KR20030051131A - Water monitoring method using algae - Google Patents

Abstract

PURPOSE: A water quality monitoring method using algae, particularly closterium ehrenbergii, is provided, thereby on-line monitoring water contamination and quantitatively analyzing the level of water contamination. CONSTITUTION: A water quality monitoring method using algae comprises the steps of: collecting a water sample; preparing a testing solution containing photosynthetic algae; mixing the water sample with the testing solution; lighting the mixture thereof and the testing solution; measuring the fluorescence intensity of the each testing solution and mixture; comparing the measured fluorescence intensity; and estimating the water contamination based on the comparison result, wherein the photosynthetic algae comprises Closterium ehrenbergii.

Description

Water Quality Monitoring Method using Algae {WATER MONITORING METHOD USING ALGAE}

The present invention relates to a water quality monitoring method using algae, and more particularly, to a method for detecting environmental stress applied to water by observing behavior patterns of photosynthetic algae such as green algae.

In general, a method of using a living organism such as fish and shellfish to monitor the leakage of harmful substances in rivers and the like is called bio monitoring. Since the introduction of fish or daphnia in Germany in the late 1970s to determine the presence of toxic substances in lakes and rivers, bio monitoring systems have been studied in countries around the world, including the Netherlands, Belgium, France, Switzerland, Austria and the United Kingdom. To date, various aquatic organisms that are suitable for the determination of toxic substances in the water system have been investigated in various angles, and various methods for monitoring water pollution have been proposed by analyzing their strange behavior patterns and biological characteristics. .

In fish-based bio-monitoring, the behavior patterns of fish such as floating, avoidance, clustering, diffusion, and mobility are continuously observed and analyzed. Then, if these observed fish show strange behavior, it is considered a sign of contamination of the water system such as incorporation of toxic substances.

However, the behavior patterns of fish are influenced not only by environmental factors such as toxic substances, but also by intrinsic factors such as fish health status and other instinctive desires. In other words, the causality rate between the behavioral pattern of fish and water pollution is not so large, and there is a problem of frequent judgment error.

In addition, in order for water pollution to affect a fish's behavioral pattern, the concentration of the pollutant must be above a certain level, and at a concentration below that, there will be no change in the behavioral pattern, and thus, the contamination cannot be detected. In addition, even if the water quality is contaminated above that limit, the behavior pattern of the fish is not immediately abnormal, and depending on the type or concentration of the pollutant, there will be a strange behavior pattern after a considerable time delay. In other words, fish are not very sensitive to water pollution.

The main challenge for biomonitoring research is to increase the causality and sensitivity.

Daphnia show improved causality and sensitivity compared to fish, but are still insufficient.

It is noteworthy that it is employ | adopted in recent years, using luminescent microorganisms.

Korean Patent No. 10-0262681, "High Sensitivity Detection Method of Environmental Damage Factors", E. I. duupand nemoa and campanie; 10-0300445, “Continuous Monitoring Method for Toxic Substances in Water Using Luminescent Microorganisms and Kits for Continuous Monitoring”, Oh, Sung-Keun; Lee, Young-Suk; 10-0305218, “Automatic Measurement of Water Toxicity Using Immobilized Luminescent Microorganisms,” Oh, Sung-Keun; Lee, Young-Soo; Korean Unexamined Patent Publication No. 2000-0024847, “Method for Measuring Toxic Substances Using Immobilized Luminescent Microorganisms and Biosensor Kit for the Same”, LSIS Co., Ltd .; 2001-0086342, “Continuous Water Toxicity Tester,” Banionia Co., Ltd., is concerned with detecting environmental pollution using luminescent microorganisms.

In bio monitoring using luminescent microorganisms, the change in the amount of emitted microorganisms is considered a sign of environmental pollution. However, the amount of luminescence of luminescent microorganisms does not appear to be changed only by environmental factors, but seems to be changed by various intrinsic factors. In other words, even in bio-monitoring using luminescent microorganisms, the causation rate for environmental pollution is not high enough.

An object of the present invention is to propose an environmental monitoring method having a high causality and sensitivity to environmental stress.

Another object of the present invention is to be sensitive to toxic substances entering the water system, to quantify and analyze the level of toxic substances introduced into the water, and to display them in numerical values, to monitor the contamination of water quality in real time, and the level of water pollution In addition to providing a quantitative analysis, the company provides a water quality monitoring method using algae that can drastically reduce related equipment and manpower requirements.

