WO2003052409A1 - Water monitoring method using algae - Google Patents

Water monitoring method using algae Download PDF

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Publication number
WO2003052409A1
WO2003052409A1 PCT/KR2002/000584 KR0200584W WO03052409A1 WO 2003052409 A1 WO2003052409 A1 WO 2003052409A1 KR 0200584 W KR0200584 W KR 0200584W WO 03052409 A1 WO03052409 A1 WO 03052409A1
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Prior art keywords
water
algae
water pollution
liquid reagent
values
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PCT/KR2002/000584
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French (fr)
Inventor
Sang Gil Kim
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Sang Gil Kim
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Application filed by Sang Gil Kim filed Critical Sang Gil Kim
Priority to AU2002246412A priority Critical patent/AU2002246412A1/en
Priority to JP2003553247A priority patent/JP2005513438A/en
Publication of WO2003052409A1 publication Critical patent/WO2003052409A1/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/18—Water
    • G01N33/186—Water using one or more living organisms, e.g. a fish
    • G01N33/1866—Water using one or more living organisms, e.g. a fish using microorganisms
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/18—Water

Definitions

  • the present invention relates to a water monitoring method using algae, and more particularly, to a method of detecting environmental stress exerted on water by observing photosynthesis of photosynthetic algae such as green algae.
  • bio monitoring a method of monitoring a harmful substance leak accident at a place such as a river by using organisms such as fishes and shellfishes. Since Germany employed a method of determining whether a toxic substance is introduced into a water system including a lake, a river, and the like by using fishes or water fleas in the late 1970's, various countries including the Netherlands, Belgium, France, Switzerland, Austria, England, and the like started to study a bio monitoring system. Heretofore, diversified searches and studies on various aquatic organisms suitable for determining whether the toxic substance is introduced into the water system have been performed. As a result, there have been proposed various methods of monitoring whether water is polluted or not by analyzing abnormal behavioral patterns or biological characteristics of the aquatic organisms.
  • bio monitoring using fishes behavioral patterns of the fishes such as floatability, abient responses, schooling, scattering, and mobility are continuously observed and analyzed. Then, if the observed fishes show abnormal behavioral patterns, it is determined as a presage of the pollution of the water system such as introduction of the toxic substances.
  • the behavioral patterns of the fishes are influenced by environmental factors such as the toxic substances as well as immanent factors such as health conditions and other instinctive desires of the fishes. That is, there is a problem in that determination errors frequently occur because little causality exists between the behavioral patterns of the fishes and the water pollution. Further, only when the concentration of pollutants should exceed a predetermined concentration, the water pollution can exert influence on the behavioral patterns of the fishes. Since there would be no changes in the behavioral patterns of the fishes below the predetermined concentration, it is impossible to detect the presence of the water pollution. Moreover, even though the water is polluted over the threshold, the fishes do not immediately show their abnormal behavioral patterns which will be shown after substantial time delay depending on the kind and concentration of the pollutants. In other words, the sensitivity of the fishes to the water pollution is relatively small.
  • Korean Patent No. 10-0300445 issued to Seong-Keun Oh and entitled “Method and kit for continuously monitoring toxic substance in water system by using luminescent microorganism”
  • Korean Patent No. 10-0305218 issued to Seong-Keun Oh and entitled
  • An object of the present invention is to provide an environmental monitoring method having a close causality and a higher sensitivity to environmental stress.
  • Another object of the present invention is to provide a water monitoring method using algae, which is sensitive to toxic substances introduced into a water system, allows the level of the toxic substances to be quantified and expressed as numerical values, and is able to monitor the presence of water pollution in real time, to perform quantitative analysis of the level of the water pollution and to remarkably reduce relevant equipment and manpower required for the water monitoring.
  • Changes in the amount of photosynthesis of specific plant cells according to variations in the regulation factors can be beforehand obtained experimentally. Meanwhile, if the number of the cells is sufficiently large, the probability that healthy cells exist according to respective environmental conditions falls within normal distribution.
