WO2014156363A1 - Procédé d'analyse de la qualité de l'eau utilisant des algues - Google Patents

Procédé d'analyse de la qualité de l'eau utilisant des algues Download PDF

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Publication number
WO2014156363A1
WO2014156363A1 PCT/JP2014/053547 JP2014053547W WO2014156363A1 WO 2014156363 A1 WO2014156363 A1 WO 2014156363A1 JP 2014053547 W JP2014053547 W JP 2014053547W WO 2014156363 A1 WO2014156363 A1 WO 2014156363A1
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light
water
algae
absorbance
green
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PCT/JP2014/053547
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English (en)
Japanese (ja)
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加来　啓憲
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栗田工業株式会社
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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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/025—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration

Definitions

  • the present invention relates to a water quality test method using algae for judging the water quality of environmental water, industrial wastewater treated water, or algae culture solution.
  • Environmental waters such as rivers, lakes, and seawater are not preferable in environments where microalgae such as blue sea bream can easily propagate.
  • treated water that has been treated from factory wastewater should not be allowed to be discharged to the outside environment with water quality that adversely affects the environment.
  • nutrient salts such as nitrogen and phosphorus
  • environmental water and factory wastewater are collected and analyzed for water quality, and the contents of pH, various heavy metals, organic carbon components, total nitrogen (TN) and phosphorus are analyzed.
  • TN total nitrogen
  • microalgae are used for industrial purposes and are grown in a culture solution for industrial use.
  • fertilizer components can be added to environmental water. is doing.
  • the current situation is that it is not possible to determine whether the culture solution is suitable for culturing actual algae without culturing the algae.
  • AGP test method which is one of sewer test methods.
  • AGP is an abbreviation for Algal Ground Potentials
  • algae is inoculated into the effluent water (treated water), and this is maintained under predetermined conditions, and the degree of growth is defined as effluent water area By comparing it with that of, the effect of this effluent on eutrophication in the effluent area is evaluated.
  • algae are measured by dry weight method, particle measurement method, absorbance method, chlorophyll method, direct measurement method and the like.
  • the present invention has been made in view of the above problems, and provides a water quality test method using algae for easily and accurately judging the water quality of environmental water, industrial wastewater treated water, or algae culture solution. With the goal.
  • the present invention adds algae to the water to be tested, and changes the color of the water to be tested containing the algae to blue light (450 to 490 nm), green light (500 to 570 nm) and red light ( 620 to 740 nm) to detect at least two or more wavelength ranges, and determine the algae productivity (AGP) or growth inhibition of the algae based on the light intensity of the two or more light wavelengths.
  • AGP algae productivity
  • the present invention adds algae to the water to be tested, and changes the color of the water to be tested containing the algae to blue light (450 to 490 nm), green light (500 to 570 nm) and red light ( 620 to 740 nm) to detect at least two or more wavelength ranges, and determine the algae productivity (AGP) or growth inhibition of the algae based on the light intensity of the two or more light wavelengths.
  • AGP algae productivity
  • the present invention adds algae to the water to be tested, and changes the color of the water to be tested containing the algae to blue light
  • the chromaticity of visible light may be detected separately for RGB, and it is inexpensive.
  • a color sensor can be applied. By applying the color sensor, online measurement is possible, and the measurement time can be greatly shortened and the measurement can be simplified. Furthermore, this color sensor can detect a wide range of color tone changes such as satellite photographs with only visible light. Thus, it is possible to determine the algae productivity (AGP) or the algae growth inhibitory property of the water under test by efficiently monitoring the culture state of the algae and comparing it with the standard environmental water or the like. .
  • the green light (500 to 570 nm) and the red light (620 to 740 nm) are decomposed and detected, and the light intensity of the green light wavelength and the light intensity of the red light wavelength are detected. From these, it is preferable to determine the algae productivity (AGP) or the growth inhibition of algae (Invention 2).
  • the absorbance of the green light is calculated from the light intensity of the green light wavelength
  • the absorbance of the red light is calculated from the light intensity of the red light wavelength
  • the red light absorbance is divided by the absorbance of the green light. It is preferable to determine the algal productivity (AGP) of the test water or the growth inhibition of the algae (Invention 3).
  • the intensity ratio between the absorbance of red light and the absorbance of green light is correlated with the culture rate of green algae performing photosynthesis, and the absorbance ratio increases as the culture rate increases. Therefore, it is possible to determine the algae productivity (AGP) or the growth inhibition of algae by comparing with the absorbance ratio of the environmental water or the like as a reference.
  • the spectrum of the water to be tested is compared by comparing the light intensity of each wavelength obtained by spectroscopy of the water to be tested containing the algae with the light intensity of each wavelength obtained by separating white light. It is preferable to calculate the absorbance of each wavelength and determine the algae productivity (AGP) or growth inhibition of the algae based on the absorbance of each light (Invention 4).