1 is a flowchart illustrating a preferred embodiment of the water quality monitoring method using algae according to the present invention,

FIG. 2 is a view showing a state in which a test solution and a mixed solution of a test solution and a sample are dropped onto a pallet to measure the amount of fluorescence while performing dimming for photosynthesis in the process of implementing the method shown in FIG. 1.

Factors regulating photosynthesis include light intensity and carbon dioxide concentration and temperature. That is, the photosynthesis amount performed in healthy photosynthetic plant cells is only affected by the extracellular regulatory factors as described above, and is hardly influenced by other factors except the health state of the cells.

Changes in the photosynthesis of certain plant cells according to changes in regulatory factors can be obtained experimentally in advance. On the other hand, if the number of cells is large enough, the probability of the presence of healthy cells for each environmental state is within the normal distribution range.

In other words, the amount of photosynthesis performed in a sufficiently large number of photosynthetic plant cells is expected to be nearly constant depending on the environmental conditions, provided that they compensate for the changes caused by the aforementioned regulatory factors.

On the other hand, it takes a considerable time for the health state to change according to environmental effects in multicellular plants, but it is observed that metabolism or health state changes almost immediately according to the environmental state.

Given these facts, it is expected that the use of photosynthesis in single-celled plants with sufficient population will enable the implementation of an environmental surveillance system with a very high causality and sensitivity to environmental stress.

According to one aspect of the present invention, there is provided an environmental monitoring method for detecting an environmental stress by observing the photosynthetic amount of photosynthetic algae.

On the other hand, looking at the flow of light energy incident on the surface of the photosynthetic object, primarily, some are emitted by the energy of reflected light reflected from the surface of the object, and only the remainder is absorbed into the body. Much of the energy absorbed into the body is used for photosynthesis, and part of it is converted into thermal energy and released out of the body, and the remainder is converted into light waves and radiated out of the body.

Light waves absorbed by the body and radiated out of the body are independent of the frequency of incident or reflected light and have a unique frequency for each organism. This light wave will be referred to herein as fluorescence.

In general, light emitting microorganisms emit and generate light waves within a frequency range including visible light by themselves, even when there is no incident light from the outside, whereas the fluorescence mentioned herein disappears as soon as the incident light disappears or with a fairly short time difference. And in most cases have a wavelength outside the visible range.

Experiments have shown that the intensity of this fluorescence is inversely proportional to the amount of photosynthesis. In other words, when the individual completes photosynthesis, the intensity of fluorescence is weakened, and when the individual's photosynthetic activity is weak due to metabolic or health conditions, the intensity of fluorescence is increased.

Therefore, in the environmental monitoring method according to the present invention, it is possible to detect the environmental stress suffered by the photosynthetic algae by observing the photosynthesis amount by measuring the amount of fluorescence.

Environmental monitoring method according to the invention can be used to detect water pollution of the water system.

The water pollution detection method includes a sampling step of collecting water to be used as a sample from the water system to detect contamination, preparing a test solution containing photosynthetic algae, preparing a mixed solution of the test solution and the sample, and the test solution And a dimming step of shining light into the mixed solution, measuring the amount of fluorescence from the test solution and the mixed solution, comparing the amount of fluorescence measured from the test solution with the measured amount of fluorescence from the mixed solution, and the comparison result in the comparison step. Assessing the extent of water pollution on the basis of

According to another aspect of the present invention, a method for monitoring the water pollution of the water system may be implemented by using the above-described evaluation of water pollution. The method further comprises the steps of preparing a reference value of water pollution to be alarmed, comparing the evaluation value obtained in the evaluation step with the reference value, and alarming according to the comparison result of the reference value and the evaluation value. It must be included.

Preferably, the method may further include converting the measured value measured in the measuring step into a digital value and storing the digital value in a database, wherein the evaluation step includes a corresponding value obtained from the digital value stored in the database. It can be evaluated by considering the measurement history of the sample taken from the water system.

In addition, after the dimming step, it is preferable to additionally include the step of observing the behavior pattern of the photosynthetic algae contained in the test solution and the photosynthetic algae contained in the mixed solution, wherein the evaluation step is observed in the observation step Evaluation can be based on the behavioral patterns of photosynthetic algae.