  • an environmental monitoring system having a close causality and a higher sensitivity to the environmental stress can be implemented by using the amount of photosynthesis performed in a sufficient population of the unicellular plants.
  • an environmental monitoring method of detecting the environmental stress by observing the amount of photosynthesis of the photosynthetic algae.
  • a portion of the light energy is first radiated as energy of reflected light which is reflected from the surfaces of the individuals and only the remainder of the light energy is absorbed into bodies of the individuals.
  • Substantial amount of the energy absorbed into the bodies of the individuals are used for the photosynthesis, a portion of the absorbed energy which has not been used for the photosynthesis is converted into thermal energy which in turn is discharged outside of the bodies of the individuals, and the remainder thereof is converted to light waves which in turn is radiated outside of the bodies of the individuals.
  • the light waves which are radiated outside of the bodies of the individuals after absorption into the bodies have no relation to the incident light or reflected light and have unique frequencies depending on respective organisms.
  • light waves are also referred to as fluorescence.
  • the intensity of fluorescence is in inverse proportion to the amount of photosynthesis. In other words, the intensity of fluorescence becomes weak when the individuals complete the photosynthesis, whereas it becomes strong when the photosynthesis activities of the individuals fail due to abnormality of the metabolism and the health conditions.
  • the environmental monitoring method of the present invention it is possible to detect the environmental stress, which is exerted on the photosynthetic algae, by observing the amount of photosynthesis through the measurement of the amount of fluorescence.
  • the environmental monitoring method of the present invention can be utilized to monitor the water pollution in the water system.
  • Such a method for detecting water pollution comprises the steps of taking water to be used as a sample from a water system in which the presence of water pollution is to be detected; preparing a liquid reagent containing photosynthetic algae; preparing a mixed liquid of the liquid reagent and the sample; illuminating the liquid reagent and the mixed liquid with light; measuring the respective amounts of fluorescence emitted from the liquid reagent and the mixed liquid; comparing the amount of fluorescence measured from the liquid reagent with that measured from the mixed liquid; and evaluating the degree of water pollution based on comparison results obtained in the comparing step.
  • the method further comprises the steps of preparing reference values for the water pollution at which a warning is raised; comparing the evaluated values obtained in the evaluating step with the reference values; and raising the warning based on results obtained through the comparison between the reference values and the evaluated values.
  • the method further comprises the steps of converting the measured values obtained in the measuring step into digitized values; and storing the digitized values in a database.
  • the evaluating step may evaluate the degree of water pollution in consideration of a measurement history of the sample, which has been taken from the relevant water system, obtained from the digitized values stored in the database.
  • the method may further comprise the step of observing behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid during the illuminating step.
  • the evaluation perfomied in the evaluating step may be made based on the behavioral patterns of the photosynthetic algae observed in the observing step.
  • the method further comprises the steps of imaging the behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid during the illuminating step; and outputting the images obtained in the imaging step to a display device.
  • the behavioral patterns of the photosynthetic algae may be observed from the images outputted to the display device.
  • FIG. 1 is a flowchart for explaining a preferred embodiment of a water monitoring method using algae according to the present invention.
  • FIG. 2 is a view showing a state where a liquid reagent and a mixed liquid of the liquid reagent and a sample are dropped onto a palette so as to measure the amount of fluorescence while performing illumination for photosynthesis in the course of implementing the method shown in FIG. 1.
  • FIG. 1 shows a schematic flowchart for explaining a water monitoring method according to this embodiment.
  • the water monitoring method comprises a reference value preparing step S00, a sampling step S10, a liquid reagent preparing step S20, a mixing step S30, an illuminating step S40, a step of measuring the amounts of fluorescence S50, a step of comparing the amount of fluorescence S60, a step of evaluating the degree of water pollution S70, a step of comparing an evaluated value S80, and a warning step S90.
  • reference values are predetermined.
  • the reference values indicates levels of the water pollution on which an administrator is required to be warned depending on uses of water systems and the kinds of pollutants.
  • water to be used as a sample is taken from a water system in which the presence of water pollution will be detected, and then, is stored in a reservoir.