  • the said white light is the reflected light from the white substance which permeate
  • the blue light, green light and red light intensity of water to be tested containing algae is compared with the light intensity of each wavelength obtained by spectrally dividing white light.
  • AGP algal productivity
  • the color of the water to be tested containing algae is detected by decomposing it into blue light, green light and red light, and the culture state of the algae is determined by comparing the light intensity of the band light of these wavelengths. Therefore, it is possible to measure the culture state of the algae on-line, and the measurement time can be greatly shortened and the measurement can be simplified. By this, the culture state of algae is efficiently monitored, and the algae productivity (AGP) or the growth inhibition of algae is determined by comparing with the standard absorbance ratio of environmental water, etc. Can do.
  • AGP algae productivity
  • the growth inhibition of algae is determined by comparing with the standard absorbance ratio of environmental water, etc. Can do.
  • the color of the water under test containing algae is detected by decomposing green light and red light, and the red light absorbance is determined from the light intensity of the band light of the green light wavelength and the light intensity of the band light of the red light wavelength.
  • the absorbance of green light is determined, the value obtained by dividing the absorbance of red light by the absorbance of green light (absorbance ratio) is calculated, and the algae productivity (AGP) of the test water of the algae is calculated from the value of this absorbance ratio. Or it becomes possible to determine the growth inhibitory property of algae.
  • algae that have been cultured in advance are added to the test water, and the color of the test water containing the algae is changed to blue light (450 to 490 nm), green light (500 to 570 nm), and red light ( 620 to 740 nm) to detect at least two or more wavelength ranges, and determine the algae productivity (AGP) or growth inhibition of the algae based on the light intensity of the two or more light wavelengths.
  • AGP algae productivity
  • the green light (500 to 570 nm) and the red light (620 to 740 nm) are decomposed and detected from the color (chromaticity) of the culture solution containing algae,
  • the algae productivity (AGP) of the water to be tested or the growth inhibition of algae is determined from the light intensity in the wavelength range of the green light and the light intensity in the wavelength range of the red light.
  • the water to be tested is not particularly limited, and environmental water such as rivers, lakes, and seawater, treated water that has been processed from factory wastewater, and culture solution for using algae for industrial purposes are tested. can do.
  • pretreatment such as filtration, pyrolysis and dilution can be performed, or fertilizer components such as nitrogen and phosphorus can be added depending on the purpose of evaluation.
  • microalgae can be used as the algae inoculated into the water to be tested, and microalgae are particularly suitable.
  • the water to be tested is fresh water, Selenastrum capricornutum, Chlorella ellipsiside, Chlorella vulgaris, Microcystis aeruginosa, f. aeruginosa Microcystis aeruginosa f.
  • Skeletonema costatum a costatum
  • Thalassiosira pseudona a compound that can be used, but not limited thereto.
  • Chattonella antique a compound that can be used, but not limited thereto.
  • These algae are preferably cultured in advance (pre-culture) in a necessary amount.
  • the above algae When the above algae is inoculated into the water to be tested, it is cultured.
  • the culture with the water to be tested is preferably performed at a temperature of 20 ⁇ 2 ° C. or 25 ⁇ 2 ° C., and a temperature adjusting mechanism may be provided in the culture tank for that purpose.
  • the algae is preferably irradiated with light having an illuminance in the range of 1,000 to 4,000 lux, and a mechanism capable of adjusting the amount of light within the above range is provided. Furthermore, it is preferable to culture while stirring and mixing the culture solution. For this reason, a shaking mechanism or an aeration mechanism is required.
  • the chromaticity of the cultured test water is detected.
  • a color sensor As a means for detecting the chromaticity of the water under test, it is preferable to use a color sensor because it is inexpensive and can separately detect green light, red light and blue light.
  • This color sensor has a mechanism in which measured colors are separated into RGB components by a color filter, and the light intensity of each color component is detected by a photo diode or the like. This color sensor can detect changes in a wide range of color tones such as satellite photographs using only visible light.
  • the color sensor is used to determine the algae culture state (growth rate) as follows. That is, first, white light is irradiated to transparent water (for example, pure water) that does not absorb light, and the transmitted light is detected by a color sensor. Since this white light is decomposed into RGB components by the color filter of the color sensor and received, the respective light intensities of the red band light (green light) R1 and the green band light (green light) G1 are measured. To do.
  • the test water in which the algae is cultured is irradiated with white light in the same manner using the same color sensor, and the transmitted light is detected with the color sensor. Since this transmitted light is separated into RGB components by the color filter of the color sensor and received, the respective light intensities of the red band light (red light) R2 and the green band light (green light) G2 are measured. To do.
  • the red band light (red light) and the green band light (green light) can be measured using, for example, a transmissive color sensor 1 as shown in FIG. 1 described in Japanese Patent Application Laid-Open No. 2010-151605. it can.