More preferably, after the dimming step, photographing the behavior pattern of the photosynthetic algae contained in the test solution and the photosynthetic algae contained in the mixed solution and outputting the image photographed in the photographing step to the image display apparatus are additionally provided. In this case, in the observation step, the behavior patterns of photosynthetic algae may be observed from an image output to the image display apparatus.

These and other features, advantages, and aspects of the present invention will become more apparent upon reading the detailed description of the preferred embodiments of the present invention described below with reference to the accompanying drawings.

This embodiment is implemented by a method of monitoring the water pollution of the water using a half moon (closterium ehrenbergii), a kind of green algae. 1 is a schematic flowchart for explaining a water quality monitoring method according to this embodiment.

Water quality monitoring method according to this embodiment, as shown in Figure 1, the standard preparation step (S00) and the sample collection step (S10) and the test liquid preparation step (S20) and mixing step (S30) and dimming step (S40) and mold It includes a light quantity measuring step (S50), a fluorescence amount comparing step (S60), a water pollution degree evaluation step (S70), an evaluation value comparison step (S80) and a warning step (S90).

In the baseline preparation step (S00), according to the use of the water system and the type of contaminants, the reference value indicating how much water pollution should be alerted to the manager is determined.

Sampling step (S10) is to take the water to be used as a sample from the water system to detect whether the contamination is stored in the reservoir.

In the preparation of the test solution (S20), prepare a culture solution of the half moon to be used as a test solution for detecting water pollution. At this time, it is preferable that the population of half-moon-lives living in the culture is kept constant within an appropriate error limit. However, if a means for correcting the measured value according to the change in the number of individuals is provided, instead of keeping the number constant, a culture solution in which the number of surviving individuals is counted may be used. The prepared culture solution is dropped in two places on the pallet 10, as shown in FIG. In this embodiment, about 1 ml of the culture solution is dropped in the first position 11 on the pallet 10, and about 2 ml of the culture solution used as the test solution is dropped in the second position 12.

In the mixing step (S30), about 1 ml of water taken as a sample is dropped on the test liquid dropped to the first position 11 on the pallet 10, and mixed.

In the dimming step (S40), the test liquid and the mixed liquid on the palette 10 are dimmed to be used for photosynthesis at the end of the moon.

In the fluorescence measurement step S50, the amount of fluorescence coming from the mixed liquid at the first position 11 on the pallet 10 and the amount of fluorescence coming from the test liquid at the second position 12 are measured. In this case, the amount of fluorescence may be measured using a commercially available fluorimeter.

In the fluorescence amount comparison step (S60), the fluorescence amount measured from the test solution and the fluorescence amount measured from the mixed solution are compared with each other.

In the water pollution evaluation step (S70), the water pollution degree is evaluated based on the comparison result in the fluorescence comparison step (S60).

On the other hand, assuming that the half moon is not present in the sampled water, the population of the half moon is present in the mixed liquid of the first position 11 mixed according to the mixing ratio in this embodiment. It will be about half of the half-moon population present in the test solution, and therefore, the amount of fluorescence measured in the fluorescence measurement step (S50) will also be about half. If a half moon is present in the sampled water, the percentage of the half moon population or the resulting amount of fluorescence will increase. However, if the results of the test using samples taken from the same water system are recorded in advance, and the evaluation results reflecting those test results at the time of evaluation in the water pollution degree evaluation step (S70), the difference of these populations is not a problem. Do not.

In the evaluation value comparison step (S80), the evaluation value obtained in the water pollution degree evaluation step (S70) is compared with the reference value prepared in the reference value preparation step (S00).

In the warning step (S90), an alarm is issued to the water management manager according to the comparison result of the reference value and the evaluation value.

Although this example illustrates the use of a half-moon culture medium as a test solution, if there is an algae culture solution that can be used as a better test solution for a material that is likely to be found as an aqueous pollutant, it may be used as a test solution. In addition, a mixture of two or more kinds of algal culture liquids may be used as a test liquid.

Optionally, the method according to this embodiment includes the measurement value digitization step S51, the measurement value storage step S52, the behavior pattern photographing step S41, the image display step S42, and the behavior pattern observation step S43. It may also include.