  • a culture solution of the closterium ehrenbergii is prepared as a liquid reagent to be used for detecting the water pollution.
  • population of the closterium elirenbergii which exists in the culture solution be kept to be constant within a proper tolerance.
  • a culture solution in which the population of the existing closterium ehrenbergii was counted may be utilized instead of maintaining the population constant.
  • the prepared culture solution is dropped onto two places in a palette 10 as shown in FIG. 2. In this embodiment, about 1 ml of the culture solution is dropped onto a first place 1 1 in the palette 10, and about 2 ml of the culture solution used as the liquid reagent is dropped onto a second place 12 in the palette 10.
  • the liquid reagent and the mixed liquid on the palette 10 are illuminated with light which can be used for the photosynthesis of the closterium ehrenbergii.
  • the step of measuring the amounts of fluorescence S50 the amount of fluorescence emitted from the mixed liquid on the first place 11 in the palette 10 and the amount of fluorescence emitted from the liquid reagent on the second place 12 in the palette 10 are measured. At this time, the amount of fluorescence can be measured by using a fluorimeter available in a market.
  • the step of comparing the amount of fluorescence S60 the amount of fluorescence measured from the liquid reagent is compared with that measured from the mixed liquid.
  • the degree of water pollution is evaluated based on the comparison results obtained in step S60.
  • the population of the closterium ehrenbergii existing in the mixed liquid on the first place 11 which is obtained through the mixing at a mixing ratio according to the present embodiment, is almost half of the population of the closterium ehrenbergii existing in the liquid reagent on the second place 12. Therefore, the amount of fluorescence measured in step S50 will also be almost half. If the closterium elirenbergii exists in the sampled water, the ratio of the populations and thus the ratio of the amounts of fluorescence will be increased.
  • step S70 if a sample taken from the same water system is beforehand tested to record test results and the test results are reflected on the evaluation in step S70, such a difference in the population is out of the question.
  • step of comparing the evaluated value S80 the evaluated value obtained in step S70 is compared with the reference value prepared in step S00.
  • the warning step S90 the administrator of the water system is warned based on the results obtained through the comparison between the reference value and the evaluated value.
  • the present embodiment has been described with respect to the case where the culture solution of the closterium ehrenbergii is used as the liquid reagent. However, if there is a culture solution of algae which can be used as a more preferable liquid reagent for a substance having a high probability of detection as a pollutant in the water system, the culture solution may be used as the liquid reagent. If necessary, a mixture of two or more culture solutions of algae may also be used as the liquid reagent.
  • the method according to the present embodiment may further comprises a step of digitizing the measured values S51 , a step of storing the measured values S52, a step of imaging behavioral patterns S41 , a step of displaying the images S42, and a step of observing the behavioral patterns S43.
  • the step of digitizing the measured values S51 the measured values obtained in the measuring step are converted into digitized values.
  • the digitized values converted as such are stored in a database in the step of storing the measured values S52.
  • the step of evaluating the degree of water pollution S70 can evaluate the degree of water pollution in consideration of a measurement history of the sample, which has been taken from the relevant water system, obtained from the digitized values stored in the database.
  • the step of imaging behavioral patterns S41 that is performed during the illuminating step S40 the behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid are imaged.
  • the step of displaying the images S42 the images obtained in step S41 are outputted to a display device.
  • the step of observing the behavioral patterns S43 the behavioral patterns of the photosynthetic algae are observed from the images outputted to the display device.
  • the evaluation performed in the evaluating step can be made based on the behavioral patterns of the photosynthetic algae observed in the observing step.
  • the behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid can be directly observed by using a microscope, an endoscope, or the like. Since the closterium ehrenbergii used in the present embodiment has a cell larger than those of the other algae, the behavioral patterns of the respective individuals can be easily observed.
  • the measured values obtained in step S50 can be quantitatively expressed as numerical values which in turn are stored so that they can be usefully utilized as data for research work related to a water quality analysis.
  • a water monitoring system for implementing the method according to the present embodiment can be automated partially or entirely in view of its operating processes so as to be controlled by a computer.