  • the transmissive color sensor 1 includes a light emitting unit 2 and a light receiving unit 3 including a color filter (not shown), and irradiates white light from the light emitting unit 2 and transmits light transmitted through the culture solution 4. Light is received by the light receiving unit 3, and the light intensity of each of the red band light (green light) and the green band light (green light) is calculated by a control mechanism (not shown).
  • a reflective color sensor 11 as shown in FIG. 2 described in JP 2010-181150 can also be used.
  • the reflective color sensor 11 includes a light receiving unit 13 including a light emitting unit 12 and a color filter (not shown), and a reflective plate 14.
  • the reflective color sensor 11 irradiates white light from the light emitting unit 12, and The light that has passed through the culture medium 15 is received by the light receiving unit 13, and the light intensity of each of the red band light (green light) and the green band light (green light) is calculated by a control mechanism (not shown).
  • Absorbance ratio: X AR / AG
  • the weight concentration of microalgae in the water to be tested is measured, and the specific growth rate is measured.
  • This specific growth rate is obtained, for example, by measuring the weight of a suspended substance in a culture solution with a glass fiber filter having a pore diameter of 1 ⁇ m and measuring the weight thereof, and calculating the weight concentration from the amount of the culture solution (test water). Then, the weight concentration C1 [mg / L] and the amount of culture solution V1 [L] at the culture day T1 [day], and the weight concentration C2 [mg / L] and the culture solution amount at the culture day T2 [day].
  • the algae is cultured in advance in the water of the discharge water area, the absorbance ratio (X) is measured, and compared with this, the treated water of the factory effluent is What is necessary is just to judge whether it shows the increase tendency of productivity or the increase tendency of the growth inhibitory property of algae.
  • the algae is cultured in advance with a reference culture solution, and the absorbance ratio (X) is measured. It is sufficient to determine whether or not it shows a tendency to increase or a growth inhibition of algae.
  • the treatment conditions will be changed in the case of treated water of factory effluent, or the components will be changed in the case of a culture solution so that the absorbance ratio (X) will have a desired tendency. You may make it take the measures.
  • the culture state of the microalgae is determined based on the green light (500 to 570 nm) and the red light (620 to 740 nm).
  • absorbance data of blue light 450 to 490 nm can be used.
  • test strain 1 that has been precultured and collected by centrifugation (1,000 G, 5 minutes) is added to a sterilized sodium hydrogen carbonate solution (15 mg / L), shaken, and sufficiently suspended. After that, it was centrifuged again to collect algal bodies.
  • AAP medium (condition 1; reference condition) using the collected algal cells of test strain 1 as simulated water for test water, AAP medium (condition 2) with a sodium nitrate concentration increased to 400 mg / L, and hypochlorous acid AAP medium (condition 3) supplemented with 0.1 mg / L of sodium was inoculated so as to have an algal body concentration of about 0.4 g / L, respectively, and cultured in a 1 L glass flat culture bottle.
  • Condition 1 was assumed to be a standard condition
  • condition 2 was a condition simulating water with high algal productivity
  • condition 3 was a condition simulating water having an algal growth inhibitory property.
  • the culture solution (test water) of this test strain 1 was irradiated with white light, and the transmitted light was measured using a detector equipped with the photosensor shown in FIGS.
  • the optical sensor received light through three color filters of RGB and measured the light intensity of each of the red band light R2 and the green band light G2.
  • the pure water produced by the pure water production apparatus WG270 manufactured by Yamato Kagaku Co., Ltd. is transparent water that does not absorb light, the pure water is irradiated with white light, and the transmitted light is similarly detected by the detector. .
  • the light intensities of the red band light R1 and the green band light G1 were measured, and the absorbance and the absorbance ratio were calculated by the following equations.
  • the weight concentration of test strain 1 in the culture solution was measured, and the specific growth rate was calculated by the following equation.
  • the specific growth rate was calculated by measuring the weight by filtering the suspended matter in the culture solution with a glass fiber filter having a pore size of 1 ⁇ m and calculating the weight concentration. Then, the weight concentration C1 [mg / L] and the amount of culture solution V1 [L] at the culture day T1 [day], and the weight concentration C2 [mg / L] and the culture solution amount at the culture day T2 [day].
  • the specific growth rate ⁇ [1 / day] was calculated from V2 [L] by the following formula.
  • Table 2 shows the measurement results of the absorbance ratio X and specific growth rate ⁇ of red band light and green band light under conditions 1, 2 and 3.
  • the color of water under test containing algae is detected by decomposing into blue light, green light and red light, and the absorbance ratio is determined from the light intensity in these wavelength regions. Calculated and compared with the absorbance ratio of environmental water, etc., as a reference, and the algae productivity (AGP) or growth inhibition of the algae is determined by the water under test, so an inexpensive detection device such as a color sensor Thus, the productivity and growth inhibition of algae can be determined with good reproducibility.
  • a color sensor can be used, it is possible to provide a water quality test method having high stability without being influenced by the proficiency level of the measurer.

