two to four 50mL ground glass bottle, number A1, A2, B1 and B2.A1, B1 use traditional microscopic counting 5 each with 0.1mL algae counting plate to count 7; A2, B2 40KHz ultrasonic treatment about 9min, then microscopic counting, each with 0.1mL algae counting plate count

8、of 7, the results in table 1. Table 1 shows: the relative standard deviation of A,B two samples by conventional microscopy is in 40% above, and the distribution of Microcystis cells in water is very uneven, only the count to 1 2 piece to determine Microcystis cell number of water samples in the pres

9、ence of large random error, not scientific and quickly to detect the true concentration. While the use of 40KHz ultrasonic treatment about 9min before Microscopic examination, because of the role of ultrasound, Microcystis community begin to disintegrate, and then make water Microcystis cell distrib

10、ution increase. After treatment, the relative standard deviation of A, B two samples is reduced to about 10%, so even if the count only 12, it can accurately reflect the true level of microcystin concentration in water samples. The ultrasonic by mechanical effecting, thermal effect and acoustic stre

11、aming effect can make the algae cell and chemical bond in material molecule rupture. Cavitation can make themicrobubble present in the liquid expand quickly then close suddenly, produce shock waves and jet that destroy the physical biological membrane, nucleus structure and configuration, and disint

12、egrate the algae cell, so Microcystis cell concentration after treatment is lower than before.EXPERIMENT RESULTS AND DISCUSSIONA. Ultrsonic Treantment Time Influence On The Counting RusultsUsing 40kHz ultrasonic to treat B sample, processing time is different, then taking the appropriate amount of s

13、amples for analysis, count of 4 times, the results are shown in table 2.TABLE.THE COMPARISON OF TRADITIONAL MICROSCOPY AND ULTRASCONIC PROCESSING METHOD Sample Packets1234567Mean ValueRelative Standard DeviationA10.671.240.591.190.992.000.891.0843.6%A22.363.461.560.984.012.141.862.3445.4%B11.171.130

14、.921.010.921.020.821.0012.4%B22.012.162.292.411.852.031.932.109.54%Note: the number of units: 108 cells / LTABLE. ULTRASONIC TREANTMENT TIME INFLUENCE ON THE COUNTING RUSULTS Parallel Sample1min3min5min9min15min20min13.550.981.962.011.771.3321.983.362.662.331.861.2130.862.462.841.861.961.4542.562.86

15、1.532.011.581.18Mean Value2.242.422.252.051.791.29Relative Standard Deviation50.24%42.45%27.17%9.65%9.01%9.55%Note: the number of units: 108 cells / L FigProcessing of Microcystis morphology FigThe effect of ultrasonic 5min Microcystis morphology FigThe effect of ultrasonic 9min Microcystis morpholo

16、gy Fig Effect of ultrasonic over 9min Microcystis morphology By the graph 1-4 visible under the ultrasonic action, large Microcystis groups gap betwwen each cell slowly expanding, part of the cells disintegrated make the uneasy counting big groups of Microcystis gradually changed into easy counting

17、small groups, thereby water Microcystis cell concentration will be more accurately draw than before.Table 2 and figure 5 show, the action time is about 9min, counting the relative deviations are smaller ( about 10% ), and cell loss is only about 10%.B. Effect of Ultrasonic Frequency on Count Result

18、InfluenceTreatment of B sample 9min with different frequency ultrasonic, it takes the right amount of samples for analysis, count of 4 times, the result is shown in table 3:From table 3 , the same treatment 9 min, when the ultrasonic frequency is about 20kHz, Microcystis cell loss is bigger, and the

19、 frequency is more than 40kHz, count of the relative deviations are smaller ( about 10% ).C. Revised DataThe ultrasonic approach can make partial Microcystis cell disintegration and loss, under the condition of 40kHz ultrasonic effect about 9min, the loss of about 10%, so after processing based on c

20、ounting results multiplied by a correlation coefficient of 1.1, it can be more close to the actual concentration of microcystin in water data.D. DiscussionTo deal with the larger samples containing Microcystis groups by ultrasonic , it can more efficiently, scientifically count out Microcystis cell

21、concentration in water; when the ultrasonic frequency is 40kHz and processing time is about 9min, it can achieve the best treatment effect, water Microcystis distribution in water is more uniform, and the relative deviations of the counting is smaller, and Microcystis cells concentration only loss of about 10%; through the counting result multiplied by correlation coefficient of 1.1 to get the data that it is closer to the actual concentration of microcystin in water.REFERENCES1 Chen Yuwei, Gao Xiyun. Taihu Microcystis growth c