  • programs for implementing a measured value analysis algorithm, a data management algorithm and the like the detailed description thereof will be omitted.
  • the aforementioned data outputting step and image displaying step are interlocked with each other to be implemented as one output device. Consequently, a user can obtain both quantified data on the degree of water pollution through the data outputting step and image information through the image outputting step even in one given place such as a laboratory and thus can monitor the degree of water pollution in the water system from all angles. Further, according to the present invention, since a time-consuming chemicophysical measurement method of determining the presence of the water pollution in the water system is not required, there is an advantage in that the presence of the water pollution can be measured and monitored in real time.
  • the degree of water pollution can be precisely expressed as numerical values.
  • the water quality measurement requires only the simple and convenient processes in which the sample is taken from the water system and the amount of fluorescence of the algae is measured by the fluorimeter, there is a further advantage in that it is not necessary to employ large-scale equipment or a number of facilities involved therewith for measuring the water quality, thereby remarkably reducing the number of relevant equipment and manpower required for the water monitoring.

Abstract

The present invention relates to a water monitoring method using algae, wherein the degree of pollution in a river can be monitored by using the algae, particularly, closterium ehrenbergii. The water monitoring method using the algae according to the present invention comprises the steps of taking raw water from a water system, adding a culture solution in which the closterium ehrenbergii has been cultivated to the taken raw water, measuring the respective amounts of fluorescence in the raw water with the culture solution added thereto and in the culture solution and comparing the amounts of fluorescence with each other, and analyzing the compared amounts of fluorescence and outputting the presence of water pollution as quantified data. Therefore, according to the present invention, there is an advantage in that the presence of the water pollution can be measured and monitored in real time. In addition, since the quantitative analysis of the degree of water pollution can be made, there is another advantage in that the degree of water pollution can be precisely expressed as numerical values. Moreover, there is a further advantage in that the number of relevant equipment and manpower required for the water monitoring can be remarkably reduced.

Description

WATER MONITORING METHOD USING ALGAE
Technical Field The present invention relates to a water monitoring method using algae, and more particularly, to a method of detecting environmental stress exerted on water by observing photosynthesis of photosynthetic algae such as green algae.
Background Art Generally, a method of monitoring a harmful substance leak accident at a place such as a river by using organisms such as fishes and shellfishes is called bio monitoring. Since Germany employed a method of determining whether a toxic substance is introduced into a water system including a lake, a river, and the like by using fishes or water fleas in the late 1970's, various countries including the Netherlands, Belgium, France, Switzerland, Austria, England, and the like started to study a bio monitoring system. Heretofore, diversified searches and studies on various aquatic organisms suitable for determining whether the toxic substance is introduced into the water system have been performed. As a result, there have been proposed various methods of monitoring whether water is polluted or not by analyzing abnormal behavioral patterns or biological characteristics of the aquatic organisms.
As for the bio monitoring using fishes, behavioral patterns of the fishes such as floatability, abient responses, schooling, scattering, and mobility are continuously observed and analyzed. Then, if the observed fishes show abnormal behavioral patterns, it is determined as a presage of the pollution of the water system such as introduction of the toxic substances.
However, the behavioral patterns of the fishes are influenced by environmental factors such as the toxic substances as well as immanent factors such as health conditions and other instinctive desires of the fishes. That is, there is a problem in that determination errors frequently occur because little causality exists between the behavioral patterns of the fishes and the water pollution. Further, only when the concentration of pollutants should exceed a predetermined concentration, the water pollution can exert influence on the behavioral patterns of the fishes. Since there would be no changes in the behavioral patterns of the fishes below the predetermined concentration, it is impossible to detect the presence of the water pollution. Moreover, even though the water is polluted over the threshold, the fishes do not immediately show their abnormal behavioral patterns which will be shown after substantial time delay depending on the kind and concentration of the pollutants. In other words, the sensitivity of the fishes to the water pollution is relatively small.
A major topic pursued in studies of the bio monitoring is to improve the causality and the sensitivity.