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Abstract

Selon l'invention, de l'eau devant être analysée est exposée à de la lumière blanche provenant d'une partie électroluminescente (2) d'un capteur de couleur en transmission (1), la lumière qui est passée à travers la solution de culture (4) est reçue par une partie de réception de lumière (3), la lumière reçue est amenée à passer dans un filtre coloré (non représenté) pour décomposer la lumière en lumière verte (500 à 570 nm) et lumière rouge (620 à 740 nm), la lumière verte et la lumière rouge sont détectées, une absorbance de la couleur verte et une absorbance de la couleur rouge sont calculées respectivement à partir d'une intensité lumineuse de la lumière verte et d'une intensité lumineuse de la lumière rouge, l'état de culture d'algues est déterminé sur la base du rapport des valeurs d'intensité lumineuse (le rapport des valeurs d'absorbance) et l'aptitude à produire des algues (le potentiel de développement d'algues) ou l'aptitude à inhiber le développement d'algues de l'eau est déterminée par comparaison du rapport de valeurs d'absorbance de l'eau avec celui d'une eau environnementale qui sert de référence. De cette manière, il devient possible de déterminer la qualité d'une eau, telle que de l'eau environnementale, de l'eau résiduaire traitée provenant d'usines ou une solution de culture d'algues, d'une manière simple et avec une grande précision.
PCT/JP2014/053547 2013-03-27 2014-02-14 Procédé d'analyse de la qualité de l'eau utilisant des algues WO2014156363A1 (fr)

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JP2013067559A JP2014187962A (ja) 2013-03-27 2013-03-27 藻類を利用した水質試験方法

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CN107796765A (zh) * 2017-10-17 2018-03-13 深圳市沃特沃德股份有限公司 饮水管理方法和装置
CN108051558A (zh) * 2017-12-13 2018-05-18 山东星火科学技术研究院 石化废水在线监测控制成套系统

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017201622A1 (fr) * 2016-05-24 2017-11-30 Real Tech Inc. Dispositif de détection de concentration d'algues utilisant une première dérivée de l'absorbance de lumière visible
EP3465148A4 (fr) * 2016-05-24 2020-01-29 Real Tech Inc. Dispositif de détection de concentration d'algues utilisant une première dérivée de l'absorbance de lumière visible
US10768117B2 (en) 2016-05-24 2020-09-08 Real Tech Holdings Inc. Device for detecting algae concentration using first derivative of visible light absorbance

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