In the measurement value digitization step S51, the measurement value measured in the measurement step is converted into a digital value, and in the measurement value storage step S52, the converted digital value is stored in a database. In this case, the water pollution degree evaluation step (S70) can evaluate the water pollution degree in consideration of the measurement history of the sample taken from the water system obtained from the digital value stored in the database.

In the behavior pattern photographing step (S41) performed after the dimming step (S40), the behavior pattern of photosynthetic algae contained in the test solution and the photosynthetic algae contained in the mixed solution are photographed, and the image display step (S42) behavior pattern photographing step The image photographed at S41 is output to the image display apparatus, and in the behavior pattern observation step S43, the behavior patterns of photosynthetic algae are observed from the image output to the image display apparatus. In this case, the evaluation step may evaluate based on the behavioral pattern of the photosynthetic algae observed in the observation step.

In the behavior pattern observation step (S43), the behavior patterns of the photosynthetic algae contained in the test solution and the photosynthetic algae contained in the mixed solution may be directly observed using a microscope or an endoscope. The half moon used in this example has a larger cell size than other birds, making it easy to observe individual behavior patterns.

Measurements measured in the fluorescence measurement step (S50) can be quantitatively quantified and stored and can be usefully used as research data related to water quality analysis.

The water quality monitoring system implementing the method according to this embodiment can be controlled by a computer by automating all or part of its operation process, and at this time, a program for implementing measurement analysis algorithms and data management algorithms can be used. However, the present invention is not described in detail.

While the invention has been described above based on the preferred embodiments, it is not intended to limit the invention but is merely exemplary. It will be apparent to those skilled in the art that changes, modifications, or adjustments from the above-described embodiments can be made without departing from the spirit of the invention. Therefore, it is intended that the appended claims cover such changes, modifications or adjustments.

Using the method according to the present invention, since the data output step and the image output step described above can be implemented as a single output device, from the user's point of view, the water pollution level through the data output step at a designated place such as a laboratory. The quantitative data and the image information through the image output step can be obtained together to monitor the level of water pollution in the water system in three dimensions. In addition, according to the present invention, since it does not require a long-term physicochemical measurement method for determining whether the water system is contaminated, it is effective to measure and monitor the water quality in real time, and to measure the amount of fluorescence of algae in response to external stimuli. As it is possible to quantitatively analyze the level of water pollution, it is possible to accurately quantify the level of water pollution.In the measurement of water quality, a simple and convenient method of measuring the amount of fluorescence of algae by taking a sample from the water system Since only a process is required, it does not require a large-scale equipment for water quality measurement or a large number of accompanying facilities, thereby significantly reducing related equipment and manpower requirements for water quality monitoring.

Claims (7)

In the method of detecting water pollution, A sampling step of collecting water to be used as a sample from the water system to detect contamination; Preparing a test solution containing photosynthetic algae, Preparing a mixed solution of the test solution and the sample; A dimming step of illuminating the test solution and the mixed solution; Measuring the amount of fluorescence emitted from the test solution and the mixed solution, Comparing the amount of fluorescence measured from the test solution with the amount of fluorescence measured from the mixed solution, and And evaluating the degree of water pollution based on the comparison result in the comparing step. The method according to claim 1, Preparing a baseline for water pollution to be alarmed; Comparing the evaluation value obtained in the evaluation step with the reference value, And alerting in response to a comparison result of the reference value and the evaluation value. The method according to claim 1, Converting the measured value measured in the measuring step into a digital value, Additionally storing the digital value in a database, The evaluation step is characterized in that the evaluation in consideration of the measurement history for the sample taken from the water system obtained from the digital value stored in the database. The method according to claim 1, After the dimming step, additionally observing the behavior pattern of the photosynthetic algae contained in the test solution and the behavior pattern of the photosynthetic algae contained in the mixed solution; The evaluation step is characterized in that the evaluation based on the behavior pattern of the photosynthetic algae observed in the observation step. The method according to claim 4, Photographing behavior patterns of the photosynthetic algae contained in the test solution and the photosynthetic algae contained in the mixed solution after the dimming step; Additionally outputting an image photographed in the photographing step to an image display device; And in the observing step, observing behavior patterns of the photosynthetic alga from the image output to the image display apparatus. The method according to any one of claims 1 to 5, And said photosynthetic algae are at least two or more species of algae each having a different causality or sensitivity depending on the type of contaminant. The method according to any one of claims 1 to 5, The photosynthetic alga is characterized in that it comprises a half moon.