Although the water fleas show an improved causality and sensitivity over the fishes, it is not yet sufficient.
A notable method which has been recently employed is to utilize luminescent microorganisms. The technology for detecting environmental pollution using the luminescent microorganisms is disclosed in Korean Patent No. 10-0262681 issued to E.I. Du Pont De
Nemours and Company and entitled "Highly sensitive method for detecting environmental insults"; Korean Patent No. 10-0300445 issued to Seong-Keun Oh and entitled "Method and kit for continuously monitoring toxic substance in water system by using luminescent microorganism"; Korean Patent No. 10-0305218 issued to Seong-Keun Oh and entitled
"Apparatus for automatically measuring water toxicity by using immobilized luminescent microorganism"; Korean Laid-Open Patent Publication No. 2000-0024847 filed in the name of LG Industrial Systems Co., Ltd. and entitled "Method of measuring toxic substance by using immobilized luminescent microorganism and bio sensor kit therefor"; Korean Laid-Open Patent Publication No. 2001-0086342 filed in the name of Bioneer
Corporation and entitled "Apparatus for continuously examining water toxicity," etc.
In the bio monitoring using the luminescent microorganisms, changes in the amount of luminescence emitted from the microorganisms are determined as a presage of the environmental pollution. However, it appears that the amount of luminescence emitted from the microorganisms is not changed due to only the environmental factors but is changed according to various immanent factors. In other words, the bio monitoring using the luminescent microorganisms also dose not have a sufficiently close causality to the environmental pollution.
Disclosure of Invention
An object of the present invention is to provide an environmental monitoring method having a close causality and a higher sensitivity to environmental stress.
Another object of the present invention is to provide a water monitoring method using algae, which is sensitive to toxic substances introduced into a water system, allows the level of the toxic substances to be quantified and expressed as numerical values, and is able to monitor the presence of water pollution in real time, to perform quantitative analysis of the level of the water pollution and to remarkably reduce relevant equipment and manpower required for the water monitoring.
As factors for regulating photosynthesis, there are the intensity of light, the concentration of carbon dioxide, and temperature. That is, the amount of photosynthesis performed in healthy photosynthetic plant cells is influenced only by such external regulation factors and hardly influenced by other factors excluding the heath conditions of the cells.
Changes in the amount of photosynthesis of specific plant cells according to variations in the regulation factors can be beforehand obtained experimentally. Meanwhile, if the number of the cells is sufficiently large, the probability that healthy cells exist according to respective environmental conditions falls within normal distribution.
In other words, it is expected that the amount of photosynthesis performed in a sufficiently large number of the plant cells is nearly constant by the respective environmental conditions if the changes according to the regulation factors are compensated.
In the meantime, it has been found that changes in the health conditions of multicelluar plants according to the environmental factors take much time, whereas the metabolism and the health conditions of unicellular plants are nearly immediately changed depending on the environmental conditions. When such facts are taken into consideration, it is expected that an environmental monitoring system having a close causality and a higher sensitivity to the environmental stress can be implemented by using the amount of photosynthesis performed in a sufficient population of the unicellular plants. According to one aspect of the present invention, there is provided an environmental monitoring method of detecting the environmental stress by observing the amount of photosynthesis of the photosynthetic algae.
Meanwhile, considering flow of light energy which has been incident on surfaces of the individuals capable of performing the photosynthesis, a portion of the light energy is first radiated as energy of reflected light which is reflected from the surfaces of the individuals and only the remainder of the light energy is absorbed into bodies of the individuals. Substantial amount of the energy absorbed into the bodies of the individuals are used for the photosynthesis, a portion of the absorbed energy which has not been used for the photosynthesis is converted into thermal energy which in turn is discharged outside of the bodies of the individuals, and the remainder thereof is converted to light waves which in turn is radiated outside of the bodies of the individuals.
The light waves which are radiated outside of the bodies of the individuals after absorption into the bodies have no relation to the incident light or reflected light and have unique frequencies depending on respective organisms. Herein, such light waves are also referred to as fluorescence.
In general luminescence of the luminescent microorganisms, light waves which fall within a frequency range including visible rays are spontaneously generated without incident light from the outside and then radiated toward the outside. On the other hand, the aforementioned fluorescence becomes extinct immediately or with a very short time interval when the incident light becomes extinct, and has a wavelength outside of the visible ray range in most cases.
It has been found from experiments that the intensity of fluorescence is in inverse proportion to the amount of photosynthesis. In other words, the intensity of fluorescence becomes weak when the individuals complete the photosynthesis, whereas it becomes strong when the photosynthesis activities of the individuals fail due to abnormality of the metabolism and the health conditions.
Consequently, according to the environmental monitoring method of the present invention, it is possible to detect the environmental stress, which is exerted on the photosynthetic algae, by observing the amount of photosynthesis through the measurement of the amount of fluorescence.
The environmental monitoring method of the present invention can be utilized to monitor the water pollution in the water system.
Such a method for detecting water pollution comprises the steps of taking water to be used as a sample from a water system in which the presence of water pollution is to be detected; preparing a liquid reagent containing photosynthetic algae; preparing a mixed liquid of the liquid reagent and the sample; illuminating the liquid reagent and the mixed liquid with light; measuring the respective amounts of fluorescence emitted from the liquid reagent and the mixed liquid; comparing the amount of fluorescence measured from the liquid reagent with that measured from the mixed liquid; and evaluating the degree of water pollution based on comparison results obtained in the comparing step.
According to another aspect of the present invention, it is possible to implement a method of monitoring water pollution in a water system by using evaluated values of the water pollution detected as such. The method further comprises the steps of preparing reference values for the water pollution at which a warning is raised; comparing the evaluated values obtained in the evaluating step with the reference values; and raising the warning based on results obtained through the comparison between the reference values and the evaluated values.
Preferably, the method further comprises the steps of converting the measured values obtained in the measuring step into digitized values; and storing the digitized values in a database. At this time, the evaluating step may evaluate the degree of water pollution in consideration of a measurement history of the sample, which has been taken from the relevant water system, obtained from the digitized values stored in the database.
Further, the method may further comprise the step of observing behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid during the illuminating step. At this time, the evaluation perfomied in the evaluating step may be made based on the behavioral patterns of the photosynthetic algae observed in the observing step.
More preferably, the method further comprises the steps of imaging the behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid during the illuminating step; and outputting the images obtained in the imaging step to a display device. At this time, in the observing step, the behavioral patterns of the photosynthetic algae may be observed from the images outputted to the display device.
Brief Description of Drawings
The above and other features, advantages and aspects of the invention will become more apparent from reading the following description of a preferred embodiment taken in connection with the accompanying drawings, in which:
FIG. 1 is a flowchart for explaining a preferred embodiment of a water monitoring method using algae according to the present invention; and
FIG. 2 is a view showing a state where a liquid reagent and a mixed liquid of the liquid reagent and a sample are dropped onto a palette so as to measure the amount of fluorescence while performing illumination for photosynthesis in the course of implementing the method shown in FIG. 1.
Best Mode for Carrying Out the Invention
A preferred embodiment of the present invention is implemented as a method of monitoring the presence of water pollution in a water system by using closterium ehrenbergii which is a kind of green alga. FIG. 1 shows a schematic flowchart for explaining a water monitoring method according to this embodiment.
As shown in FIG. 1 , the water monitoring method according to the embodiment comprises a reference value preparing step S00, a sampling step S10, a liquid reagent preparing step S20, a mixing step S30, an illuminating step S40, a step of measuring the amounts of fluorescence S50, a step of comparing the amount of fluorescence S60, a step of evaluating the degree of water pollution S70, a step of comparing an evaluated value S80, and a warning step S90.
In the reference value preparing step SOO, reference values are predetermined. The reference values indicates levels of the water pollution on which an administrator is required to be warned depending on uses of water systems and the kinds of pollutants. In the sampling step S10, water to be used as a sample is taken from a water system in which the presence of water pollution will be detected, and then, is stored in a reservoir.
In the liquid reagent preparing step S20, a culture solution of the closterium ehrenbergii is prepared as a liquid reagent to be used for detecting the water pollution. At this time, it is preferred that population of the closterium elirenbergii which exists in the culture solution be kept to be constant within a proper tolerance. However, if there is provided a means for compensating measured values in accordance with changes in the population, a culture solution in which the population of the existing closterium ehrenbergii was counted may be utilized instead of maintaining the population constant. The prepared culture solution is dropped onto two places in a palette 10 as shown in FIG. 2. In this embodiment, about 1 ml of the culture solution is dropped onto a first place 1 1 in the palette 10, and about 2 ml of the culture solution used as the liquid reagent is dropped onto a second place 12 in the palette 10.
In the mixing step S30, about 1 mβ of the water which has been taken as the sample is dropped onto and mixed with the liquid reagent which has been already dropped onto the first place 11 in the palette 10.
In the illuminating step S40, the liquid reagent and the mixed liquid on the palette 10 are illuminated with light which can be used for the photosynthesis of the closterium ehrenbergii. In the step of measuring the amounts of fluorescence S50, the amount of fluorescence emitted from the mixed liquid on the first place 11 in the palette 10 and the amount of fluorescence emitted from the liquid reagent on the second place 12 in the palette 10 are measured. At this time, the amount of fluorescence can be measured by using a fluorimeter available in a market. In the step of comparing the amount of fluorescence S60, the amount of fluorescence measured from the liquid reagent is compared with that measured from the mixed liquid.
In the step of evaluating the degree of water pollution S70, the degree of water pollution is evaluated based on the comparison results obtained in step S60. Meanwhile, assuming that the closterium ehrenbergii does not exist in the sampled water, the population of the closterium ehrenbergii existing in the mixed liquid on the first place 11, which is obtained through the mixing at a mixing ratio according to the present embodiment, is almost half of the population of the closterium ehrenbergii existing in the liquid reagent on the second place 12. Therefore, the amount of fluorescence measured in step S50 will also be almost half. If the closterium elirenbergii exists in the sampled water, the ratio of the populations and thus the ratio of the amounts of fluorescence will be increased. However, if a sample taken from the same water system is beforehand tested to record test results and the test results are reflected on the evaluation in step S70, such a difference in the population is out of the question. In the step of comparing the evaluated value S80, the evaluated value obtained in step S70 is compared with the reference value prepared in step S00.
Tn the warning step S90, the administrator of the water system is warned based on the results obtained through the comparison between the reference value and the evaluated value. The present embodiment has been described with respect to the case where the culture solution of the closterium ehrenbergii is used as the liquid reagent. However, if there is a culture solution of algae which can be used as a more preferable liquid reagent for a substance having a high probability of detection as a pollutant in the water system, the culture solution may be used as the liquid reagent. If necessary, a mixture of two or more culture solutions of algae may also be used as the liquid reagent.
Alternatively, the method according to the present embodiment may further comprises a step of digitizing the measured values S51 , a step of storing the measured values S52, a step of imaging behavioral patterns S41 , a step of displaying the images S42, and a step of observing the behavioral patterns S43. In the step of digitizing the measured values S51 , the measured values obtained in the measuring step are converted into digitized values. The digitized values converted as such are stored in a database in the step of storing the measured values S52. In such a case, the step of evaluating the degree of water pollution S70 can evaluate the degree of water pollution in consideration of a measurement history of the sample, which has been taken from the relevant water system, obtained from the digitized values stored in the database.
In the step of imaging behavioral patterns S41 that is performed during the illuminating step S40, the behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid are imaged. In the step of displaying the images S42, the images obtained in step S41 are outputted to a display device. Tn the step of observing the behavioral patterns S43, the behavioral patterns of the photosynthetic algae are observed from the images outputted to the display device. In such a case, the evaluation performed in the evaluating step can be made based on the behavioral patterns of the photosynthetic algae observed in the observing step. In the step of observing the behavioral patterns S43, the behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid can be directly observed by using a microscope, an endoscope, or the like. Since the closterium ehrenbergii used in the present embodiment has a cell larger than those of the other algae, the behavioral patterns of the respective individuals can be easily observed. The measured values obtained in step S50 can be quantitatively expressed as numerical values which in turn are stored so that they can be usefully utilized as data for research work related to a water quality analysis.
A water monitoring system for implementing the method according to the present embodiment can be automated partially or entirely in view of its operating processes so as to be controlled by a computer. At this time, although it is possible to use programs for implementing a measured value analysis algorithm, a data management algorithm and the like, the detailed description thereof will be omitted.
According to the method of the present invention, the aforementioned data outputting step and image displaying step are interlocked with each other to be implemented as one output device. Consequently, a user can obtain both quantified data on the degree of water pollution through the data outputting step and image information through the image outputting step even in one given place such as a laboratory and thus can monitor the degree of water pollution in the water system from all angles. Further, according to the present invention, since a time-consuming chemicophysical measurement method of determining the presence of the water pollution in the water system is not required, there is an advantage in that the presence of the water pollution can be measured and monitored in real time. In addition, since the quantitative analysis of the degree of water pollution can be made through the fluorescence measurement of the algae which respond to external stimuli, there is another advantage in that the degree of water pollution can be precisely expressed as numerical values. Furthermore, since the water quality measurement requires only the simple and convenient processes in which the sample is taken from the water system and the amount of fluorescence of the algae is measured by the fluorimeter, there is a further advantage in that it is not necessary to employ large-scale equipment or a number of facilities involved therewith for measuring the water quality, thereby remarkably reducing the number of relevant equipment and manpower required for the water monitoring.
Although the present invention has been described in connection with the preferred embodiment thereof, the description is not limitative but only illustrative. It will be apparent to those skilled in the art that various modifications, changes or adjustments can be made thereto without departing from the technical spirit and scope of the invention. Therefore, the appended claims intend to include such various modifications, changes or adjustments.

Claims

1. A method of detecting water pollution, comprising the steps of: taking water to be used as a sample from a water system in which the presence of water pollution is to be detected; preparing a liquid reagent containing photosynthetic algae; preparing a mixed liquid of the liquid reagent and the sample; illuminating the liquid reagent and the mixed liquid with light; measuring the respective amounts of fluorescence emitted from the liquid reagent and the mixed liquid; comparing the amount of fluorescence measured from the liquid reagent with that measured from the mixed liquid; and evaluating the degree of water pollution based on comparison results obtained in the comparing step.
2. The method as claimed in claim 1, further comprising the steps of: preparing reference values for the water pollution at which a warning is raised; comparing evaluated values obtained in the evaluating step with the reference values; and raising the warning based on results obtained through the comparison between the reference values and the evaluated values.
3. The method as claimed in claim 1, further comprising the steps of: converting the measured values obtained in the measuring step into digitized values; and storing the digitized values in a database, wherein in the evaluating step, the evaluation on the degree of water pollution is made in consideration of a measurement history of samples, which have been taken from the relevant water system, obtained from the digitized values stored in the database.
4. The method as claimed in claim 1 , further comprising the step of: observing behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid during the illuminating step, wherein the evaluation performed in the evaluating step is made based on the behavioral patterns of the photosynthetic algae observed in the observing step.
5. The method as claimed in claim 4, further comprising the steps of: imaging the behavioral patterns of the respective photosynthetic algae contained in the liquid reagent and the mixed liquid during the illuminating step; and outputting the images obtained in the imaging step to a display device, wherein in the observing step, the behavioral patterns of the photosynthetic algae are observed from the images outputted to the display device.
6. The method as claimed in any one of claims 1 to 5, wherein the photosynthetic algae comprise at least two kinds of algae having different causalities or sensitivities according to the kinds of pollutants.
7. The method as claimed in any one of claims 1 to 5, wherein the photosynthetic algae comprise closterium elirenbergii.
PCT/KR2002/000584 2001-12-14 2002-04-03 Water monitoring method using algae WO2003052409A1 (en)

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